# Patent

## Title
Methods for treating hypercholesterolemia

## Abstract

Disclosed herein are antisense compounds and methods for decreasing LDL-C in an individual having elevated LDL-C. Additionally disclosed are antisense compounds and methods for treating, preventing, or ameliorating hypercholesterolemia and/or atherosclerosis. Further disclosed are antisense compounds and methods for decreasing coronary heart disease risk. Such methods include administering to an individual in need of treatment an antisense compound targeted to a PCSK9 nucleic acid. The antisense compounds administered include gapmer antisense oligonucleotides.

## Background
REFERENCE TO RELATED APPLICATIONS
This application is a continuation application of U.S. application Ser. No. 14/152,825, filed Jan. 10, 2014; which is a continuation application of U.S. application Ser. No. 13/302,070, filed Nov. 22, 2011, now issued as U.S. Pat. No. 8,664,190; which is a continuation application of U.S. application Ser. No. 12/478,488, filed Jun. 4, 2009, now issued as U.S. Pat. No. 8,093,222; which is a continuation-in-part of U.S. application Ser. No. 12/516,457, filed Jan. 20, 2010, now issued as U.S. Pat. No. 8,084,437; which is a National Stage application of International PCT Application No. PCT/US07/024369, filed Nov. 27, 2007, which claims the benefit of U.S. Provisional Application No. 60/988,074, filed Nov. 14, 2007, U.S. Provisional Application No. 60/986,286, filed Nov. 7, 2007, U.S. Provisional Application No. 60/912,892, filed Apr. 19, 2007, and U.S. Provisional Application No. 60/867,395, filed Nov. 27, 2006, which all are herein incorporated by reference in their entirety. U.S. application Ser. No. 12/478,488 claims the benefit of U.S. Provisional Application No. 61/059,169, filed Jun. 5, 2008, and U.S. Provisional Application No. 61/058,707, filed Jun. 4, 2008, which are herein incorporated by reference in their entirety.


SEQUENCE LISTING
The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled BIOL0090USC3SEQ_ST25.txt, created on Nov. 11, 2014 which is 144 Kb in size. The information in the electronic format of the sequence listing is incorporated herein by reference in its entirety.
FIELD OF THE INVENTION
Methods are provided for lowering LDL-C levels in an individual having elevated LDL-C levels. Such methods are further useful to treat hypercholesterolemia, and to reduce the risk of coronary heart disease.
BACKGROUND
Atherosclerosis is a complex, polygenic disease of arterial degeneration that can lead to coronary heart disease (CHD). In Western societies, complications arising from atherosclerosis are the most common causes of death. Several risk factors for CHD have been well-established, and include elevated low density lipoprotein-cholesterol (LDL-C), low levels of high density lipoprotein cholesterol (HDL-C), cigarette smoking, hypertension, age, and family history of early CHD. Given the abundant clinical data and epidemiological studies indicating that lowering LDL-C is beneficial for the prevention of adverse coronary events, the primary target of pharmacological intervention in the treatment and prevention of CHD is lowered LDL-C.
In individuals with autosomal dominant hypercholesterolemia (ADH), elevated LDL-C levels have been linked to mutations in the genes encoding LDL-receptor (LDL-R), apolipoprotein B (apoB), or proprotein convertase subtilisin/kexin type 9 (PCSK9) (Abifadel et al., Nat. Genet., 2003, 34:154-156). The PCSK9 gene (also known as FH3; NARC1; NARC-1; and HCHOLA3) was mapped to Chromosome 1 at location 1p32.3. PCSK9 was identified as a third locus associated with ADH when gain-of-function mutations in PCSK9 were found to be linked to elevated LDL-C levels. ApoB-100 participates in the intracellular assembly and secretion of triglyceride-rich lipoproteins and is a ligand for the LDL-R. PCSK9 is proposed to reduce LDL-R expression levels in the liver. Reduced LDL-R expression results in reduced hepatic uptake of circulating apoB-100-containing lipoproteins, which in turn leads to elevated cholesterol.
Familial hypercholesterolemia (FH) is caused by hundreds of different mutations in the LDL-R, and is phenotypically characterized by elevated plasma LDL-C levels and deposits of LDL-C in tendons, skin and arteries, leading to premature cardiovascular disease. Homozygous and heterozygous mutations in the LDL-R are associated with FH. Likewise, heterozygous and homozygous mutations in the ligand-binding domain of PCSK9 are associated with the FH phenotype. Mutations in this gene have been associated with a third form of autosomal dominant FH.
In mice genetically deficient in PCSK9, a marked increase in LDL-R expression and increased plasma LDL clearance rate were observed. Conversely, overexpression in mice of wild-type PCSK9, or certain PCSK9 mutants, promotes decreased LDL-R expression and elevated LDL-C levels.
SUMMARY OF THE INVENTION
Provided herein are methods for the treatment of hypercholesterolemia and/or atherosclerosis, as well as methods for the reduction of elevated LDL-C levels and CHD risk. Methods for treating hypercholesterolemia comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. In such methods, an antisense compound targeted to a PCSK9 nucleic acid may be targeted to sequences as set forth in GENBANK® Accession No. NM_174936.2, nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_032977.8, or GENBANK® Accession No. AK124635.1.
Methods for treating and/or preventing atherosclerosis comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Methods for reducing LDL-C levels comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Methods for reducing CHD risk comprise administering to an individual an antisense compound targeted to a PCSK9 nucleic acid. Such methods may comprise the administration of a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid.
Methods are provided for reducing LDL-C levels in an individual having elevated LDL-C levels, comprising administering to the individual a therapeutically effective amount of an antisense oligonucleotide targeted to a PCSK9 nucleic acid, thereby reducing LDL-C levels. Also provided are methods for reducing LDL-C levels in an individual comprising selecting an individual having elevated LDL-C levels, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and additionally monitoring LDL-cholesterol levels. Further provided are methods for treating atherosclerosis in an individual, comprising selecting an individual diagnosed with atherosclerosis, administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and monitoring atherosclerosis. Also provided are methods for reducing coronary heart disease risk, comprising selecting an individual having elevated LDL-C levels and one or more additional indicators of coronary heart disease, administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid, and monitoring LDL-C levels. Additionally provided are methods for treating hypercholesterolemia, comprising administering to an individual diagnosed with hypercholesterolemia a therapeutically effective amount of an antisense oligonucleotide targeted to a PCSK9 nucleic acid, thereby reducing cholesterol levels.
Further provided is a use of an effective amount of one or more of the disclosed antisense compounds for treating atherosclerosis or hypercholesterolemia in an individual in need of such treatment. Also provided is a use of an effective amount of one or more of the disclosed antisense compounds for reducing LDL-C levels or reducing CHD risk in an individual in need of such treatment. Also provided is the use of one or more of the disclosed antisense compounds in the manufacture of a medicament for the treatment of atherosclerosis or hypercholesterolemia. Further provided is the use of one or more of the disclosed antisense compounds in the manufacture of a medicament for reducing LDL-C levels or for reducing CHD risk.
In any of the methods provided, a PCSK9 nucleic acid may be the sequence set forth in SEQ ID NO: 1. Thus, the antisense compound may be targeted to a PCSK9 nucleic acid as set forth in SEQ ID NO: 1.
Any of the methods provided herein may further comprise monitoring LDL-C levels.
An individual may be selected for administration of an antisense compound targeted to a PCSK9 nucleic acid when the individual exhibits an LDL-C level above 100 mg/dL, above 130 mg/dL, above 160 mg/dL, or above 190 mg/dL. Administration of an antisense compound targeted to a PCSK9 nucleic acid may result in an LDL-C level below 190 mg/dL, below 160 mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50 mg/dL.
In any of the aforementioned methods, administration of the antisense compound may comprise parenteral administration. The parenteral administration may further comprise subcutaneous or intravenous administration.
In any of the methods provided herein, the antisense compound may have least 80%, at least 90%, or at least 95% complementarity to SEQ ID NO: 1. Alternatively, the antisense compound may have 100% complementarity to SEQ ID NO: 1.
The antisense compounds provided herein and employed in any of the described methods may be 8 to 80 subunits in length, 12 to 50 subunits in length, 12 to 30 subunits in length, 15 to 30 subunits in length, 18 to 24 subunits in length, 19 to 22 subunits in length, or 20 subunits in length. Further, the antisense compounds employed in any of the described methods may be antisense oligonucleotides 8 to 80 nucleotides in length, 12 to 50 nucleotides in length, 12 to 30 nucleotides in length 15 to 30 nucleotides in length, 18 to 24 nucleotides in length, 19 to 22 nucleotides in length, or 20 nucleotides in length.
In any of the methods provided, the antisense compound may be an antisense oligonucleotide. Moreover, the antisense oligonucleotide may be a gapmer antisense oligonucleotide. The gapmer antisense oligonucleotide may comprise a gap segment of ten 2′-deoxynucleotides positioned between wing segments of five 2′-MOE nucleotides. The gapmer antisense oligonucleotide may be a gap-widened antisense oligonucleotide, comprising a gap segment of fourteen 2′-deoxynucleotides positioned between wing segments of three 2′-MOE nucleotides.
In any of the methods provided, the antisense compounds may have at least one modified internucleoside linkage. Additionally, each internucleoside linkage may be a phosphorothioate internucleoside linkage. Each cytosine may be a 5-methyl cytosine.
Also provided herein are antisense compounds targeted to a PCSK9 nucleic acid. Further provided are antisense oligonucleotides targeted to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may be targeted to PCSK9 nucleic acids, which include the sequences as set forth in GENBANK® Accession No. NM_174936.2, nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_032977.8, or GENBANK® Accession No. AK124635.1. The antisense compounds, including antisense oligonucleotides, may have at least 70%, at least 80%, at least 90%, or at least 95% complementarity to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may have 99% complementarity to a PCSK9 nucleic acid. The antisense compounds, including antisense oligonucleotides, may have 100% complementarity to a PCSK9 nucleic acid. For any of the antisense compounds provided, including antisense oligonucleotides, the PCSK9 nucleic acid may be the sequence set forth in SEQ ID NO: 1.
Antisense compounds targeted to a PCSK9 nucleic acid may be 8 to 80 subunits in length, 12 to 50 subunits in length, 12 to 30 subunits in length, 15 to 30 subunits in length, 18 to 24 subunits in length, 19 to 22 subunits in length, or 20 subunits in length. Antisense oligonucleotides targeted to a PCSK9 nucleic acid may be 8 to 80 nucleotides in length, 12 to 50 nucleotides in length, 12 to 30 nucleotides in length, 15 to 30 nucleotides in length, 18 to 24 nucleotides in length, 19 to 22 nucleotides in length, or 20 nucleotides in length.
Moreover, the antisense compound may be a gapmer antisense oligonucleotide. The gapmer antisense oligonucleotide may comprise a gap segment of ten 2′-deoxynucleotides positioned between wing segments of five 2′-MOE nucleotides. The gapmer antisense oligonucleotide may be a gap-widened antisense oligonucleotide, comprising a gap segment of fourteen 2′-deoxynucleotides positioned between wing segments of three 2′-MOE nucleotides.
Antisense compounds, including antisense oligonucleotides, targeted to a PCSK9 nucleic acid may have at least one modified internucleoside linkage. Additionally, each internucleoside linkage may be a phosphorothioate internucleoside linkage. Each cytosine may be a 5-methyl cytosine.
Antisense compounds, including antisense oligonucleotides, targeted to a PCSK9 nucleic acid may have at least one modified nucleoside. In certain embodiments, a modified nucleoside is a sugar-modified nucleoside. In certain such embodiments, the sugar-modified nucleosides can further comprise a natural or modified heterocyclic base moiety and/or a natural or modified internucleoside linkage and may include further modifications independent from the sugar modification. In certain embodiments, a sugar modified nucleoside is a 2′-modified nucleoside, wherein the sugar ring is modified at the 2′ carbon from natural ribose or 2′-deoxy-ribose.
In certain embodiments, a 2′ modified nucleoside has a 2′-F, 2′-OCH2 (2′-OMe) or a 2′-O(CH2)2—OCH3 (2′-O-methoxyethyl or 2′-MOE) substituent group
In certain embodiments, a 2′-modified nucleoside has a bicyclic sugar moiety. In certain such embodiments, the bicyclic sugar moiety is a D sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar moiety is a D sugar in the beta configuration. In certain such embodiments, the bicyclic sugar moiety is an L sugar in the alpha configuration. In certain such embodiments, the bicyclic sugar moiety is an L sugar in the beta configuration.
In certain embodiments, the bicyclic sugar moiety comprises a bridge group between the 2′ and the 4′-carbon atoms. In certain such embodiments, the bridge group comprises from 1 to 8 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises from 1 to 4 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises 2 or 3 linked biradical groups. In certain embodiments, the bicyclic sugar moiety comprises 2 linked biradical groups. In certain embodiments, a linked biradical group is selected from —O—, —S—, —N(R1)-, —C(R1)(R2)-, —C(R1)=C(R1)-, —C(R1)=N—, —C(═NR1)-, —Si(R1)(R2)-, —S(═O)2-, —S(═O)—, —C(═O)— and —C(═S)—; where each R1 and R2 is, independently, H, hydroxyl, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, C5-C20 aryl, substituted C5-C20 aryl, a heterocycle radical, a substituted hetero-cycle radical, heteroaryl, substituted heteroaryl, C5-C7 alicyclic radical, substituted C5-C7 alicyclic radical, halogen, substituted oxy (—O—), amino, substituted amino, azido, carboxyl, substituted carboxyl, acyl, substituted acyl, CN, thiol, substituted thiol, sulfonyl (S(═O)2-H), substituted sulfonyl, sulfoxyl (S(═O)—H) or substituted sulfoxyl; and each substituent group is, independently, halogen, C1-C12 alkyl, substituted C1-C12 alkyl, C2-C12 alkenyl, substituted C2-C12 alkenyl, C2-C12 alkynyl, substituted C2-C12 alkynyl, amino, substituted amino, acyl, substituted acyl, C1-C12 amino alkyl, C1-C12 aminoalkoxy, substituted C1-C12 aminoalkyl, substituted C1-C12 aminoalkoxy or a protecting group.
In some embodiments, the bicyclic sugar moiety is bridged between the 2′ and 4′ carbon atoms with a biradical group selected from —O—(CH2)p-, —O—CH2-, —O—CH2CH2-, —O—CH(alkyl)-, —NH—(CH2)p-, —N(alkyl)-(CH2)p-, —O—CH(alkyl)-, —(CH(alkyl))-(CH2)p-, —NH—O—(CH2)p-, —N(alkyl)-O—(CH2)p-, or —O—N(alkyl)-(CH2)p-, wherein p is 1, 2, 3, 4 or 5 and each alkyl group can be further substituted. In certain embodiments, p is 1, 2 or 3.
In one aspect, each of said bridges is, independently, —[C(R1)(R2)]n-, —[C(R1)(R2)]n-O—, —C(R1R2)-N(R1)-O— or —C(R1R2)-O—N(R1)-. In another aspect, each of said bridges is, independently, 4′-(CH2)3-2′, 4′-(CH2)2-2′, 4′-CH2-O-2′, 4′-(CH2)2-O-2′, 4′-CH2-O—N(R1)-2′ and 4′-CH2-N(R1)-O-2′- wherein each R1 is, independently, H, a protecting group or C1-C12 alkyl.
The present disclosure further discloses subsets of the antisense compounds above that show superior properties relating to pharmacodynamics and/or pharmacokinetics, among other properties. Exemplary antisense compounds reduce PCSK9 expression in a cultured cell model system, e.g., Hep3B cells, cynomolgus monkey hepatocytes, or human primary hepatocytes. In particular embodiments, antisense compounds reduce PCSK9 expression in a dose-dependent manner in a cell culture system, where the antisense compounds have an IC50 in the nanomolar range.
Exemplary antisense compounds also reduce the expression of a PCSK9 nucleic acid in an animal model, such as a transgenic mouse that expresses a human PCSK9 nucleic acid (HuPCSK9Tg mice). In particular embodiments, antisense compounds include those that reduce PCSK9 express in an animal model with a physiology that correlates closely with humans, such as monkey, e.g., the cynomolgus monkey.
Exemplary antisense compounds display minimal side effects. Side effects include responses to the administration of the antisense compound unrelated to the targeting of a PCSK9 nucleic acid, such as an inflammatory response in the host individual. Exemplary antisense compounds are well tolerated by the host individual. Tolerability may be determined though histological analysis of various cell types, and includes examination of such markers as cytoplasmic swelling from multifocal apoptosis in liver cells, follicular hyperplasia in spleen cells, macrophage infiltration, and the like. In particular embodiments, antisense compounds produce minimal signs of host intolerance.
Exemplary antisense compounds further display favorable pharmacokinetics. In particular embodiments, antisense compounds accumulate at a relatively high ratio in the liver versus other sensitive organs, such as kidneys. In particular embodiments, antisense compounds exhibit relatively high half-lives in relevant biological fluids or tissues, e.g., liver tissue, reflecting higher stability and resistance to nucleases, for example.
In particular embodiments, antisense compounds target identical sequences in human and animal PCSK9 sequences. In other embodiments, antisense compounds target PCSK9 sequences without a known single nucleotide polymorphism (SNP), have a high relative G-content, have minimal secondary structure, have greatly reduced effects on the host's serum chemistry, body weight, or histopathology, compared to other antisense compounds, and/or induce a minimal adverse immunohistocompatibility reaction.
In a particular embodiment, the antisense compound is selected from the group of Isis 405881, Isis 399819, Isis 395165, Isis 405879, Isis 406008, Isis 405891, Isis 395186, Isis 405988, Isis 405994, Isis 406023, Isis 395187, Isis 395185, Isis 406033, Isis 405923, Isis 399900, Isis 405995, Isis 405991, Isis 406005, Isis 399793, and Isis 395152. The aforementioned antisense compounds effectively repress PCSK9 expression in a Hep3B cell culture system. In another embodiment, antisense compounds effectively inhibit PCSK9 expression with a low IC50 in a battery of cell culture systems. Exemplary antisense compounds include those selected from the group of Isis 395165, Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405881, Isis 405891, Isis 405988, Isis 405994, and Isis 406008. In yet another embodiment, antisense compounds exhibit minimal or no adverse histopathological effects when administered at particularly high doses to an individual. Exemplary antisense compounds include those selected from the group consisting of Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405891, and Isis 405988. In yet another embodiment, antisense compounds affect therapeutic end-points, e.g., reduction of plasma LDL-C, liver TG, or liver PCSK9 mRNA, in an animal model. Exemplary antisense compounds include Isis 405879. In yet another embodiment, antisense compounds display combinations of the characteristics above and reduce liver PCSK9 mRNA expression in an animal model with high efficiency, e.g., Isis 405879.
Definitions
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by on of skill in the art to which the invention(s) belong. Unless specific definitions are provided, the nomenclature utilized in connection with, and the procedures and techniques of, analytical chemistry, synthetic organic chemistry, and medicinal and pharmaceutical chemistry described herein are those well known and commonly used in the art. Standard techniques may be used for chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of subjects. Certain such techniques and procedures may be found for example in “Antisense Drug Technology: Principles, Strategies, and Applications.” by Stanley Crooke, Boca Raton: Taylor & Francis Group, 2008; “Carbohydrate Modifications in Antisense Research” Edited by Sangvi and Cook, American Chemical Society, Washington D.C., 1994; and “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., 18th edition, 1990; and which is hereby incorporated by reference for any purpose. Where permitted, all patents, patent applications, published applications and publications, GENBANK sequences, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety. All of the GENBANK® Accession Nos. along with their associated sequence and structural data pertaining to such sequences including gene organization and structural elements and SNP information that may be found in sequence databases such as the National Center for Biotechnology Information (NCBI) are incorporated herein by reference in their entirety. In the event that there is a plurality of definitions for terms herein, those in this section prevail. Where reference is made to a URL or other such identifier or address, it is understood that such identifiers can change and particular information on the internet can come and go, but equivalent information can be found by searching the internet. Reference thereto evidences the availability and public dissemination of such information.
A “pharmaceutical composition” means a mixture of substances suitable for administering to an individual. For example, a pharmaceutical composition may comprise an antisense oligonucleotide and a sterile aqueous solution.
“Administering” means providing a pharmaceutical agent to an individual, and includes, but is not limited to administering by a medical professional and self-administering.
“Individual” means a human or non-human animal selected for treatment or therapy.
“Duration” means the period of time during which an activity or event continues. In certain embodiments, the duration of treatment is the period of time during which doses of a pharmaceutical agent are administered.
“Parenteral administration,” means administration through injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, or intramuscular administration.
“Subcutaneous administration” means administration just below the skin. “Intravenous administration” means administration into a vein.
“Dose” means a specified quantity of a pharmaceutical agent provided in a single administration. In certain embodiments, a dose may be administered in two or more boluses, tablets, or injections. For example, in certain embodiments, where subcutaneous administration is desired, the desired dose requires a volume not easily accommodated by a single injection. In such embodiments, two or more injections may be used to achieve the desired dose. In certain embodiments, a dose may be administered in two or more injections to minimize injection site reaction in a individual.
“Dosage unit” means a form in which a pharmaceutical agent is provided. In certain embodiments, a dosage unit is a vial containing lyophilized antisense oligonucleotide. In certain embodiments, a dosage unit is a vial containing reconstituted antisense oligonucleotide.
“Pharmaceutical agent” means a substance provides a therapeutic benefit when administered to a individual. For example, in certain embodiments, an antisense oligonucleotide targeted to PCSK9 is pharmaceutical agent.
“Diluent” means an ingredient in a composition that lacks pharmacological activity, but is pharmaceutically necessary or desirable. For example, in drugs that are injected the diluent may be a liquid, e.g., saline solution.
“Active pharmaceutical ingredient” means the substance in a pharmaceutical composition that provides a desired effect.
“Pharmaceutically acceptable carrier” means a medium or diluent that does not interfere with the structure of the oligonucleotide. Certain of such carriers enable pharmaceutical compositions to be formulated as, for example, tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspension and lozenges for the oral ingestion by a subject.
“Therapeutically effective amount” means an amount of a pharmaceutical agent that provides a therapeutic benefit to an individual. In certain embodiments, a therapeutically effective amount of antisense compound targeted to a PCSK9 nucleic acid is an amount that decreases LDL-C in the individual.
“Hypercholesterolemia” means a condition characterized by elevated serum cholesterol.
“Hyperlipidemia” means a condition characterized by elevated serum lipids.
“Hypertriglyceridemia” means a condition characterized by elevated serum triglyceride levels.
“Non-familial hypercholesterolemia” means a condition characterized by elevated serum cholesterol that is not the result of a single gene mutation.
“Polygenic hypercholesterolemia” means a condition characterized by elevated cholesterol that results from the influence of a variety of genetic factors. In certain embodiments, polygenic hypercholesterolemia may be exacerbated by dietary intake of lipids.
“Familial hypercholesterolemia (FH)” means an autosomal dominant metabolic disorder characterized by a mutation in the LDL-receptor (LDL-R) gene, markedly elevated LDL-C and premature onset of atherosclerosis. A diagnosis of familial hypercholesterolemia is made when an individual meets one or more of the following criteria: genetic testing confirming 2 mutated LDL-receptor genes; genetic testing confirming one mutated LDL-receptor gene; document history of untreated serum LDL-cholesterol greater than 500 mg/dL; tendinous and/or cutaneous xanthoma prior to age 10 years; or, both parents have documented elevated serum LDL-cholesterol prior to lipid-lowering therapy consistent with heterozygous familial hypercholesterolemia.
“Homozygous familial hypercholesterolemia” or “HoFH” means a condition characterized by a mutation in both maternal and paternal LDL-R genes.
“Heterozygous familial hypercholesterolemia” or “HeFH” means a condition characterized by a mutation in either the maternal or paternal LDL-R gene.
“Mixed dyslipidemia” means a condition characterized by elevated serum cholesterol and elevated serum triglycerides.
“Diabetic dyslipidemia” or “Type II diabetes with dyslipidemia” means a condition characterized by Type II diabetes, reduced HDL-C, elevated serum triglycerides, and elevated small, dense LDL particles.
“CHD risk equivalents,” means indicators of clinical atherosclerotic disease that confer a high risk for coronary heart disease, and include clinical coronary heart disease, symptomatic carotid artery disease, peripheral arterial disease, and/or abdominal aortic aneurysm.
“Metabolic syndrome” means a condition characterized by a clustering of lipid and non-lipid cardiovascular risk factors of metabolic origin. In certain embodiments, metabolic syndrome is identified by the presence of any 3 of the following factors: waist circumference of greater than 102 cm in men or greater than 88 cm in women; serum triglyceride of at least 150 mg/dL; HDL-C less than 40 mg/dL in men or less than 50 mg/dL in women; blood pressure of at least 130/85 mmHg; and fasting glucose of at least 110 mg/dL.
“Non-alcoholic fatty liver disease (NAFLD)” means a condition characterized by fatty inflammation of the liver that is not due to excessive alcohol use (for example, alcohol consumption of over 20 g/day). In certain embodiments, NAFLD is related to insulin resistance and the metabolic syndrome.
“Non-alcoholic steatohepatitis (NASH)” means a condition characterized by inflammation and the accumulation of fat and fibrous tissue in the liver, that is not due to excessive alcohol use. NASH is an extreme form of NAFLD.
“Major risk factors” mean factors that contribute to a high risk for coronary heart disease, and include without limitation cigarette smoking, hypertension, low HDL-C, family history of coronary heart disease, and age.
“CHD risk factors” mean CHD risk equivalents and major risk factors.
“Coronary heart disease (CHD)” means a narrowing of the small blood vessels that supply blood and oxygen to the heart, which is often a result of atherosclerosis.
“Reduced coronary heart disease risk” means a reduction in the likelihood that an individual will develop coronary heart disease. In certain embodiments, a reduction in coronary heart disease risk is measured by an improvement in one or more CHD risk factors, for example, a decrease in LDL-C levels.
“Atherosclerosis” means a hardening of the arteries affecting large and medium-sized arteries and is characterized by the presence of fatty deposits. The fatty deposits are called “atheromas” or “plaques,” which consist mainly of cholesterol and other fats, calcium and scar tissue, and damage the lining of arteries.
“History of coronary heart disease” means the occurrence of clinically evident coronary heart disease in the medical history of an individual or a individual's family member.
“Early onset coronary heart disease” means a diagnosis of coronary heart disease prior to age 50.
“Statin intolerant individual” means an individual who as a result of statin therapy experiences one or more of creatine kinase increases, liver function test abnormalities, muscle aches, or central nervous system side effects.
“Efficacy” means the ability to produce a desired effect. For example, efficacy of a lipid-lowering therapy may be reduction in the concentration of one or more of LDL-C, VLDL-C, IDL-C, non-HDL-C, ApoB, lipoprotein(a), or triglycerides.
“Acceptable safety profile” means a pattern of side effects that is within clinically acceptable limits.
“Side effects” means physiological responses attributable to a treatment other than desired effects. In certain embodiments, side effects include, without limitation, injection site reactions, liver function test abnormalities, renal function abnormalities, liver toxicity, renal toxicity, central nervous system abnormalities, and myopathies. For example, increased aminotransferase levels in serum may indicate liver toxicity or liver function abnormality. For example, increased bilirubin may indicate liver toxicity or liver function abnormality.
“Injection site reaction” means inflammation or abnormal redness of skin at a site of injection in an individual.
“Individual compliance” means adherence to a recommended or prescribed therapy by an individual.
“Lipid-lowering therapy” means a therapeutic regimen provided to an individual to reduce one or more lipids in a individual. In certain embodiments, a lipid-lowering therapy is provide to reduce one or more of ApoB, total cholesterol, LDL-C, VLDL-C, IDL-C, non-HDL-C, triglycerides, small dense LDL particles, and Lp(a) in a individual.
“Lipid-lowering agent” means a pharmaceutical agent provided to an individual to achieve a lowering of lipids in the individual. For example, in certain embodiments, a lipid-lowering agent is provided to an individual to reduce one or more of ApoB, LDL-C, total cholesterol, and triglycerides.
“LDL-C target” means an LDL-C level that is desired following lipid-lowering therapy.
“Comply” means the adherence with a recommended therapy by a individual.
“Recommended therapy” means a therapeutic regimen recommended by a medical professional for the treatment, amelioration, or prevention of a disease.
“Low LDL-receptor activity” means LDL-receptor activity that is not sufficiently high to maintain clinically acceptable levels of LDL-C in the bloodstream.
“Cardiovascular outcome” means the occurrence of major adverse cardiovascular events.
“Improved cardiovascular outcome” means a reduction in the occurrence of major adverse cardiovascular events, or the risk thereof. Examples of major adverse cardiovascular events include, without limitation, death, reinfarction, stroke, cardiogenic shock, pulmonary edema, cardiac arrest, and atrial dysrhythmia.
“Surrogate markers of cardiovascular outcome” means indirect indicators of cardiovascular events, or the risk thereof. For example, surrogate markers of cardiovascular outcome include carotid intimal media thickness (CIMT). Another example of a surrogate marker of cardiovascular outcome includes atheroma size. Atheroma size may be determined by intravascular ultrasound (IVUS). Surrogate markers also include increased HDL-cholesterol, or any combination of the markers above.
“Increased HDL-C” means an increase in serum HDL-C in an individualover time.
“Lipid-lowering” means a reduction in one or more serum lipids in an individual over time.
“Co-administration” means administration of two or more pharmaceutical agents to a individual. The two or more pharmaceutical agents may be in a single pharmaceutical composition, or may be in separate pharmaceutical compositions. Each of the two or more pharmaceutical agents may be administered through the same or different routes of administration. Co-administration encompasses administration in parallel or sequentially.
“Administered concomitantly” refers to the administration of two agents at the same therapeutic time frame, in any manner in which the pharmacological effects of both are manifest in the patient at the same time. Concomitant administration does not require that both agents be administered in a single pharmaceutical composition, in the same dosage form, or by the same route of administration.
“Therapeutic lifestyle change” means dietary and lifestyle changes intended to lower cholesterol and reduce the risk of developing heart disease, and includes recommendations for dietary intake of total daily calories, total fat, saturated fat, polyunsaturated fat, monounsaturated fat, carbohydrate, protein, cholesterol, insoluble fiber, as well as recommendations for physical activity.
“Statin” means a pharmaceutical agent that inhibits the activity of HMG-CoA reductase.
“HMG-CoA reductase inhibitor” means a pharmaceutical agent that acts through the inhibition of the enzyme HMG-CoA reductase.
“Cholesterol absorption inhibitor” means a pharmaceutical agent that inhibits the absorption of exogenous cholesterol obtained from diet.
“LDL apheresis” means a form of apheresis by which LDL-C is removed from blood. Typically, a individual's blood is removed from a vein, and separated into red cells and plasma. LDL-C is filtered out of the plasma prior to return of the plasma and red blood cells to the individual.
“MTP inhibitor” means a pharmaceutical agent that inhibits the enzyme microsomal triglyceride transfer protein.
“Low density lipoprotein-cholesterol (LDL-C)” means cholesterol carried in low density lipoprotein particles. Concentration of LDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum LDL-C” and “plasma LDL-C” mean LDL-C in the serum and plasma, respectively.
“Very low density lipoprotein-cholesterol (VLDL-C)” means cholesterol associated with very low density lipoprotein particles. Concentration of VLDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum VLDL-C” and “plasma VLDL-C” mean VLDL-C in the serum or plasma, respectively.
“Intermediate low density lipoprotein-cholesterol (IDL-C)” means cholesterol associated with intermediate density lipoprotein. Concentration of IDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum IDL-C” and “plasma IDL-C” mean IDL-C in the serum or plasma, respectively.
“Non-high density lipoprotein-cholesterol (Non-HDL-C)” means cholesterol associated with lipoproteins other than high density lipoproteins, and includes, without limitation, LDL-C, VLDL-C, and IDL-C.
“High density lipoprotein-C(HDL-C)” means cholesterol associated with high density lipoprotein particles. Concentration of HDL-C in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum HDL-C” and “plasma HDL-C” mean HDL-C in the serum and plasma, respectively.
“Total cholesterol” means all types of cholesterol, including, but not limited to, LDL-C, HDL-C, IDL-C and VLDL-C. Concentration of total cholesterol in serum (or plasma) is typically quantified in mg/dL or nmol/L.
“Lipoprotein(a)” or “Lp(a)” means a lipoprotein particle that is comprised of LDL-C, an apolipoprotein(a) particle, and an apolipoproteinB-100 particle.
“ApoA1” means apolipoprotein-A1 protein in serum. Concentration of ApoA1 in serum is typically quantified in mg/dL or nmol/L.
“ApoB:ApoA1 ratio” means the ratio of ApoB concentration to ApoA1 concentration.
“ApoB-containing lipoprotein” means any lipoprotein that has apolipoprotein B as its protein component, and is understood to include LDL, VLDL, IDL, and lipoprotein(a).
“Small LDL particle” means a subclass of LDL particles characterized by a smaller, denser size compared to other LDL particles. In certain embodiments, large LDL particles are 23-27 nm in diameter. In certain embodiments, intermediate LDL particles are 21.2-23 nm in diameter. In certain embodiments, small LDL particles are 18-21.2 nm in diameter. In certain embodiments, particle size is measured by nuclear magnetic resonance analysis.
“Small VLDL particle” means a subclass of VLDL particles characterized by a smaller, denser size compared to other VLDL particles. In certain embodiments, large VLDL particles are greater than 60 nm in diameter. In certain embodiments, medium VLDL particles are 35-60 nm in diameter. In certain embodiments, small VLDL particles are 27-35 nm in diameter. In certain embodiments, particle size is measured by nuclear magnetic resonance analysis.
“Triglycerides” means lipids that are the triesters of glycerol. “Serum triglycerides” mean triglycerides present in serum. “Liver triglycerides” mean triglycerides present in liver tissue.
“Serum lipids” mean cholesterol and triglycerides in the serum.
“Elevated total cholesterol” means total cholesterol at a concentration in an individual at which lipid-lowering therapy is recommended, and includes, without limitation, elevated LDL-C”, “elevated VLDL-C,” “elevated IDL-C,” and “elevated non-HDL-C.” In certain embodiments, total cholesterol concentrations of less than 200 mg/dL, 200-239 mg/dL, and greater than 240 mg/dL are considered desirable, borderline high, and high, respectively. In certain embodiments, LDL-C concentrations of 100 mg/dL, 100-129 mg/dL, 130-159 mg/dL, 160-189 mg/dL, and greater than 190 mg/dL are considered optimal, near optimal/above optimal, borderline high, high, and very high, respectively.
“Elevated triglyceride” means concentrations of triglyceride in the serum or liver at which lipid-lowering therapy is recommended, and includes “elevated serum triglyceride” and “elevated liver triglyceride.” In certain embodiments, serum triglyceride concentration of 150-199 mg/dL, 200-499 mg/dL, and greater than or equal to 500 mg/dL is considered borderline high, high, and very high, respectively.
“Elevated small LDL particles” means a concentration of small LDL particles in an individual at which lipid-lowering therapy is recommended.
“Elevated small VLDL particles” means a concentration of small VLDL particles in an individual at which lipid-lowering therapy is recommended.
“Elevated lipoprotein(a)” means a concentration of lipoprotein(a) in an individual at which lipid-lowering therapy is recommended.
“Low HDL-C” means a concentration of HDL-C in an individual at which lipid-lowering therapy is recommended. In certain embodiments lipid-lowering therapy is recommended when low HDL-C is accompanied by elevations in non-HDL-C and/or elevations in triglyceride. In certain embodiments, HDL-C concentrations of less than 40 mg/dL are considered low. In certain embodiments, HDL-C concentrations of less than 50 mg/dL are considered low.
“ApoB” means apolipoprotein B-100 protein. Concentration of ApoB in serum (or plasma) is typically quantified in mg/dL or nmol/L. “Serum ApoB” and “plasma ApoB” mean ApoB in the serum and plasma, respectively.
“LDL/HDL ratio” means the ratio of LDL-C to HDL-C.
“Oxidized-LDL” or “Ox-LDL-C” means LDL-C that is oxidized following exposure to free radicals.
“Individual having elevated LDL-C levels” means an individual who has been identified by a medical professional (e.g., a physician) as having LDL-C levels near or above the level at which therapeutic intervention is recommended, according to guidelines recognized by medical professionals. Such an individual may also be considered “in need of treatment” to decrease LDL-C levels.
“Individual having elevated apoB-100 levels” means an individual who has been identified as having apoB-100 levels near or above the level at which therapeutic intervention is recommended, according to guidelines recognized by medical professionals. Such an individual may also be considered “in need of treatment” to decrease apoB-100 levels.
“Treatment of elevated LDL-C levels” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual having elevated LDL-C levels.
“Treatment of atherosclerosis” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual who, based upon a physician's assessment, has or is likely to have atherosclerosis. “Prevention of atherosclerosis” means administration of an antisense compound targeted to a PCSK9 nucleic acid to an individual who, based upon a physician's assessment, is susceptible to atherosclerosis.
“Ameliorate” means to make better or improve a detrimental condition in an individual.
“Target nucleic acid,” “target RNA,” “target RNA transcript” and “nucleic acid target” all mean any nucleic acid capable of being targeted by antisense compounds.
“PCSK9 nucleic acid” means any nucleic acid encoding PCSK9. For example, in certain embodiments, a PCSK9 nucleic acid includes, without limitation, a DNA sequence encoding PCSK9, an RNA sequence transcribed from DNA encoding PCSK9, and an mRNA sequence encoding PCSK9. “PCSK9 mRNA” means an mRNA encoding a PCSK9 protein.
“Targeting” means the process of design and selection of an antisense compound that will specifically hybridize to a target nucleic acid and induce a desired effect.
“Targeted” means having a nucleobase sequence that will allow specific hybridization of an antisense compound to a target nucleic acid to induce a desired effect. In certain embodiments, a desired effect is reduction of a target nucleic acid. In certain such embodiments, a desired effect is reduction of PCSK9 mRNA.
“Antisense inhibition” means reduction of a target nucleic acid levels in the presence of an antisense compound complementary to a target nucleic acid compared to target nucleic acid levels in the absence of the antisense compound.
“Target region” means a fragment of a target nucleic acid to which one or more antisense compounds is targeted.
“Target segment” means the sequence of nucleotides of a target nucleic acid to which an antisense compound is targeted. “5′ target site” refers to the 5′-most nucleotide of a target segment. “3′ target site” refers to the 3′-most nucleotide of a target segment.
“Active target region” means a target region to which one or more active antisense compounds is targeted. “Active antisense compounds” means antisense compounds that reduce target nucleic acid levels.
“Hybridization” means the annealing of complementary nucleic acid molecules. In certain embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense compound and a nucleic acid target. In certain such embodiments, complementary nucleic acid molecules include, but are not limited to, an antisense oligonucleotide and a nucleic acid target
“Stringent hybridization conditions” means conditions under which a nucleic acid molecule, such as an antisense compound, will hybridize to a target nucleic acid sequence, but to a minimal number of other sequences.
“Specifically hybridizable” means an antisense compound that hybridizes to a target nucleic acid to induce a desired effect, while exhibiting minimal or no effects on non-target nucleic acids.
“Complementarity” means the capacity for pairing between nucleobases of a first nucleic acid and a second nucleic acid.
“Fully complementary” means each nucleobase of a first nucleic acid is capable of pairing with each nucleobase of a second nucleic acid. In certain embodiments, a first nucleic acid is an antisense compound and a target nucleic acid is a second nucleic acid. In certain such embodiments, an antisense oligonucleotide is a first nucleic acid and a target nucleic acid is a second nucleic acid.
“Non-complementary nucleobase” means a nucleobase of first nucleic acid that is not capable of pairing with the corresponding nucleobase of a target nucleic acid. “Mismatch” a nucleobase of first nucleic acid that is not capable of pairing with the corresponding nucleobase of a target nucleic acid.
“Portion” means a defined number of contiguous (i.e., linked) nucleobases. In certain embodiments, a portion is a defined number of contiguous nucleobases of a target nucleic acid. In certain embodiments, a portion is a defined number of contiguous nucleobases of an antisense compound.
“Oligomeric compound” means a polymer of linked monomeric subunits which is capable of hybridizing to at least a region of an RNA molecule.
“Antisense compound” means an oligomeric compound that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.
“Oligonucleotide” means a polymer of linked nucleosides each of which can be modified or unmodified, independent one from another.
“Oligonucleoside” means an oligonucleotide in which the internucleoside linkages do not contain a phosphorus atom.
“Unmodified nucleotide” means a nucleotide composed of naturally occurring nucleobases, sugar moieties and internucleoside linkages. In certain embodiments, an unmodified nucleotide is an RNA nucleotide (i.e., β-D-ribonucleosides) or a DNA nucleotide (i.e., β-D-deoxyribonucleoside).
“Antisense oligonucleotide” means a single-stranded oligonucleotide having a nucleobase sequence that permits hybridization to a corresponding region of a target nucleic acid.
“Motif” means the pattern of unmodified and modified nucleosides in an antisense compound.
“Chimeric antisense compound” means an antisense compound that has at least 2 chemically distinct regions, each region having a plurality of subunits.
A “gapmer” means an antisense compound in which an internal position having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having one or more nucleotides that are chemically distinct from the nucleosides of the internal region.
A “gap segment” means the plurality of nucleotides that make up the internal region of a gapmer.
A “wing segment” means the external region of a gapmer.
“Gap-widened” means an antisense compound has a gap segment of 12 or more contiguous 2′-deoxyribonucleotides positioned between 5′ and 3′ wing segments having from one to six nucleotides having modified sugar moieties.
“Nucleoside” means a nucleobase linked to a sugar.
“Nucleobase” means a heterocyclic base moiety capable of pairing with a base of another nucleic acid.
“Nucleotide” means a nucleoside having a phosphate group covalently linked to the sugar portion of the nucleoside.
“Nucleobase sequence” means the order of contiguous nucleobases independent of any sugar, linkage, and/or nucleobase modification.
“Modified nucleotide” means a nucleotide having, independently, a modified sugar moiety, modified internucleoside linkage, or modified nucleobase.
“Modified nucleoside” means a nucleotide having, independently, a modified sugar moiety or modified nucleobase.
“Contiguous nucleobases” means nucleobases immediately adjacent to each other.
“Modified oligonucleotide” means an oligonucleotide comprising a modified internucleoside linkage, a modified sugar, and/or a modified nucleobase.
“Single-stranded modified oligonucleotide” means a modified oligonucleotide which is not hybridized to a complementary strand.
“Internucleoside linkage” refers to the chemical bond between nucleosides.
“Linked nucleoside” means adjacent nucleosides which are bonded together.
“Linked deoxynucleoside” means a nucleic acid base (A, G, C, T, U) substituted by deoxyribose linked by a phosphate ester to form a nucleotide.
“Naturally occurring internucleoside linkage” means a 3′ to 5′ phosphodiester linkage.
“Modified internucleoside linkage” means substitution and/or any change from a naturally occurring internucleoside bond. In certain instances, the modified internucleoside linkage refers to a phosphodiester internucleoside bond.
“Phosphorothioate linkage” means a linkage between nucleosides where the phosphodiester bond is modified by replacing one of the non-bridging oxygen atoms with a sulfur atom. A phosphorothioate linkage is a modified internucleoside linkage.
“Modified sugar moiety” means substitution and/or any change from a natural sugar moiety. For the purposes of this disclosure, a “natural sugar moiety” is a sugar moiety found in DNA (2′-H) or RNA (2′-OH).
“Modified sugar” refers to a substitution and/or any change from a natural sugar.
“Bicyclic sugar” means a furosyl ring modified by the bridging of the two non-geminal ring atoms. A bicyclic sugar is a modified sugar.
“Modified nucleobase” means any nucleobase other than adenine, cytosine, guanine, thymidine, or uracil. An “unmodified nucleobase” means the purine bases adenine (A) and guanine (G), and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).
“Modified sugar moiety” means a sugar moiety having a substitution and/or any change from a natural sugar moiety.
“Natural sugar moiety” means a sugar moiety found in DNA (2′-H) or RNA (2′-OH).
“2′-O-methoxyethyl sugar moiety” means a 2′-substituted furosyl ring having a 2′-O(CH2)2-OCH3 (2′-O-methoxyethyl or 2′-MOE) substituent group.
“2′-O-methoxyethyl nucleotide” means a nucleotide comprising a 2′-O-methoxyethyl modified sugar moiety.
“2′-O-methoxyethyl” refers to an O-methoxy-ethyl modification of the 2′ position of a furosyl ring. A 2′-O-methoxyethyl modified sugar is a modified sugar.
“Bicyclic nucleic acid sugar moiety” means a furosyl ring modified by the bridging of two non-geminal ring atoms.
“5-methylcytosine” means a cytosine modified with a methyl group attached to the 5′ position. A 5-methylcytosine is a modified nucleobase.
“Prodrug” means a therapeutic agent that is prepared in an inactive form that is converted to an active form (i.e., drug) within the body or cells thereof by the action of endogenous enzymes or other chemicals and/or conditions.
“Pharmaceutically acceptable salts” means physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.
“Salts” refers to physiologically and pharmaceutically acceptable salts of antisense compounds, i.e., salts that retain the desired biological activity of the parent oligonucleotide and do not impart undesired toxicological effects thereto.
“Cures” means a method or course that restores health or a prescribed treatment for an illness.
“Slows progression” means decrease in the development of the said disease.
“Cap structure” or “terminal cap moiety” means chemical modifications, which have been incorporated at either terminus of an antisense compound.
“ISIS 301012” means a lipid-lowering agent that is an antisense oligonucleotide having the sequence “GCCTCAGTCTGCTTCGCACC” (SEQ ID NO: 457), where each internucleoside linkage is a phosphorothioate internucleoside linkage, each cytosine is a 5-methylcytosine, nucleotides 6-15 are 2′-deoxynucleotides, and nucleotides 1-5 and 16-20 are 2′-O-methoxyethyl nucleotides. ISIS 301012 does not target PCSK9. It is used herein as a non-PCSK9 control.



BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 depicts the location of PCSK9 mRNA target sequences of exemplary antisense compounds.
FIG. 2 depicts the results of a five-point dose response experiment with exemplary antisense compounds in HEK-293 cells. The antisense compounds are identified by Isis number. The bars from left to right represent PCSK9 expression in relative units at 0 nM, 25 nM, 50 nM, 100 nM, 200 nM, and 300 nM antisense oligonucleotides.
FIG. 3 depicts the results of a five-point dose response experiment with exemplary antisense compounds in cyno primary hepatocytes. The antisense compounds are identified by Isis number. The bars from left to right represent PCSK9 expression in relative units at 300 nM, 150 nM, 50 nM, 25 nM, and 10 nM antisense oligonucleotides. Control values are shown in the rightmost two bars.
FIG. 4 depicts a reduction of murine PCSK9 mRNA expression (relative units) in the liver of h-apoB/CETP transgenic mice when administered Isis 394816. Isis 394816 was administered by intraperitoneal injection for 6 weeks at a dosing regimen of 20 mg/kg/wk or 50 mg/kg/wk. Isis 329993 was administered at 50 mg/kg/wk as a control.
FIG. 5, Panel A, depicts inhibition of plasma LDL-C (mg/dL) in mice administered Isis 394816 at 20 mg/kg/wk or 50 mg/kg/wk. FIG. 5, Panel B, depicts inhibition of liver TG (mg/gram) in mice administered Isis 394816 at 20 mg/kg/wk or 50 mg/kg/wk.
FIG. 6 depicts the dose-dependent inhibition (relative units) of liver PCSK9 mRNA, plasma LDL-C, and liver TG in response to the indicated doses of Isis 394816.
FIG. 7 depicts a correlation between LDL-C lowering and PCSK9 mRNA inhibition, using the data from FIG. 6.
FIG. 8 depicts the accumulation of exemplary antisense compounds in rat tissues (m/g tissue) following administration of antisense compounds. The antisense compounds are identified by Isis number. The bars from left to right represent antisense compound accumulation in kidney and liver.
FIG. 9 depicts the accumulation of the identified antisense compounds in tissues (m/g tissue) following administration to a cyno monkey. Accumulation at each dosing concentration (15 mg/kg and 30 mg/kg) is shown in cyno kidney (left side bar) and cyno liver (right side bar).
FIG. 10 depicts PCSK9 mRNA levels (relative units) following administration of the identified antisense compounds to a cyno monkey. Results are shown at each dosing concentration (15 mg/kg and 30 mg/kg).
FIG. 11 depicts hepatic PCSK9 mRNA levels (relative units) detected by Northern analysis. Cyno monkeys were administered Isis 405879 (“BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg.
FIG. 12 depicts plasma LDL-C levels (mg/dL) following administration of Isis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg. Left side bars represent baseline levels of LDL-C in control animals; right side bars represent LDL-C following administration of antisense compounds.
FIG. 13 depicts the ratio of plasma HDL/LDL following administration of Isis 405879 (labeled “BMS-844421”) at a dose of 15 mg/kg or 30 mg/kg. Left side bars represent baseline HDL/LDL in control animals; right side bars represent HDL/LDL following administration of antisense compounds.
FIG. 14 depicts the effect of BMS-844421 and control ASOs on liver human PCSK9 mRNA in transgenic mice (6 wk).
FIG. 15 depicts the effect of BMS-844421 and control ASOs on plasma levels of human PCSK9 protein in transgenic mice (6 wk).
FIG. 16 depicts plasma hPCSK9 at baseline (FIG. 16A), 3 wk (FIG. 16B), and 6 wk (FIG. 16C) treatment.
FIG. 17 depicts the BMS-844421 dose response for plasma hPCSK9 levels in transgenic mice at 6 weeks (wk).
FIG. 18 depicts the BMS-844421 dose response for liver mRNA and plasma protein (hPCSK9).
FIG. 19 depicts the dose response for plasma hPCSK9 in male and female transgenic mice.
FIG. 20 depicts the dose response for liver hPCSK9 mRNA in male versus female transgenic mice.
FIG. 21 depicts the effect of the ASOs on liver endogenous PCSK9 mRNA in the transgenic mouse after 6 weeks of administration. The numbers following the hyphens provide the mg ASO/kg body weight dose of each ASO. For example, “844421-30” is the data for BMS-84421 administered to test animals at 30 mg/kg.
FIG. 22 depicts the effect of ASOs on liver endogenous HMGCoA reductase mRNA in the transgenic mouse after 6 weeks of administration.
FIG. 23 depicts LDLR protein concentration following a 6 wk ASO treatment, as assayed by a Western blot.
FIG. 24 depicts the quantification of the Western blot shown in FIG. 23. The LDLR band is normalized to a transferring receptor (TR) band, as observed using anti-LDLR and anti-TR antibodies, respectively.
FIG. 25 depicts the Liver PCSK9 mRNA concentration in cyno monkeys after 13 weeks of treatment. The numbers in parentheses are administered doses in mg/kg.
FIG. 26 depicts the concentration of BMS-844421 in the liver of cyno monkeys after 13 weeks of treatment.
FIG. 27 depicts the average serum LDL-C levels in the cyno monkey study. The numbers in parentheses are administered doses in mg/kg.
FIG. 28A depicts the pharmacodynamic responses in serum lipid assays for the cyno 2.5 and 5 wk ASO studies, as displayed in a scatterplot of individual animal data. FIG. 28B depicts the average ratio of HDL/LDL in BMS-844421-treated cynos at 2.5 and 5 wk. The numbers in parentheses in FIG. 28B are administered doses in mg/kg.
FIG. 29 depicts downregulation of PCSK9 target in cyno monkeys at 13 wk in the toxicology study of liver mRNA and plasma PCSK9.



DETAILED DESCRIPTION
Overview
Elevated levels of LDL-cholesterol (HDL-C) are recognized as a major independent risk factor for coronary heart disease (CHD). Even in individuals undergoing aggressive treatment with currently available cholesterol-lowering agents to reduce LDL-cholesterol (LDL-C) levels, coronary events still occur, and elevated LDL-C levels remain a major risk factor for coronary heart disease in these individuals. Furthermore, many individuals undergoing LDL-lowering therapy do not reach their target LDL-C levels, and thus remain at risk for CHD. Accordingly, there is a need for additional LDL-C lowering agents.
The antisense compounds and methods provided herein are useful for the treatment of hypercholesterolemia. Treatment of hypercholesterolemia encompasses a therapeutic regimen that results in a clinically desirable outcome. For example, the antisense compounds and methods provided herein are useful for the treatment of elevated cholesterol, such as elevated LDL-C. In addition, the antisense compounds and methods provided herein may be used to reduce the risk of CHD, in individuals exhibiting one or more risk factors for CHD. Furthermore, the antisense compounds and methods provided herein may be used to treat and/or prevent atherosclerosis.
As illustrated herein, administration of an antisense oligonucleotide targeted to PCSK9 to animals fed a high-fat diet (an experimental model of hyperlipidemia) resulted in antisense inhibition of PCSK9, upregulation of the LDL-R, reduction of LDL-C levels, and reduction of liver triglycerides. Thus, it is demonstrated that in an experimental model of hyperlipidemia, antisense inhibition of PCSK9 results in lowering LDL-C levels. Accordingly, provided herein are methods for the treatment of reducing LDL-C levels through the administration of an antisense compound targeted to a PCSK9 nucleic acid. Increased LDL-C levels are considered a risk factor for CHD, and are also linked to atherosclerosis. Accordingly, also provided herein are methods for the reduction of CHD risk, and for the prevention and/or treatment of atherosclerosis. Also provided herein are methods for the treatment of conditions characterized by elevated liver triglycerides, such as hepatic steatosis.
In a particular embodiment, methods comprise the use of antisense compounds targeted to particularly advantageous sequences within a PCSK9 nucleic acid. The PCSK9 target sequences are selected on the basis of superior results shown in one or more selection criteria. The presently disclosed antisense compounds all exhibit high in vitro efficacy, determined using a battery of cell models to measure antisense regulation of PCSK9 expression. Further, preferred antisense compounds exhibit relatively high in vivo efficacy, determined using a transgenic mouse model that expresses human PCSK9 or a cynomolgus (“cyno”) monkey model. The antisense compound ISIS 405879 (SEQ ID NO: 248) displays superior results in suppressing PCSK9 mRNA expression in the liver of a host individual.
The present antisense compounds are tolerated to different extents when administered to a host individual. Increased tolerability can depend on a number of factors, including, but not limited to, the nucleotide sequence of the antisense compound, chemical modifications to the nucleotides, the particular motif of unmodified and modified nucleosides in the antisense compound, or combinations thereof. Antisense compounds that exhibit increased or superior tolerability are particularly preferred in the present methods. The present antisense compounds also can show different pharmacokinetic properties, depending on their nucleotide sequence, chemical modifications, motifs, or combinations thereof. Particularly preferred antisense compounds also exhibit favorable pharmacokinetics when administered to a host individual.
Certain Indications
In certain embodiments, the invention provides methods of treating an individual comprising administering one or more pharmaceutical compositions of the present invention. In certain embodiments, the individual has hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, acute coronary syndrome, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.
Guidelines for lipid-lowering therapy were established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). The guidelines include obtaining a complete lipoprotein profile, typically after a 9 to 12 hour fast, for determination of LDL-C, total cholesterol, and HDL-C levels. According to the most recently established guidelines, LDL-C levels of 130-159 mg/dL, 160-189 mg/dL, and greater than or equal to 190 mg/dL are considered borderline high, high, and very high, respectively. Total cholesterol levels of 200-239 and greater than or equal to 240 mg/dL are considered borderline high and high, respectively. HDL-C levels of less than 40 mg/dL are considered low.
In certain embodiments, the individual has been identified as in need of lipid-lowering therapy. In certain such embodiments, the individual has been identified as in need of lipid-lowering therapy according to the guidelines established in 2001 by Adult Treatment Panel III (ATP III) of the National Cholesterol Education Program (NCEP), and updated in 2004 (Grundy et al., Circulation, 2004, 110, 227-239). In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 190 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 130 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy has LDL-C above 100 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 160 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 130 mg/dL. In certain such embodiments, the individual in need of lipid-lowering therapy should maintain LDL-C below 100 mg/dL. In certain such embodiments, the individual should maintain LDL-C below 70 mg/dL or even below 50 mg/dL.
In certain embodiments, the invention provides methods for reducing ApoB in an individual. In certain embodiments, the invention provides methods for reducing ApoB-containing lipoprotein in an individual. In certain embodiments, the invention provides methods for reducing LDL-C in an individual. In certain embodiments, the invention provides methods for reducing VLDL-C in an individual. In certain embodiments, the invention provides methods for reducing IDL-C in an individual. In certain embodiments, the invention provides methods for reducing non-HDL-C in an individual. In certain embodiments the invention provides methods for reducing Lp(a) in an individual. In certain embodiments, the invention provides methods for reducing serum triglyceride in an individual. In certain embodiments, the invention provides methods for reducing liver triglyceride in an individual. In certain embodiments, the invention provides methods for reducing Ox-LDL-C in an individual. In certain embodiments, the invention provides methods for reducing small LDL particles in an individual. In certain embodiments, the invention provides methods for reducing small VLDL particles in an individual. In certain embodiments, the invention provides methods for reducing phospholipids in an individual. In certain embodiments, the invention provides methods for reducing oxidized phospholipids in an individual.
In certain embodiments, the methods provided by the present invention do not lower HDL-C. In certain embodiments, the methods provided by the present invention do not result in accumulation of lipids in the liver.
In one embodiment are methods for decreasing LDL-C levels, or alternatively methods for treating hypercholesterolemia, by administering to an individual suffering from elevated LDL-C levels a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid. In another embodiment, a method of decreasing LDL-C levels comprises selecting an individual in need of a decrease in LDL-C levels, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid. In a further embodiment, a method of reducing coronary heart disease risk includes selecting an individual having elevated LDL-C levels and one or more additional indicators of coronary heart disease risk, and administering to the individual a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid.
In other embodiments, the LDL-C level is from 100-129 mg/dL, from 130 to 159 mg/dL, from 160-189 mg/dL, or greater than or equal to 190 mg/dL.
In one embodiment, administration of a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is accompanied by monitoring of LDL-C levels in the serum of an individual, to determine an individual's response to administration of the antisense compound. An individual's response to administration of the antisense compound is used by a physician to determine the amount and duration of therapeutic intervention.
In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid results in LDL-C levels below 190 mg/dL, below 160 mg/dL, below 130 mg/dL, below 100 mg/dL, below 70 mg/dL, or below 50 mg/dL. In another embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid decreases LDL-C by at least 15%, by at least 25%, by at least 50%, by at least 60%, by at least 70%, by at least 75%, by at least 80%, by at least 85%, by at least 90%, or by at least 95%.
An individual having elevated LDL-C levels may also exhibit reduced HDL-C levels and/or elevated total cholesterol levels. Accordingly, in one embodiment a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having elevated LDL-C levels, who also has reduced HDL-C levels and/or elevated total cholesterol levels.
Individuals having elevated LDL-C levels may also exhibit elevated triglyceride levels. Accordingly, in one embodiment a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having elevated LDL-C levels, and also having elevated triglyceride levels.
Atherosclerosis can lead to coronary heart disease, stroke, or peripheral vascular disease. Elevated LDL-C levels are considered a risk factor in the development and progression of atherosclerosis. Accordingly, in one embodiment, a therapeutically effective amount of an antisense compound targeted to a PCSK9 nucleic acid is administered to an individual having atherosclerosis. In a further embodiment, a therapeutically effective amount of antisense compound targeted to a PCSK9 nucleic acid is administered to an individual susceptible to atherosclerosis. Atherosclerosis is assessed directly through routine imaging techniques such as, for example, ultrasound imaging techniques that reveal carotid intimomedial thickness. Accordingly, treatment and/or prevention of atherosclerosis further include monitoring atherosclerosis through routine imaging techniques. In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid leads to a lessening of the severity of atherosclerosis, as indicated by, for example, a reduction of carotid intimomedial thickness in arteries.
Measurements of cholesterol, lipoproteins and triglycerides are obtained using serum or plasma collected from an individual. Methods of obtaining serum or plasma samples are routine, as are methods of preparation of the serum samples for analysis of cholesterol, triglycerides, and other serum markers.
A physician may determine the need for therapeutic intervention for individuals in cases where more or less aggressive LDL-lowering therapy is needed. The practice of the methods herein may be applied to any altered guidelines provided by the NCEP, or other entities that establish guidelines for physicians used in treating any of the diseases or conditions listed herein, for determining coronary heart disease risk and diagnosing metabolic syndrome.
In one embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid is parenteral administration. Parenteral administration may be intravenous or subcutaneous administration. Accordingly, in another embodiment, administration of an antisense compound targeted to a PCSK9 nucleic acid is intravenous or subcutaneous administration. Administration may include multiple doses of an antisense compound targeted to a PCSK9 nucleic acid.
In certain embodiments, a pharmaceutical composition comprising an antisense compound targeted to PCSK9 is for use in therapy. In certain embodiments, the therapy is the reduction of LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments, the therapy is the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, acute coronary syndrome, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease. In additional embodiments, the therapy is the reduction of CHD risk. In certain aspects, the therapy is prevention of atherosclerosis. In certain embodiments, the therapy is the prevention of coronary heart disease.
In certain embodiments, a pharmaceutical composition comprising an antisense compound targeted to PCSK9 is used for the preparation of a medicament for reducing LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp(a), serum triglyceride, liver triglyceride, Ox-LDL-C, small LDL particles, small VLDL, phospholipids, or oxidized phospholipids in an individual. In certain embodiments pharmaceutical composition comprising an antisense compound targeted to PCKS9 is used for the preparation of a medicament for reducing coronary heart disease risk. In certain embodiments, an antisense compound targeted to PCSK9 is used for the preparation of a medicament for the treatment of hypercholesterolemia, mixed dyslipidemia, atherosclerosis, a risk of developing atherosclerosis, coronary heart disease, a history of coronary heart disease, early onset coronary heart disease, one or more risk factors for coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipidemia, hyperfattyacidemia, hepatic steatosis, non-alcoholic steatohepatitis, or non-alcoholic fatty liver disease.
Certain Combination Therapies
In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with one or more other pharmaceutical agents. In certain embodiments, such one or more other pharmaceutical agents are designed to treat the same disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat a different disease or condition as the one or more pharmaceutical compositions of the present invention. In certain embodiments, such one or more other pharmaceutical agents are designed to treat an undesired effect of one or more pharmaceutical compositions of the present invention. In certain embodiments, one or more pharmaceutical compositions of the present invention are co-administered with another pharmaceutical agent to treat an undesired effect of that other pharmaceutical agent. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at the same time. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are administered at different times. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared together in a single formulation. In certain embodiments, one or more pharmaceutical compositions of the present invention and one or more other pharmaceutical agents are prepared separately. For example, a composition may comprise a pharmaceutical agent for separate, sequential, or simultaneous administration with an antisense compound.
In certain embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include lipid-lowering agents or LXR agonists. In certain such embodiments, pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to atorvastatin, simvastatin, rosuvastatin, and ezetimibe. In certain such embodiments, the lipid-lowering agent is administered prior to administration of a pharmaceutical composition of the present invention. In certain such embodiments, the lipid-lowering agent is administered following administration of a pharmaceutical composition of the present invention. In certain such embodiments the lipid-lowering agent is administered at the same time as a pharmaceutical composition of the present invention. In certain such embodiments the dose of a co-administered lipid-lowering agent is the same as the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is lower than the dose that would be administered if the lipid-lowering agent was administered alone. In certain such embodiments the dose of a co-administered lipid-lowering agent is greater than the dose that would be administered if the lipid-lowering agent was administered alone.
In certain embodiments, a co-administered lipid-lowering agent is a HMG-CoA reductase inhibitor. In certain such embodiments the HMG-CoA reductase inhibitor is a statin. In certain such embodiments, the statin is selected from, for example, atorvastatin, simvastatin, pravastatin, fluvastatin, and rosuvastatin.
In certain embodiments, a co-administered lipid-lowering agent is a cholesterol absorption inhibitor. In certain such embodiments, cholesterol absorption inhibitor is ezetimibe.
In certain embodiments, a co-administered lipid-lowering agent is a co-formulated HMG-CoA reductase inhibitor and cholesterol absorption inhibitor. In certain such embodiments the co-formulated lipid-lowering agent is ezetimibe/simvastatin.
In certain embodiments, a co-administered lipid-lowering agent is a microsomal triglyceride transfer protein inhibitor (MTP inhibitor).
In certain embodiments, a co-administered lipid-lowering agent is an oligonucleotide targeted to ApoB.
In certain embodiments, a co-administered pharmaceutical agent is a bile acid sequestrant. In certain such embodiments, the bile acid sequestrant is selected from cholestyramine, colestipol, and colesevelam.
In certain embodiments, a co-administered pharmaceutical agent is a nicotinic acid. In certain such embodiments, the nicotinic acid is selected from immediate release nicotinic acid, extended release nicotinic acid, and sustained release nicotinic acid.
In certain embodiments, a co-administered pharmaceutical agent is a fibric acid. In certain such embodiments, a fibric acid is selected from gemfibrozil, fenofibrate, clofibrate, bezafibrate, and ciprofibrate.
Further examples of pharmaceutical agents that may be co-administered with a pharmaceutical composition of the present invention include, but are not limited to, corticosteroids, including but not limited to prednisone; LXR agonists; immunoglobulins, including, but not limited to intravenous immunoglobulin (IVIg); analgesics (e.g., acetaminophen); anti-inflammatory agents, including, but not limited to non-steroidal anti-inflammatory drugs (e.g., ibuprofen, COX-1 inhibitors, and COX-2, inhibitors); salicylates; antibiotics; antivirals; antifungal agents; antidiabetic agents (e.g., biguanides, glucosidase inhibitors, insulins, sulfonylureas, and thiazolidenediones); adrenergic modifiers; diuretics; hormones (e.g., anabolic steroids, androgen, estrogen, calcitonin, progestin, somatostan, and thyroid hormones); immunomodulators; muscle relaxants; antihistamines; osteoporosis agents (e.g., biphosphonates, calcitonin, and estrogens); prostaglandins, antineoplastic agents; psychotherapeutic agents; sedatives; poison oak or poison sumac products; antibodies; and vaccines.
In certain embodiments, the pharmaceutical compositions of the present invention may be administered in conjunction with a lipid-lowering therapy. In certain such embodiments, a lipid-lowering therapy is therapeutic lifestyle change. In certain such embodiments, a lipid-lowering therapy is LDL apheresis.
Antisense Compounds
Oligomeric compounds include, but are not limited to, oligonucleotides, oligonucleosides, oligonucleotide analogs, oligonucleotide mimetics, antisense compounds, antisense oligonucleotides, and siRNAs. An oligomeric compound may be “antisense” to a target nucleic acid, meaning that is capable of undergoing hybridization to a target nucleic acid through hydrogen bonding.
In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted. In certain such embodiments, an antisense oligonucleotide has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 8 to 80, 12 to 50, 12 to 30 or 15 to 30 subunits in length. In other words, antisense compounds are from 8 to 80, 12 to 50, 12 to 30 or 15 to 30 linked subunits. In certain such embodiments, the antisense compounds are 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 subunits in length.
In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 12 to 30 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.
In certain embodiment, an antisense compound targeted to a PCSK9 nucleic acid is 15 to 30 subunits in length. In other words, antisense compounds are from 15 to 30 linked subunits. In certain such embodiments, the antisense compounds are 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 subunits in length.
In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 15 to 30 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 18 to 24 subunits in length. In other words, antisense compounds are from 18 to 24 linked subunits. In one embodiment, the antisense compounds are 18, 19, 20, 21, 22, 23, or 24 subunits in length.
In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 18 to 24 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 18, 19, 20, 21, 22, 23, or 24 nucleotides in length.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 19 to 22 subunits in length. In other words, antisense compounds are from 19 to 22 linked subunits. This embodies antisense compounds of 19, 20, 21, or 22 subunits in length.
In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 19 to 22 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 19, 20, 21, or 22 nucleotides in length.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid is 20 subunits in length. In certain such embodiments, antisense compounds are 20 linked subunits in length.
In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid is 20 nucleotides in length. In certain such embodiments, an antisense oligonucleotide targeted to an PCSK9 nucleic acid is 20 linked nucleotides in length.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence target a region identified herein.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, 3727-3751, or 3798-3824.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, 3727-3751, or 3798-3824.
In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid may target the following nucleotide regions of SEQ ID NO: 1: 320-405, 441-445, 527-544, 582-590, 744-781, 811-820, 918-922, 953-959, 1034-1036, 1152-1153, 1174-1199, 1251-1272, 1445-1464, 1603-1604, 1625-1627, 1707-1734, 1766-1811, 1832-1848, 1880-1904, 1956-1961, 1982-1939, 2030-2039, 2060-2099, 2140-2149, 2170-2186, 2207-2304, 2355-2409, 2435-2503, 2529-2581, 2626-2648, 2669-2749, 2770-2827, 2856-2876, 2891-2899, 2930-2982, 3014-3226, 3253-3436, 3457-3471, or 3497-3542.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 320-405, 441-445, 527-544, 582-590, 744-781, 811-820, 918-922, 953-959, 1034-1036, 1152-1153, 1174-1199, 1251-1272, 1445-1464, 1603-1604, 1625-1627, 1707-1734, 1766-1811, 1832-1848, 1880-1904, 1956-1961, 1982-1939, 2030-2039, 2060-2099, 2140-2149, 2170-2186, 2207-2304, 2355-2409, 2435-2503, 2529-2581, 2626-2648, 2669-2749, 2770-2827, 2856-2876, 2891-2899, 2930-2982, 3014-3226, 3253-3436, 3457-3471, or 3497-3542.
In certain embodiments, a shortened or truncated antisense compound targeted to a PCSK9 nucleic acid has a single subunit deleted from the 5′ end (5′ truncation), or alternatively from the 3′ end (3′ truncation). A shortened or truncated antisense compound targeted to a PCSK9 nucleic acid may have two subunits deleted from the 5′ end, or alternatively may have two subunits deleted from the 3′ end, of the antisense compound. Alternatively, the deleted nucleosides may be dispersed throughout the antisense compound, for example, in an antisense compound having one nucleoside deleted from the 5′ end and one nucleoside deleted from the 3′ end.
When a single additional subunit is present in a lengthened antisense compound, the additional subunit may be located at the 5′ or 3′ end of the antisense compound. When two are more additional subunits are present, the added subunits may be adjacent to each other, for example, in an antisense compound having two subunits added to the 5′ end (5′ addition), or alternatively to the 3′ end (3′ addition), of the antisense compound. Alternatively, the added subunits may be dispersed throughout the antisense compound, for example, in an antisense compound having one subunit added to the 5′ end and one subunit added to the 3′ end.
It is possible to increase or decrease the length of an antisense compound, such as an antisense oligonucleotide, and/or introduce mismatch bases without eliminating activity. For example, in Woolf et al. (Proc. Natl. Acad. Sci. USA 89:7305-7309, 1992), a series of antisense oligonucleotides 13-25 nucleobases in length were tested for their ability to induce cleavage of a target RNA in an oocyte injection model. Antisense oligonucleotides 25 nucleobases in length with 8 or 11 mismatch bases near the ends of the antisense oligonucleotides were able to direct specific cleavage of the target mRNA, albeit to a lesser extent than the antisense oligonucleotides that contained no mismatches. Similarly, target specific cleavage was achieved using 13 nucleobase antisense oligonucleotides, including those with 1 or 3 mismatches.
Gautschi et al (J. Natl. Cancer Inst. 93:463-471, March 2001) demonstrated the ability of an oligonucleotide having 100% complementarity to the bcl-2 mRNA and having 3 mismatches to the bcl-xL mRNA to reduce the expression of both bcl-2 and bcl-xL in vitro and in vivo. Furthermore, this oligonucleotide demonstrated potent anti-tumor activity in vivo.
Maher and Dolnick (Nuc. Acid. Res. 16:3341-3358, 1988) tested a series of tandem 14 nucleobase antisense oligonucleotides, and a 28 and 42 nucleobase antisense oligonucleotides comprised of the sequence of two or three of the tandem antisense oligonucleotides, respectively, for their ability to arrest translation of human DHFR in a rabbit reticulocyte assay. Each of the three 14 nucleobase antisense oligonucleotides alone was able to inhibit translation, albeit at a more modest level than the 28 or 42 nucleobase antisense oligonucleotides.
PCT/US2007/068404 describes incorporation of chemically-modified high-affinity nucleotides into short antisense compounds about 8-16 nucleobases in length and that such compounds are useful in the reduction of target RNAs in animals with increased potency and improved therapeutic index.
In certain embodiments, antisense compounds targeted to a PCSK9 nucleic acid are short antisense compounds. In certain embodiments, such short antisense compounds are oligonucleotide compounds. In certain embodiments, such short antisense compounds are about 8 to 16, preferably 9 to 15, more preferably 9 to 14, more preferably 10 to 14 nucleotides in length and comprises a gap region flanked on each side by a wing, wherein each wing independently consists of 1 to 3 nucleotides. Preferred motifs include but are not limited to wing-deoxy gap-wing motifs selected from 3-10-3, 2-10-3, 2-10-2, 1-10-1, 2-8-2, 1-8-1, 3-6-3 or 1-6-1.
Antisense compounds targeted to a PCSK9 nucleic acid are synthesized in vitro and do not include antisense compositions of biological origin, or genetic vector constructs designed to direct the in vivo synthesis of antisense molecules.
In certain embodiments, an antisense compound is targeted to a region of a PCSK9 nucleic acid that does not contain a single nucleotide polymorphism (SNPs). In certain embodiments, an antisense compound is targeted to a region of a PCSK9 nucleic acid that does contain a single nucleotide polymorph (SNPs). A single nucleotide polymorphism refers to polymorphisms that are the result of a single nucleotide alteration or the existence of two or more alternative sequences which can be, for example, different allelic forms of a gene. A polymorphism may comprise one or more base changes including, for example, an insertion, a repeat, or a deletion. In certain embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid overlaps with a SNP at the following positions: 428, 432, 449, 996, 1011, 1044, 1317, 1565, 1617, 1618, 1671, 1711, 1722, 1836, 1911. In certain embodiments, the compounds provided herein that target a region of a PCSK9 nucleic acid that contains one or more SNPs will contain the appropriate base substitution, insertion, repeat or deletion such that the compound is fully complementary to the altered PCSK9 nucleic acid sequence.
A subgroup of these antisense compounds were selected for further characterization, based on the ability of the selected antisense compounds to inhibit PCSK9 expression selectively and effectively in cell culture assays, set forth in the Examples. The antisense compounds selected for further evaluation are listed in Table 1. The 5′ and 3′ boundaries of the PCSK9 target sequence are shown for each antisense compound, with reference to the nucleotide positions of SEQ ID NO: 1.
“Motif” in Table 1 refers to the use of chemically modified nucleosides at the 5′ and 3′ ends of the antisense compounds. In a “5-10-5” motif, for example, five chemical modified nucleosides at both the 5′ and 3′ ends flank ten unmodified nucleosides in the center of the antisense compound. As disclosed in more detail below, modified nucleosides may make the antisense compounds more resistant to nucleases, among other things, which generally improves the pharmacodynamic and pharmacokinetic properties of the antisense compounds when administered to an individual.
Table 1 further discloses the nucleobase sequence of the antisense compounds. In certain embodiments, an antisense compound has a nucleobase sequence that, when written in the 5′ to 3′ direction, comprises the reverse complement of the target segment of a target nucleic acid to which it is targeted.












TABLE 1






5′
3′


SEQ


Isis
Target
Target


ID


No.
Site
Site
Sequence 5′-3′
Motif
NO







405881
1005
1024
CACCCTTGGCCACGCCGGCA
5-10-5
250


 


399819
2509
2528
CCCACTCAAGGGCCAGGCCA
5-10-5
 65


 


395165
1004
1023
ACCCTTGGCCACGCCGGCAT
5-10-5
 28


 


405879
1002
1021
CCTTGGCCACGCCGGCATCC
5-10-5
248


 


406008
 602
 621
CTTGGTGAGGTATCCCCGGC
5-10-5
188


 


405891
1567
1586
TCCTCAGGGAACCAGGCCTC
5-10-5
352


 


395186
2100
2119
CTTTGCATTCCAGACCTGGG
5-10-5
 60


 


405988
1854
1873
GGCAGCACCTGGCAATGGCG
5-10-5
381


 


405994
1928
1947
GCAGTGGACACGGGTCCCCA
5-10-5
400


 


406023
 787
 806
TGGTATTCATCCGCCCGGTA
5-10-5
212


 


395187
2310
2329
GGCAGCAGATGGCAACGGCT
5-10-5
 62


 


395185
1920
1939
CACGGGTCCCCATGCTGGCC
5-10-5
 59


 


406033
 967
 986
CCTGCCAGGTGGGTGCCATG
5-10-5
237


 


405923
1295
1314
GGCATTGGTGGCCCCAACTG
5-10-5
288


 


399900
1569
1588
GGTCCTCAGGGAACCAGGCC
3-14-3
 50


 


405995
1930
1949
TGGCAGTGGACACGGGTCCC
5-10-5
402


 


405991
1922
1941
GACACGGGTCCCCATGCTGG
5-10-5
394


 


406005
 559
 578
CGGGCAGTGCGCTCTGACTG
5-10-5
180


 


399793
 417
 436
CCTCGGAACGCAAGGCTAGC
5-10-5
  8


 


395152
 410
 429
ACGCAAGGCTAGCACCAGCT
5-10-5
  7









Antisense Compound Motifs

In certain embodiments, antisense compounds targeted to a PCSK9 nucleic acid have chemically modified subunits arranged in patterns, or motifs, to confer to the antisense compounds properties such as enhanced the inhibitory activity, increased binding affinity for a target nucleic acid, or resistance to degradation by in vivo nucleases.
Chimeric antisense compounds typically contain at least one region modified so as to confer increased resistance to nuclease degradation, increased cellular uptake, increased binding affinity for the target nucleic acid, and/or increased inhibitory activity. A second region of a chimeric antisense compound may serve as a substrate for the cellular endonuclease RNase H, which cleaves the RNA strand of an RNA:DNA duplex.
Antisense compounds having a gapmer motif are considered chimeric antisense compounds. In a gapmer an internal position having a plurality of nucleotides that supports RNaseH cleavage is positioned between external regions having a plurality of nucleotides that are chemically distinct from the nucleosides of the internal region. In the case of an antisense oligonucleotide having a gapmer motif, the gap segment generally serves as the substrate for endonuclease cleavage, while the wing segments comprise modified nucleosides. The regions of a gapmer are differentiated by the types of sugar moieties comprising each distinct region. The types of sugar moieties that are used to differentiate the regions of a gapmer may in some embodiments include β-D-ribonucleosides, β-D-deoxyribonucleosides, 2′-modified nucleosides (such 2′-modified nucleosides may include 2′-MOE, and 2′-O—CH3, among others), and bicyclic sugar modified nucleosides (such bicyclic sugar modified nucleosides may include those having a 4′-(CH2)n-O-2′ bridge, where n=1 or n=2). In general, each distinct region comprises uniform sugar moieties. The wing-gap-wing motif is frequently described as “X—Y—Z”, where “X” represents the length of the 5′ wing region, “Y” represents the length of the gap region, and “Z” represents the length of the 3′ wing region.
In some embodiments, an antisense compound targeted to a PCSK9 nucleic acid has a gap-widened motif. In other embodiments, an antisense oligonucleotide targeted to a PCSK9 nucleic acid has a gap-widened motif.
The gap-widened antisense oligonucleotides described herein may have various wing-gap-wing motifs selected from: 1-16-1, 2-15-1, 1-15-2, 1-14-3, 3-14-1, 2-14-2, 1-13-4, 4-13-1, 2-13-3, 3-13-2, 1-12-5, 5-12-1, 2-12-4, 4-12-2, 3-12-3, 1-11-6, 6-11-1, 2-11-5, 5-11-2, 3-11-4, 4-11-3, 1-17-1, 2-16-1, 1-16-2, 1-15-3, 3-15-1, 2-15-2, 1-14-4, 4-14-1, 2-14-3, 3-14-2, 1-13-5, 5-13-1, 2-13-4, 4-13-2, 3-13-3, 1-12-6, 6-12-1, 2-12-5, 5-12-2, 3-12-4, 4-12-3, 1-11-7, 7-11-1, 2-11-6, 6-11-2, 3-11-5, 5-11-3, 4-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 1-16-3, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 5-14-1, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4, 1-18-1, 1-17-2, 2-17-1, 1-16-3, 3-16-1, 2-16-2, 1-15-4, 4-15-1, 2-15-3, 3-15-2, 1-14-5, 2-14-4, 4-14-2, 3-14-3, 1-13-6, 6-13-1, 2-13-5, 5-13-2, 3-13-4, 4-13-3, 1-12-7, 7-12-1, 2-12-6, 6-12-2, 3-12-5, 5-12-3, 4-12-4, 1-11-8, 8-11-1, 2-11-7, 7-11-2, 3-11-6, 6-11-3, 4-11-5, 5-11-4, 1-19-1, 1-18-2, 2-18-1, 1-17-3, 3-17-1, 2-17-2, 1-16-4, 4-16-1, 2-16-3, 3-16-2, 1-15-5, 2-15-4, 4-15-2, 3-15-3, 1-14-6, 6-14-1, 2-14-5, 5-14-2, 3-14-4, 4-14-3, 1-13-7, 7-13-1, 2-13-6, 6-13-2, 3-13-5, 5-13-3, 4-13-4, 1-12-8, 8-12-1, 2-12-7, 7-12-2, 3-12-6, 6-12-3, 4-12-5, 5-12-4, 2-11-8, 8-11-2, 3-11-7, 7-11-3, 4-11-6, 6-11-4, 5-11-5, 1-20-1, 1-19-2, 2-19-1, 1-18-3, 3-18-1, 2-18-2, 1-17-4, 4-17-1, 2-17-3, 3-17-2, 1-16-5, 2-16-4, 4-16-2, 3-16-3, 1-15-6, 6-15-1, 2-15-5, 5-15-2, 3-15-4, 4-15-3, 1-14-7, 7-14-1, 2-14-6, 6-14-2, 3-14-5, 5-14-3, 4-14-4, 1-13-8, 8-13-1, 2-13-7, 7-13-2, 3-13-6, 6-13-3, 4-13-5, 5-13-4, 2-12-8, 8-12-2, 3-12-7, 7-12-3, 4-12-6, 6-12-4, 5-12-5, 3-11-8, 8-11-3, 4-11-7, 7-11-4, 5-11-6, 6-11-5, 1-21-1, 1-20-2, 2-20-1, 1-20-3, 3-19-1, 2-19-2, 1-18-4, 4-18-1, 2-18-3, 3-18-2, 1-17-5, 2-17-4, 4-17-2, 3-17-3, 1-16-6, 6-16-1, 2-16-5, 5-16-2, 3-16-4, 4-16-3, 1-15-7, 7-15-1, 2-15-6, 6-15-2, 3-15-5, 5-15-3, 4-15-4, 1-14-8, 8-14-1, 2-14-7, 7-14-2, 3-14-6, 6-14-3, 4-14-5, 5-14-4, 2-13-8, 8-13-2, 3-13-7, 7-13-3, 4-13-6, 6-13-4, 5-13-5, 1-12-10, 10-12-1, 2-12-9, 9-12-2, 3-12-8, 8-12-3, 4-12-7, 7-12-4, 5-12-6, 6-12-5, 4-11-8, 8-11-4, 5-11-7, 7-11-5, 6-11-6, 1-22-1, 1-21-2, 2-21-1, 1-21-3, 3-20-1, 2-20-2, 1-19-4, 4-19-1, 2-19-3, 3-19-2, 1-18-5, 2-18-4, 4-18-2, 3-18-3, 1-17-6, 6-17-1, 2-17-5, 5-17-2, 3-17-4, 4-17-3, 1-16-7, 7-16-1, 2-16-6, 6-16-2, 3-16-5, 5-16-3, 4-16-4, 1-15-8, 8-15-1, 2-15-7, 7-15-2, 3-15-6, 6-15-3, 4-15-5, 5-15-4, 2-14-8, 8-14-2, 3-14-7, 7-14-3, 4-14-6, 6-14-4, 5-14-5, 3-13-8, 8-13-3, 4-13-7, 7-13-4, 5-13-6, 6-13-5, 4-12-8, 8-12-4, 5-12-7, 7-12-5, 6-12-6, 5-11-8, 8-11-5, 6-11-7, or 7-11-6. In certain preferred embodiments, a gap-widened motif includes, but is not limited to, 2-13-5, 3-14-3, 3-14-4 gapmer motif.
In one embodiment, a gap-widened antisense oligonucleotide targeted to a PCSK9 nucleic acid has a gap segment of fourteen 2′-deoxyribonucleotides positioned between wing segments of three chemically modified nucleosides. In one embodiment, the chemical modification comprises a 2′-sugar modification. In another embodiment, the chemical modification comprises a 2′-MOE sugar modification.
In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid possess a 5-10-5 gapmer motif.
Target Nucleic Acids, Target Regions and Nucleotide Sequences
Nucleotide sequences that encode PCSK9 include, without limitation, the following: GENBANK® Accession No. NM_174936.2, first deposited with GENBANK® on Jun. 1, 2003, and incorporated herein as SEQ ID NO: 1; nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT_032977.8, first deposited with GENBANK® on Feb. 26, 2006, and incorporated herein as SEQ ID NO: 2; and GENBANK® Accession No. AK124635.1, first deposited with GENBANK® on Sep. 8, 2003, and incorporated herein as SEQ ID NO: 3.
It is noted that some portions of these nucleotide sequences share identical sequence. For example, portions of SEQ ID NO: 1 are identical to portions of SEQ ID NO: 2; portions of SEQ ID NO: 1 are identical to portions of SEQ ID NO: 3; and portions of SEQ ID NO: 2 are identical to portions of SEQ ID NO: 3. Accordingly, antisense compounds targeted to SEQ ID NO: 1 may also target SEQ ID NO: 2 and/or SEQ ID NO: 3; antisense compounds targeted to SEQ ID NO: 2 may also target SEQ ID NO: 1 and/or SEQ ID NO: 3; and antisense compounds targeted to SEQ ID NO: 3 may also target SEQ ID NO: 1 and/or SEQ ID NO: 2. Examples of such antisense compounds are shown in the following tables.
In certain embodiments, antisense compounds target a PCSK9 nucleic acid having the sequence of GENBANK® Accession No. NM_174936.2, first deposited with GENBANK® on Jun. 1, 2003, and incorporated herein as SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide targets SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is at least 90% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is at least 95% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. In certain embodiments, an antisense oligonucleotide targeted to SEQ ID NO: 1 comprises a nucleotide sequence selected from the nucleotide sequences set forth in Table 2.







TABLE 2







Nucleotide sequences targeted to 


NM_174936.2 (SEQ ID NO: 1)











5′
3′




Target
Target



SEQ
Site on
Site on



ID
SEQ ID
SEQ ID



NO
NO: 1
NO: 1
Sequence (5′-3′)





  4
 135
 154
GCGCGGAATCCTGGCTGGGA


 


  5
 242
 261
GAGGAGACCTAGAGGCCGTG


 


159
 294
 313
GCCTGGAGCTGACGGTGCCC


 


160
 298
 317
GACCGCCTGGAGCTGACGGT


 


  6
 300
 319
AGGACCGCCTGGAGCTGACG


 


162
 406
 425
AAGGCTAGCACCAGCTCCTC


 


163
 407
 426
CAAGGCTAGCACCAGCTCCT


 


164
 408
 427
GCAAGGCTAGCACCAGCTCC


 


165
 409
 428
CGCAAGGCTAGCACCAGCTC


 


  7
 410
 429
ACGCAAGGCTAGCACCAGCT


 


166
 411
 430
AACGCAAGGCTAGCACCAGC


 


167
 412
 431
GAACGCAAGGCTAGCACCAG


 


168
 413
 432
GGAACGCAAGGCTAGCACCA


 


169
 414
 433
CGGAACGCAAGGCTAGCACC


 


  8
 417
 436
CCTCGGAACGCAAGGCTAGC


 


170
 421
 440
TCCTCCTCGGAACGCAAGGC


 


171
 446
 465
GTGCTCGGGTGCTTCGGCCA


 


172
 466
 485
TGGAAGGTGGCTGTGGTTCC


 


  9
 480
 499
CCTTGGCGCAGCGGTGGAAG


 


173
 482
 501
ATCCTTGGCGCAGCGGTGGA


 


174
 484
 503
GGATCCTTGGCGCAGCGGTG


 


175
 488
 507
CCACGGATCCTTGGCGCAGC


 


176
 507
 526
CGTAGGTGCCAGGCAACCTC


 


177
 545
 564
TGACTGCGAGAGGTGGGTCT


 


178
 555
 574
CAGTGCGCTCTGACTGCGAG


 


179
 557
 576
GGCAGTGCGCTCTGACTGCG


 


180
 559
 578
CGGGCAGTGCGCTCTGACTG


 


 10
 561
 580
GGCGGGCAGTGCGCTCTGAC


 


181
 562
 581
CGGCGGGCAGTGCGCTCTGA


 


182
 591
 610
ATCCCCGGCGGGCAGCCTGG


 


183
 595
 614
AGGTATCCCCGGCGGGCAGC


 


184
 597
 616
TGAGGTATCCCCGGCGGGCA


 


185
 598
 617
GTGAGGTATCCCCGGCGGGC


 


186
 599
 618
GGTGAGGTATCCCCGGCGGG


 


 11
 600
 619
TGGTGAGGTATCCCCGGCGG


 


187
 601
 620
TTGGTGAGGTATCCCCGGCG


 


188
 602
 621
CTTGGTGAGGTATCCCCGGC


 


189
 603
 622
TCTTGGTGAGGTATCCCCGG


 


190
 604
 623
ATCTTGGTGAGGTATCCCCG


 


191
 605
 624
GATCTTGGTGAGGTATCCCC


 


 12
 606
 625
GGATCTTGGTGAGGTATCCC


 


192
 607
 626
AGGATCTTGGTGAGGTATCC


 


193
 609
 628
GCAGGATCTTGGTGAGGTAT


 


194
 611
 630
ATGCAGGATCTTGGTGAGGT


 


195
 613
 632
ACATGCAGGATCTTGGTGAG


 


 13
 615
 634
AGACATGCAGGATCTTGGTG


 


196
 617
 636
GAAGACATGCAGGATCTTGG


 


 14
 620
 639
ATGGAAGACATGCAGGATCT


 


197
 628
 647
AGAAGGCCATGGAAGACATG


 


198
 638
 657
GAAGCCAGGAAGAAGGCCAT


 


 15
 646
 665
TTCACCAGGAAGCCAGGAAG


 


199
 648
 667
TCTTCACCAGGAAGCCAGGA


 


 16
 651
 670
TCATCTTCACCAGGAAGCCA


 


200
 653
 672
ACTCATCTTCACCAGGAAGC


 


201
 655
 674
CCACTCATCTTCACCAGGAA


 


202
 657
 676
CGCCACTCATCTTCACCAGG


 


203
 659
 678
GTCGCCACTCATCTTCACCA


 


204
 661
 680
AGGTCGCCACTCATCTTCAC


 


205
 663
 682
GCAGGTCGCCACTCATCTTC


 


206
 665
 684
CAGCAGGTCGCCACTCATCT


 


207
 667
 686
TCCAGCAGGTCGCCACTCAT


 


208
 685
 704
GGCAACTTCAAGGCCAGCTC


 


 17
 705
 724
CCTCGATGTAGTCGACATGG


 


209
 724
 743
GCAAAGACAGAGGAGTCCTC


 


210
 782
 801
TTCATCCGCCCGGTACCGTG


 


211
 784
 803
TATTCATCCGCCCGGTACCG


 


 18
 785
 804
GTATTCATCCGCCCGGTACC


 


212
 787
 806
TGGTATTCATCCGCCCGGTA


 


213
 789
 808
GCTGGTATTCATCCGCCCGG


 


214
 791
 810
GGGCTGGTATTCATCCGCCC


 


215
 821
 840
ATACACCTCCACCAGGCTGC


 


216
 832
 851
GTGTCTAGGAGATACACCTC


 


 19
 835
 854
CTGGTGTCTAGGAGATACAC


 


217
 837
 856
TGCTGGTGTCTAGGAGATAC


 


 20
 840
 859
GTATGCTGGTGTCTAGGAGA


 


 21
 860
 879
GATTTCCCGGTGGTCACTCT


 


218
 862
 881
TCGATTTCCCGGTGGTCACT


 


219
 863
 882
CTCGATTTCCCGGTGGTCAC


 


220
 864
 883
CCTCGATTTCCCGGTGGTCA


 


221
 865
 884
CCCTCGATTTCCCGGTGGTC


 


 22
 866
 885
GCCCTCGATTTCCCGGTGGT


 


222
 867
 886
TGCCCTCGATTTCCCGGTGG


 


223
 868
 887
CTGCCCTCGATTTCCCGGTG


 


224
 869
 888
CCTGCCCTCGATTTCCCGGT


 


225
 870
 889
CCCTGCCCTCGATTTCCCGG


 


226
 874
 893
ATGACCCTGCCCTCGATTTC


 


227
 876
 895
CCATGACCCTGCCCTCGATT


 


228
 878
 897
GACCATGACCCTGCCCTCGA


 


 23
 880
 899
GTGACCATGACCCTGCCCTC


 


229
 882
 901
CGGTGACCATGACCCTGCCC


 


230
 884
 903
GTCGGTGACCATGACCCTGC


 


231
 886
 905
AAGTCGGTGACCATGACCCT


 


232
 888
 907
CGAAGTCGGTGACCATGACC


 


 24
 890
 909
CTCGAAGTCGGTGACCATGA


 


233
 898
 917
GGCACATTCTCGAAGTCGGT


 


 25
 923
 942
GTGGAAGCGGGTCCCGTCCT


 


234
 933
 952
TGGCCTGTCTGTGGAAGCGG


 


235
 960
 979
GGTGGGTGCCATGACTGTCA


 


236
 963
 982
CCAGGTGGGTGCCATGACTG


 


237
 967
 986
CCTGCCAGGTGGGTGCCATG


 


 26
 970
 989
ACCCCTGCCAGGTGGGTGCC


 


238
 972
 991
CCACCCCTGCCAGGTGGGTG


 


 27
 975
 994
TGACCACCCCTGCCAGGTGG


 


239
 977
 996
GCTGACCACCCCTGCCAGGT


 


240
 985
1004
TCCCGGCCGCTGACCACCCC


 


241
 989
1008
GGCATCCCGGCCGCTGACCA


 


242
 992
1011
GCCGGCATCCCGGCCGCTGA


 


243
 997
1016
GCCACGCCGGCATCCCGGCC


 


244
 998
1017
GGCCACGCCGGCATCCCGGC


 


245
 999
1018
TGGCCACGCCGGCATCCCGG


 


246
1000
1019
TTGGCCACGCCGGCATCCCG


 


247
1001
1020
CTTGGCCACGCCGGCATCCC


 


248
1002
1021
CCTTGGCCACGCCGGCATCC


 


249
1003
1022
CCCTTGGCCACGCCGGCATC


 


 28
1004
1023
ACCCTTGGCCACGCCGGCAT


 


447
1004
1023
ACCCTTGGTCACGCCGGCAT


 


250
1005
1024
CACCCTTGGCCACGCCGGCA


 


251
1006
1025
GCACCCTTGGCCACGCCGGC


 


252
1007
1026
GGCACCCTTGGCCACGCCGG


 


253
1008
1027
TGGCACCCTTGGCCACGCCG


 


254
1009
1028
CTGGCACCCTTGGCCACGCC


 


255
1010
1029
GCTGGCACCCTTGGCCACGC


 


256
1015
1034
CGCATGCTGGCACCCTTGGC


 


257
1036
1055
CAGTTGAGCACGCGCAGGCT


 


258
1038
1057
GGCAGTTGAGCACGCGCAGG


 


 29
1040
1059
TTGGCAGTTGAGCACGCGCA


 


259
1042
1061
CCTTGGCAGTTGAGCACGCG


 


 30
1045
1064
TTCCCTTGGCAGTTGAGCAC


 


260
1047
1066
CCTTCCCTTGGCAGTTGAGC


 


261
1051
1070
GTGCCCTTCCCTTGGCAGTT


 


262
1053
1072
CCGTGCCCTTCCCTTGGCAG


 


263
1064
1083
GGTGCCGCTAACCGTGCCCT


 


264
1076
1095
CAGGCCTATGAGGGTGCCGC


 


 31
1077
1096
CCAGGCCTATGAGGGTGCCG


 


458
1079
1092
GCCTATGAGGGTGC


 


459
1084
1097
TCCAGGCCTATGAG


 


265
1088
1107
CCGAATAAACTCCAGGCCTA


 


266
1096
1115
TGGCTTTTCCGAATAAACTC


 


 32
1098
1117
GCTGGCTTTTCCGAATAAAC


 


267
1100
1119
CAGCTGGCTTTTCCGAATAA


 


268
1102
1121
ACCAGCTGGCTTTTCCGAAT


 


269
1104
1123
GGACCAGCTGGCTTTTCCGA


 


270
1108
1127
GGCTGGACCAGCTGGCTTTT


 


271
1119
1138
GTGGCCCCACAGGCTGGACC


 


272
1132
1151
AGCAGCACCACCAGTGGCCC


 


273
1154
1173
GCTGTACCCACCCGCCAGGG


 


274
1200
1219
CGACCCCAGCCCTCGCCAGG


 


 33
1210
1229
GTGACCAGCACGACCCCAGC


 


275
1212
1231
CGGTGACCAGCACGACCCCA


 


276
1214
1233
AGCGGTGACCAGCACGACCC


 


277
1216
1235
GCAGCGGTGACCAGCACGAC


 


278
1218
1237
CGGCAGCGGTGACCAGCACG


 


279
1219
1238
CCGGCAGCGGTGACCAGCAC


 


280
1222
1241
TTGCCGGCAGCGGTGACCAG


 


281
1224
1243
AGTTGCCGGCAGCGGTGACC


 


282
1226
1245
GAAGTTGCCGGCAGCGGTGA


 


283
1228
1247
CGGAAGTTGCCGGCAGCGGT


 


284
1230
1249
CCCGGAAGTTGCCGGCAGCG


 


285
1232
1251
GTCCCGGAAGTTGCCGGCAG


 


286
1273
1292
ATGACCTCGGGAGCTGAGGC


 


287
1283
1302
CCCAACTGTGATGACCTCGG


 


288
1295
1314
GGCATTGGTGGCCCCAACTG


 


149
1297
1316
TGGGCATTGGTGGCCCCAAC


 


289
1305
1324
GCTGGTCTTGGGCATTGGTG


 


290
1318
1337
CCCAGGGTCACCGGCTGGTC


 


291
1320
1339
TCCCCAGGGTCACCGGCTGG


 


292
1322
1341
AGTCCCCAGGGTCACCGGCT


 


293
1324
1343
AAAGTCCCCAGGGTCACCGG


 


 34
1326
1345
CCAAAGTCCCCAGGGTCACC


 


294
1328
1347
CCCCAAAGTCCCCAGGGTCA


 


128
1330
1349
GTCCCCAAAGTCCCCAGGGT


 


295
1333
1352
TTGGTCCCCAAAGTCCCCAG


 


 35
1335
1354
AGTTGGTCCCCAAAGTCCCC


 


296
1337
p<
AAAGTTGGTCCCCAAAGTCC


 


 36
1340
1359
GCCAAAGTTGGTCCCCAAAG


 


297
1342
1361
CGGCCAAAGTTGGTCCCCAA


 


298
1344
1363
AGCGGCCAAAGTTGGTCCCC


 


299
1346
1365
ACAGCGGCCAAAGTTGGTCC


 


300
1348
1367
ACACAGCGGCCAAAGTTGGT


 


301
1350
1369
CCACACAGCGGCCAAAGTTG


 


 37
1352
1371
GTCCACACAGCGGCCAAAGT


 


302
1354
1373
AGGTCCACACAGCGGCCAAA


 


303
1356
1375
AGAGGTCCACACAGCGGCCA


 


304
1358
1377
AAAGAGGTCCACACAGCGGC


 


 38
1361
1380
GGCAAAGAGGTCCACACAGC


 


305
1380
1399
CAATGATGTCCTCCCCTGGG


 


306
1387
1406
GAGGCACCAATGATGTCCTC


 


 39
1389
1408
TGGAGGCACCAATGATGTCC


 


307
1391
1410
GCTGGAGGCACCAATGATGT


 


308
1393
1412
TCGCTGGAGGCACCAATGAT


 


309
1395
1414
AGTCGCTGGAGGCACCAATG


 


310
1397
1416
GCAGTCGCTGGAGGCACCAA


 


 40
1400
1419
GCTGCAGTCGCTGGAGGCAC


 


311
1402
1421
GTGCTGCAGTCGCTGGAGGC


 


312
1404
1423
AGGTGCTGCAGTCGCTGGAG


 


313
1406
1425
GCAGGTGCTGCAGTCGCTGG


 


314
1407
1426
AGCAGGTGCTGCAGTCGCTG


 


315
1409
1428
AAAGCAGGTGCTGCAGTCGC


 


 41
1411
1430
ACAAAGCAGGTGCTGCAGTC


 


316
1413
1432
ACACAAAGCAGGTGCTGCAG


 


317
1415
1434
TGACACAAAGCAGGTGCTGC


 


318
1425
1444
TCCCACTCTGTGACACAAAG


 


101
1465
1484
ATGGCTGCAATGCCAGCCAC


 


319
1467
1486
TCATGGCTGCAATGCCAGCC


 


 42
1470
1489
GCATCATGGCTGCAATGCCA


 


320
1472
1491
CAGCATCATGGCTGCAATGC


 


321
1474
1493
GACAGCATCATGGCTGCAAT


 


322
1476
1495
CAGACAGCATCATGGCTGCA


 


 43
1478
1497
GGCAGACAGCATCATGGCTG


 


323
1480
1499
TCGGCAGACAGCATCATGGC


 


324
1482
1501
GCTCGGCAGACAGCATCATG


 


325
1484
1503
CGGCTCGGCAGACAGCATCA


 


326
1486
1505
TCCGGCTCGGCAGACAGCAT


 


327
1500
1519
CGGCCAGGGTGAGCTCCGGC


 


328
1513
1532
CTCTGCCTCAACTCGGCCAG


 


329
1515
1534
GTCTCTGCCTCAACTCGGCC


 


330
1517
1536
CAGTCTCTGCCTCAACTCGG


 


331
1519
1538
ATCAGTCTCTGCCTCAACTC


 


332
1521
1540
GGATCAGTCTCTGCCTCAAC


 


333
1523
1542
GTGGATCAGTCTCTGCCTCA


 


334
1525
1544
AAGTGGATCAGTCTCTGCCT


 


 44
1526
1545
GAAGTGGATCAGTCTCTGCC


 


335
1528
1547
GAGAAGTGGATCAGTCTCTG


 


336
1530
1549
CAGAGAAGTGGATCAGTCTC


 


337
1532
1551
GGCAGAGAAGTGGATCAGTC


 


 45
1534
1553
TTGGCAGAGAAGTGGATCAG


 


338
1536
1555
CTTTGGCAGAGAAGTGGATC


 


 46
1539
1558
CATCTTTGGCAGAGAAGTGG


 


339
1541
1560
GACATCTTTGGCAGAGAAGT


 


340
1543
1562
ATGACATCTTTGGCAGAGAA


 


 47
1545
1564
TGATGACATCTTTGGCAGAG


 


341
1547
1566
ATTGATGACATCTTTGGCAG


 


342
1549
1568
TCATTGATGACATCTTTGGC


 


 48
1552
1571
GCCTCATTGATGACATCTTT


 


343
1554
1573
AGGCCTCATTGATGACATCT


 


344
1556
1575
CCAGGCCTCATTGATGACAT


 


345
1558
1577
AACCAGGCCTCATTGATGAC


 


346
1560
1579
GGAACCAGGCCTCATTGATG


 


347
1561
1580
GGGAACCAGGCCTCATTGAT


 


348
1562
1581
AGGGAACCAGGCCTCATTGA


 


349
1563
1582
CAGGGAACCAGGCCTCATTG


 


 49
1564
1583
TCAGGGAACCAGGCCTCATT


 


 49
1564
1583
TCAGGGAACCAGGCCTCATT


 


350
1565
1584
CTCAGGGAACCAGGCCTCAT


 


351
1566
1585
CCTCAGGGAACCAGGCCTCA


 


352
1567
1586
TCCTCAGGGAACCAGGCCTC


 


353
1568
1587
GTCCTCAGGGAACCAGGCCT


 


 50
1569
1588
GGTCCTCAGGGAACCAGGCC


 


354
1570
1589
TGGTCCTCAGGGAACCAGGC


 


355
1571
1590
CTGGTCCTCAGGGAACCAGG


 


356
1572
1591
GCTGGTCCTCAGGGAACCAG


 


357
1573
1592
CGCTGGTCCTCAGGGAACCA


 


 87
1576
1595
ACCCGCTGGTCCTCAGGGAA


 


358
1578
1597
GTACCCGCTGGTCCTCAGGG


 


359
1580
1599
CAGTACCCGCTGGTCCTCAG


 


 51
1583
1602
GGTCAGTACCCGCTGGTCCT


 


119
1605
1624
GCAGGGCGGCCACCAGGTTG


 


360
1628
1647
ACCTGCCCCATGGGTGCTGG


 


 52
1640
1659
AAACAGCTGCCAACCTGCCC


 


361
1642
1661
CAAAACAGCTGCCAACCTGC


 


 53
1645
1664
CTGCAAAACAGCTGCCAACC


 


362
1647
1666
TCCTGCAAAACAGCTGCCAA


 


363
1649
1668
AGTCCTGCAAAACAGCTGCC


 


364
1660
1679
GCTGACCATACAGTCCTGCA


 


365
1672
1691
GGCCCCGAGTGTGCTGACCA


 


 54
1675
1694
GTAGGCCCCGAGTGTGCTGA


 


366
1677
1696
GTGTAGGCCCCGAGTGTGCT


 


367
1679
1698
CCGTGTAGGCCCCGAGTGTG


 


368
1681
1700
ATCCGTGTAGGCCCCGAGTG


 


369
1683
1702
CCATCCGTGTAGGCCCCGAG


 


370
1685
1704
GGCCATCCGTGTAGGCCCCG


 


371
1687
1706
GTGGCCATCCGTGTAGGCCC


 


372
1735
1754
CTGGAGCAGCTCAGCAGCTC


 


460
1735
1748
CAGCTCAGCAGCTC


 


373
1737
1756
AACTGGAGCAGCTCAGCAGC


 


 55
1740
1759
AGAAACTGGAGCAGCTCAGC


 


374
1742
1761
GGAGAAACTGGAGCAGCTCA


 


375
1744
1763
CTGGAGAAACTGGAGCAGCT


 


 56
1746
1765
TCCTGGAGAAACTGGAGCAG


 


 57
1812
1831
CGTTGTGGGCCCGGCAGACC


 


376
1849
1868
CACCTGGCAATGGCGTAGAC


 


377
1850
1869
GCACCTGGCAATGGCGTAGA


 


378
1851
1870
AGCACCTGGCAATGGCGTAG


 


379
1852
1871
CAGCACCTGGCAATGGCGTA


 


380
1853
1872
GCAGCACCTGGCAATGGCGT


 


381
1854
1873
GGCAGCACCTGGCAATGGCG


 


382
1855
1874
AGGCAGCACCTGGCAATGGC


 


383
1856
1875
CAGGCAGCACCTGGCAATGG


 


384
1857
1876
GCAGGCAGCACCTGGCAATG


 


 58
1858
1877
AGCAGGCAGCACCTGGCAAT


 


385
1859
1878
TAGCAGGCAGCACCTGGCAA


 


386
1860
1879
GTAGCAGGCAGCACCTGGCA


 


387
1905
1924
TGGCCTCAGCTGGTGGAGCT


 


388
1915
1934
GTCCCCATGCTGGCCTCAGC


 


389
1916
1935
GGTCCCCATGCTGGCCTCAG


 


390
1917
1936
GGGTCCCCATGCTGGCCTCA


 


391
1918
1937
CGGGTCCCCATGCTGGCCTC


 


392
1919
1938
ACGGGTCCCCATGCTGGCCT


 


 59
1920
1939
CACGGGTCCCCATGCTGGCC


 


 59
1920
1939
CACGGGTCCCCATGCTGGCC


 


393
1921
1940
ACACGGGTCCCCATGCTGGC


 


394
1922
1941
GACACGGGTCCCCATGCTGG


 


395
1923
1942
GGACACGGGTCCCCATGCTG


 


396
1924
1943
TGGACACGGGTCCCCATGCT


 


397
1925
1944
GTGGACACGGGTCCCCATGC


 


398
1926
1945
AGTGGACACGGGTCCCCATG


 


399
1927
1946
CAGTGGACACGGGTCCCCAT


 


400
1928
1947
GCAGTGGACACGGGTCCCCA


 


401
1929
1948
GGCAGTGGACACGGGTCCCC


 


402
1930
1949
TGGCAGTGGACACGGGTCCC


 


403
1931
1950
GTGGCAGTGGACACGGGTCC


 


404
1932
1951
GGTGGCAGTGGACACGGGTC


 


405
1933
1952
TGGTGGCAGTGGACACGGGT


 


406
1936
1955
TGTTGGTGGCAGTGGACACG


 


407
1962
1981
AGCTGCAGCCTGTGAGGACG


 


408
1990
2009
GTGCCAAGGTCCTCCACCTC


 


409
2010
2029
TCAGCACAGGCGGCTTGTGG


 


410
2040
2059
CCACGCACTGGTTGGGCTGA


 


 60
2100
2119
CTTTGCATTCCAGACCTGGG


 


411
2101
2120
ACTTTGCATTCCAGACCTGG


 


412
2102
2121
GACTTTGCATTCCAGACCTG


 


413
2103
2122
TGACTTTGCATTCCAGACCT


 


414
2104
2123
TTGACTTTGCATTCCAGACC


 


 61
2105
2124
CTTGACTTTGCATTCCAGAC


 


415
2107
2126
TCCTTGACTTTGCATTCCAG


 


416
2120
2139
CGGGATTCCATGCTCCTTGA


 


417
2150
2169
GCAGGCCACGGTCACCTGCT


 


418
2187
2206
GGAGGGCACTGCAGCCAGTC


 


419
2305
2324
CAGATGGCAACGGCTGTCAC


 


420
2306
2325
GCAGATGGCAACGGCTGTCA


 


421
2307
2326
AGCAGATGGCAACGGCTGTC


 


422
2308
2327
CAGCAGATGGCAACGGCTGT


 


423
2309
2328
GCAGCAGATGGCAACGGCTG


 


 62
2310
2329
GGCAGCAGATGGCAACGGCT


 


424
2311
2330
CGGCAGCAGATGGCAACGGC


 


425
2312
2331
CCGGCAGCAGATGGCAACGG


 


426
2313
2332
TCCGGCAGCAGATGGCAACG


 


427
2314
2333
CTCCGGCAGCAGATGGCAAC


 


428
2315
2334
GCTCCGGCAGCAGATGGCAA


 


429
2325
2344
CCAGGTGCCGGCTCCGGCAG


 


430
2335
2354
GAGGCCTGCGCCAGGTGCCG


 


154
2410
2429
TTTTAAAGCTCAGCCCCAGC


 


 63
2415
2434
AACCATTTTAAAGCTCAGCC


 


 64
2504
2523
TCAAGGGCCAGGCCAGCAGC


 


 65
2509
2528
CCCACTCAAGGGCCAGGCCA


 


122
2582
2601
GGAGGGAGCTTCCTGGCACC


 


 66
2597
2616
ATGCCCCACAGTGAGGGAGG


 


 67
2606
2625
AATGGTGAAATGCCCCACAG


 


153
2649
2668
TTGGGAGCAGCTGGCAGCAC


 


 68
2750
2769
CATGGGAAGAATCCTGCCTC


 


431
2828
2847
ATGAGGGCCATCAGCACCTT


 


432
2829
2848
GATGAGGGCCATCAGCACCT


 


433
2830
2849
AGATGAGGGCCATCAGCACC


 


434
2831
2850
GAGATGAGGGCCATCAGCAC


 


 69
2832
2851
GGAGATGAGGGCCATCAGCA


 


435
2833
2852
TGGAGATGAGGGCCATCAGC


 


436
2834
2853
CTGGAGATGAGGGCCATCAG


 


437
2835
2854
GCTGGAGATGAGGGCCATCA


 


438
2836
2855
AGCTGGAGATGAGGGCCATC


 


461
2877
2890
TTAATCAGGGAGCC


 


 70
2900
2919
TAGATGCCATCCAGAAAGCT


 


439
2902
2921
GCTAGATGCCATCCAGAAAG


 


440
2903
2922
GGCTAGATGCCATCCAGAAA


 


441
2904
2923
TGGCTAGATGCCATCCAGAA


 


442
2905
2924
CTGGCTAGATGCCATCCAGA


 


 71
2906
2925
TCTGGCTAGATGCCATCCAG


 


443
2907
2926
CTCTGGCTAGATGCCATCCA


 


444
2908
2927
CCTCTGGCTAGATGCCATCC


 


445
2909
2928
GCCTCTGGCTAGATGCCATC


 


446
2910
2929
AGCCTCTGGCTAGATGCCAT


 


 72
2983
3002
GGCATAGAGCAGAGTAAAGG


 


 73
2988
3007
AGCCTGGCATAGAGCAGAGT


 


135
2994
3013
TAGCACAGCCTGGCATAGAG


 


112
3227
3246
GAAGAGGCTTGGCTTCAGAG


 


 74
3233
3252
AAGTAAGAAGAGGCTTGGCT


 


 75
3437
3456
GCTCAAGGAGGGACAGTTGT


 


 76
3472
3491
AAAGATAAATGTCTGCTTGC


 


 77
3477
3496
ACCCAAAAGATAAATGTCTG


 


 78
3543
3562
TCTTCAAGTTACAAAAGCAA


 


 99
3550
3569
ATAAATATCTTCAAGTTACA









In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 5-10-5 gapmer motif. Table 3 illustrates gapmer antisense compounds targeted to SEQ ID NO: 1, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 3







Gapmer antisense compounds having a 5-10-5 motif targeted


to SEQ ID NO: 1
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 1
NO: 1
Mismatches
Sequence (5′-3′)





395149
5-10-5
  4
 135
 154
0
GCGCGGAATCCTGGCTGGGA


 


395150
5-10-5
  5
 242
 261
0
GAGGAGACCTAGAGGCCGTG


 


395151
5-10-5
  6
 300
 319
0
AGGACCGCCTGGAGCTGACG


 


395152
5-10-5
  7
 410
 429
0
ACGCAAGGCTAGCACCAGCT


 


399793
5-10-5
  8
 417
 436
0
CCTCGGAACGCAAGGCTAGC


 


395153
5-10-5
  9
 480
 499
0
CCTTGGCGCAGCGGTGGAAG


 


395154
5-10-5
 10
 561
 580
0
GGCGGGCAGTGCGCTCTGAC


 


395155
5-10-5
 11
 600
 619
0
TGGTGAGGTATCCCCGGCGG


 


399794
5-10-5
 12
 606
 625
0
GGATCTTGGTGAGGTATCCC


 


399795
5-10-5
 13
 615
 634
0
AGACATGCAGGATCTTGGTG


 


395156
5-10-5
 14
 620
 639
0
ATGGAAGACATGCAGGATCT


 


395157
5-10-5
 15
 646
 665
0
TTCACCAGGAAGCCAGGAAG


 


399796
5-10-5
 16
 651
 670
0
TCATCTTCACCAGGAAGCCA


 


395158
5-10-5
 17
 705
 724
0
CCTCGATGTAGTCGACATGG


 


395159
5-10-5
 18
 785
 804
0
GTATTCATCCGCCCGGTACC


 


395160
5-10-5
 19
 835
 854
0
CTGGTGTCTAGGAGATACAC


 


399797
5-10-5
 20
 840
 859
0
GTATGCTGGTGTCTAGGAGA


 


395161
5-10-5
 21
 860
 879
0
GATTTCCCGGTGGTCACTCT


 


399798
5-10-5
 22
 866
 885
0
GCCCTCGATTTCCCGGTGGT


 


399799
5-10-5
 23
 880
 899
0
GTGACCATGACCCTGCCCTC


 


395162
5-10-5
 24
 890
 909
0
CTCGAAGTCGGTGACCATGA


 


395163
5-10-5
 25
 923
 942
0
GTGGAAGCGGGTCCCGTCCT


 


395164
5-10-5
 26
 970
 989
0
ACCCCTGCCAGGTGGGTGCC


 


399800
5-10-5
 27
 975
 994
0
TGACCACCCCTGCCAGGTGG


 


395165
5-10-5
 28
1004
1023
0
ACCCTTGGCCACGCCGGCAT


 


395166
5-10-5
 29
1040
1059
0
TTGGCAGTTGAGCACGCGCA


 


399801
5-10-5
 30
1045
1064
0
TTCCCTTGGCAGTTGAGCAC


 


395167
5-10-5
 31
1077
1096
0
CCAGGCCTATGAGGGTGCCG


 


395168
5-10-5
 32
1098
1117
0
GCTGGCTTTTCCGAATAAAC


 


395169
5-10-5
 33
1210
1229
0
GTGACCAGCACGACCCCAGC


 


395170
5-10-5
149
1297
1316
0
TGGGCATTGGTGGCCCCAAC


 


395171
5-10-5
 34
1326
1345
0
CCAAAGTCCCCAGGGTCACC


 


395172
5-10-5
128
1330
1349
0
GTCCCCAAAGTCCCCAGGGT


 


399802
5-10-5
 35
1335
1354
0
AGTTGGTCCCCAAAGTCCCC


 


395173
5-10-5
 36
1340
1359
0
GCCAAAGTTGGTCCCCAAAG


 


399803
5-10-5
 37
1352
1371
0
GTCCACACAGCGGCCAAAGT


 


395174
5-10-5
 38
1361
1380
0
GGCAAAGAGGTCCACACAGC


 


395175
5-10-5
 39
1389
1408
0
TGGAGGCACCAATGATGTCC


 


399804
5-10-5
 40
1400
1419
0
GCTGCAGTCGCTGGAGGCAC


 


399805
5-10-5
 41
1411
1430
0
ACAAAGCAGGTGCTGCAGTC


 


395176
5-10-5
101
1465
1484
0
ATGGCTGCAATGCCAGCCAC


 


399806
5-10-5
 42
1470
1489
0
GCATCATGGCTGCAATGCCA


 


399807
5-10-5
 43
1478
1497
0
GGCAGACAGCATCATGGCTG


 


399808
5-10-5
 44
1526
1545
0
GAAGTGGATCAGTCTCTGCC


 


395177
5-10-5
 45
1534
1553
0
TTGGCAGAGAAGTGGATCAG


 


399809
5-10-5
 46
1539
1558
0
CATCTTTGGCAGAGAAGTGG


 


399810
5-10-5
 47
1545
1564
0
TGATGACATCTTTGGCAGAG


 


399811
5-10-5
 48
1552
1571
0
GCCTCATTGATGACATCTTT


 


399812
5-10-5
 49
1564
1583
0
TCAGGGAACCAGGCCTCATT


 


395178
5-10-5
 50
1569
1588
0
GGTCCTCAGGGAACCAGGCC


 


395179
5-10-5
 87
1576
1595
0
ACCCGCTGGTCCTCAGGGAA


 


399813
5-10-5
 51
1583
1602
0
GGTCAGTACCCGCTGGTCCT


 


395180
5-10-5
119
1605
1624
0
GCAGGGCGGCCACCAGGTTG


 


395181
5-10-5
 52
1640
1659
0
AAACAGCTGCCAACCTGCCC


 


399814
5-10-5
 53
1645
1664
0
CTGCAAAACAGCTGCCAACC


 


395182
5-10-5
 54
1675
1694
0
GTAGGCCCCGAGTGTGCTGA


 


399815
5-10-5
 55
1740
1759
0
AGAAACTGGAGCAGCTCAGC


 


399816
5-10-5
 56
1746
1765
0
TCCTGGAGAAACTGGAGCAG


 


395183
5-10-5
 57
1812
1831
0
CGTTGTGGGCCCGGCAGACC


 


395184
5-10-5
 58
1858
1877
0
AGCAGGCAGCACCTGGCAAT


 


395185
5-10-5
 59
1920
1939
0
CACGGGTCCCCATGCTGGCC


 


395186
5-10-5
 60
2100
2119
0
CTTTGCATTCCAGACCTGGG


 


399817
5-10-5
 61
2105
2124
0
CTTGACTTTGCATTCCAGAC


 


395187
5-10-5
 62
2310
2329
0
GGCAGCAGATGGCAACGGCT


 


395188
5-10-5
154
2410
2429
0
TTTTAAAGCTCAGCCCCAGC


 


399818
5-10-5
 63
2415
2434
0
AACCATTTTAAAGCTCAGCC


 


395189
5-10-5
 64
2504
2523
0
TCAAGGGCCAGGCCAGCAGC


 


399819
5-10-5
 65
2509
2528
0
CCCACTCAAGGGCCAGGCCA


 


399820
5-10-5
122
2582
2601
0
GGAGGGAGCTTCCTGGCACC


 


395190
5-10-5
 66
2597
2616
0
ATGCCCCACAGTGAGGGAGG


 


395191
5-10-5
 67
2606
2625
0
AATGGTGAAATGCCCCACAG


 


395192
5-10-5
153
2649
2668
0
TTGGGAGCAGCTGGCAGCAC


 


395193
5-10-5
 68
2750
2769
0
CATGGGAAGAATCCTGCCTC


 


395194
5-10-5
 69
2832
2851
0
GGAGATGAGGGCCATCAGCA


 


395195
5-10-5
 70
2900
2919
0
TAGATGCCATCCAGAAAGCT


 


399821
5-10-5
 71
2906
2925
0
TCTGGCTAGATGCCATCCAG


 


395196
5-10-5
 72
2983
3002
0
GGCATAGAGCAGAGTAAAGG


 


399822
5-10-5
 73
2988
3007
0
AGCCTGGCATAGAGCAGAGT


 


399823
5-10-5
135
2994
3013
0
TAGCACAGCCTGGCATAGAG


 


395197
5-10-5
112
3227
3246
0
GAAGAGGCTTGGCTTCAGAG


 


399824
5-10-5
 74
3233
3252
0
AAGTAAGAAGAGGCTTGGCT


 


395198
5-10-5
 75
3437
3456
0
GCTCAAGGAGGGACAGTTGT


 


395199
5-10-5
 76
3472
3491
0
AAAGATAAATGTCTGCTTGC


 


399825
5-10-5
 77
3477
3496
0
ACCCAAAAGATAAATGTCTG


 


395200
5-10-5
 78
3543
3562
0
TCTTCAAGTTACAAAAGCAA


 


399826
5-10-5
 99
3550
3569
0
ATAAATATCTTCAAGTTACA


 


405861
5-10-5
162
 406
 425
0
AAGGCTAGCACCAGCTCCTC


 


405862
5-10-5
163
 407
 426
0
CAAGGCTAGCACCAGCTCCT


 


405863
5-10-5
164
 408
 427
0
GCAAGGCTAGCACCAGCTCC


 


405864
5-10-5
165
 409
 428
0
CGCAAGGCTAGCACCAGCTC


 


405865
5-10-5
166
 411
 430
0
AACGCAAGGCTAGCACCAGC


 


405866
5-10-5
167
 412
 431
0
GAACGCAAGGCTAGCACCAG


 


405867
5-10-5
168
 413
 432
0
GGAACGCAAGGCTAGCACCA


 


405868
5-10-5
169
 414
 433
0
CGGAACGCAAGGCTAGCACC


 


405869
5-10-5
218
 862
 881
0
TCGATTTCCCGGTGGTCACT


 


405870
5-10-5
219
 863
 882
0
CTCGATTTCCCGGTGGTCAC


 


405871
5-10-5
220
 864
 883
0
CCTCGATTTCCCGGTGGTCA


 


405872
5-10-5
221
 865
 884
0
CCCTCGATTTCCCGGTGGTC


 


405873
5-10-5
222
 867
 886
0
TGCCCTCGATTTCCCGGTGG


 


405874
5-10-5
223
 868
 887
0
CTGCCCTCGATTTCCCGGTG


 


405875
5-10-5
224
 869
 888
0
CCTGCCCTCGATTTCCCGGT


 


405876
5-10-5
225
 870
 889
0
CCCTGCCCTCGATTTCCCGG


 


405877
5-10-5
246
1000
1019
0
TTGGCCACGCCGGCATCCCG


 


405878
5-10-5
247
1001
1020
0
CTTGGCCACGCCGGCATCCC


 


405879
5-10-5
248
1002
1021
0
CCTTGGCCACGCCGGCATCC


 


405880
5-10-5
249
1003
1022
0
CCCTTGGCCACGCCGGCATC


 


405881
5-10-5
250
1005
1024
0
CACCCTTGGCCACGCCGGCA


 


405882
5-10-5
251
1006
1025
0
GCACCCTTGGCCACGCCGGC


 


405883
5-10-5
252
1007
1026
0
GGCACCCTTGGCCACGCCGG


 


405884
5-10-5
253
1008
1027
0
TGGCACCCTTGGCCACGCCG


 


405885
5-10-5
346
1560
1579
0
GGAACCAGGCCTCATTGATG


 


405886
5-10-5
347
1561
1580
0
GGGAACCAGGCCTCATTGAT


 


405887
5-10-5
348
1562
1581
0
AGGGAACCAGGCCTCATTGA


 


405888
5-10-5
349
1563
1582
0
CAGGGAACCAGGCCTCATTG


 


405889
5-10-5
350
1565
1584
0
CTCAGGGAACCAGGCCTCAT


 


405890
5-10-5
351
1566
1585
0
CCTCAGGGAACCAGGCCTCA


 


405891
5-10-5
352
1567
1586
0
TCCTCAGGGAACCAGGCCTC


 


405892
5-10-5
353
1568
1587
0
GTCCTCAGGGAACCAGGCCT


 


405893
5-10-5
431
2828
2847
0
ATGAGGGCCATCAGCACCTT


 


405894
5-10-5
432
2829
2848
0
GATGAGGGCCATCAGCACCT


 


405895
5-10-5
433
2830
2849
0
AGATGAGGGCCATCAGCACC


 


405896
5-10-5
434
2831
2850
0
GAGATGAGGGCCATCAGCAC


 


405897
5-10-5
435
2833
2852
0
TGGAGATGAGGGCCATCAGC


 


405898
5-10-5
436
2834
2853
0
CTGGAGATGAGGGCCATCAG


 


405899
5-10-5
437
2835
2854
0
GCTGGAGATGAGGGCCATCA


 


405900
5-10-5
438
2836
2855
0
AGCTGGAGATGAGGGCCATC


 


405901
5-10-5
439
2902
2921
0
GCTAGATGCCATCCAGAAAG


 


405902
5-10-5
440
2903
2922
0
GGCTAGATGCCATCCAGAAA


 


405903
5-10-5
441
2904
2923
0
TGGCTAGATGCCATCCAGAA


 


405904
5-10-5
442
2905
2924
0
CTGGCTAGATGCCATCCAGA


 


405905
5-10-5
443
2907
2926
0
CTCTGGCTAGATGCCATCCA


 


405906
5-10-5
444
2908
2927
0
CCTCTGGCTAGATGCCATCC


 


405907
5-10-5
445
2909
2928
0
GCCTCTGGCTAGATGCCATC


 


405908
5-10-5
446
2910
2929
0
AGCCTCTGGCTAGATGCCAT


 


405909
5-10-5
267
1100
1119
0
CAGCTGGCTTTTCCGAATAA


 


405910
5-10-5
268
1102
1121
0
ACCAGCTGGCTTTTCCGAAT


 


405911
5-10-5
269
1104
1123
0
GGACCAGCTGGCTTTTCCGA


 


405912
5-10-5
270
1108
1127
0
GGCTGGACCAGCTGGCTTTT


 


405913
5-10-5
275
1212
1231
0
CGGTGACCAGCACGACCCCA


 


405914
5-10-5
276
1214
1233
0
AGCGGTGACCAGCACGACCC


 


405915
5-10-5
277
1216
1235
0
GCAGCGGTGACCAGCACGAC


 


405916
5-10-5
278
1218
1237
0
CGGCAGCGGTGACCAGCACG


 


405917
5-10-5
280
1222
1241
0
TTGCCGGCAGCGGTGACCAG


 


405918
5-10-5
281
1224
1243
0
AGTTGCCGGCAGCGGTGACC


 


405919
5-10-5
282
1226
1245
0
GAAGTTGCCGGCAGCGGTGA


 


405920
5-10-5
283
1228
1247
0
CGGAAGTTGCCGGCAGCGGT


 


405921
5-10-5
284
1230
1249
0
CCCGGAAGTTGCCGGCAGCG


 


405922
5-10-5
285
1232
1251
0
GTCCCGGAAGTTGCCGGCAG


 


405923
5-10-5
288
1295
1314
0
GGCATTGGTGGCCCCAACTG


 


405924
5-10-5
290
1318
1337
0
CCCAGGGTCACCGGCTGGTC


 


405925
5-10-5
292
1322
1341
0
AGTCCCCAGGGTCACCGGCT


 


405926
5-10-5
293
1324
1343
0
AAAGTCCCCAGGGTCACCGG


 


405927
5-10-5
294
1328
1347
0
CCCCAAAGTCCCCAGGGTCA


 


405928
5-10-5
295
1333
1352
0
TTGGTCCCCAAAGTCCCCAG


 


405929
5-10-5
296
1337
1356
0
AAAGTTGGTCCCCAAAGTCC


 


405930
5-10-5
297
1342
1361
0
CGGCCAAAGTTGGTCCCCAA


 


405931
5-10-5
298
1344
1363
0
AGCGGCCAAAGTTGGTCCCC


 


405932
5-10-5
299
1346
1365
0
ACAGCGGCCAAAGTTGGTCC


 


405933
5-10-5
300
1348
1367
0
ACACAGCGGCCAAAGTTGGT


 


405934
5-10-5
301
1350
1369
0
CCACACAGCGGCCAAAGTTG


 


405935
5-10-5
302
1354
1373
0
AGGTCCACACAGCGGCCAAA


 


405936
5-10-5
304
1358
1377
0
AAAGAGGTCCACACAGCGGC


 


405937
5-10-5
306
1387
1406
0
GAGGCACCAATGATGTCCTC


 


405938
5-10-5
307
1391
1410
0
GCTGGAGGCACCAATGATGT


 


405939
5-10-5
308
1393
1412
0
TCGCTGGAGGCACCAATGAT


 


405940
5-10-5
309
1395
1414
0
AGTCGCTGGAGGCACCAATG


 


405941
5-10-5
310
1397
1416
0
GCAGTCGCTGGAGGCACCAA


 


405942
5-10-5
311
1402
1421
0
GTGCTGCAGTCGCTGGAGGC


 


405943
5-10-5
312
1404
1423
0
AGGTGCTGCAGTCGCTGGAG


 


405944
5-10-5
314
1407
1426
0
AGCAGGTGCTGCAGTCGCTG


 


405945
5-10-5
315
1409
1428
0
AAAGCAGGTGCTGCAGTCGC


 


405946
5-10-5
316
1413
1432
0
ACACAAAGCAGGTGCTGCAG


 


405947
5-10-5
317
1415
1434
0
TGACACAAAGCAGGTGCTGC


 


405948
5-10-5
319
1467
1486
0
TCATGGCTGCAATGCCAGCC


 


405949
5-10-5
320
1472
1491
0
CAGCATCATGGCTGCAATGC


 


405950
5-10-5
321
1474
1493
0
GACAGCATCATGGCTGCAAT


 


405951
5-10-5
322
1476
1495
0
CAGACAGCATCATGGCTGCA


 


405952
5-10-5
323
1480
1499
0
TCGGCAGACAGCATCATGGC


 


405953
5-10-5
325
1484
1503
0
CGGCTCGGCAGACAGCATCA


 


405954
5-10-5
326
1486
1505
0
TCCGGCTCGGCAGACAGCAT


 


405955
5-10-5
328
1513
1532
0
CTCTGCCTCAACTCGGCCAG


 


405956
5-10-5
329
1515
1534
0
GTCTCTGCCTCAACTCGGCC


 


405957
5-10-5
330
1517
1536
0
CAGTCTCTGCCTCAACTCGG


 


405958
5-10-5
331
1519
1538
0
ATCAGTCTCTGCCTCAACTC


 


405959
5-10-5
332
1521
1540
0
GGATCAGTCTCTGCCTCAAC


 


405960
5-10-5
333
1523
1542
0
GTGGATCAGTCTCTGCCTCA


 


405961
5-10-5
334
1525
1544
0
AAGTGGATCAGTCTCTGCCT


 


405962
5-10-5
335
1528
1547
0
GAGAAGTGGATCAGTCTCTG


 


405963
5-10-5
337
1532
1551
0
GGCAGAGAAGTGGATCAGTC


 


405964
5-10-5
338
1536
1555
0
CTTTGGCAGAGAAGTGGATC


 


405965
5-10-5
339
1541
1560
0
GACATCTTTGGCAGAGAAGT


 


405966
5-10-5
340
1543
1562
0
ATGACATCTTTGGCAGAGAA


 


405967
5-10-5
341
1547
1566
0
ATTGATGACATCTTTGGCAG


 


405968
5-10-5
342
1549
1568
0
TCATTGATGACATCTTTGGC


 


405969
5-10-5
343
1554
1573
0
AGGCCTCATTGATGACATCT


 


405970
5-10-5
344
1556
1575
0
CCAGGCCTCATTGATGACAT


 


405971
5-10-5
355
1571
1590
0
CTGGTCCTCAGGGAACCAGG


 


405972
5-10-5
357
1573
1592
0
CGCTGGTCCTCAGGGAACCA


 


405973
5-10-5
358
1578
1597
0
GTACCCGCTGGTCCTCAGGG


 


405974
5-10-5
359
1580
1599
0
CAGTACCCGCTGGTCCTCAG


 


405975
5-10-5
361
1642
1661
0
CAAAACAGCTGCCAACCTGC


 


405976
5-10-5
362
1647
1666
0
TCCTGCAAAACAGCTGCCAA


 


405977
5-10-5
363
1649
1668
0
AGTCCTGCAAAACAGCTGCC


 


405978
5-10-5
365
1672
1691
0
GGCCCCGAGTGTGCTGACCA


 


405979
5-10-5
366
1677
1696
0
GTGTAGGCCCCGAGTGTGCT


 


405980
5-10-5
367
1679
1698
0
CCGTGTAGGCCCCGAGTGTG


 


405981
5-10-5
368
1681
1700
0
ATCCGTGTAGGCCCCGAGTG


 


405982
5-10-5
370
1685
1704
0
GGCCATCCGTGTAGGCCCCG


 


405983
5-10-5
371
1687
1706
0
GTGGCCATCCGTGTAGGCCC


 


405984
5-10-5
372
1735
1754
0
CTGGAGCAGCTCAGCAGCTC


 


405985
5-10-5
373
1737
1756
0
AACTGGAGCAGCTCAGCAGC


 


405986
5-10-5
374
1742
1761
0
GGAGAAACTGGAGCAGCTCA


 


405987
5-10-5
375
1744
1763
0
CTGGAGAAACTGGAGCAGCT


 


405988
5-10-5
381
1854
1873
0
GGCAGCACCTGGCAATGGCG


 


405989
5-10-5
383
1856
1875
0
CAGGCAGCACCTGGCAATGG


 


405990
5-10-5
386
1860
1879
0
GTAGCAGGCAGCACCTGGCA


 


405991
5-10-5
394
1922
1941
0
GACACGGGTCCCCATGCTGG


 


405992
5-10-5
396
1924
1943
0
TGGACACGGGTCCCCATGCT


 


405993
5-10-5
398
1926
1945
0
AGTGGACACGGGTCCCCATG


 


405994
5-10-5
400
1928
1947
0
GCAGTGGACACGGGTCCCCA


 


405995
5-10-5
402
1930
1949
0
TGGCAGTGGACACGGGTCCC


 


405996
5-10-5
412
2102
2121
0
GACTTTGCATTCCAGACCTG


 


405997
5-10-5
415
2107
2126
0
TCCTTGACTTTGCATTCCAG


 


405998
5-10-5
426
2313
2332
0
TCCGGCAGCAGATGGCAACG


 


405999
5-10-5
160
 298
 317
0
GACCGCCTGGAGCTGACGGT


 


406000
5-10-5
173
 482
 501
0
ATCCTTGGCGCAGCGGTGGA


 


406001
5-10-5
174
 484
 503
0
GGATCCTTGGCGCAGCGGTG


 


406002
5-10-5
175
 488
 507
0
CCACGGATCCTTGGCGCAGC


 


406003
5-10-5
178
 555
 574
0
CAGTGCGCTCTGACTGCGAG


 


406004
5-10-5
179
 557
 576
0
GGCAGTGCGCTCTGACTGCG


 


406005
5-10-5
180
 559
 578
0
CGGGCAGTGCGCTCTGACTG


 


406006
5-10-5
181
 562
 581
0
CGGCGGGCAGTGCGCTCTGA


 


406007
5-10-5
183
 595
 614
0
AGGTATCCCCGGCGGGCAGC


 


406008
5-10-5
188
 602
 621
0
CTTGGTGAGGTATCCCCGGC


 


406009
5-10-5
190
 604
 623
0
ATCTTGGTGAGGTATCCCCG


 


406010
5-10-5
193
 609
 628
0
GCAGGATCTTGGTGAGGTAT


 


406011
5-10-5
194
 611
 630
0
ATGCAGGATCTTGGTGAGGT


 


406012
5-10-5
195
 613
 632
0
ACATGCAGGATCTTGGTGAG


 


406013
5-10-5
196
 617
 636
0
GAAGACATGCAGGATCTTGG


 


406014
5-10-5
199
 648
 667
0
TCTTCACCAGGAAGCCAGGA


 


406015
5-10-5
200
 653
 672
0
ACTCATCTTCACCAGGAAGC


 


406016
5-10-5
201
 655
 674
0
CCACTCATCTTCACCAGGAA


 


406017
5-10-5
203
 659
 678
0
GTCGCCACTCATCTTCACCA


 


406018
5-10-5
204
 661
 680
0
AGGTCGCCACTCATCTTCAC


 


406019
5-10-5
205
 663
 682
0
GCAGGTCGCCACTCATCTTC


 


406020
5-10-5
206
 665
 684
0
CAGCAGGTCGCCACTCATCT


 


406021
5-10-5
210
 782
 801
0
TTCATCCGCCCGGTACCGTG


 


406022
5-10-5
211
 784
 803
0
TATTCATCCGCCCGGTACCG


 


406023
5-10-5
212
 787
 806
0
TGGTATTCATCCGCCCGGTA


 


406024
5-10-5
213
 789
 808
0
GCTGGTATTCATCCGCCCGG


 


406025
5-10-5
216
 832
 851
0
GTGTCTAGGAGATACACCTC


 


406026
5-10-5
217
 837
 856
0
TGCTGGTGTCTAGGAGATAC


 


406027
5-10-5
226
 874
 893
0
ATGACCCTGCCCTCGATTTC


 


406028
5-10-5
227
 876
 895
0
CCATGACCCTGCCCTCGATT


 


406029
5-10-5
228
 878
 897
0
GACCATGACCCTGCCCTCGA


 


406030
5-10-5
229
 882
 901
0
CGGTGACCATGACCCTGCCC


 


406031
5-10-5
230
 884
 903
0
GTCGGTGACCATGACCCTGC


 


406032
5-10-5
232
 888
 907
0
CGAAGTCGGTGACCATGACC


 


406033
5-10-5
237
 967
 986
0
CCTGCCAGGTGGGTGCCATG


 


406034
5-10-5
238
 972
 991
0
CCACCCCTGCCAGGTGGGTG


 


406035
5-10-5
239
 977
 996
0
GCTGACCACCCCTGCCAGGT


 


406036
5-10-5
241
 989
1008
0
GGCATCCCGGCCGCTGACCA


 


406037
5-10-5
242
 992
1011
0
GCCGGCATCCCGGCCGCTGA


 


406038
5-10-5
245
 999
1018
0
TGGCCACGCCGGCATCCCGG


 


406039
5-10-5
257
1036
1055
0
CAGTTGAGCACGCGCAGGCT


 


406040
5-10-5
258
1038
1057
0
GGCAGTTGAGCACGCGCAGG


 


406041
5-10-5
259
1042
1061
0
CCTTGGCAGTTGAGCACGCG


 


406042
5-10-5
260
1047
1066
0
CCTTCCCTTGGCAGTTGAGC


 


406043
5-10-5
261
1051
1070
0
GTGCCCTTCCCTTGGCAGTT


 


406044
5-10-5
262
1053
1072
0
CCGTGCCCTTCCCTTGGCAG


 


406045
5-10-5
266
1096
1115
0
TGGCTTTTCCGAATAAACTC


 


408642
5-10-5
264
1076
1095
0
CAGGCCTATGAGGGTGCCGC


 


408653
5-10-5
354
1570
1589
0
TGGTCCTCAGGGAACCAGGC


 


409126
5-10-5
447
1004
1023
1
ACCCTTGGTCACGCCGGCAT


 


410529
5-10-5
184
 597
 616
0
TGAGGTATCCCCGGCGGGCA


 


410530
5-10-5
185
 598
 617
0
GTGAGGTATCCCCGGCGGGC


 


410531
5-10-5
186
 599
 618
0
GGTGAGGTATCCCCGGCGGG


 


410532
5-10-5
187
 601
 620
0
TTGGTGAGGTATCCCCGGCG


 


410533
5-10-5
189
 603
 622
0
TCTTGGTGAGGTATCCCCGG


 


410534
5-10-5
191
 605
 624
0
GATCTTGGTGAGGTATCCCC


 


410535
5-10-5
192
 607
 626
0
AGGATCTTGGTGAGGTATCC


 


410536
5-10-5
243
 997
1016
0
GCCACGCCGGCATCCCGGCC


 


410537
5-10-5
244
 998
1017
0
GGCCACGCCGGCATCCCGGC


 


410538
5-10-5
254
1009
1028
0
CTGGCACCCTTGGCCACGCC


 


410539
5-10-5
255
1010
1029
0
GCTGGCACCCTTGGCCACGC


 


410540
5-10-5
356
1572
1591
0
GCTGGTCCTCAGGGAACCAG


 


410541
5-10-5
376
1849
1868
0
CACCTGGCAATGGCGTAGAC


 


410542
5-10-5
377
1850
1869
0
GCACCTGGCAATGGCGTAGA


 


410543
5-10-5
378
1851
1870
0
AGCACCTGGCAATGGCGTAG


 


410544
5-10-5
379
1852
1871
0
CAGCACCTGGCAATGGCGTA


 


410545
5-10-5
380
1853
1872
0
GCAGCACCTGGCAATGGCGT


 


410546
5-10-5
382
1855
1874
0
AGGCAGCACCTGGCAATGGC


 


410547
5-10-5
384
1857
1876
0
GCAGGCAGCACCTGGCAATG


 


410548
5-10-5
385
1859
1878
0
TAGCAGGCAGCACCTGGCAA


 


410549
5-10-5
388
1915
1934
0
GTCCCCATGCTGGCCTCAGC


 


410550
5-10-5
389
1916
1935
0
GGTCCCCATGCTGGCCTCAG


 


410551
5-10-5
390
1917
1936
0
GGGTCCCCATGCTGGCCTCA


 


410552
5-10-5
391
1918
1937
0
CGGGTCCCCATGCTGGCCTC


 


410553
5-10-5
392
1919
1938
0
ACGGGTCCCCATGCTGGCCT


 


410554
5-10-5
393
1921
1940
0
ACACGGGTCCCCATGCTGGC


 


410555
5-10-5
395
1923
1942
0
GGACACGGGTCCCCATGCTG


 


410556
5-10-5
397
1925
1944
0
GTGGACACGGGTCCCCATGC


 


410557
5-10-5
399
1927
1946
0
CAGTGGACACGGGTCCCCAT


 


410558
5-10-5
401
1929
1948
0
GGCAGTGGACACGGGTCCCC


 


410559
5-10-5
403
1931
1950
0
GTGGCAGTGGACACGGGTCC


 


410560
5-10-5
404
1932
1951
0
GGTGGCAGTGGACACGGGTC


 


410561
5-10-5
405
1933
1952
0
TGGTGGCAGTGGACACGGGT


 


410562
5-10-5
411
2101
2120
0
ACTTTGCATTCCAGACCTGG


 


410563
5-10-5
413
2103
2122
0
TGACTTTGCATTCCAGACCT


 


410564
5-10-5
414
2104
2123
0
TTGACTTTGCATTCCAGACC


 


410565
5-10-5
419
2305
2324
0
CAGATGGCAACGGCTGTCAC


 


410566
5-10-5
420
2306
2325
0
GCAGATGGCAACGGCTGTCA


 


410567
5-10-5
421
2307
2326
0
AGCAGATGGCAACGGCTGTC


 


410568
5-10-5
422
2308
2327
0
CAGCAGATGGCAACGGCTGT


 


410569
5-10-5
423
2309
2328
0
GCAGCAGATGGCAACGGCTG


 


410570
5-10-5
424
2311
2330
0
CGGCAGCAGATGGCAACGGC


 


410571
5-10-5
425
2312
2331
0
CCGGCAGCAGATGGCAACGG


 


410572
5-10-5
427
2314
2333
0
CTCCGGCAGCAGATGGCAAC


 


410573
5-10-5
428
2315
2334
0
GCTCCGGCAGCAGATGGCAA


 


410730
5-10-5
202
 657
 676
0
CGCCACTCATCTTCACCAGG


 


410731
5-10-5
207
 667
 686
0
TCCAGCAGGTCGCCACTCAT


 


410732
5-10-5
214
 791
 810
0
GGGCTGGTATTCATCCGCCC


 


410733
5-10-5
231
 886
 905
0
AAGTCGGTGACCATGACCCT


 


410734
5-10-5
279
1219
1238
0
CCGGCAGCGGTGACCAGCAC


 


410735
5-10-5
291
1320
1339
0
TCCCCAGGGTCACCGGCTGG


 


410736
5-10-5
303
1356
1375
0
AGAGGTCCACACAGCGGCCA


 


410737
5-10-5
313
1406
1425
0
GCAGGTGCTGCAGTCGCTGG


 


410738
5-10-5
324
1482
1501
0
GCTCGGCAGACAGCATCATG


 


410739
5-10-5
336
1530
1549
0
CAGAGAAGTGGATCAGTCTC


 


410740
5-10-5
345
1558
1577
0
AACCAGGCCTCATTGATGAC


 


410741
5-10-5
369
1683
1702
0
CCATCCGTGTAGGCCCCGAG


 


410742
5-10-5
159
 294
 313
0
GCCTGGAGCTGACGGTGCCC


 


410743
5-10-5
170
 421
 440
0
TCCTCCTCGGAACGCAAGGC


 


410744
5-10-5
171
 446
 465
0
GTGCTCGGGTGCTTCGGCCA


 


410745
5-10-5
172
 466
 485
0
TGGAAGGTGGCTGTGGTTCC


 


410746
5-10-5
176
 507
 526
0
CGTAGGTGCCAGGCAACCTC


 


410747
5-10-5
177
 545
 564
0
TGACTGCGAGAGGTGGGTCT


 


410748
5-10-5
182
 591
 610
0
ATCCCCGGCGGGCAGCCTGG


 


410749
5-10-5
197
 628
 647
0
AGAAGGCCATGGAAGACATG


 


410750
5-10-5
198
 638
 657
0
GAAGCCAGGAAGAAGGCCAT


 


410751
5-10-5
208
 685
 704
0
GGCAACTTCAAGGCCAGCTC


 


410752
5-10-5
209
 724
 743
0
GCAAAGACAGAGGAGTCCTC


 


410753
5-10-5
215
 821
 840
0
ATACACCTCCACCAGGCTGC


 


410754
5-10-5
233
 898
 917
0
GGCACATTCTCGAAGTCGGT


 


410755
5-10-5
234
 933
 952
0
TGGCCTGTCTGTGGAAGCGG


 


410756
5-10-5
235
 960
 979
0
GGTGGGTGCCATGACTGTCA


 


410757
5-10-5
240
 985
1004
0
TCCCGGCCGCTGACCACCCC


 


410758
5-10-5
256
1015
1034
0
CGCATGCTGGCACCCTTGGC


 


410759
5-10-5
263
1064
1083
0
GGTGCCGCTAACCGTGCCCT


 


410760
5-10-5
265
1088
1107
0
CCGAATAAACTCCAGGCCTA


 


410761
5-10-5
271
1119
1138
0
GTGGCCCCACAGGCTGGACC


 


410762
5-10-5
272
1132
1151
0
AGCAGCACCACCAGTGGCCC


 


410763
5-10-5
273
1154
1173
0
GCTGTACCCACCCGCCAGGG


 


410764
5-10-5
274
1200
1219
0
CGACCCCAGCCCTCGCCAGG


 


410765
5-10-5
286
1273
1292
0
ATGACCTCGGGAGCTGAGGC


 


410766
5-10-5
287
1283
1302
0
CCCAACTGTGATGACCTCGG


 


410767
5-10-5
289
1305
1324
0
GCTGGTCTTGGGCATTGGTG


 


410768
5-10-5
305
1380
1399
0
CAATGATGTCCTCCCCTGGG


 


410769
5-10-5
318
1425
1444
0
TCCCACTCTGTGACACAAAG


 


410770
5-10-5
327
1500
1519
0
CGGCCAGGGTGAGCTCCGGC


 


410771
5-10-5
360
1628
1647
0
ACCTGCCCCATGGGTGCTGG


 


410772
5-10-5
364
1660
1679
0
GCTGACCATACAGTCCTGCA


 


410773
5-10-5
387
1905
1924
0
TGGCCTCAGCTGGTGGAGCT


 


410774
5-10-5
406
1936
1955
0
TGTTGGTGGCAGTGGACACG


 


410775
5-10-5
407
1962
1981
0
AGCTGCAGCCTGTGAGGACG


 


410776
5-10-5
408
1990
2009
0
GTGCCAAGGTCCTCCACCTC


 


410777
5-10-5
409
2010
2029
0
TCAGCACAGGCGGCTTGTGG


 


410778
5-10-5
410
2040
2059
0
CCACGCACTGGTTGGGCTGA


 


410779
5-10-5
416
2120
2139
0
CGGGATTCCATGCTCCTTGA


 


410780
5-10-5
417
2150
2169
0
GCAGGCCACGGTCACCTGCT


 


410781
5-10-5
418
2187
2206
0
GGAGGGCACTGCAGCCAGTC


 


410782
5-10-5
429
2325
2344
0
CCAGGTGCCGGCTCCGGCAG


 


410783
5-10-5
430
2335
2354
0
GAGGCCTGCGCCAGGTGCCG









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 3-14-3 gap-widened motif. Table 4 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 4







Gapmer antisense compounds having a 3-14-3 motif


targeted to SEQ ID NO: 1
















5′
3′







Target
Target






SEQ
Site to
Site to




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 1
NO: 1
Mismatches
Sequence (5′-3′)





399871
3-14-3
  4
 135
 154
0
GCGCGGAATCCTGGCTGGGA


 


399872
3-14-3
  5
 242
 261
0
GAGGAGACCTAGAGGCCGTG


 


399873
3-14-3
  6
 300
 319
0
AGGACCGCCTGGAGCTGACG


 


399874
3-14-3
  7
 410
 429
0
ACGCAAGGCTAGCACCAGCT


 


399949
3-14-3
  8
 417
 436
0
CCTCGGAACGCAAGGCTAGC


 


399875
3-14-3
  9
 480
 499
0
CCTTGGCGCAGCGGTGGAAG


 


399876
3-14-3
 10
 561
 580
0
GGCGGGCAGTGCGCTCTGAC


 


399877
3-14-3
 11
 600
 619
0
TGGTGAGGTATCCCCGGCGG


 


399950
3-14-3
 12
 606
 625
0
GGATCTTGGTGAGGTATCCC


 


399951
3-14-3
 13
 615
 634
0
AGACATGCAGGATCTTGGTG


 


399878
3-14-3
 14
 620
 639
0
ATGGAAGACATGCAGGATCT


 


399879
3-14-3
 15
 646
 665
0
TTCACCAGGAAGCCAGGAAG


 


399952
3-14-3
 16
 651
 670
0
TCATCTTCACCAGGAAGCCA


 


399880
3-14-3
 17
 705
 724
0
CCTCGATGTAGTCGACATGG


 


399881
3-14-3
 18
 785
 804
0
GTATTCATCCGCCCGGTACC


 


399882
3-14-3
 19
 835
 854
0
CTGGTGTCTAGGAGATACAC


 


399953
3-14-3
 20
 840
 859
0
GTATGCTGGTGTCTAGGAGA


 


399883
3-14-3
 21
 860
 879
0
GATTTCCCGGTGGTCACTCT


 


399954
3-14-3
 22
 866
 885
0
GCCCTCGATTTCCCGGTGGT


 


399955
3-14-3
 23
 880
 899
0
GTGACCATGACCCTGCCCTC


 


399884
3-14-3
 24
 890
 909
0
CTCGAAGTCGGTGACCATGA


 


399885
3-14-3
 25
 923
 942
0
GTGGAAGCGGGTCCCGTCCT


 


399886
3-14-3
 26
 970
 989
0
ACCCCTGCCAGGTGGGTGCC


 


399956
3-14-3
 27
 975
 994
0
TGACCACCCCTGCCAGGTGG


 


399887
3-14-3
 28
1004
1023
0
ACCCTTGGCCACGCCGGCAT


 


399888
3-14-3
 29
1040
1059
0
TTGGCAGTTGAGCACGCGCA


 


399957
3-14-3
 30
1045
1064
0
TTCCCTTGGCAGTTGAGCAC


 


399889
3-14-3
 31
1077
1096
0
CCAGGCCTATGAGGGTGCCG


 


399890
3-14-3
 32
1098
1117
0
GCTGGCTTTTCCGAATAAAC


 


399891
3-14-3
 33
1210
1229
0
GTGACCAGCACGACCCCAGC


 


399892
3-14-3
149
1297
1316
0
TGGGCATTGGTGGCCCCAAC


 


399893
3-14-3
 34
1326
1345
0
CCAAAGTCCCCAGGGTCACC


 


399894
3-14-3
128
1330
1349
0
GTCCCCAAAGTCCCCAGGGT


 


399958
3-14-3
 35
1335
1354
0
AGTTGGTCCCCAAAGTCCCC


 


399895
3-14-3
 36
1340
1359
0
GCCAAAGTTGGTCCCCAAAG


 


399959
3-14-3
 37
1352
1371
0
GTCCACACAGCGGCCAAAGT


 


399896
3-14-3
 38
1361
1380
0
GGCAAAGAGGTCCACACAGC


 


399897
3-14-3
 39
1389
1408
0
TGGAGGCACCAATGATGTCC


 


399960
3-14-3
 40
1400
1419
0
GCTGCAGTCGCTGGAGGCAC


 


399961
3-14-3
 41
1411
1430
0
ACAAAGCAGGTGCTGCAGTC


 


399898
3-14-3
101
1465
1484
0
ATGGCTGCAATGCCAGCCAC


 


399962
3-14-3
 42
1470
1489
0
GCATCATGGCTGCAATGCCA


 


399963
3-14-3
 43
1478
1497
0
GGCAGACAGCATCATGGCTG


 


399964
3-14-3
 44
1526
1545
0
GAAGTGGATCAGTCTCTGCC


 


399899
3-14-3
 45
1534
1553
0
TTGGCAGAGAAGTGGATCAG


 


399965
3-14-3
 46
1539
1558
0
CATCTTTGGCAGAGAAGTGG


 


399966
3-14-3
 47
1545
1564
0
TGATGACATCTTTGGCAGAG


 


399967
3-14-3
 48
1552
1571
0
GCCTCATTGATGACATCTTT


 


399968
3-14-3
 49
1564
1583
0
TCAGGGAACCAGGCCTCATT


 


399900
3-14-3
 50
1569
1588
0
GGTCCTCAGGGAACCAGGCC


 


399901
3-14-3
 87
1576
1595
0
ACCCGCTGGTCCTCAGGGAA


 


399969
3-14-3
 51
1583
1602
0
GGTCAGTACCCGCTGGTCCT


 


399902
3-14-3
119
1605
1624
0
GCAGGGCGGCCACCAGGTTG


 


399903
3-14-3
 52
1640
1659
0
AAACAGCTGCCAACCTGCCC


 


399970
3-14-3
 53
1645
1664
0
CTGCAAAACAGCTGCCAACC


 


399904
3-14-3
 54
1675
1694
0
GTAGGCCCCGAGTGTGCTGA


 


399971
3-14-3
 55
1740
1759
0
AGAAACTGGAGCAGCTCAGC


 


399972
3-14-3
 56
1746
1765
0
TCCTGGAGAAACTGGAGCAG


 


399905
3-14-3
 57
1812
1831
0
CGTTGTGGGCCCGGCAGACC


 


399906
3-14-3
 58
1858
1877
0
AGCAGGCAGCACCTGGCAAT


 


399907
3-14-3
 59
1920
1939
0
CACGGGTCCCCATGCTGGCC


 


399908
3-14-3
 60
2100
2119
0
CTTTGCATTCCAGACCTGGG


 


399973
3-14-3
 61
2105
2124
0
CTTGACTTTGCATTCCAGAC


 


399909
3-14-3
 62
2310
2329
0
GGCAGCAGATGGCAACGGCT


 


399910
3-14-3
154
2410
2429
0
TTTTAAAGCTCAGCCCCAGC


 


399974
3-14-3
 63
2415
2434
0
AACCATTTTAAAGCTCAGCC


 


399911
3-14-3
 64
2504
2523
0
TCAAGGGCCAGGCCAGCAGC


 


399975
3-14-3
 65
2509
2528
0
CCCACTCAAGGGCCAGGCCA


 


399976
3-14-3
122
2582
2601
0
GGAGGGAGCTTCCTGGCACC


 


399912
3-14-3
 66
2597
2616
0
ATGCCCCACAGTGAGGGAGG


 


399913
3-14-3
 67
2606
2625
0
AATGGTGAAATGCCCCACAG


 


399914
3-14-3
153
2649
2668
0
TTGGGAGCAGCTGGCAGCAC


 


399915
3-14-3
 68
2750
2769
0
CATGGGAAGAATCCTGCCTC


 


399916
3-14-3
 69
2832
2851
0
GGAGATGAGGGCCATCAGCA


 


399917
3-14-3
 70
2900
2919
0
TAGATGCCATCCAGAAAGCT


 


399977
3-14-3
 71
2906
2925
0
TCTGGCTAGATGCCATCCAG


 


399918
3-14-3
 72
2983
3002
0
GGCATAGAGCAGAGTAAAGG


 


399978
3-14-3
 73
2988
3007
0
AGCCTGGCATAGAGCAGAGT


 


399979
3-14-3
135
2994
3013
0
TAGCACAGCCTGGCATAGAG


 


399919
3-14-3
112
3227
3246
0
GAAGAGGCTTGGCTTCAGAG


 


399980
3-14-3
 74
3233
3252
0
AAGTAAGAAGAGGCTTGGCT


 


399920
3-14-3
 75
3437
3456
0
GCTCAAGGAGGGACAGTTGT


 


399921
3-14-3
 76
3472
3491
0
AAAGATAAATGTCTGCTTGC


 


399981
3-14-3
 77
3477
3496
0
ACCCAAAAGATAAATGTCTG


 


399922
3-14-3
 78
3543
3562
0
TCTTCAAGTTACAAAAGCAA


 


399982
3-14-3
 99
3550
3569
0
ATAAATATCTTCAAGTTACA


 


410574
3-14-3
184
 597
 616
0
TGAGGTATCCCCGGCGGGCA


 


410575
3-14-3
185
 598
 617
0
GTGAGGTATCCCCGGCGGGC


 


410576
3-14-3
186
 599
 618
0
GGTGAGGTATCCCCGGCGGG


 


410577
3-14-3
187
 601
 620
0
TTGGTGAGGTATCCCCGGCG


 


410578
3-14-3
188
 602
 621
0
CTTGGTGAGGTATCCCCGGC


 


410579
3-14-3
189
 603
 622
0
TCTTGGTGAGGTATCCCCGG


 


410580
3-14-3
190
 604
 623
0
ATCTTGGTGAGGTATCCCCG


 


410581
3-14-3
191
 605
 624
0
GATCTTGGTGAGGTATCCCC


 


410582
3-14-3
192
 607
 626
0
AGGATCTTGGTGAGGTATCC


 


405604
3-14-3
236
 963
 982
0
CCAGGTGGGTGCCATGACTG


 


410583
3-14-3
243
 997
1016
0
GCCACGCCGGCATCCCGGCC


 


410584
3-14-3
244
 998
1017
0
GGCCACGCCGGCATCCCGGC


 


410585
3-14-3
245
 999
1018
0
TGGCCACGCCGGCATCCCGG


 


410586
3-14-3
246
1000
1019
0
TTGGCCACGCCGGCATCCCG


 


410587
3-14-3
247
1001
1020
0
CTTGGCCACGCCGGCATCCC


 


410588
3-14-3
248
1002
1021
0
CCTTGGCCACGCCGGCATCC


 


410589
3-14-3
249
1003
1022
0
CCCTTGGCCACGCCGGCATC


 


410590
3-14-3
250
1005
1024
0
CACCCTTGGCCACGCCGGCA


 


410591
3-14-3
251
1006
1025
0
GCACCCTTGGCCACGCCGGC


 


410592
3-14-3
252
1007
1026
0
GGCACCCTTGGCCACGCCGG


 


410593
3-14-3
253
1008
1027
0
TGGCACCCTTGGCCACGCCG


 


410594
3-14-3
254
1009
1028
0
CTGGCACCCTTGGCCACGCC


 


410595
3-14-3
255
1010
1029
0
GCTGGCACCCTTGGCCACGC


 


410596
3-14-3
348
1562
1581
0
AGGGAACCAGGCCTCATTGA


 


410597
3-14-3
349
1563
1582
0
CAGGGAACCAGGCCTCATTG


 


410598
3-14-3
350
1565
1584
0
CTCAGGGAACCAGGCCTCAT


 


410599
3-14-3
351
1566
1585
0
CCTCAGGGAACCAGGCCTCA


 


410600
3-14-3
352
1567
1586
0
TCCTCAGGGAACCAGGCCTC


 


410601
3-14-3
353
1568
1587
0
GTCCTCAGGGAACCAGGCCT


 


410602
3-14-3
354
1570
1589
0
TGGTCCTCAGGGAACCAGGC


 


410603
3-14-3
355
1571
1590
0
CTGGTCCTCAGGGAACCAGG


 


410604
3-14-3
356
1572
1591
0
GCTGGTCCTCAGGGAACCAG


 


405641
3-14-3
373
1737
1756
0
AACTGGAGCAGCTCAGCAGC


 


410605
3-14-3
376
1849
1868
0
CACCTGGCAATGGCGTAGAC


 


410606
3-14-3
377
1850
1869
0
GCACCTGGCAATGGCGTAGA


 


410607
3-14-3
378
1851
1870
0
AGCACCTGGCAATGGCGTAG


 


410608
3-14-3
379
1852
1871
0
CAGCACCTGGCAATGGCGTA


 


410609
3-14-3
380
1853
1872
0
GCAGCACCTGGCAATGGCGT


 


410610
3-14-3
381
1854
1873
0
GGCAGCACCTGGCAATGGCG


 


410611
3-14-3
382
1855
1874
0
AGGCAGCACCTGGCAATGGC


 


410612
3-14-3
383
1856
1875
0
CAGGCAGCACCTGGCAATGG


 


410613
3-14-3
384
1857
1876
0
GCAGGCAGCACCTGGCAATG


 


410614
3-14-3
385
1859
1878
0
TAGCAGGCAGCACCTGGCAA


 


410615
3-14-3
388
1915
1934
0
GTCCCCATGCTGGCCTCAGC


 


410616
3-14-3
389
1916
1935
0
GGTCCCCATGCTGGCCTCAG


 


410617
3-14-3
390
1917
1936
0
GGGTCCCCATGCTGGCCTCA


 


410618
3-14-3
391
1918
1937
0
CGGGTCCCCATGCTGGCCTC


 


410619
3-14-3
392
1919
1938
0
ACGGGTCCCCATGCTGGCCT


 


410620
3-14-3
393
1921
1940
0
ACACGGGTCCCCATGCTGGC


 


410621
3-14-3
394
1922
1941
0
GACACGGGTCCCCATGCTGG


 


410622
3-14-3
395
1923
1942
0
GGACACGGGTCCCCATGCTG


 


410623
3-14-3
396
1924
1943
0
TGGACACGGGTCCCCATGCT


 


410624
3-14-3
397
1925
1944
0
GTGGACACGGGTCCCCATGC


 


410625
3-14-3
398
1926
1945
0
AGTGGACACGGGTCCCCATG


 


410626
3-14-3
399
1927
1946
0
CAGTGGACACGGGTCCCCAT


 


410627
3-14-3
400
1928
1947
0
GCAGTGGACACGGGTCCCCA


 


410628
3-14-3
401
1929
1948
0
GGCAGTGGACACGGGTCCCC


 


410629
3-14-3
402
1930
1949
0
TGGCAGTGGACACGGGTCCC


 


410630
3-14-3
403
1931
1950
0
GTGGCAGTGGACACGGGTCC


 


410631
3-14-3
404
1932
1951
0
GGTGGCAGTGGACACGGGTC


 


410632
3-14-3
405
1933
1952
0
TGGTGGCAGTGGACACGGGT


 


410633
3-14-3
411
2101
2120
0
ACTTTGCATTCCAGACCTGG


 


410634
3-14-3
412
2102
2121
0
GACTTTGCATTCCAGACCTG


 


410635
3-14-3
413
2103
2122
0
TGACTTTGCATTCCAGACCT


 


410636
3-14-3
414
2104
2123
0
TTGACTTTGCATTCCAGACC


 


410637
3-14-3
419
2305
2324
0
CAGATGGCAACGGCTGTCAC


 


410638
3-14-3
420
2306
2325
0
GCAGATGGCAACGGCTGTCA


 


410639
3-14-3
421
2307
2326
0
AGCAGATGGCAACGGCTGTC


 


410640
3-14-3
422
2308
2327
0
CAGCAGATGGCAACGGCTGT


 


410641
3-14-3
423
2309
2328
0
GCAGCAGATGGCAACGGCTG


 


410642
3-14-3
424
2311
2330
0
CGGCAGCAGATGGCAACGGC


 


410643
3-14-3
425
2312
2331
0
CCGGCAGCAGATGGCAACGG


 


410644
3-14-3
426
2313
2332
0
TCCGGCAGCAGATGGCAACG


 


410645
3-14-3
427
2314
2333
0
CTCCGGCAGCAGATGGCAAC


 


410646
3-14-3
428
2315
2334
0
GCTCCGGCAGCAGATGGCAA









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 2-13-5 gap-widened motif. Table 5 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 5







Gapmer antisense compounds having a 2-13-5 motif


targeted to SEQ ID NO: 1
















5′
3′







start
Start






SEQ
site to
Site to




ISIS

ID
SEQ ID
SEQ ID




No.
Motif
NO
NO: 1
NO: 1
Mismatches
Sequences (5′ to 3′)





410647
2-13-5
184
 597
 616
0
TGAGGTATCCCCGGCGGGCA


 


410648
2-13-5
185
 598
 617
0
GTGAGGTATCCCCGGCGGGC


 


410649
2-13-5
186
 599
 618
0
GGTGAGGTATCCCCGGCGGG


 


410650
2-13-5
 11
 600
 619
0
TGGTGAGGTATCCCCGGCGG


 


410651
2-13-5
187
 601
 620
0
TTGGTGAGGTATCCCCGGCG


 


410652
2-13-5
188
 602
 621
0
CTTGGTGAGGTATCCCCGGC


 


410653
2-13-5
189
 603
 622
0
TCTTGGTGAGGTATCCCCGG


 


410654
2-13-5
190
 604
 623
0
ATCTTGGTGAGGTATCCCCG


 


410655
2-13-5
191
 605
 624
0
GATCTTGGTGAGGTATCCCC


 


410656
2-13-5
 12
 606
 625
0
GGATCTTGGTGAGGTATCCC


 


410657
2-13-5
192
 607
 626
0
AGGATCTTGGTGAGGTATCC


 


410658
2-13-5
243
 997
1016
0
GCCACGCCGGCATCCCGGCC


 


410659
2-13-5
244
 998
1017
0
GGCCACGCCGGCATCCCGGC


 


410660
2-13-5
245
 999
1018
0
TGGCCACGCCGGCATCCCGG


 


410661
2-13-5
246
1000
1019
0
TTGGCCACGCCGGCATCCCG


 


410662
2-13-5
247
1001
1020
0
CTTGGCCACGCCGGCATCCC


 


410663
2-13-5
248
1002
1021
0
CCTTGGCCACGCCGGCATCC


 


410664
2-13-5
249
1003
1022
0
CCCTTGGCCACGCCGGCATC


 


410665
2-13-5
 28
1004
1023
0
ACCCTTGGCCACGCCGGCAT


 


410666
2-13-5
250
1005
1024
0
CACCCTTGGCCACGCCGGCA


 


410667
2-13-5
251
1006
1025
0
GCACCCTTGGCCACGCCGGC


 


410668
2-13-5
252
1007
1026
0
GGCACCCTTGGCCACGCCGG


 


410669
2-13-5
253
1008
1027
0
TGGCACCCTTGGCCACGCCG


 


410670
2-13-5
254
1009
1028
0
CTGGCACCCTTGGCCACGCC


 


410671
2-13-5
255
1010
1029
0
GCTGGCACCCTTGGCCACGC


 


410672
2-13-5
348
1562
1581
0
AGGGAACCAGGCCTCATTGA


 


410673
2-13-5
349
1563
1582
0
CAGGGAACCAGGCCTCATTG


 


410674
2-13-5
 49
1564
1583
0
TCAGGGAACCAGGCCTCATT


 


410675
2-13-5
350
1565
1584
0
CTCAGGGAACCAGGCCTCAT


 


410676
2-13-5
351
1566
1585
0
CCTCAGGGAACCAGGCCTCA


 


410677
2-13-5
352
1567
1586
0
TCCTCAGGGAACCAGGCCTC


 


410678
2-13-5
353
1568
1587
0
GTCCTCAGGGAACCAGGCCT


 


410679
2-13-5
 50
1569
1588
0
GGTCCTCAGGGAACCAGGCC


 


410680
2-13-5
354
1570
1589
0
TGGTCCTCAGGGAACCAGGC


 


410681
2-13-5
355
1571
1590
0
CTGGTCCTCAGGGAACCAGG


 


410682
2-13-5
356
1572
1591
0
GCTGGTCCTCAGGGAACCAG


 


410683
2-13-5
376
1849
1868
0
CACCTGGCAATGGCGTAGAC


 


410684
2-13-5
377
1850
1869
0
GCACCTGGCAATGGCGTAGA


 


410685
2-13-5
378
1851
1870
0
AGCACCTGGCAATGGCGTAG


 


410686
2-13-5
379
1852
1871
0
CAGCACCTGGCAATGGCGTA


 


410687
2-13-5
380
1853
1872
0
GCAGCACCTGGCAATGGCGT


 


410688
2-13-5
381
1854
1873
0
GGCAGCACCTGGCAATGGCG


 


410689
2-13-5
382
1855
1874
0
AGGCAGCACCTGGCAATGGC


 


410690
2-13-5
383
1856
1875
0
CAGGCAGCACCTGGCAATGG


 


410691
2-13-5
384
1857
1876
0
GCAGGCAGCACCTGGCAATG


 


410692
2-13-5
 58
1858
1877
0
AGCAGGCAGCACCTGGCAAT


 


410693
2-13-5
385
1859
1878
0
TAGCAGGCAGCACCTGGCAA


 


410694
2-13-5
388
1915
1934
0
GTCCCCATGCTGGCCTCAGC


 


410695
2-13-5
389
1916
1935
0
GGTCCCCATGCTGGCCTCAG


 


410696
2-13-5
390
1917
1936
0
GGGTCCCCATGCTGGCCTCA


 


410697
2-13-5
391
1918
1937
0
CGGGTCCCCATGCTGGCCTC


 


410698
2-13-5
392
1919
1938
0
ACGGGTCCCCATGCTGGCCT


 


410699
2-13-5
 59
1920
1939
0
CACGGGTCCCCATGCTGGCC


 


410700
2-13-5
393
1921
1940
0
ACACGGGTCCCCATGCTGGC


 


410701
2-13-5
394
1922
1941
0
GACACGGGTCCCCATGCTGG


 


410702
2-13-5
395
1923
1942
0
GGACACGGGTCCCCATGCTG


 


410703
2-13-5
396
1924
1943
0
TGGACACGGGTCCCCATGCT


 


410704
2-13-5
397
1925
1944
0
GTGGACACGGGTCCCCATGC


 


410705
2-13-5
398
1926
1945
0
AGTGGACACGGGTCCCCATG


 


410706
2-13-5
399
1927
1946
0
CAGTGGACACGGGTCCCCAT


 


410707
2-13-5
400
1928
1947
0
GCAGTGGACACGGGTCCCCA


 


410708
2-13-5
401
1929
1948
0
GGCAGTGGACACGGGTCCCC


 


410709
2-13-5
402
1930
1949
0
TGGCAGTGGACACGGGTCCC


 


410710
2-13-5
403
1931
1950
0
GTGGCAGTGGACACGGGTCC


 


410711
2-13-5
404
1932
1951
0
GGTGGCAGTGGACACGGGTC


 


410712
2-13-5
405
1933
1952
0
TGGTGGCAGTGGACACGGGT


 


410713
2-13-5
 60
2100
2119
0
CTTTGCATTCCAGACCTGGG


 


410714
2-13-5
411
2101
2120
0
ACTTTGCATTCCAGACCTGG


 


410715
2-13-5
412
2102
2121
0
GACTTTGCATTCCAGACCTG


 


410716
2-13-5
413
2103
2122
0
TGACTTTGCATTCCAGACCT


 


410717
2-13-5
414
2104
2123
0
TTGACTTTGCATTCCAGACC


 


410718
2-13-5
 61
2105
2124
0
CTTGACTTTGCATTCCAGAC


 


410719
2-13-5
419
2305
2324
0
CAGATGGCAACGGCTGTCAC


 


410720
2-13-5
420
2306
2325
0
GCAGATGGCAACGGCTGTCA


 


410721
2-13-5
421
2307
2326
0
AGCAGATGGCAACGGCTGTC


 


410722
2-13-5
422
2308
2327
0
CAGCAGATGGCAACGGCTGT


 


410723
2-13-5
423
2309
2328
0
GCAGCAGATGGCAACGGCTG


 


410724
2-13-5
62
2310
2329
0
GGCAGCAGATGGCAACGGCT


 


410725
2-13-5
424
2311
2330
0
CGGCAGCAGATGGCAACGGC


 


410726
2-13-5
425
2312
2331
0
CCGGCAGCAGATGGCAACGG


 


410727
2-13-5
426
2313
2332
0
TCCGGCAGCAGATGGCAACG


 


410728
2-13-5
427
2314
2333
0
CTCCGGCAGCAGATGGCAAC


 


410729
2-13-5
428
2315
2334
0
GCTCCGGCAGCAGATGGCAA









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 1. In certain such embodiments, the nucleotide sequences illustrated in Table 2 have a 3-13-4 gap-widened motif. Table 6 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 1, having a 3-13-4 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 6







Gapmer antisense compounds having a 3-13-4 motif


targeted to SEQ ID NO: 1















SEQ
5′
3′






ID
start
Start




Oligo
Motif
NO:
site
Site
Mismatches
OligoSeq





405526
3-13-4
236
 963
 982
0
CCAGGTGGGTGCCATGACTG


 


405557
3-13-4
 50
1569
1588
0
GGTCCTCAGGGAACCAGGCC


 


405564
3-13-4
373
1737
1756
0
AACTGGAGCAGCTCAGCAGC









The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 1: 294-317, 406-440, 406-526, 410-436, 410-499, 446-526, 545-581, 591-619, 591-704, 591-743, 595-622, 600-626, 600-639, 600-670, 601-628, 602-628, 603-630, 611-636, 620-647, 638-665, 648-674, 657-684, 705-743, 782-810, 821-859, 835-859, 835-917, 835-942, 860-887, 860-899, 860-909, 860-917, 869-895, 878-905, 888-909, 923-952, 960-1034, 960-1173, 960-986, 967-991, 970-1023, 970-1064, 970-1117, 970-996, 977-1004, 985-1011, 989-1016, 992-1019, 997-1024, 997-1024, 998-1025, 999-1026, 1000-1027, 1001-1028, 1002-1021, 1002-1029, 1003-1029, 1004-1029, 1005-1029, 1006-1029, 1007-1034, 1036-1061, 1045-1072, 1076-1096, 1088-1115, 1098-1123, 1200-1251, 1210-1237, 1219-1245, 1228-1251, 1273-1444, 1295-1316, 1318-1345, 1328-1354, 1337-1361, 1344-1371, 1354-1377, 1380-1406, 1389-1416, 1400-1426, 1409-1434, 1465-1491, 1465-1602, 1474-1499, 1482-1519, 1513-1540, 1523-1549, 1526-1602, 1526-1624, 1532-1558, 1541-1568, 1552-1579, 1560-1587, 1561-1589, 1564-1591, 1565-1592, 1566-1592, 1567-1592, 1570-1597, 1571-1599, 1605-1706, 1628-1706, 1640-1666, 1672-1698, 1681-1706, 1735-1761, 1735-1765, 1740-1765, 1849-1876, 1849-1879, 1850-1877, 1851-1877, 1852-1878, 1852-1879, 1853-1879, 1854-1879, 1905-1955, 1915-1942, 1916-1943, 1917-1944, 1918-1945, 1919-1946, 1920-1939, 1920-1947, 1921-1948, 1922-1949, 1923-1950, 1924-1951, 1925-1952, 1926-1952, 1927-1952, 1928-1955, 1962-2059, 2040-2126, 2100-2126, 2100-2139, 2100-2206, 2101-2126, 2305-2332, 2305-2354, 2306-2333, 2307-2334, 2308-2334, 2309-2334, 2310-2334, 2410-2434, 2504-2528, 2509-2528, 2582-2625, 2606-2668, 2828-2855, 2832-2851, 2900-2927, 2900-2929, 2902-2927, 2983-3007, 2983-3013, 3227-3252, 3227-3456, 3472-3496, or 3543-3569.
In certain embodiments, a target region is nucleotides 294-317 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 294-317 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 159 or 160. In certain such embodiments, an antisense compound targeted to nucleotides 294-317 of SEQ ID NO: 1 is selected from ISIS NOs: 410742 or 405999.
In certain embodiments, a target region is nucleotides 406-440 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 406-440 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 162, 163, 164, 165, 166, 167, 168, 169 or 170. In certain such embodiments, an antisense compound targeted to nucleotides 406-440 of SEQ ID NO: 1 is selected from ISIS NOs: 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399949, or 410743.
In certain embodiments, a target region is nucleotides 406-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 406-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, or 176. In certain such embodiments, an antisense compound targeted to nucleotides 406-526 of SEQ ID NO: 1 is selected from ISIS NOs: 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399950, 410743, 410744, 410745, 395153, 399875, 406000, 406001, 406002 or 410746.
In certain embodiments, a target region is nucleotides 410-436 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 410-436 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168, 169 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 410-436 of SEQ ID NO: 1 is selected from ISIS NOs: 395152, 399874, 405865, 405866, 405867, 405868, or 399793.
In certain embodiments, a target region is nucleotides 410-499 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 410-499 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 9, 166, 167, 169, 170, 171 or 172. In certain such embodiments, an antisense compound targeted to nucleotides 410-499 of SEQ ID NO: 1 is selected from ISIS NOs: 395152, 399874, 405865, 405866, 405867, 405868, 399793, 399949, 410743, 410744, 410745, 395153, or 399875.
In certain embodiments, a target region is nucleotides 446-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 446-526 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 9, 171, 172, 173, 174, 175 or 176. In certain such embodiments, an antisense compound targeted to nucleotides 446-526 of SEQ ID NO: 1 is selected from ISIS NOs: 410744, 410745, 395153, 399875, 406000, 406001, 406002, or 410746.
In certain embodiments, a target region is nucleotides 545-581 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 545-581 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 545-581 of SEQ ID NO: 1 is selected from ISIS NOs: 410747, 406003, 406004, 406005, 395154, 399876, or 406006.
In certain embodiments, a target region is nucleotides 591-619 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-619 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 182, 183, 184, 185 or 186. In certain such embodiments, an antisense compound targeted to nucleotides 591-619 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, or 410650.
In certain embodiments, a target region is nucleotides 591-704 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-704 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, or 208. In certain such embodiments, an antisense compound targeted to nucleotides 591-704 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, or 410751.
In certain embodiments, a target region is nucleotides 591-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 591-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 17, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, or 209. In certain such embodiments, an antisense compound targeted to nucleotides 591-743 of SEQ ID NO: 1 is selected from ISIS NOs: 410748, 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, or 410752.
In certain embodiments, a target region is nucleotides 595-622 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 595-622 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 183, 184, 185, 186, 187, 188, or 189. In certain such embodiments, an antisense compound targeted to nucleotides 595-622 of SEQ ID NO: 1 is selected from ISIS NOs: 406007, 410529, 410574, 410647, 410530, 410575, 410648, 410531, 410576, 410649, 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, or 410653.
In certain embodiments, a target region is nucleotides 600-626 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-626 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 87, 188, 189, 190, 191, or 192. In certain such embodiments, an antisense compound targeted to nucleotides 600-626 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, or 410657.
In certain embodiments, a target region is nucleotides 600-639 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-639 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 187, 188, 189, 190, 191, 192, 193, 194, 195, or 196. In certain such embodiments, an antisense compound targeted to nucleotides 600-639 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, 406011, 406012, 399795, 399951, 406013, 395156, or 399878.
In certain embodiments, a target region is nucleotides 600-670 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 600-670 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 14, 15, 16, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, or 199. In certain such embodiments, an antisense compound targeted to nucleotides 600-670 of SEQ ID NO: 1 is selected from ISIS NOs: 395155, 399877, 410650, 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, 406011, 406012, 399795, 399951, 406013, 395156, 399878, or 399952.
In certain embodiments, a target region is nucleotides 601-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 601-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 601-628 of SEQ ID NO: 1 is selected from ISIS NOs: 410532, 410577, 410651, 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, or 406010.
In certain embodiments, a target region is nucleotides 602-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 602-628 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 602-628 of SEQ ID NO: 1 is selected from ISIS NOs: 406008, 410578, 410652, 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, or 406010.
In certain embodiments, a target region is nucleotides 603-630 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 603-630 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 603-630 of SEQ ID NO: 1 is selected from ISIS NOs: 410533, 410579, 410653, 406009, 410580, 410654, 410534, 410581, 410655, 399794, 399950, 410656, 410535, 410582, 410657, 406010, or 406011.
In certain embodiments, a target region is nucleotides 611-636 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 611-636 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 13, 194, 195 or 196. In certain such embodiments, an antisense compound targeted to nucleotides 611-636 of SEQ ID NO: 1 is selected from ISIS NOs: 406011, 406012, 399795, 399951, or 406013.
In certain embodiments, a target region is nucleotides 620-647 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 620-647 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 14 or 197. In certain such embodiments, an antisense compound targeted to nucleotides 620-647 of SEQ ID NO: 1 is selected from ISIS NOs: 395156, 399878, or 410749.
In certain embodiments, a target region is nucleotides 638-665 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 638-665 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 638-665 of SEQ ID NO: 1 is selected from ISIS NOs: 410750, 395157, or 399879.
In certain embodiments, a target region is nucleotides 648-674 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 648-674 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 16, 199, 200, or 201. In certain such embodiments, an antisense compound targeted to nucleotides 648-674 of SEQ ID NO: 1 is selected from ISIS NOs: 406014, 399796, 399952, 406015, or 406016.
In certain embodiments, a target region is nucleotides 657-684 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 657-684 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205, or 206. In certain such embodiments, an antisense compound targeted to nucleotides 657-684 of SEQ ID NO: 1 is selected from ISIS NOs: 410730, 406017, 406018, 406019, or 406020.
In certain embodiments, a target region is nucleotides 705-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 705-743 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17 or 209. In certain such embodiments, an antisense compound targeted to nucleotides 705-743 of SEQ ID NO: 1 is selected from ISIS NOs: 395158, 399880, or 410752.
In certain embodiments, a target region is nucleotides 782-810 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 782-810 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213, or 214. In certain such embodiments, an antisense compound targeted to nucleotides 782-810 of SEQ ID NO: 1 is selected from ISIS NOs: 406021, 406022, 395159, 399881, 406023, 406024, or 410732.
In certain embodiments, a target region is nucleotides 821-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 821-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 821-859 of SEQ ID NO: 1 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797, or 399953.
In certain embodiments, a target region is nucleotides 835-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-859 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 835-859 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, or 399953.
In certain embodiments, a target region is nucleotides 835-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 835-917 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, or 410754.
In certain embodiments, a target region is nucleotides 835-942 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 835-942 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 835-942 of SEQ ID NO: 1 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, 410754, 395163, or 399885.
In certain embodiments, a target region is nucleotides 860-887 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-887 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220, 221, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 860-887 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, or 405874.
In certain embodiments, a target region is nucleotides 860-899 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-899 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227 or 228. In certain such embodiments, an antisense compound targeted to nucleotides 860-899 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, or 399955.
In certain embodiments, a target region is nucleotides 860-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 860-909 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.
In certain embodiments, a target region is nucleotides 860-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 860-917 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 860-917 of SEQ ID NO: 1 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884, or 410754.
In certain embodiments, a target region is nucleotides 869-895 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 869-895 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226, or 227. In certain such embodiments, an antisense compound targeted to nucleotides 869-895 of SEQ ID NO: 1 is selected from ISIS NOs: 405875, 405876, 406027, or 406028.
In certain embodiments, a target region is nucleotides 878-905 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 878-905 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, or 231. In certain such embodiments, an antisense compound targeted to nucleotides 878-905 of SEQ ID NO: 1 is selected from ISIS NOs: 406029, 399799, 399955, 406030, 406031, or 410733.
In certain embodiments, a target region is nucleotides 888-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 888-909 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 888-909 of SEQ ID NO: 1 is selected from ISIS NOs: 406032, 395162, or 399884.
In certain embodiments, a target region is nucleotides 923-952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 923-952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain such embodiments, an antisense compound targeted to nucleotides 923-952 of SEQ ID NO: 1 is selected from ISIS NOs: 395163, 399885, or 410755.
In certain embodiments, a target region is nucleotides 960-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 960-1034 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, or 410758.
In certain embodiments, a target region is nucleotides 960-1173 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-1173 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, or 273. In certain such embodiments, an antisense compound targeted to nucleotides 960-1173 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, or 410763.
In certain embodiments, a target region is nucleotides 960-986 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 960-986 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. In certain such embodiments, an antisense compound targeted to nucleotides 960-986 of SEQ ID NO: 1 is selected from ISIS NOs: 410756, 405526, 405604, or 406033.
In certain embodiments, a target region is nucleotides 967-991 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 967-991 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 237 or 238. In certain such embodiments, an antisense compound targeted to nucleotides 967-991 of SEQ ID NO: 1 is selected from ISIS NOs: 406033 or 406034.
In certain embodiments, a target region is nucleotides 970-1023 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1023 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, or 249. In certain such embodiments, an antisense compound targeted to nucleotides 970-1023 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 410665.
In certain embodiments, a target region is nucleotides 970-1064 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1064 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, or 263. In certain such embodiments, an antisense compound targeted to nucleotides 970-1064 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 399957.
In certain embodiments, a target region is nucleotides 970-1117 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-1117 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 29, 30, 31, 32, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 149, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261, 262, 263, 264, 265, or 266. In certain such embodiments, an antisense compound targeted to nucleotides 970-1117 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 399890.
In certain embodiments, a target region is nucleotides 970-996 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 970-996 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, or 238. In certain such embodiments, an antisense compound targeted to nucleotides 970-996 of SEQ ID NO: 1 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, or 406035.
In certain embodiments, a target region is nucleotides 977-1004 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 977-1004 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 239 or 240. In certain such embodiments, an antisense compound targeted to nucleotides 977-1004 of SEQ ID NO: 1 is selected from ISIS NOs: 406035 or 410757.
In certain embodiments, a target region is nucleotides 985-1011 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 985-1011 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. In certain such embodiments, an antisense compound targeted to nucleotides 985-1011 of SEQ ID NO: 1 is selected from ISIS NOs: 410757, 406036, or 406037.
In certain embodiments, a target region is nucleotides 989-1016 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 989-1016 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 241, 242, or 243. In certain such embodiments, an antisense compound targeted to nucleotides 989-1016 of SEQ ID NO: 1 is selected from ISIS NOs: 406036, 406037, 410536, 410583, or 410658.
In certain embodiments, a target region is nucleotides 992-1019 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 992-1019 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 242, 243, 244, 245, or 246. In certain such embodiments, an antisense compound targeted to nucleotides 992-1019 of SEQ ID NO: 1 is selected from ISIS NOs: 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, or 410661.
In certain embodiments, a target region is nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, or 410666.
In certain embodiments, a target region is nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 997-1024 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 997-1024 of SEQ ID NO: 1 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, or 410666.
In certain embodiments, a target region is nucleotides 998-1025 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 998-1025 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247, 248, 249, 250, or 251. In certain such embodiments, an antisense compound targeted to nucleotides 998-1025 of SEQ ID NO: 1 is selected from ISIS NOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, or 410667.
In certain embodiments, a target region is nucleotides 999-1026 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 999-1026 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248, 249, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 999-1026 of SEQ ID NO: 1 is selected from ISIS NOs: 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, or 410668.
In certain embodiments, a target region is nucleotides 1000-1027 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1000-1027 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, or 253. In certain such embodiments, an antisense compound targeted to nucleotides 1000-1027 of SEQ ID NO: 1 is selected from ISIS NOs: 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, or 410669.
In certain embodiments, a target region is nucleotides 1001-1028 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1001-1028 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 251, 252, 253, or 254. In certain such embodiments, an antisense compound targeted to nucleotides 1001-1028 of SEQ ID NO: 1 is selected from ISIS NOs: 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, 410669, 410538, 410594, or 410670.
In certain embodiments, a target region is nucleotides 1002-1021 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1002-1021 of SEQ ID NO: 1. In one such embodiment, an antisense compound targeted to nucleotides 1002-1021 of SEQ ID NO: 1 is ISIS NO: 405879.
In certain embodiments, a target region is nucleotides 1002-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1002-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1002-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405833, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1003-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1003-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1003-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405880, 410589, 410664, 395165, 399887, 409126, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1004-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1004-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1004-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 395165, 399887, 409126, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1005-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1005-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1005-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1006-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1006-1029 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1006-1029 of SEQ ID NO: 1 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1007-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1007-1034 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1007-1034 of SEQ ID NO: 1 is selected from ISIS NOs: 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, 410671, or 410758.
In certain embodiments, a target region is nucleotides 1036-1061 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1036-1061 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 257, 258, or 259. In certain such embodiments, an antisense compound targeted to nucleotides 1036-1061 of SEQ ID NO: 1 is selected from ISIS NOs: 406039, 406040, 395166, 399888, or 406041.
In certain embodiments, a target region is nucleotides 1045-1072 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1045-1072 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 30, 260, 261, or 262. In certain such embodiments, an antisense compound targeted to nucleotides 1045-1072 of SEQ ID NO: 1 is selected from ISIS NOs: 399801, 399957, 406042, 406043, or 406044.
In certain embodiments, a target region is nucleotides 1076-1096 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1076-1096 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 31 or 264. In certain such embodiments, an antisense compound targeted to nucleotides 1076-1096 of SEQ ID NO: 1 is selected from ISIS NOs: 408642, 395167, or 399889.
In certain embodiments, a target region is nucleotides 1088-1115 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1088-1115 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 265 or 266. In certain such embodiments, an antisense compound targeted to nucleotides 1088-1115 of SEQ ID NO: 1 is selected from ISIS NOs: 410760 or 406045.
In certain embodiments, a target region is nucleotides 1098-1123 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1098-1123 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 267, 268, or 269. In certain such embodiments, an antisense compound targeted to nucleotides 1098-1123 of SEQ ID NO: 1 is selected from ISIS NOs: 395168, 399890, 405909, 405910, or 405911.
In certain embodiments, a target region is nucleotides 1200-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1200-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1200-1251 of SEQ ID NO: 1 is selected from ISIS NOs: 410764, 395169, 399891, 405913, 405914, 405915, 405916, 410734, 405917, 405918, 405919, 405920, 405921, or 405922.
In certain embodiments, a target region is nucleotides 1210-1237 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1210-1237 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278. In certain such embodiments, an antisense compound targeted to nucleotides 1210-1237 of SEQ ID NO: 1 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, or 405916.
In certain embodiments, a target region is nucleotides 1219-1245 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1219-1245 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. In certain such embodiments, an antisense compound targeted to nucleotides 1219-1245 of SEQ ID NO: 1 is selected from ISIS NOs: 410734, 405917, 405918, or 405919.
In certain embodiments, a target region is nucleotides 1228-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1228-1251 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1228-1251 of SEQ ID NO: 1 is selected from ISIS NOs: 405920, 405921, or 405922.
In certain embodiments, a target region is nucleotides 1273-1444 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1273-1444 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39, 40, 41, 286, 287, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 316, and 318. In certain such embodiments, an antisense compound targeted to nucleotides 1273-1444 of SEQ ID NO: 1 is selected from ISIS NOs: 410765, 410766, 405923, 395170, 399892, 410767, 405924, 410735, 405925, 405926, 395171, 399893, 405927, 395172, 399894, 405928, 399802, 399958, 405929, 395173, 399895, 405930, 405931, 405932, 405933, 405934, 399803, 399959, 405935, 410736, 405936, 395174, or 410769.
In certain embodiments, a target region is nucleotides 1295-1316 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1295-1316 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 149 or 288. In certain such embodiments, an antisense compound targeted to nucleotides 1295-1316 of SEQ ID NO: 1 is selected from ISIS NOs: 405923, 395170, or 399892.
In certain embodiments, a target region is nucleotides 1318-1345 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1318-1345 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293. In certain such embodiments, an antisense compound targeted to nucleotides 1318-1345 of SEQ ID NO: 1 is selected from ISIS NOs: 405924, 410735, 405925, 405926, 395171, or 399893.
In certain embodiments, a target region is nucleotides 1328-1354 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1328-1354 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 35, 128, 294 or 295. In certain such embodiments, an antisense compound targeted to nucleotides 1328-1354 of SEQ ID NO: 1 is selected from ISIS NOs: 405927, 395172, 399894, 405928, 399802, or 399958.
In certain embodiments, a target region is nucleotides 1337-1361 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1337-1361 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 36, 296, or 297. In certain such embodiments, an antisense compound targeted to nucleotides 1337-1361 of SEQ ID NO: 1 is selected from ISIS NOs: 405929, 395173, 399895, or 405930.
In certain embodiments, a target region is nucleotides 1344-1371 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1344-1371 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 37, 298, 299, 300, or 301. In certain such embodiments, an antisense compound targeted to nucleotides 1344-1371 of SEQ ID NO: 1 is selected from ISIS NOs: 405931, 405932, 405933, 405934, 399803, or 399959.
In certain embodiments, a target region is nucleotides 1354-1377 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1354-1377 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 302, 303, or 304. In certain such embodiments, an antisense compound targeted to nucleotides 1354-1377 of SEQ ID NO: 1 is selected from ISIS NOs: 405935, 410736, or 405936.
In certain embodiments, a target region is nucleotides 1380-1406 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1380-1406 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 305 or 306. In certain such embodiments, an antisense compound targeted to nucleotides 1380-1406 of SEQ ID NO: 1 is selected from ISIS NOs: 410768 or 405937.
In certain embodiments, a target region is nucleotides 1389-1416 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1389-1416 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310. In certain such embodiments, an antisense compound targeted to nucleotides 1389-1416 of SEQ ID NO: 1 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940, or 405941.
In certain embodiments, a target region is nucleotides 1400-1426 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1400-1426 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313, or 314. In certain such embodiments, an antisense compound targeted to nucleotides 1400-1426 of SEQ ID NO: 1 is selected from ISIS NOs: 399804, 399960, 405942, 405943, 410737, or 405944.
In certain embodiments, a target region is nucleotides 1409-1434 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1409-1434 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 315, 316, or 317. In certain such embodiments, an antisense compound targeted to nucleotides 1409-1434 of SEQ ID NO: 1 is selected from ISIS NOs: 405945, 399805, 399961, 405946, or 405947.
In certain embodiments, a target region is nucleotides 1465-1491 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1465-1491 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. In certain such embodiments, an antisense compound targeted to nucleotides 1465-1491 of SEQ ID NO: 1 is selected from ISIS NOs: 395176, 399898, 405948, 399806, 399962, or 405949.
In certain embodiments, a target region is nucleotides 1465-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1465-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1465-1602 of SEQ ID NO: 1 is selected from ISIS NOs: 395176, 399898, 405948, 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, 410770, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, or 399969.
In certain embodiments, a target region is nucleotides 1474-1499 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1474-1499 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 321, 322, or 323. In certain such embodiments, an antisense compound targeted to nucleotides 1474-1499 of SEQ ID NO: 1 is selected from ISIS NOs: 405950, 405951, 399807, 399963, or 405952.
In certain embodiments, a target region is nucleotides 1482-1519 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1482-1519 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 324, 325, 326, or 327. In certain such embodiments, an antisense compound targeted to nucleotides 1482-1519 of SEQ ID NO: 1 is selected from ISIS NOs: 410738, 405953, 405954, or 410770.
In certain embodiments, a target region is nucleotides 1513-1540 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1513-1540 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331, or 332. In certain such embodiments, an antisense compound targeted to nucleotides 1513-1540 of SEQ ID NO: 1 is selected from ISIS NOs: 405955, 405956, 405957, 405958, or 405959.
In certain embodiments, a target region is nucleotides 1523-1549 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1523-1549 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335, or 336. In certain such embodiments, an antisense compound targeted to nucleotides 1523-1549 of SEQ ID NO: 1 is selected from ISIS NOs: 405960, 405961, 399808, 399964, 405962, or 410739.
In certain embodiments, a target region is nucleotides 1526-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1526-1602 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1526-1602 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399969.
In certain embodiments, a target region is nucleotides 1526-1624 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1526-1624 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1526-1624 of SEQ ID NO: 1 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399902.
In certain embodiments, a target region is nucleotides 1532-1558 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1558 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 46, 337, or 338. In certain such embodiments, an antisense compound targeted to nucleotides 1532-1558 of SEQ ID NO: 1 is selected from ISIS NOs: 405963, 395177, 399899, 405964, 399809, or 399965.
In certain embodiments, a target region is nucleotides 1541-1568 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1541-1568 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 47, 340, 341, or 342. In certain such embodiments, an antisense compound targeted to nucleotides 1541-1568 of SEQ ID NO: 1 is selected from ISIS NOs: 405965, 405966, 399810, 399966, 405967, or 405968.
In certain embodiments, a target region is nucleotides 1552-1579 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1552-1579 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 343, 344, 345, or 346. In certain such embodiments, an antisense compound targeted to nucleotides 1552-1579 of SEQ ID NO: 1 is selected from ISIS NOs: 399811, 399967, 405969, 405970, 410740, or 405885.
In certain embodiments, a target region is nucleotides 1560-1587 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1560-1587 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, 351, 352, or 353. In certain such embodiments, an antisense compound targeted to nucleotides 1560-1587 of SEQ ID NO: 1 is selected from ISIS NOs: 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, or 410678.
In certain embodiments, a target region is nucleotides 1561-1589 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1561-1589 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 346, 347, 348, 349, 350, 351, 352, 353, or 354. In certain such embodiments, an antisense compound targeted to nucleotides 1561-1589 of SEQ ID NO: 1 is selected from ISIS NOs: 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, or 410680.
In certain embodiments, a target region is nucleotides 1564-1591 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1564-1591 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355, or 356. In certain such embodiments, an antisense compound targeted to nucleotides 1564-1591 of SEQ ID NO: 1 is selected from ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, or 410682.
In certain embodiments, a target region is nucleotides 1565-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1565-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1565-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 1566-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 1567-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1567-1592 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1567-1592 of SEQ ID NO: 1 is selected from ISIS NOs: 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 405557, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 1570-1597 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1570-1597 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 354, 355, 356, 357 or 358. In certain such embodiments, an antisense compound targeted to nucleotides 1570-1597 of SEQ ID NO: 1 is selected from ISIS NOs: 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, 405972 or 405973.
In certain embodiments, a target region is nucleotides 1571-1599 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1571-1599 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 87, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1571-1599 of SEQ ID NO: 1 is selected from ISIS NOs: 405971, 410603, 410681, 410540, 410604, 410682, 405972, 395179, 399901, 405973, or 405974.
In certain embodiments, a target region is nucleotides 1605-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1605-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 119, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1605-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 395180, 399902, 410771, 395181, 399903, 405975, 399814, 399970, 405976, 405977, 410772, 405978, 395182, 399904, 405979, 405980, 405981, 410741, 405982, or 405983.
In certain embodiments, a target region is nucleotides 1628-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1628-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1628-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 410771, 395181, 399903, 405975, 399814, 399970, 405976, 405977, 410772, 405978, 395182, 399904, 405979, 405980, 405981, 410741, 405982, or 405983.
In certain embodiments, a target region is nucleotides 1640-1666 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1640-1666 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. In certain such embodiments, an antisense compound targeted to nucleotides 1640-1666 of SEQ ID NO: 1 is selected from ISIS NOs: 395181, 399903, 405975, 399814, 399970, or 405976.
In certain embodiments, a target region is nucleotides 1672-1698 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1672-1698 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. In certain such embodiments, an antisense compound targeted to nucleotides 1672-1698 of SEQ ID NO: 1 is selected from ISIS NOs: 405978, 395182, 399904, 405979, or 405980.
In certain embodiments, a target region is nucleotides 1681-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1681-1706 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 1681-1706 of SEQ ID NO: 1 is selected from ISIS NOs: 405981, 410741, 405982, or 405983.
In certain embodiments, a target region is nucleotides 1735-1761 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1735-1761 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 372, 373, or 374. In certain such embodiments, an antisense compound targeted to nucleotides 1735-1761 of SEQ ID NO: 1 is selected from ISIS NOs: 405984, 405564, 405641, 405985, 399815, 399971, or 405986.
In certain embodiments, a target region is nucleotides 1735-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1735-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 1735-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 405984, 405564, 405641, 405985, 399815, 399971, 405986, 405987, 399816 or 399972.
In certain embodiments, a target region is nucleotides 1740-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1740-1765 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 1740-1765 of SEQ ID NO: 1 is selected from ISIS NOs: 399815, 399971, 405986, 405987, 399816, or 399972.
In certain embodiments, a target region is nucleotides 1849-1876 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1849-1876 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380, 381, 382, 383, or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1849-1876 of SEQ ID NO: 1 is selected from ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, or 410691.
In certain embodiments, a target region is nucleotides 1849-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1849-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1849-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410541, 410605, 410683, 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, or 405990.
In certain embodiments, a target region is nucleotides 1850-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1850-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1850-1877 of SEQ ID NO: 1 is selected from ISIS NOs: 410542, 410606, 410684, 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, or 410692.
In certain embodiments, a target region is nucleotides 1851-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1851-1877 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381, 382, 383, or 384. In certain such embodiments, an antisense compound targeted to nucleotides 1851-1877 of SEQ ID NO: 1 is selected from ISIS NOs: 410543, 410607, 410685, 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, or 410692.
In certain embodiments, a target region is nucleotides 1852-1878 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1852-1878 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382, 383, 384, or 385. In certain such embodiments, an antisense compound targeted to nucleotides 1852-1878 of SEQ ID NO: 1 is selected from ISIS NOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, or 410693.
In certain embodiments, a target region is nucleotides 1852-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1852-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1852-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410544, 410608, 410686, 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.
In certain embodiments, a target region is nucleotides 1853-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1853-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1853-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 410545, 410609, 410687, 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.
In certain embodiments, a target region is nucleotides 1854-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1854-1879 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384, 385, or 386. In certain such embodiments, an antisense compound targeted to nucleotides 1854-1879 of SEQ ID NO: 1 is selected from ISIS NOs: 405988, 410610, 410688, 410546, 410611, 410689, 405989, 410612, 410690, 410547, 410613, 395184, 410691, 399906, 410692, 410548, 410614, 410693, or 405990.
In certain embodiments, a target region is nucleotides 1905-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1905-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, or 406. In certain such embodiments, an antisense compound targeted to nucleotides 1905-1955 of SEQ ID NO: 1 is selected from ISIS NOs: 410773, 410549, 410615, 410694, 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, or 410774.
In certain embodiments, a target region is nucleotides 1915-1942 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1915-1942 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394, or 395. In certain such embodiments, an antisense compound targeted to nucleotides 1915-1942 of SEQ ID NO: 1 is selected from ISIS NOs: 410549, 410615, 410694, 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, or 410702.
In certain embodiments, a target region is nucleotides 1916-1943 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1916-1943 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395, or 396. In certain such embodiments, an antisense compound targeted to nucleotides 1916-1943 of SEQ ID NO: 1 is selected from ISIS NOs: 410550, 410616, 410695, 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, or 410703.
In certain embodiments, a target region is nucleotides 1917-1944 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1917-1944 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393, 394, 395, 396, or 397. In certain such embodiments, an antisense compound targeted to nucleotides 1917-1944 of SEQ ID NO: 1 is selected from ISIS NOs: 410551, 410617, 410696, 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, or 410704.
In certain embodiments, a target region is nucleotides 1918-1945 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1918-1945 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394, 395, 396, 397, or 398. In certain such embodiments, an antisense compound targeted to nucleotides 1918-1945 of SEQ ID NO: 1 is selected from ISIS NOs: 410552, 410618, 410697, 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, or 410705.
In certain embodiments, a target region is nucleotides 1919-1946 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1919-1946 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395, 396, 397, or 399. In certain such embodiments, an antisense compound targeted to nucleotides 1919-1946 of SEQ ID NO: 1 is selected from ISIS NOs: 410553, 410619, 410698, 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, or 410706.
In certain embodiments, a target region is nucleotides 1920-1939 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1920-1939 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59. In certain such embodiments, an antisense compound targeted to nucleotides 1920-1939 of SEQ ID NO: 1 is selected from ISIS NOs: 395185, 399907, or 410699.
In certain embodiments, a target region is nucleotides 1920-1947 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1920-1947 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396, 397, 398, 399, or 400. In certain such embodiments, an antisense compound targeted to nucleotides 1920-1947 of SEQ ID NO: 1 is selected from ISIS NOs: 395185, 399907, 410699, 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, or 410707.
In certain embodiments, a target region is nucleotides 1921-1948 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1921-1948 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397, 398, 399, 400, or 401. In certain such embodiments, an antisense compound targeted to nucleotides 1921-1948 of SEQ ID NO: 1 is selected from ISIS NOs: 410554, 410620, 410700, 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, or 410708.
In certain embodiments, a target region is nucleotides 1922-1949 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1922-1949 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398, 399, 400, 401, or 402. In certain such embodiments, an antisense compound targeted to nucleotides 1922-1949 of SEQ ID NO: 1 is selected from ISIS NOs: 405991, 410621, 410701, 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, 410708, 405995, 410629, or 410709.
In certain embodiments, a target region is nucleotides 1923-1950 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1923-1950 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399, 400, 401, 402, or 403. In certain such embodiments, an antisense compound targeted to nucleotides 1923-1950 of SEQ ID NO: 1 is selected from ISIS NOs: 410555, 410622, 410702, 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405944, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, or 410710.
In certain embodiments, a target region is nucleotides 1924-1951 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1924-1951 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400, 401, 402, 403, or 404. In certain such embodiments, an antisense compound targeted to nucleotides 1924-1951 of SEQ ID NO: 1 is selected from ISIS NOs: 405992, 410623, 410703, 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, or 410711.
In certain embodiments, a target region is nucleotides 1925-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1925-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1925-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 410556, 410624, 410704, 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.
In certain embodiments, a target region is nucleotides 1926-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1926-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1926-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 405993, 410625, 410705, 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.
In certain embodiments, a target region is nucleotides 1927-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1927-1952 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403, 404, or 405. In certain such embodiments, an antisense compound targeted to nucleotides 1927-1952 of SEQ ID NO: 1 is selected from ISIS NOs: 410557, 410626, 410706, 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, or 410712.
In certain embodiments, a target region is nucleotides 1928-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1928-1955 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404, 405, or 406. In certain such embodiments, an antisense compound targeted to nucleotides 1928-1955 of SEQ ID NO: 1 is selected from ISIS NOs: 405994, 410627, 410707, 410558, 410628, 410708, 405995, 410629, 410709, 410559, 410630, 410710, 410560, 410631, 410711, 410561, 410632, 410712, or 410774.
In certain embodiments, a target region is nucleotides 1962-2059 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 1962-2059 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 407, 408, 409, or 410. In certain such embodiments, an antisense compound targeted to nucleotides 1962-2059 of SEQ ID NO: 1 is selected from ISIS NOs: 410775, 410776, 410777, or 410778.
In certain embodiments, a target region is nucleotides 2040-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2040-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2040-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 410778, 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, or 405997.
In certain embodiments, a target region is nucleotides 2100-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718 or 405997.
In certain embodiments, a target region is nucleotides 2100-2139 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2139 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2139 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, 405997 or 410779.
In certain embodiments, a target region is nucleotides 2100-2206 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2100-2206 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, 416, 417, or 418. In certain such embodiments, an antisense compound targeted to nucleotides 2100-2206 of SEQ ID NO: 1 is selected from ISIS NOs: 395186, 399908, 410713, 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, 405997, 405997, 410780, or 410781.
In certain embodiments, a target region is nucleotides 2101-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2101-2126 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2101-2126 of SEQ ID NO: 1 is selected from ISIS NOs: 410562, 410633, 410714, 405996, 410634, 410715, 410563, 410635, 410716, 410564, 410636, 410717, 399817, 399973, 410718, or 405997.
In certain embodiments, a target region is nucleotides 2305-2332 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2305-2332 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, or 426. In certain such embodiments, an antisense compound targeted to nucleotides 2305-2332 of SEQ ID NO: 1 is selected from ISIS NOs: 410565, 410637, 410719, 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, or 410727.
In certain embodiments, a target region is nucleotides 2305-2354 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2305-2354 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments, an antisense compound targeted to nucleotides 2305-2354 of SEQ ID NO: 1 is selected from ISIS NOs: 410565, 410637, 410719, 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410783.
In certain embodiments, a target region is nucleotides 2306-2333 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2306-2333 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423, 424, 425, 426, or 427. In certain such embodiments, an antisense compound targeted to nucleotides 2306-2333 of SEQ ID NO: 1 is selected from ISIS NOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, or 410728.
In certain embodiments, a target region is nucleotides 2307-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2307-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2307-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2308-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2308-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2308-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2309-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2309-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2309-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2310-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2310-2334 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2310-2334 of SEQ ID NO: 1 is selected from ISIS NOs: 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2410-2434 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2410-2434 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 2410-2434 of SEQ ID NO: 1 is selected from ISIS NOs: 395188, 399910, 399818, or 399974.
In certain embodiments, a target region is nucleotides 2504-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2504-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 2504-2528 of SEQ ID NO: 1 is selected from ISIS NOs: 395189, 399911, 399819, or 399975.
In certain embodiments, a target region is nucleotides 2509-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2509-2528 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 65. In certain such embodiments, an antisense compound targeted to nucleotides 2509-2528 of SEQ ID NO: 1 is selected from ISIS NOs: 399819 or 399975.
In certain embodiments, a target region is nucleotides 2582-2625 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2582-2625 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certain such embodiments, an antisense compound targeted to nucleotides 2582-2625 of SEQ ID NO: 1 is selected from ISIS NOs: 399820, 399976, 395190, 399912, 395191, or 399913.
In certain embodiments, a target region is nucleotides 2606-2668 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2606-2668 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 67 or 153. In certain such embodiments, an antisense compound targeted to nucleotides 2606-2668 of SEQ ID NO: 1 is selected from ISIS NOs: 395191, 399913, 395192, or 399914.
In certain embodiments, a target region is nucleotides 2828-2855 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2828-2855 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434, 435, 436, 437 or 438. In certain such embodiments, an antisense compound targeted to nucleotides 2828-2855 of SEQ ID NO: 1 is selected from ISIS NOs: 405893, 405894, 405895, 405896, 395194, 399916, 405897, 405898, 405899, or 405900.
In certain embodiments, a target region is nucleotides 2832-2851 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2832-2851 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69. In certain such embodiments, an antisense compound targeted to nucleotides 2832-2851 of SEQ ID NO: 1 is selected from ISIS NOs: 395194, or 399916.
In certain embodiments, a target region is nucleotides 2900-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2900-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 2900-2927 of SEQ ID NO: 1 is selected from ISIS NOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821, 399977, 405905, or 405906.
In certain embodiments, a target region is nucleotides 2900-2929 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2900-2929 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 446 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 2900-2929 of SEQ ID NO: 1 is selected from ISIS NOs: 395195, 399917, 405901, 405902, 405903, 405904, 399821, 399977, 405905, 405906, 405907, or 405908.
In certain embodiments, a target region is nucleotides 2902-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2902-2927 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 2902-2927 of SEQ ID NO: 1 is selected from ISIS NOs: 405901, 405902, 405903, 405904, 399821, 399977, 405905, or 405906.
In certain embodiments, a target region is nucleotides 2983-3007 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2983-3007 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72 or 73. In certain such embodiments, an antisense compound targeted to nucleotides 2983-3007 of SEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918, 399822, or 399978.
In certain embodiments, a target region is nucleotides 2983-3013 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 2983-3013 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certain such embodiments, an antisense compound targeted to nucleotides 2983-3013 of SEQ ID NO: 1 is selected from ISIS NOs: 395196, 399918, 399822, 399978, 399823, or 399979.
In certain embodiments, a target region is nucleotides 3227-3252 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3227-3252 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3227-3252 of SEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919, 399824, or 399980.
In certain embodiments, a target region is nucleotides 3227-3456 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3227-3456 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3227-3456 of SEQ ID NO: 1 is selected from ISIS NOs: 395197, 399919, 399824, 399980, 395198, or 399920.
In certain embodiments, a target region is nucleotides 3472-3496 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3472-3496 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 3472-3496 of SEQ ID NO: 1 is selected from ISIS NOs: 395199, 399921, 399825 or 399981.
In certain embodiments, a target region is nucleotides 3543-3569 of SEQ ID NO: 1. In certain embodiments, an antisense compound is targeted to nucleotides 3543-3569 of SEQ ID NO: 1. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 3543-3569 of SEQ ID NO: 1 is selected from ISIS NOs: 395200, 399922, 399826 or 399982.
In certain embodiments, antisense compound target a PCSK9 nucleic acid having the sequence of nucleotides 25475000 to 25504000 of GENBANK® Accession No. NT 032977.8, first deposited with GENBANK® on Feb. 26, 2006, and incorporated herein as SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide is targeted to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 2 is at least 90% complementary to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 2 is at least 95% complementary to SEQ ID NO: 2. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 1 is 100% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide comprises a nucleotide sequence selected from a nucleotide sequence set forth in Table 7.







TABLE 7







Antisense sequences targeted to


NT_032977.8 (SEQ ID NO: 2)











5′
3′




Start
Start



SEQ
Site to
Site to



ID
SEQ ID
SEQ ID



NO
NO: 2
NO: 2
Sequence (5′ to 3′)





  4
 2274
 2293
GCGCGGAATCCTGGCTGGGA


 


  5
 2381
 2400
GAGGAGACCTAGAGGCCGTG


 


159
 2433
 2452
GCCTGGAGCTGACGGTGCCC


 


160
 2437
 2456
GACCGCCTGGAGCTGACGGT


 


  6
 2439
 2458
AGGACCGCCTGGAGCTGACG


 


162
 2545
 2564
AAGGCTAGCACCAGCTCCTC


 


163
 2546
 2565
CAAGGCTAGCACCAGCTCCT


 


164
 2547
 2566
GCAAGGCTAGCACCAGCTCC


 


165
 2548
 2567
CGCAAGGCTAGCACCAGCTC


 


  7
 2549
 2568
ACGCAAGGCTAGCACCAGCT


 


166
 2550
 2569
AACGCAAGGCTAGCACCAGC


 


167
 2551
 2570
GAACGCAAGGCTAGCACCAG


 


168
 2552
 2571
GGAACGCAAGGCTAGCACCA


 


169
 2553
 2572
CGGAACGCAAGGCTAGCACC


 


  8
 2556
 2575
CCTCGGAACGCAAGGCTAGC


 


170
 2560
 2579
TCCTCCTCGGAACGCAAGGC


 


171
 2585
 2604
GTGCTCGGGTGCTTCGGCCA


 


172
 2605
 2624
TGGAAGGTGGCTGTGGTTCC


 


  9
 2619
 2638
CCTTGGCGCAGCGGTGGAAG


 


107
 3056
 3075
CCCACTATAATGGCAAGCCC


 


 80
 4306
 4325
AACCCAGTTCTAATGCACCT


 


106
 5140
 5159
CCAGTCAGAGTAGAACAGAG


 


102
 5590
 5609
ATGTGCAGAGATCAATCACA


 


121
 5599
 5618
GGAGCCTACATGTGCAGAGA


 


 94
 5667
 5686
AGCATGGCACCAGCATCTGC


 


176
 6444
 6463
CGTAGGTGCCAGGCAACCTC


 


177
 6482
 6501
TGACTGCGAGAGGTGGGTCT


 


178
 6492
 6511
CAGTGCGCTCTGACTGCGAG


 


179
 6494
 6513
GGCAGTGCGCTCTGACTGCG


 


180
 6496
 6515
CGGGCAGTGCGCTCTGACTG


 


 10
 6498
 6517
GGCGGGCAGTGCGCTCTGAC


 


181
 6499
 6518
CGGCGGGCAGTGCGCTCTGA


 


182
 6528
 6547
ATCCCCGGCGGGCAGCCTGG


 


183
 6532
 6551
AGGTATCCCCGGCGGGCAGC


 


184
 6534
 6553
TGAGGTATCCCCGGCGGGCA


 


185
 6535
 6554
GTGAGGTATCCCCGGCGGGC


 


186
 6536
 6555
GGTGAGGTATCCCCGGCGGG


 


 11
 6537
 6556
TGGTGAGGTATCCCCGGCGG


 


187
 6538
 6557
TTGGTGAGGTATCCCCGGCG


 


188
 6539
 6558
CTTGGTGAGGTATCCCCGGC


 


189
 6540
 6559
TCTTGGTGAGGTATCCCCGG


 


190
 6541
 6560
ATCTTGGTGAGGTATCCCCG


 


191
 6542
 6561
GATCTTGGTGAGGTATCCCC


 


 12
 6543
 6562
GGATCTTGGTGAGGTATCCC


 


192
 6544
 6563
AGGATCTTGGTGAGGTATCC


 


193
 6546
 6565
GCAGGATCTTGGTGAGGTAT


 


194
 6548
 6567
ATGCAGGATCTTGGTGAGGT


 


195
 6550
 6569
ACATGCAGGATCTTGGTGAG


 


 13
 6552
 6571
AGACATGCAGGATCTTGGTG


 


196
 6554
 6573
GAAGACATGCAGGATCTTGG


 


 14
 6557
 6576
ATGGAAGACATGCAGGATCT


 


197
 6565
 6584
AGAAGGCCATGGAAGACATG


 


198
 6575
 6594
GAAGCCAGGAAGAAGGCCAT


 


 15
 6583
 6602
TTCACCAGGAAGCCAGGAAG


 


199
 6585
 6604
TCTTCACCAGGAAGCCAGGA


 


 16
 6588
 6607
TCATCTTCACCAGGAAGCCA


 


200
 6590
 6609
ACTCATCTTCACCAGGAAGC


 


201
 6592
 6611
CCACTCATCTTCACCAGGAA


 


202
 6594
 6613
CGCCACTCATCTTCACCAGG


 


203
 6596
 6615
GTCGCCACTCATCTTCACCA


 


204
 6598
 6617
AGGTCGCCACTCATCTTCAC


 


205
 6600
 6619
GCAGGTCGCCACTCATCTTC


 


206
 6602
 6621
CAGCAGGTCGCCACTCATCT


 


207
 6604
 6623
TCCAGCAGGTCGCCACTCAT


 


108
 6652
 6671
CCCAGCCCTATCAGGAAGTG


 


144
 7099
 7118
TGACATCCAGGAGGGAGGAG


 


 91
 7556
 7575
AGACTGATGGAAGGCATTGA


 


131
 7565
 7584
GTGTTGAGCAGACTGATGGA


 


145
 8836
 8855
TGACATCTTGTCTGGGAGCC


 


 90
 8948
 8967
AGACTAGGAGCCTGAGTTTT


 


125
 9099
 9118
GGCCTGCAGAAGCCAGAGAG


 


 17
 9130
 9149
CCTCGATGTAGTCGACATGG


 


209
 9149
 9168
GCAAAGACAGAGGAGTCCTC


 


210
 9207
 9226
TTCATCCGCCCGGTACCGTG


 


211
 9209
 9228
TATTCATCCGCCCGGTACCG


 


 18
 9210
 9229
GTATTCATCCGCCCGGTACC


 


212
 9212
 9231
TGGTATTCATCCGCCCGGTA


 


213
 9214
 9233
GCTGGTATTCATCCGCCCGG


 


214
 9216
 9235
GGGCTGGTATTCATCCGCCC


 


148
10252
10271
TGGCAGCAACTCAGACATAT


 


127
10633
10652
GGTGGTAATTTGTCACAGCA


 


 84
11308
11327
AAGGTCACACAGTTAAGAGT


 


 79
11472
11491
AAATGCAGGGCTAAAATCAC


 


 88
12715
12734
ACTGGATACATTGGCAGACA


 


111
12928
12947
CTAGAGGAACCACTAGATAT


 


 85
13681
13700
ACAAATTCCCAGACTCAGCA


 


100
13746
13765
ATCTCAGGACAGGTGAGCAA


 


116
13760
13779
GAGTAGAGATTCTCATCTCA


 


129
13816
13835
GTGCCATCTGAACAGCACCT


 


117
13828
13847
GAGTCTTCTGAAGTGCCATC


 


 81
13903
13922
AAGCAGGGCCTCAGGTGGAA


 


110
13926
13945
CCTGGAACCCCTGCAGCCAG


 


152
13977
13996
TTCAGGCAGGTTGCTGCTAG


 


 83
13986
14005
AAGGAAGACTTCAGGCAGGT


 


140
13998
14017
TCAGCCAGGCCAAAGGAAGA


 


137
14112
14131
TAGGGAGAGCTCACAGATGC


 


136
14122
14141
TAGGAGAAAGTAGGGAGAGC


 


132
14179
14198
TAAAAGCTGCAAGAGACTCA


 


139
14267
14286
TCAGAGAAAACAGTCACCGA


 


 92
14397
14416
AGAGACAGGAAGCTGCAGCT


 


142
14404
14423
TCATTTTAGAGACAGGAAGC


 


113
14441
14460
GAATAACAGTGATGTCTGGC


 


138
14494
14513
TCACAGCTCACCGAGTCTGC


 


 98
14524
14543
AGTGTAAAATAAAGCCCCTA


 


 96
14601
14620
AGGACCCAAGTCATCCTGCT


 


124
14631
14650
GGCCATCAGCTGGCAATGCT


 


 82
14670
14689
AAGGAAAGGGAGGCCTAGAG


 


133
14675
14694
TAGACAAGGAAAGGGAGGCC


 


103
14681
14700
ATTTCATAGACAAGGAAAGG


 


155
14801
14820
CTTATAGTTAACACACAGAA


 


156
14809
14828
AAGTCAACCTTATAGTTAAC


 


215
14877
14896
ATACACCTCCACCAGGCTGC


 


216
14888
14907
GTGTCTAGGAGATACACCTC


 


 19
14891
14910
CTGGTGTCTAGGAGATACAC


 


217
14893
14912
TGCTGGTGTCTAGGAGATAC


 


 20
14896
14915
GTATGCTGGTGTCTAGGAGA


 


 21
14916
14935
GATTTCCCGGTGGTCACTCT


 


218
14918
14937
TCGATTTCCCGGTGGTCACT


 


219
14919
14938
CTCGATTTCCCGGTGGTCAC


 


220
14920
14939
CCTCGATTTCCCGGTGGTCA


 


221
14921
14940
CCCTCGATTTCCCGGTGGTC


 


 22
14922
14941
GCCCTCGATTTCCCGGTGGT


 


222
14923
14942
TGCCCTCGATTTCCCGGTGG


 


223
14924
14943
CTGCCCTCGATTTCCCGGTG


 


224
14925
14944
CCTGCCCTCGATTTCCCGGT


 


225
14926
14945
CCCTGCCCTCGATTTCCCGG


 


226
14930
14949
ATGACCCTGCCCTCGATTTC


 


227
14932
14951
CCATGACCCTGCCCTCGATT


 


228
14934
14953
GACCATGACCCTGCCCTCGA


 


 23
14936
14955
GTGACCATGACCCTGCCCTC


 


229
14938
14957
CGGTGACCATGACCCTGCCC


 


230
14940
14959
GTCGGTGACCATGACCCTGC


 


231
14942
14961
AAGTCGGTGACCATGACCCT


 


232
14944
14963
CGAAGTCGGTGACCATGACC


 


 24
14946
14965
CTCGAAGTCGGTGACCATGA


 


233
14954
14973
GGCACATTCTCGAAGTCGGT


 


 25
14979
14998
GTGGAAGCGGGTCCCGTCCT


 


235
15254
15273
GGTGGGTGCCATGACTGTCA


 


236
15257
15276
CCAGGTGGGTGCCATGACTG


 


237
15261
15280
CCTGCCAGGTGGGTGCCATG


 


 26
15264
15283
ACCCCTGCCAGGTGGGTGCC


 


238
15266
15285
CCACCCCTGCCAGGTGGGTG


 


 27
15269
15288
TGACCACCCCTGCCAGGTGG


 


239
15271
15290
GCTGACCACCCCTGCCAGGT


 


240
15279
15298
TCCCGGCCGCTGACCACCCC


 


241
15283
15302
GGCATCCCGGCCGCTGACCA


 


242
15286
15305
GCCGGCATCCCGGCCGCTGA


 


243
15291
15310
GCCACGCCGGCATCCCGGCC


 


244
15292
15311
GGCCACGCCGGCATCCCGGC


 


245
15293
15312
TGGCCACGCCGGCATCCCGG


 


246
15294
15313
TTGGCCACGCCGGCATCCCG


 


247
15295
15314
CTTGGCCACGCCGGCATCCC


 


248
15296
15315
CCTTGGCCACGCCGGCATCC


 


249
15297
15316
CCCTTGGCCACGCCGGCATC


 


 28
15298
15317
ACCCTTGGCCACGCCGGCAT


 


447
15298
15317
ACCCTTGGTCACGCCGGCAT


 


250
15299
15318
CACCCTTGGCCACGCCGGCA


 


251
15300
15319
GCACCCTTGGCCACGCCGGC


 


252
15301
15320
GGCACCCTTGGCCACGCCGG


 


253
15302
15321
TGGCACCCTTGGCCACGCCG


 


254
15303
15322
CTGGCACCCTTGGCCACGCC


 


255
15304
15323
GCTGGCACCCTTGGCCACGC


 


256
15309
15328
CGCATGCTGGCACCCTTGGC


 


257
15330
15349
CAGTTGAGCACGCGCAGGCT


 


258
15332
15351
GGCAGTTGAGCACGCGCAGG


 


 29
15334
15353
TTGGCAGTTGAGCACGCGCA


 


259
15336
15355
CCTTGGCAGTTGAGCACGCG


 


 30
15339
15358
TTCCCTTGGCAGTTGAGCAC


 


260
15341
15360
CCTTCCCTTGGCAGTTGAGC


 


261
15345
15364
GTGCCCTTCCCTTGGCAGTT


 


262
15347
15366
CCGTGCCCTTCCCTTGGCAG


 


263
15358
15377
GGTGCCGCTAACCGTGCCCT


 


 86
15471
15490
ACAGCATTCTTGGTTAGGAG


 


 97
16134
16153
AGTCAAGCTGCTGCCCAGAG


 


120
16668
16687
GCTAGTTATTAAGCACCTGC


 


150
17267
17286
TGTGAGCTCTGGCCCAGTGG


 


115
18377
18396
GAGTAAGGCAGGTTACTCTC


 


134
18408
18427
TAGATGTGACTAACATTTAA


 


157
18561
18580
AGGAACAAAGCCAAGGTCAC


 


266
18591
18610
TGGCTTTTCCGAATAAACTC


 


 32
18593
18612
GCTGGCTTTTCCGAATAAAC


 


267
18595
18614
CAGCTGGCTTTTCCGAATAA


 


268
18597
18616
ACCAGCTGGCTTTTCCGAAT


 


269
18599
18618
GGACCAGCTGGCTTTTCCGA


 


270
18603
18622
GGCTGGACCAGCTGGCTTTT


 


271
18614
18633
GTGGCCCCACAGGCTGGACC


 


272
18627
18646
AGCAGCACCACCAGTGGCCC


 


273
18649
18668
GCTGTACCCACCCGCCAGGG


 


274
18695
18714
CGACCCCAGCCCTCGCCAGG


 


 33
18705
18724
GTGACCAGCACGACCCCAGC


 


275
18707
18726
CGGTGACCAGCACGACCCCA


 


276
18709
18728
AGCGGTGACCAGCACGACCC


 


277
18711
18730
GCAGCGGTGACCAGCACGAC


 


278
18713
18732
CGGCAGCGGTGACCAGCACG


 


279
18714
18733
CCGGCAGCGGTGACCAGCAC


 


280
18717
18736
TTGCCGGCAGCGGTGACCAG


 


281
18719
18738
AGTTGCCGGCAGCGGTGACC


 


282
18721
18740
GAAGTTGCCGGCAGCGGTGA


 


283
18723
18742
CGGAAGTTGCCGGCAGCGGT


 


284
18725
18744
CCCGGAAGTTGCCGGCAGCG


 


285
18727
18746
GTCCCGGAAGTTGCCGGCAG


 


105
19203
19222
CACATTAGCCTTGCTCAAGT


 


151
19913
19932
TGTGATGACCTGGAAAGGTG


 


288
19931
19950
GGCATTGGTGGCCCCAACTG


 


149
19933
19952
TGGGCATTGGTGGCCCCAAC


 


289
19941
19960
GCTGGTCTTGGGCATTGGTG


 


290
19954
19973
CCCAGGGTCACCGGCTGGTC


 


291
19956
19975
TCCCCAGGGTCACCGGCTGG


 


292
19958
19977
AGTCCCCAGGGTCACCGGCT


 


293
19960
19979
AAAGTCCCCAGGGTCACCGG


 


 34
19962
19981
CCAAAGTCCCCAGGGTCACC


 


294
19964
19983
CCCCAAAGTCCCCAGGGTCA


 


128
19966
19985
GTCCCCAAAGTCCCCAGGGT


 


295
19969
19988
TTGGTCCCCAAAGTCCCCAG


 


 35
19971
19990
AGTTGGTCCCCAAAGTCCCC


 


296
19973
19992
AAAGTTGGTCCCCAAAGTCC


 


 36
19976
19995
GCCAAAGTTGGTCCCCAAAG


 


297
19978
19997
CGGCCAAAGTTGGTCCCCAA


 


298
19980
19999
AGCGGCCAAAGTTGGTCCCC


 


299
19982
20001
ACAGCGGCCAAAGTTGGTCC


 


300
19984
20003
ACACAGCGGCCAAAGTTGGT


 


301
19986
20005
CCACACAGCGGCCAAAGTTG


 


 37
19988
20007
GTCCACACAGCGGCCAAAGT


 


302
19990
20009
AGGTCCACACAGCGGCCAAA


 


303
19992
20011
AGAGGTCCACACAGCGGCCA


 


304
19994
20013
AAAGAGGTCCACACAGCGGC


 


 38
19997
20016
GGCAAAGAGGTCCACACAGC


 


305
20016
20035
CAATGATGTCCTCCCCTGGG


 


306
20023
20042
GAGGCACCAATGATGTCCTC


 


 39
20025
20044
TGGAGGCACCAATGATGTCC


 


307
20027
20046
GCTGGAGGCACCAATGATGT


 


308
20029
20048
TCGCTGGAGGCACCAATGAT


 


309
20031
20050
AGTCGCTGGAGGCACCAATG


 


310
20033
20052
GCAGTCGCTGGAGGCACCAA


 


 40
20036
20055
GCTGCAGTCGCTGGAGGCAC


 


311
20038
20057
GTGCTGCAGTCGCTGGAGGC


 


312
20040
20059
AGGTGCTGCAGTCGCTGGAG


 


313
20042
20061
GCAGGTGCTGCAGTCGCTGG


 


314
20043
20062
AGCAGGTGCTGCAGTCGCTG


 


315
20045
20064
AAAGCAGGTGCTGCAGTCGC


 


 41
20047
20066
ACAAAGCAGGTGCTGCAGTC


 


316
20049
20068
ACACAAAGCAGGTGCTGCAG


 


317
20051
20070
TGACACAAAGCAGGTGCTGC


 


318
20061
20080
TCCCACTCTGTGACACAAAG


 


158
20100
20119
GTGGTGACTTACCAGCCACG


 


109
20188
20207
CCCCTGCACAGAGCCTGGCA


 


141
20624
20643
TCATGGCTGCAATGCCTGGT


 


320
20629
20648
CAGCATCATGGCTGCAATGC


 


321
20631
20650
GACAGCATCATGGCTGCAAT


 


322
20633
20652
CAGACAGCATCATGGCTGCA


 


 43
20635
20654
GGCAGACAGCATCATGGCTG


 


323
20637
20656
TCGGCAGACAGCATCATGGC


 


324
20639
20658
GCTCGGCAGACAGCATCATG


 


325
20641
20660
CGGCTCGGCAGACAGCATCA


 


326
20643
20662
TCCGGCTCGGCAGACAGCAT


 


327
20657
20676
CGGCCAGGGTGAGCTCCGGC


 


328
20670
20689
CTCTGCCTCAACTCGGCCAG


 


329
20672
20691
GTCTCTGCCTCAACTCGGCC


 


330
20674
20693
CAGTCTCTGCCTCAACTCGG


 


331
20676
20695
ATCAGTCTCTGCCTCAACTC


 


332
20678
20697
GGATCAGTCTCTGCCTCAAC


 


333
20680
20699
GTGGATCAGTCTCTGCCTCA


 


334
20682
20701
AAGTGGATCAGTCTCTGCCT


 


 44
20683
20702
GAAGTGGATCAGTCTCTGCC


 


335
20685
20704
GAGAAGTGGATCAGTCTCTG


 


336
20687
20706
CAGAGAAGTGGATCAGTCTC


 


337
20689
20708
GGCAGAGAAGTGGATCAGTC


 


 45
20691
20710
TTGGCAGAGAAGTGGATCAG


 


338
20693
20712
CTTTGGCAGAGAAGTGGATC


 


 46
20696
20715
CATCTTTGGCAGAGAAGTGG


 


339
20698
20717
GACATCTTTGGCAGAGAAGT


 


340
20700
20719
ATGACATCTTTGGCAGAGAA


 


 47
20702
20721
TGATGACATCTTTGGCAGAG


 


341
20704
20723
ATTGATGACATCTTTGGCAG


 


342
20706
20725
TCATTGATGACATCTTTGGC


 


 48
20709
20728
GCCTCATTGATGACATCTTT


 


343
20711
20730
AGGCCTCATTGATGACATCT


 


344
20713
20732
CCAGGCCTCATTGATGACAT


 


345
20715
20734
AACCAGGCCTCATTGATGAC


 


346
20717
20736
GGAACCAGGCCTCATTGATG


 


347
20718
20737
GGGAACCAGGCCTCATTGAT


 


348
20719
20738
AGGGAACCAGGCCTCATTGA


 


349
20720
20739
CAGGGAACCAGGCCTCATTG


 


 49
20721
20740
TCAGGGAACCAGGCCTCATT


 


350
20722
20741
CTCAGGGAACCAGGCCTCAT


 


351
20723
20742
CCTCAGGGAACCAGGCCTCA


 


352
20724
20743
TCCTCAGGGAACCAGGCCTC


 


353
20725
20744
GTCCTCAGGGAACCAGGCCT


 


 50
20726
20745
GGTCCTCAGGGAACCAGGCC


 


354
20727
20746
TGGTCCTCAGGGAACCAGGC


 


355
20728
20747
CTGGTCCTCAGGGAACCAGG


 


356
20729
20748
GCTGGTCCTCAGGGAACCAG


 


357
20730
20749
CGCTGGTCCTCAGGGAACCA


 


 87
20733
20752
ACCCGCTGGTCCTCAGGGAA


 


358
20735
20754
GTACCCGCTGGTCCTCAGGG


 


359
20737
20756
CAGTACCCGCTGGTCCTCAG


 


 51
20740
20759
GGTCAGTACCCGCTGGTCCT


 


119
20762
20781
GCAGGGCGGCCACCAGGTTG


 


360
20785
20804
ACCTGCCCCATGGGTGCTGG


 


 93
20995
21014
AGAGAGGAGGGCTTAAAGAA


 


 95
21082
21101
AGCTGCCAACCTGCAAAAAG


 


361
21088
21107
CAAAACAGCTGCCAACCTGC


 


 53
21091
21110
CTGCAAAACAGCTGCCAACC


 


362
21093
21112
TCCTGCAAAACAGCTGCCAA


 


363
21095
21114
AGTCCTGCAAAACAGCTGCC


 


364
21106
21125
GCTGACCATACAGTCCTGCA


 


365
21118
21137
GGCCCCGAGTGTGCTGACCA


 


 54
21121
21140
GTAGGCCCCGAGTGTGCTGA


 


366
21123
21142
GTGTAGGCCCCGAGTGTGCT


 


367
21125
21144
CCGTGTAGGCCCCGAGTGTG


 


368
21127
21146
ATCCGTGTAGGCCCCGAGTG


 


369
21129
21148
CCATCCGTGTAGGCCCCGAG


 


370
21131
21150
GGCCATCCGTGTAGGCCCCG


 


371
21133
21152
GTGGCCATCCGTGTAGGCCC


 


372
21181
21200
CTGGAGCAGCTCAGCAGCTC


 


460
21181
21194
CAGCTCAGCAGCTC


 


373
21183
21202
AACTGGAGCAGCTCAGCAGC


 


 55
21186
21205
AGAAACTGGAGCAGCTCAGC


 


374
21188
21207
GGAGAAACTGGAGCAGCTCA


 


375
21190
21209
CTGGAGAAACTGGAGCAGCT


 


 56
21192
21211
TCCTGGAGAAACTGGAGCAG


 


143
21481
21500
TGAAAATCCATCCAGCACTG


 


 89
21589
21608
AGAACCATGGAGCACCTGAG


 


448
21692
21711
CTGCCCTTCCACCAAAATGC


 


449
21693
21712
ACTGCCCTTCCACCAAAATG


 


450
21694
21713
CACTGCCCTTCCACCAAAAT


 


451
21695
21714
GCACTGCCCTTCCACCAAAA


 


123
21696
21715
GGCACTGCCCTTCCACCAAA


 


452
21697
21716
GGGCACTGCCCTTCCACCAA


 


453
21698
21717
TGGGCACTGCCCTTCCACCA


 


454
21699
21718
CTGGGCACTGCCCTTCCACC


 


455
21700
21719
GCTGGGCACTGCCCTTCCAC


 


 57
22096
22115
CGTTGTGGGCCCGGCAGACC


 


376
22133
22152
CACCTGGCAATGGCGTAGAC


 


377
22134
22153
GCACCTGGCAATGGCGTAGA


 


378
22135
22154
AGCACCTGGCAATGGCGTAG


 


379
22136
22155
CAGCACCTGGCAATGGCGTA


 


380
22137
22156
GCAGCACCTGGCAATGGCGT


 


381
22138
22157
GGCAGCACCTGGCAATGGCG


 


382
22139
22158
AGGCAGCACCTGGCAATGGC


 


383
22140
22159
CAGGCAGCACCTGGCAATGG


 


384
22141
22160
GCAGGCAGCACCTGGCAATG


 


 58
22142
22161
AGCAGGCAGCACCTGGCAAT


 


385
22143
22162
TAGCAGGCAGCACCTGGCAA


 


386
22144
22163
GTAGCAGGCAGCACCTGGCA


 


387
22189
22208
TGGCCTCAGCTGGTGGAGCT


 


388
22199
22218
GTCCCCATGCTGGCCTCAGC


 


389
22200
22219
GGTCCCCATGCTGGCCTCAG


 


390
22201
22220
GGGTCCCCATGCTGGCCTCA


 


391
22202
22221
CGGGTCCCCATGCTGGCCTC


 


392
22203
22222
ACGGGTCCCCATGCTGGCCT


 


 59
22204
22223
CACGGGTCCCCATGCTGGCC


 


393
22205
22224
ACACGGGTCCCCATGCTGGC


 


394
22206
22225
GACACGGGTCCCCATGCTGG


 


395
22207
22226
GGACACGGGTCCCCATGCTG


 


396
22208
22227
TGGACACGGGTCCCCATGCT


 


397
22209
22228
GTGGACACGGGTCCCCATGC


 


398
22210
22229
AGTGGACACGGGTCCCCATG


 


399
22211
22230
CAGTGGACACGGGTCCCCAT


 


400
22212
22231
GCAGTGGACACGGGTCCCCA


 


401
22213
22232
GGCAGTGGACACGGGTCCCC


 


402
22214
22233
TGGCAGTGGACACGGGTCCC


 


403
22215
22234
GTGGCAGTGGACACGGGTCC


 


404
22216
22235
GGTGGCAGTGGACACGGGTC


 


405
22217
22236
TGGTGGCAGTGGACACGGGT


 


406
22220
22239
TGTTGGTGGCAGTGGACACG


 


126
22292
22311
GGTGCATAAGGAGAAAGAGA


 


408
23985
24004
GTGCCAAGGTCCTCCACCTC


 


409
24005
24024
TCAGCACAGGCGGCTTGTGG


 


410
24035
24054
CCACGCACTGGTTGGGCTGA


 


 60
24095
24114
CTTTGCATTCCAGACCTGGG


 


411
24096
24115
ACTTTGCATTCCAGACCTGG


 


412
24097
24116
GACTTTGCATTCCAGACCTG


 


413
24098
24117
TGACTTTGCATTCCAGACCT


 


414
24099
24118
TTGACTTTGCATTCCAGACC


 


 61
24100
24119
CTTGACTTTGCATTCCAGAC


 


415
24102
24121
TCCTTGACTTTGCATTCCAG


 


416
24115
24134
CGGGATTCCATGCTCCTTGA


 


147
24858
24877
TGAGTTCATTTAAGAGTGGA


 


118
24907
24926
GCACCATCCAGACCAGAATC


 


114
25413
25432
GAGAGGTTCAGATCCAGGCC


 


418
25994
26013
GGAGGGCACTGCAGCCAGTC


 


419
26112
26131
CAGATGGCAACGGCTGTCAC


 


420
26113
26132
GCAGATGGCAACGGCTGTCA


 


421
26114
26133
AGCAGATGGCAACGGCTGTC


 


422
26115
26134
CAGCAGATGGCAACGGCTGT


 


423
26116
26135
GCAGCAGATGGCAACGGCTG


 


 62
26117
26136
GGCAGCAGATGGCAACGGCT


 


424
26118
26137
CGGCAGCAGATGGCAACGGC


 


425
26119
26138
CCGGCAGCAGATGGCAACGG


 


426
26120
26139
TCCGGCAGCAGATGGCAACG


 


427
26121
26140
CTCCGGCAGCAGATGGCAAC


 


428
26122
26141
GCTCCGGCAGCAGATGGCAA


 


429
26132
26151
CCAGGTGCCGGCTCCGGCAG


 


430
26142
26161
GAGGCCTGCGCCAGGTGCCG


 


154
26217
26236
TTTTAAAGCTCAGCCCCAGC


 


 63
26222
26241
AACCATTTTAAAGCTCAGCC


 


 64
26311
26330
TCAAGGGCCAGGCCAGCAGC


 


 65
26316
26335
CCCACTCAAGGGCCAGGCCA


 


122
26389
26408
GGAGGGAGCTTCCTGGCACC


 


 66
26404
26423
ATGCCCCACAGTGAGGGAGG


 


 67
26413
26432
AATGGTGAAATGCCCCACAG


 


153
26456
26475
TTGGGAGCAGCTGGCAGCAC


 


 68
26557
26576
CATGGGAAGAATCCTGCCTC


 


431
26635
26654
ATGAGGGCCATCAGCACCTT


 


432
26636
26655
GATGAGGGCCATCAGCACCT


 


433
26637
26656
AGATGAGGGCCATCAGCACC


 


434
26638
26657
GAGATGAGGGCCATCAGCAC


 


 69
26639
26658
GGAGATGAGGGCCATCAGCA


 


435
26640
26659
TGGAGATGAGGGCCATCAGC


 


436
26641
26660
CTGGAGATGAGGGCCATCAG


 


437
26642
26661
GCTGGAGATGAGGGCCATCA


 


438
26643
26662
AGCTGGAGATGAGGGCCATC


 


461
26684
26697
TTAATCAGGGAGCC


 


 70
26707
26726
TAGATGCCATCCAGAAAGCT


 


439
26709
26728
GCTAGATGCCATCCAGAAAG


 


440
26710
26729
GGCTAGATGCCATCCAGAAA


 


441
26711
26730
TGGCTAGATGCCATCCAGAA


 


442
26712
26731
CTGGCTAGATGCCATCCAGA


 


 71
26713
26732
TCTGGCTAGATGCCATCCAG


 


443
26714
26733
CTCTGGCTAGATGCCATCCA


 


444
26715
26734
CCTCTGGCTAGATGCCATCC


 


445
26716
26735
GCCTCTGGCTAGATGCCATC


 


446
26717
26736
AGCCTCTGGCTAGATGCCAT


 


 72
26790
26809
GGCATAGAGCAGAGTAAAGG


 


 73
26795
26814
AGCCTGGCATAGAGCAGAGT


 


135
26801
26820
TAGCACAGCCTGGCATAGAG


 


112
27034
27053
GAAGAGGCTTGGCTTCAGAG


 


 74
27040
27059
AAGTAAGAAGAGGCTTGGCT


 


 75
27244
27263
GCTCAAGGAGGGACAGTTGT


 


 76
27279
27298
AAAGATAAATGTCTGCTTGC


 


 77
27284
27303
ACCCAAAAGATAAATGTCTG


 


 78
27350
27369
TCTTCAAGTTACAAAAGCAA


 


 99
27357
27376
ATAAATATCTTCAAGTTACA









In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 5-10-5 gapmer motif. Table 8 illustrates gapmer antisense compounds targeted to SEQ ID NO: 2, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 8







Gapmer antisense compounds having a 5-10-5 motif


targeted to SEQ ID NO: 2
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 2
NO: 2
Mismatches
Sequence (5′-3′)





395149
5-10-5
  4
 2274
 2293
0
GCGCGGAATCCTGGCTGGGA


 


395150
5-10-5
  5
 2381
 2400
0
GAGGAGACCTAGAGGCCGTG


 


395151
5-10-5
  6
 2439
 2458
0
AGGACCGCCTGGAGCTGACG


 


395152
5-10-5
  7
 2549
 2568
0
ACGCAAGGCTAGCACCAGCT


 


399793
5-10-5
  8
 2556
 2575
0
CCTCGGAACGCAAGGCTAGC


 


395153
5-10-5
  9
 2619
 2638
0
CCTTGGCGCAGCGGTGGAAG


 


399837
5-10-5
107
 3056
 3075
0
CCCACTATAATGGCAAGCCC


 


399838
5-10-5
 80
 4306
 4325
0
AACCCAGTTCTAATGCACCT


 


399839
5-10-5
106
 5140
 5159
0
CCAGTCAGAGTAGAACAGAG


 


395221
5-10-5
102
 5590
 5609
0
ATGTGCAGAGATCAATCACA


 


399840
5-10-5
121
 5599
 5618
0
GGAGCCTACATGTGCAGAGA


 


399841
5-10-5
 94
 5667
 5686
0
AGCATGGCACCAGCATCTGC


 


395154
5-10-5
 10
 6498
 6517
0
GGCGGGCAGTGCGCTCTGAC


 


395155
5-10-5
 11
 6537
 6556
0
TGGTGAGGTATCCCCGGCGG


 


399794
5-10-5
 12
 6543
 6562
0
GGATCTTGGTGAGGTATCCC


 


399795
5-10-5
 13
 6552
 6571
0
AGACATGCAGGATCTTGGTG


 


395156
5-10-5
 14
 6557
 6576
0
ATGGAAGACATGCAGGATCT


 


395157
5-10-5
 15
 6583
 6602
0
TTCACCAGGAAGCCAGGAAG


 


399796
5-10-5
 16
 6588
 6607
0
TCATCTTCACCAGGAAGCCA


 


399842
5-10-5
108
 6652
 6671
0
CCCAGCCCTATCAGGAAGTG


 


399843
5-10-5
144
 7099
 7118
0
TGACATCCAGGAGGGAGGAG


 


399844
5-10-5
 91
 7556
 7575
0
AGACTGATGGAAGGCATTGA


 


399845
5-10-5
131
 7565
 7584
0
GTGTTGAGCAGACTGATGGA


 


399846
5-10-5
145
 8836
 8855
0
TGACATCTTGTCTGGGAGCC


 


399847
5-10-5
 90
 8948
 8967
0
AGACTAGGAGCCTGAGTTTT


 


399848
5-10-5
125
 9099
 9118
0
GGCCTGCAGAAGCCAGAGAG


 


395158
5-10-5
 17
 9130
 9149
0
CCTCGATGTAGTCGACATGG


 


395159
5-10-5
 18
 9210
 9229
0
GTATTCATCCGCCCGGTACC


 


399849
5-10-5
148
10252
10271
0
TGGCAGCAACTCAGACATAT


 


395222
5-10-5
127
10633
10652
0
GGTGGTAATTTGTCACAGCA


 


395223
5-10-5
 84
11308
11327
0
AAGGTCACACAGTTAAGAGT


 


399850
5-10-5
 79
11472
11491
0
AAATGCAGGGCTAAAATCAC


 


399851
5-10-5
 88
12715
12734
0
ACTGGATACATTGGCAGACA


 


399852
5-10-5
111
12928
12947
0
CTAGAGGAACCACTAGATAT


 


395201
5-10-5
 85
13681
13700
0
ACAAATTCCCAGACTCAGCA


 


399827
5-10-5
100
13746
13765
0
ATCTCAGGACAGGTGAGCAA


 


399828
5-10-5
116
13760
13779
0
GAGTAGAGATTCTCATCTCA


 


395202
5-10-5
129
13816
13835
0
GTGCCATCTGAACAGCACCT


 


399829
5-10-5
117
13828
13847
0
GAGTCTTCTGAAGTGCCATC


 


399830
5-10-5
 81
13903
13922
0
AAGCAGGGCCTCAGGTGGAA


 


395203
5-10-5
110
13926
13945
0
CCTGGAACCCCTGCAGCCAG


 


395204
5-10-5
152
13977
13996
0
TTCAGGCAGGTTGCTGCTAG


 


399831
5-10-5
 83
13986
14005
0
AAGGAAGACTTCAGGCAGGT


 


395205
5-10-5
140
13998
14017
0
TCAGCCAGGCCAAAGGAAGA


 


399832
5-10-5
137
14112
14131
0
TAGGGAGAGCTCACAGATGC


 


395206
5-10-5
136
14122
14141
0
TAGGAGAAAGTAGGGAGAGC


 


395207
5-10-5
132
14179
14198
0
TAAAAGCTGCAAGAGACTCA


 


395208
5-10-5
139
14267
14286
0
TCAGAGAAAACAGTCACCGA


 


399833
5-10-5
 92
14397
14416
0
AGAGACAGGAAGCTGCAGCT


 


395209
5-10-5
142
14404
14423
0
TCATTTTAGAGACAGGAAGC


 


395210
5-10-5
113
14441
14460
0
GAATAACAGTGATGTCTGGC


 


395211
5-10-5
138
14494
14513
0
TCACAGCTCACCGAGTCTGC


 


395212
5-10-5
 98
14524
14543
0
AGTGTAAAATAAAGCCCCTA


 


395213
5-10-5
 96
14601
14620
0
AGGACCCAAGTCATCCTGCT


 


395214
5-10-5
124
14631
14650
0
GGCCATCAGCTGGCAATGCT


 


399834
5-10-5
 82
14670
14689
0
AAGGAAAGGGAGGCCTAGAG


 


395215
5-10-5
133
14675
14694
0
TAGACAAGGAAAGGGAGGCC


 


395216
5-10-5
103
14681
14700
0
ATTTCATAGACAAGGAAAGG


 


395217
5-10-5
155
14801
14820
0
CTTATAGTTAACACACAGAA


 


399835
5-10-5
156
14809
14828
0
AAGTCAACCTTATAGTTAAC


 


395160
5-10-5
 19
14891
14910
0
CTGGTGTCTAGGAGATACAC


 


399797
5-10-5
 20
14896
14915
0
GTATGCTGGTGTCTAGGAGA


 


395161
5-10-5
 21
14916
14935
0
GATTTCCCGGTGGTCACTCT


 


399798
5-10-5
 22
14922
14941
0
GCCCTCGATTTCCCGGTGGT


 


399799
5-10-5
 23
14936
14955
0
GTGACCATGACCCTGCCCTC


 


395162
5-10-5
 24
14946
14965
0
CTCGAAGTCGGTGACCATGA


 


395163
5-10-5
 25
14979
14998
0
GTGGAAGCGGGTCCCGTCCT


 


395164
5-10-5
 26
15264
15283
0
ACCCCTGCCAGGTGGGTGCC


 


399800
5-10-5
 27
15269
15288
0
TGACCACCCCTGCCAGGTGG


 


395165
5-10-5
 28
15298
15317
0
ACCCTTGGCCACGCCGGCAT


 


395166
5-10-5
 29
15334
15353
0
TTGGCAGTTGAGCACGCGCA


 


399801
5-10-5
 30
15339
15358
0
TTCCCTTGGCAGTTGAGCAC


 


399853
5-10-5
 86
15471
15490
0
ACAGCATTCTTGGTTAGGAG


 


399854
5-10-5
 97
16134
16153
0
AGTCAAGCTGCTGCCCAGAG


 


399855
5-10-5
120
16668
16687
0
GCTAGTTATTAAGCACCTGC


 


399856
5-10-5
150
17267
17286
0
TGTGAGCTCTGGCCCAGTGG


 


399857
5-10-5
115
18377
18396
0
GAGTAAGGCAGGTTACTCTC


 


399858
5-10-5
134
18408
18427
0
TAGATGTGACTAACATTTAA


 


395224
5-10-5
157
18561
18580
0
AGGAACAAAGCCAAGGTCAC


 


395168
5-10-5
 32
18593
18612
0
GCTGGCTTTTCCGAATAAAC


 


395169
5-10-5
 33
18705
18724
0
GTGACCAGCACGACCCCAGC


 


399859
5-10-5
105
19203
19222
0
CACATTAGCCTTGCTCAAGT


 


399860
5-10-5
151
19913
19932
0
TGTGATGACCTGGAAAGGTG


 


395170
5-10-5
149
19933
19952
0
TGGGCATTGGTGGCCCCAAC


 


395171
5-10-5
 34
19962
19981
0
CCAAAGTCCCCAGGGTCACC


 


395172
5-10-5
128
19966
19985
0
GTCCCCAAAGTCCCCAGGGT


 


399802
5-10-5
 35
19971
19990
0
AGTTGGTCCCCAAAGTCCCC


 


395173
5-10-5
 36
19976
19995
0
GCCAAAGTTGGTCCCCAAAG


 


399803
5-10-5
 37
19988
20007
0
GTCCACACAGCGGCCAAAGT


 


395174
5-10-5
 38
19997
20016
0
GGCAAAGAGGTCCACACAGC


 


395175
5-10-5
 39
20025
20044
0
TGGAGGCACCAATGATGTCC


 


399804
5-10-5
 40
20036
20055
0
GCTGCAGTCGCTGGAGGCAC


 


399805
5-10-5
 41
20047
20066
0
ACAAAGCAGGTGCTGCAGTC


 


399861
5-10-5
158
20100
20119
0
GTGGTGACTTACCAGCCACG


 


399862
5-10-5
109
20188
20207
0
CCCCTGCACAGAGCCTGGCA


 


399863
5-10-5
141
20624
20643
0
TCATGGCTGCAATGCCTGGT


 


399807
5-10-5
 43
20635
20654
0
GGCAGACAGCATCATGGCTG


 


399808
5-10-5
 44
20683
20702
0
GAAGTGGATCAGTCTCTGCC


 


395177
5-10-5
 45
20691
20710
0
TTGGCAGAGAAGTGGATCAG


 


399809
5-10-5
 46
20696
20715
0
CATCTTTGGCAGAGAAGTGG


 


399810
5-10-5
 47
20702
20721
0
TGATGACATCTTTGGCAGAG


 


399811
5-10-5
 48
20709
20728
0
GCCTCATTGATGACATCTTT


 


399812
5-10-5
 49
20721
20740
0
TCAGGGAACCAGGCCTCATT


 


395178
5-10-5
 50
20726
20745
0
GGTCCTCAGGGAACCAGGCC


 


395179
5-10-5
 87
20733
20752
0
ACCCGCTGGTCCTCAGGGAA


 


399813
5-10-5
 51
20740
20759
0
GGTCAGTACCCGCTGGTCCT


 


395180
5-10-5
119
20762
20781
0
GCAGGGCGGCCACCAGGTTG


 


399864
5-10-5
 93
20995
21014
0
AGAGAGGAGGGCTTAAAGAA


 


399865
5-10-5
 95
21082
21101
0
AGCTGCCAACCTGCAAAAAG


 


399814
5-10-5
 53
21091
21110
0
CTGCAAAACAGCTGCCAACC


 


395182
5-10-5
 54
21121
21140
0
GTAGGCCCCGAGTGTGCTGA


 


399815
5-10-5
 55
21186
21205
0
AGAAACTGGAGCAGCTCAGC


 


399816
5-10-5
 56
21192
21211
0
TCCTGGAGAAACTGGAGCAG


 


399866
5-10-5
143
21481
21500
0
TGAAAATCCATCCAGCACTG


 


399867
5-10-5
 89
21589
21608
0
AGAACCATGGAGCACCTGAG


 


399868
5-10-5
123
21696
21715
0
GGCACTGCCCTTCCACCAAA


 


395183
5-10-5
 57
22096
22115
0
CGTTGTGGGCCCGGCAGACC


 


395184
5-10-5
 58
22142
22161
0
AGCAGGCAGCACCTGGCAAT


 


395185
5-10-5
 59
22204
22223
0
CACGGGTCCCCATGCTGGCC


 


395225
5-10-5
126
22292
22311
0
GGTGCATAAGGAGAAAGAGA


 


395186
5-10-5
 60
24095
24114
0
CTTTGCATTCCAGACCTGGG


 


399817
5-10-5
 61
24100
24119
0
CTTGACTTTGCATTCCAGAC


 


395226
5-10-5
147
24858
24877
0
TGAGTTCATTTAAGAGTGGA


 


399869
5-10-5
118
24907
24926
0
GCACCATCCAGACCAGAATC


 


399870
5-10-5
114
25413
25432
0
GAGAGGTTCAGATCCAGGCC


 


395187
5-10-5
 62
26117
26136
0
GGCAGCAGATGGCAACGGCT


 


395188
5-10-5
154
26217
26236
0
TTTTAAAGCTCAGCCCCAGC


 


399818
5-10-5
 63
26222
26241
0
AACCATTTTAAAGCTCAGCC


 


395189
5-10-5
 64
26311
26330
0
TCAAGGGCCAGGCCAGCAGC


 


399819
5-10-5
 65
26316
26335
0
CCCACTCAAGGGCCAGGCCA


 


399820
5-10-5
122
26389
26408
0
GGAGGGAGCTTCCTGGCACC


 


395190
5-10-5
 66
26404
26423
0
ATGCCCCACAGTGAGGGAGG


 


395191
5-10-5
 67
26413
26432
0
AATGGTGAAATGCCCCACAG


 


395192
5-10-5
153
26456
26475
0
TTGGGAGCAGCTGGCAGCAC


 


395193
5-10-5
 68
26557
26576
0
CATGGGAAGAATCCTGCCTC


 


395194
5-10-5
 69
26639
26658
0
GGAGATGAGGGCCATCAGCA


 


395195
5-10-5
 70
26707
26726
0
TAGATGCCATCCAGAAAGCT


 


399821
5-10-5
 71
26713
26732
0
TCTGGCTAGATGCCATCCAG


 


395196
5-10-5
 72
26790
26809
0
GGCATAGAGCAGAGTAAAGG


 


399822
5-10-5
 73
26795
26814
0
AGCCTGGCATAGAGCAGAGT


 


399823
5-10-5
135
26801
26820
0
TAGCACAGCCTGGCATAGAG


 


395197
5-10-5
112
27034
27053
0
GAAGAGGCTTGGCTTCAGAG


 


399824
5-10-5
 74
27040
27059
0
AAGTAAGAAGAGGCTTGGCT


 


395198
5-10-5
 75
27244
27263
0
GCTCAAGGAGGGACAGTTGT


 


395199
5-10-5
 76
27279
27298
0
AAAGATAAATGTCTGCTTGC


 


399825
5-10-5
 77
27284
27303
0
ACCCAAAAGATAAATGTCTG


 


395200
5-10-5
 78
27350
27369
0
TCTTCAAGTTACAAAAGCAA


 


399826
5-10-5
 99
27357
27376
0
ATAAATATCTTCAAGTTACA


 


405861
5-10-5
162
 2545
 2564
0
AAGGCTAGCACCAGCTCCTC


 


405862
5-10-5
163
 2546
 2565
0
CAAGGCTAGCACCAGCTCCT


 


405863
5-10-5
164
 2547
 2566
0
GCAAGGCTAGCACCAGCTCC


 


405864
5-10-5
165
 2548
 2567
0
CGCAAGGCTAGCACCAGCTC


 


405865
5-10-5
166
 2550
 2569
0
AACGCAAGGCTAGCACCAGC


 


405866
5-10-5
167
 2551
 2570
0
GAACGCAAGGCTAGCACCAG


 


405867
5-10-5
168
 2552
 2571
0
GGAACGCAAGGCTAGCACCA


 


405868
5-10-5
169
 2553
 2572
0
CGGAACGCAAGGCTAGCACC


 


405869
5-10-5
218
14918
14937
0
TCGATTTCCCGGTGGTCACT


 


405870
5-10-5
219
14919
14938
0
CTCGATTTCCCGGTGGTCAC


 


405871
5-10-5
220
14920
14939
0
CCTCGATTTCCCGGTGGTCA


 


405872
5-10-5
221
14921
14940
0
CCCTCGATTTCCCGGTGGTC


 


405873
5-10-5
222
14923
14942
0
TGCCCTCGATTTCCCGGTGG


 


405874
5-10-5
223
14924
14943
0
CTGCCCTCGATTTCCCGGTG


 


405875
5-10-5
224
14925
14944
0
CCTGCCCTCGATTTCCCGGT


 


405876
5-10-5
225
14926
14945
0
CCCTGCCCTCGATTTCCCGG


 


405877
5-10-5
246
15294
15313
0
TTGGCCACGCCGGCATCCCG


 


405878
5-10-5
247
15295
15314
0
CTTGGCCACGCCGGCATCCC


 


405879
5-10-5
248
15296
15315
0
CCTTGGCCACGCCGGCATCC


 


405880
5-10-5
249
15297
15316
0
CCCTTGGCCACGCCGGCATC


 


405881
5-10-5
250
15299
15318
0
CACCCTTGGCCACGCCGGCA


 


405882
5-10-5
251
15300
15319
0
GCACCCTTGGCCACGCCGGC


 


405883
5-10-5
252
15301
15320
0
GGCACCCTTGGCCACGCCGG


 


405884
5-10-5
253
15302
15321
0
TGGCACCCTTGGCCACGCCG


 


405885
5-10-5
346
20717
20736
0
GGAACCAGGCCTCATTGATG


 


405886
5-10-5
347
20718
20737
0
GGGAACCAGGCCTCATTGAT


 


405887
5-10-5
348
20719
20738
0
AGGGAACCAGGCCTCATTGA


 


405888
5-10-5
349
20720
20739
0
CAGGGAACCAGGCCTCATTG


 


405889
5-10-5
350
20722
20741
0
CTCAGGGAACCAGGCCTCAT


 


405890
5-10-5
351
20723
20742
0
CCTCAGGGAACCAGGCCTCA


 


405891
5-10-5
352
20724
20743
0
TCCTCAGGGAACCAGGCCTC


 


405892
5-10-5
353
20725
20744
0
GTCCTCAGGGAACCAGGCCT


 


405893
5-10-5
431
26635
26654
0
ATGAGGGCCATCAGCACCTT


 


405894
5-10-5
432
26636
26655
0
GATGAGGGCCATCAGCACCT


 


405895
5-10-5
433
26637
26656
0
AGATGAGGGCCATCAGCACC


 


405896
5-10-5
434
26638
26657
0
GAGATGAGGGCCATCAGCAC


 


405897
5-10-5
435
26640
26659
0
TGGAGATGAGGGCCATCAGC


 


405898
5-10-5
436
26641
26660
0
CTGGAGATGAGGGCCATCAG


 


405899
5-10-5
437
26642
26661
0
GCTGGAGATGAGGGCCATCA


 


405900
5-10-5
438
26643
26662
0
AGCTGGAGATGAGGGCCATC


 


405901
5-10-5
439
26709
26728
0
GCTAGATGCCATCCAGAAAG


 


405902
5-10-5
440
26710
26729
0
GGCTAGATGCCATCCAGAAA


 


405903
5-10-5
441
26711
26730
0
TGGCTAGATGCCATCCAGAA


 


405904
5-10-5
442
26712
26731
0
CTGGCTAGATGCCATCCAGA


 


405905
5-10-5
443
26714
26733
0
CTCTGGCTAGATGCCATCCA


 


405906
5-10-5
444
26715
26734
0
CCTCTGGCTAGATGCCATCC


 


405907
5-10-5
445
26716
26735
0
GCCTCTGGCTAGATGCCATC


 


405908
5-10-5
446
26717
26736
0
AGCCTCTGGCTAGATGCCAT


 


405909
5-10-5
267
18595
18614
0
CAGCTGGCTTTTCCGAATAA


 


405910
5-10-5
268
18597
18616
0
ACCAGCTGGCTTTTCCGAAT


 


405911
5-10-5
269
18599
18618
0
GGACCAGCTGGCTTTTCCGA


 


405912
5-10-5
270
18603
18622
0
GGCTGGACCAGCTGGCTTTT


 


405913
5-10-5
275
18707
18726
0
CGGTGACCAGCACGACCCCA


 


405914
5-10-5
276
18709
18728
0
AGCGGTGACCAGCACGACCC


 


405915
5-10-5
277
18711
18730
0
GCAGCGGTGACCAGCACGAC


 


405916
5-10-5
278
18713
18732
0
CGGCAGCGGTGACCAGCACG


 


405917
5-10-5
280
18717
18736
0
TTGCCGGCAGCGGTGACCAG


 


405918
5-10-5
281
18719
18738
0
AGTTGCCGGCAGCGGTGACC


 


405919
5-10-5
282
18721
18740
0
GAAGTTGCCGGCAGCGGTGA


 


405920
5-10-5
283
18723
18742
0
CGGAAGTTGCCGGCAGCGGT


 


405921
5-10-5
284
18725
18744
0
CCCGGAAGTTGCCGGCAGCG


 


405922
5-10-5
285
18727
18746
0
GTCCCGGAAGTTGCCGGCAG


 


405923
5-10-5
288
19931
19950
0
GGCATTGGTGGCCCCAACTG


 


405924
5-10-5
290
19954
19973
0
CCCAGGGTCACCGGCTGGTC


 


405925
5-10-5
292
19958
19977
0
AGTCCCCAGGGTCACCGGCT


 


405926
5-10-5
293
19960
19979
0
AAAGTCCCCAGGGTCACCGG


 


405927
5-10-5
294
19964
19983
0
CCCCAAAGTCCCCAGGGTCA


 


405928
5-10-5
295
19969
19988
0
TTGGTCCCCAAAGTCCCCAG


 


405929
5-10-5
296
19973
19992
0
AAAGTTGGTCCCCAAAGTCC


 


405930
5-10-5
297
19978
19997
0
CGGCCAAAGTTGGTCCCCAA


 


405931
5-10-5
298
19980
19999
0
AGCGGCCAAAGTTGGTCCCC


 


405932
5-10-5
299
19982
20001
0
ACAGCGGCCAAAGTTGGTCC


 


405933
5-10-5
300
19984
20003
0
ACACAGCGGCCAAAGTTGGT


 


405934
5-10-5
301
19986
20005
0
CCACACAGCGGCCAAAGTTG


 


405935
5-10-5
302
19990
20009
0
AGGTCCACACAGCGGCCAAA


 


405936
5-10-5
304
19994
20013
0
AAAGAGGTCCACACAGCGGC


 


405937
5-10-5
306
20023
20042
0
GAGGCACCAATGATGTCCTC


 


405938
5-10-5
307
20027
20046
0
GCTGGAGGCACCAATGATGT


 


405939
5-10-5
308
20029
20048
0
TCGCTGGAGGCACCAATGAT


 


405940
5-10-5
309
20031
20050
0
AGTCGCTGGAGGCACCAATG


 


405941
5-10-5
310
20033
20052
0
GCAGTCGCTGGAGGCACCAA


 


405942
5-10-5
311
20038
20057
0
GTGCTGCAGTCGCTGGAGGC


 


405943
5-10-5
312
20040
20059
0
AGGTGCTGCAGTCGCTGGAG


 


405944
5-10-5
314
20043
20062
0
AGCAGGTGCTGCAGTCGCTG


 


405945
5-10-5
315
20045
20064
0
AAAGCAGGTGCTGCAGTCGC


 


405946
5-10-5
316
20049
20068
0
ACACAAAGCAGGTGCTGCAG


 


405947
5-10-5
317
20051
20070
0
TGACACAAAGCAGGTGCTGC


 


405949
5-10-5
320
20629
20648
0
CAGCATCATGGCTGCAATGC


 


405950
5-10-5
321
20631
20650
0
GACAGCATCATGGCTGCAAT


 


405951
5-10-5
322
20633
20652
0
CAGACAGCATCATGGCTGCA


 


405952
5-10-5
323
20637
20656
0
TCGGCAGACAGCATCATGGC


 


405953
5-10-5
325
20641
20660
0
CGGCTCGGCAGACAGCATCA


 


405954
5-10-5
326
20643
20662
0
TCCGGCTCGGCAGACAGCAT


 


405955
5-10-5
328
20670
20689
0
CTCTGCCTCAACTCGGCCAG


 


405956
5-10-5
329
20672
20691
0
GTCTCTGCCTCAACTCGGCC


 


405957
5-10-5
330
20674
20693
0
CAGTCTCTGCCTCAACTCGG


 


405958
5-10-5
331
20676
20695
0
ATCAGTCTCTGCCTCAACTC


 


405959
5-10-5
332
20678
20697
0
GGATCAGTCTCTGCCTCAAC


 


405960
5-10-5
333
20680
20699
0
GTGGATCAGTCTCTGCCTCA


 


405961
5-10-5
334
20682
20701
0
AAGTGGATCAGTCTCTGCCT


 


405962
5-10-5
335
20685
20704
0
GAGAAGTGGATCAGTCTCTG


 


405963
5-10-5
337
20689
20708
0
GGCAGAGAAGTGGATCAGTC


 


405964
5-10-5
338
20693
20712
0
CTTTGGCAGAGAAGTGGATC


 


405965
5-10-5
339
20698
20717
0
GACATCTTTGGCAGAGAAGT


 


405966
5-10-5
340
20700
20719
0
ATGACATCTTTGGCAGAGAA


 


405967
5-10-5
341
20704
20723
0
ATTGATGACATCTTTGGCAG


 


405968
5-10-5
342
20706
20725
0
TCATTGATGACATCTTTGGC


 


405969
5-10-5
343
20711
20730
0
AGGCCTCATTGATGACATCT


 


405970
5-10-5
344
20713
20732
0
CCAGGCCTCATTGATGACAT


 


405971
5-10-5
355
20728
20747
0
CTGGTCCTCAGGGAACCAGG


 


405972
5-10-5
357
20730
20749
0
CGCTGGTCCTCAGGGAACCA


 


405973
5-10-5
358
20735
20754
0
GTACCCGCTGGTCCTCAGGG


 


405974
5-10-5
359
20737
20756
0
CAGTACCCGCTGGTCCTCAG


 


405975
5-10-5
361
21088
21107
0
CAAAACAGCTGCCAACCTGC


 


405976
5-10-5
362
21093
21112
0
TCCTGCAAAACAGCTGCCAA


 


405977
5-10-5
363
21095
21114
0
AGTCCTGCAAAACAGCTGCC


 


405978
5-10-5
365
21118
21137
0
GGCCCCGAGTGTGCTGACCA


 


405979
5-10-5
366
21123
21142
0
GTGTAGGCCCCGAGTGTGCT


 


405980
5-10-5
367
21125
21144
0
CCGTGTAGGCCCCGAGTGTG


 


405981
5-10-5
368
21127
21146
0
ATCCGTGTAGGCCCCGAGTG


 


405982
5-10-5
370
21131
21150
0
GGCCATCCGTGTAGGCCCCG


 


405983
5-10-5
371
21133
21152
0
GTGGCCATCCGTGTAGGCCC


 


405984
5-10-5
372
21181
21200
0
CTGGAGCAGCTCAGCAGCTC


 


405985
5-10-5
373
21183
21202
0
AACTGGAGCAGCTCAGCAGC


 


405986
5-10-5
374
21188
21207
0
GGAGAAACTGGAGCAGCTCA


 


405987
5-10-5
375
21190
21209
0
CTGGAGAAACTGGAGCAGCT


 


405988
5-10-5
381
22138
22157
0
GGCAGCACCTGGCAATGGCG


 


405989
5-10-5
383
22140
22159
0
CAGGCAGCACCTGGCAATGG


 


405990
5-10-5
386
22144
22163
0
GTAGCAGGCAGCACCTGGCA


 


405991
5-10-5
394
22206
22225
0
GACACGGGTCCCCATGCTGG


 


405992
5-10-5
396
22208
22227
0
TGGACACGGGTCCCCATGCT


 


405993
5-10-5
398
22210
22229
0
AGTGGACACGGGTCCCCATG


 


405994
5-10-5
400
22212
22231
0
GCAGTGGACACGGGTCCCCA


 


405995
5-10-5
402
22214
22233
0
TGGCAGTGGACACGGGTCCC


 


405996
5-10-5
412
24097
24116
0
GACTTTGCATTCCAGACCTG


 


405997
5-10-5
415
24102
24121
0
TCCTTGACTTTGCATTCCAG


 


405998
5-10-5
426
26120
26139
0
TCCGGCAGCAGATGGCAACG


 


405999
5-10-5
160
 2437
 2456
0
GACCGCCTGGAGCTGACGGT


 


406003
5-10-5
178
 6492
 6511
0
CAGTGCGCTCTGACTGCGAG


 


406004
5-10-5
179
 6494
 6513
0
GGCAGTGCGCTCTGACTGCG


 


406005
5-10-5
180
 6496
 6515
0
CGGGCAGTGCGCTCTGACTG


 


406006
5-10-5
181
 6499
 6518
0
CGGCGGGCAGTGCGCTCTGA


 


406007
5-10-5
183
 6532
 6551
0
AGGTATCCCCGGCGGGCAGC


 


406008
5-10-5
188
 6539
 6558
0
CTTGGTGAGGTATCCCCGGC


 


406009
5-10-5
190
 6541
 6560
0
ATCTTGGTGAGGTATCCCCG


 


406010
5-10-5
193
 6546
 6565
0
GCAGGATCTTGGTGAGGTAT


 


406011
5-10-5
194
 6548
 6567
0
ATGCAGGATCTTGGTGAGGT


 


406012
5-10-5
195
 6550
 6569
0
ACATGCAGGATCTTGGTGAG


 


406013
5-10-5
196
 6554
 6573
0
GAAGACATGCAGGATCTTGG


 


406014
5-10-5
199
 6585
 6604
0
TCTTCACCAGGAAGCCAGGA


 


406015
5-10-5
200
 6590
 6609
0
ACTCATCTTCACCAGGAAGC


 


406016
5-10-5
201
 6592
 6611
0
CCACTCATCTTCACCAGGAA


 


406017
5-10-5
203
 6596
 6615
0
GTCGCCACTCATCTTCACCA


 


406018
5-10-5
204
 6598
 6617
0
AGGTCGCCACTCATCTTCAC


 


406019
5-10-5
205
 6600
 6619
0
GCAGGTCGCCACTCATCTTC


 


406020
5-10-5
206
 6602
 6621
0
CAGCAGGTCGCCACTCATCT


 


406021
5-10-5
210
 9207
 9226
0
TTCATCCGCCCGGTACCGTG


 


406022
5-10-5
211
 9209
 9228
0
TATTCATCCGCCCGGTACCG


 


406023
5-10-5
212
 9212
 9231
0
TGGTATTCATCCGCCCGGTA


 


406024
5-10-5
213
 9214
 9233
0
GCTGGTATTCATCCGCCCGG


 


406025
5-10-5
216
14888
14907
0
GTGTCTAGGAGATACACCTC


 


406026
5-10-5
217
14893
14912
0
TGCTGGTGTCTAGGAGATAC


 


406027
5-10-5
226
14930
14949
0
ATGACCCTGCCCTCGATTTC


 


406028
5-10-5
227
14932
14951
0
CCATGACCCTGCCCTCGATT


 


406029
5-10-5
228
14934
14953
0
GACCATGACCCTGCCCTCGA


 


406030
5-10-5
229
14938
14957
0
CGGTGACCATGACCCTGCCC


 


406031
5-10-5
230
14940
14959
0
GTCGGTGACCATGACCCTGC


 


406032
5-10-5
232
14944
14963
0
CGAAGTCGGTGACCATGACC


 


406033
5-10-5
237
15261
15280
0
CCTGCCAGGTGGGTGCCATG


 


406034
5-10-5
238
15266
15285
0
CCACCCCTGCCAGGTGGGTG


 


406035
5-10-5
239
15271
15290
0
GCTGACCACCCCTGCCAGGT


 


406036
5-10-5
241
15283
15302
0
GGCATCCCGGCCGCTGACCA


 


406037
5-10-5
242
15286
15305
0
GCCGGCATCCCGGCCGCTGA


 


406038
5-10-5
245
15293
15312
0
TGGCCACGCCGGCATCCCGG


 


406039
5-10-5
257
15330
15349
0
CAGTTGAGCACGCGCAGGCT


 


406040
5-10-5
258
15332
15351
0
GGCAGTTGAGCACGCGCAGG


 


406041
5-10-5
259
15336
15355
0
CCTTGGCAGTTGAGCACGCG


 


406042
5-10-5
260
15341
15360
0
CCTTCCCTTGGCAGTTGAGC


 


406043
5-10-5
261
15345
15364
0
GTGCCCTTCCCTTGGCAGTT


 


406044
5-10-5
262
15347
15366
0
CCGTGCCCTTCCCTTGGCAG


 


406045
5-10-5
266
18591
18610
0
TGGCTTTTCCGAATAAACTC


 


406478
5-10-5
448
21692
21711
0
CTGCCCTTCCACCAAAATGC


 


406479
5-10-5
449
21693
21712
0
ACTGCCCTTCCACCAAAATG


 


406480
5-10-5
450
21694
21713
0
CACTGCCCTTCCACCAAAAT


 


406481
5-10-5
451
21695
21714
0
GCACTGCCCTTCCACCAAAA


 


406482
5-10-5
452
21697
21716
0
GGGCACTGCCCTTCCACCAA


 


406483
5-10-5
453
21698
21717
0
TGGGCACTGCCCTTCCACCA


 


406484
5-10-5
454
21699
21718
0
CTGGGCACTGCCCTTCCACC


 


406485
5-10-5
455
21700
21719
0
GCTGGGCACTGCCCTTCCAC


 


408653
5-10-5
354
20727
20746
0
TGGTCCTCAGGGAACCAGGC


 


409126
5-10-5
447
15298
15317
1
ACCCTTGGTCACGCCGGCAT


 


410529
5-10-5
184
 6534
 6553
0
TGAGGTATCCCCGGCGGGCA


 


410530
5-10-5
185
 6535
 6554
0
GTGAGGTATCCCCGGCGGGC


 


410531
5-10-5
186
 6536
 6555
0
GGTGAGGTATCCCCGGCGGG


 


410532
5-10-5
187
 6538
 6557
0
TTGGTGAGGTATCCCCGGCG


 


410533
5-10-5
189
 6540
 6559
0
TCTTGGTGAGGTATCCCCGG


 


410534
5-10-5
191
 6542
 6561
0
GATCTTGGTGAGGTATCCCC


 


410535
5-10-5
192
 6544
 6563
0
AGGATCTTGGTGAGGTATCC


 


410536
5-10-5
243
15291
15310
0
GCCACGCCGGCATCCCGGCC


 


410537
5-10-5
244
15292
15311
0
GGCCACGCCGGCATCCCGGC


 


410538
5-10-5
254
15303
15322
0
CTGGCACCCTTGGCCACGCC


 


410539
5-10-5
255
15304
15323
0
GCTGGCACCCTTGGCCACGC


 


410540
5-10-5
356
20729
20748
0
GCTGGTCCTCAGGGAACCAG


 


410541
5-10-5
376
22133
22152
0
CACCTGGCAATGGCGTAGAC


 


410542
5-10-5
377
22134
22153
0
GCACCTGGCAATGGCGTAGA


 


410543
5-10-5
378
22135
22154
0
AGCACCTGGCAATGGCGTAG


 


410544
5-10-5
379
22136
22155
0
CAGCACCTGGCAATGGCGTA


 


410545
5-10-5
380
22137
22156
0
GCAGCACCTGGCAATGGCGT


 


410546
5-10-5
382
22139
22158
0
AGGCAGCACCTGGCAATGGC


 


410547
5-10-5
384
22141
22160
0
GCAGGCAGCACCTGGCAATG


 


410548
5-10-5
385
22143
22162
0
TAGCAGGCAGCACCTGGCAA


 


410549
5-10-5
388
22199
22218
0
GTCCCCATGCTGGCCTCAGC


 


410550
5-10-5
389
22200
22219
0
GGTCCCCATGCTGGCCTCAG


 


410551
5-10-5
390
22201
22220
0
GGGTCCCCATGCTGGCCTCA


 


410552
5-10-5
391
22202
22221
0
CGGGTCCCCATGCTGGCCTC


 


410553
5-10-5
392
22203
22222
0
ACGGGTCCCCATGCTGGCCT


 


410554
5-10-5
393
22205
22224
0
ACACGGGTCCCCATGCTGGC


 


410555
5-10-5
395
22207
22226
0
GGACACGGGTCCCCATGCTG


 


410556
5-10-5
397
22209
22228
0
GTGGACACGGGTCCCCATGC


 


410557
5-10-5
399
22211
22230
0
CAGTGGACACGGGTCCCCAT


 


410558
5-10-5
401
22213
22232
0
GGCAGTGGACACGGGTCCCC


 


410559
5-10-5
403
22215
22234
0
GTGGCAGTGGACACGGGTCC


 


410560
5-10-5
404
22216
22235
0
GGTGGCAGTGGACACGGGTC


 


410561
5-10-5
405
22217
22236
0
TGGTGGCAGTGGACACGGGT


 


410562
5-10-5
411
24096
24115
0
ACTTTGCATTCCAGACCTGG


 


410563
5-10-5
413
24098
24117
0
TGACTTTGCATTCCAGACCT


 


410564
5-10-5
414
24099
24118
0
TTGACTTTGCATTCCAGACC


 


410565
5-10-5
419
26112
26131
0
CAGATGGCAACGGCTGTCAC


 


410566
5-10-5
420
26113
26132
0
GCAGATGGCAACGGCTGTCA


 


410567
5-10-5
421
26114
26133
0
AGCAGATGGCAACGGCTGTC


 


410568
5-10-5
422
26115
26134
0
CAGCAGATGGCAACGGCTGT


 


410569
5-10-5
423
26116
26135
0
GCAGCAGATGGCAACGGCTG


 


410570
5-10-5
424
26118
26137
0
CGGCAGCAGATGGCAACGGC


 


410571
5-10-5
425
26119
26138
0
CCGGCAGCAGATGGCAACGG


 


410572
5-10-5
427
26121
26140
0
CTCCGGCAGCAGATGGCAAC


 


410573
5-10-5
428
26122
26141
0
GCTCCGGCAGCAGATGGCAA


 


410730
5-10-5
202
 6594
 6613
0
CGCCACTCATCTTCACCAGG


 


410731
5-10-5
207
 6604
 6623
0
TCCAGCAGGTCGCCACTCAT


 


410732
5-10-5
214
 9216
 9235
0
GGGCTGGTATTCATCCGCCC


 


410733
5-10-5
231
14942
14961
0
AAGTCGGTGACCATGACCCT


 


410734
5-10-5
279
18714
18733
0
CCGGCAGCGGTGACCAGCAC


 


410735
5-10-5
291
19956
19975
0
TCCCCAGGGTCACCGGCTGG


 


410736
5-10-5
303
19992
20011
0
AGAGGTCCACACAGCGGCCA


 


410737
5-10-5
313
20042
20061
0
GCAGGTGCTGCAGTCGCTGG


 


410738
5-10-5
324
20639
20658
0
GCTCGGCAGACAGCATCATG


 


410739
5-10-5
336
20687
20706
0
CAGAGAAGTGGATCAGTCTC


 


410740
5-10-5
345
20715
20734
0
AACCAGGCCTCATTGATGAC


 


410741
5-10-5
369
21129
21148
0
CCATCCGTGTAGGCCCCGAG


 


410742
5-10-5
159
 2433
 2452
0
GCCTGGAGCTGACGGTGCCC


 


410743
5-10-5
170
 2560
 2579
0
TCCTCCTCGGAACGCAAGGC


 


410744
5-10-5
171
 2585
 2604
0
GTGCTCGGGTGCTTCGGCCA


 


410745
5-10-5
172
 2605
 2624
0
TGGAAGGTGGCTGTGGTTCC


 


410746
5-10-5
176
 6444
 6463
0
CGTAGGTGCCAGGCAACCTC


 


410747
5-10-5
177
 6482
 6501
0
TGACTGCGAGAGGTGGGTCT


 


410748
5-10-5
182
 6528
 6547
0
ATCCCCGGCGGGCAGCCTGG


 


410749
5-10-5
197
 6565
 6584
0
AGAAGGCCATGGAAGACATG


 


410750
5-10-5
198
 6575
 6594
0
GAAGCCAGGAAGAAGGCCAT


 


410752
5-10-5
209
 9149
 9168
0
GCAAAGACAGAGGAGTCCTC


 


410753
5-10-5
215
14877
14896
0
ATACACCTCCACCAGGCTGC


 


410754
5-10-5
233
14954
14973
0
GGCACATTCTCGAAGTCGGT


 


410756
5-10-5
235
15254
15273
0
GGTGGGTGCCATGACTGTCA


 


410757
5-10-5
240
15279
15298
0
TCCCGGCCGCTGACCACCCC


 


410758
5-10-5
256
15309
15328
0
CGCATGCTGGCACCCTTGGC


 


410759
5-10-5
263
15358
15377
0
GGTGCCGCTAACCGTGCCCT


 


410761
5-10-5
271
18614
18633
0
GTGGCCCCACAGGCTGGACC


 


410762
5-10-5
272
18627
18646
0
AGCAGCACCACCAGTGGCCC


 


410763
5-10-5
273
18649
18668
0
GCTGTACCCACCCGCCAGGG


 


410764
5-10-5
274
18695
18714
0
CGACCCCAGCCCTCGCCAGG


 


410767
5-10-5
289
19941
19960
0
GCTGGTCTTGGGCATTGGTG


 


410768
5-10-5
305
20016
20035
0
CAATGATGTCCTCCCCTGGG


 


410769
5-10-5
318
20061
20080
0
TCCCACTCTGTGACACAAAG


 


410770
5-10-5
327
20657
20676
0
CGGCCAGGGTGAGCTCCGGC


 


410771
5-10-5
360
20785
20804
0
ACCTGCCCCATGGGTGCTGG


 


410772
5-10-5
364
21106
21125
0
GCTGACCATACAGTCCTGCA


 


410773
5-10-5
387
22189
22208
0
TGGCCTCAGCTGGTGGAGCT


 


410774
5-10-5
406
22220
22239
0
TGTTGGTGGCAGTGGACACG


 


410776
5-10-5
408
23985
24004
0
GTGCCAAGGTCCTCCACCTC


 


410777
5-10-5
409
24005
24024
0
TCAGCACAGGCGGCTTGTGG


 


410778
5-10-5
410
24035
24054
0
CCACGCACTGGTTGGGCTGA


 


410779
5-10-5
416
24115
24134
0
CGGGATTCCATGCTCCTTGA


 


410781
5-10-5
418
25994
26013
0
GGAGGGCACTGCAGCCAGTC


 


410782
5-10-5
429
26132
26151
0
CCAGGTGCCGGCTCCGGCAG


 


410783
5-10-5
430
26142
26161
0
GAGGCCTGCGCCAGGTGCCG









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 3-14-3 gap-widened motif. Table 9 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 9







Gapmer antisense compounds having a 3-14-3 motif


targeted to SEQ ID NO: 2
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 2
NO: 2
Mismatches
Sequence (5′-3′)





399871
3-14-3
  4
 2274
 2293
0
GCGCGGAATCCTGGCTGGGA


 


399872
3-14-3
  5
 2381
 2400
0
GAGGAGACCTAGAGGCCGTG


 


399873
3-14-3
  6
 2439
 2458
0
AGGACCGCCTGGAGCTGACG


 


399874
3-14-3
  7
 2549
 2568
0
ACGCAAGGCTAGCACCAGCT


 


399875
3-14-3
  9
 2619
 2638
0
CCTTGGCGCAGCGGTGGAAG


 


399876
3-14-3
 10
 6498
 6517
0
GGCGGGCAGTGCGCTCTGAC


 


399877
3-14-3
 11
 6537
 6556
0
TGGTGAGGTATCCCCGGCGG


 


399878
3-14-3
 14
 6557
 6576
0
ATGGAAGACATGCAGGATCT


 


399879
3-14-3
 15
 6583
 6602
0
TTCACCAGGAAGCCAGGAAG


 


399880
3-14-3
 17
 9130
 9149
0
CCTCGATGTAGTCGACATGG


 


399881
3-14-3
 18
 9210
 9229
0
GTATTCATCCGCCCGGTACC


 


399882
3-14-3
 19
14891
14910
0
CTGGTGTCTAGGAGATACAC


 


399883
3-14-3
 21
14916
14935
0
GATTTCCCGGTGGTCACTCT


 


399884
3-14-3
 24
14946
14965
0
CTCGAAGTCGGTGACCATGA


 


399885
3-14-3
 25
14979
14998
0
GTGGAAGCGGGTCCCGTCCT


 


399886
3-14-3
 26
15264
15283
0
ACCCCTGCCAGGTGGGTGCC


 


399887
3-14-3
 28
15298
15317
0
ACCCTTGGCCACGCCGGCAT


 


399888
3-14-3
 29
15334
15353
0
TTGGCAGTTGAGCACGCGCA


 


399890
3-14-3
 32
18593
18612
0
GCTGGCTTTTCCGAATAAAC


 


399891
3-14-3
 33
18705
18724
0
GTGACCAGCACGACCCCAGC


 


399892
3-14-3
149
19933
19952
0
TGGGCATTGGTGGCCCCAAC


 


399893
3-14-3
 34
19962
19981
0
CCAAAGTCCCCAGGGTCACC


 


399894
3-14-3
128
19966
19985
0
GTCCCCAAAGTCCCCAGGGT


 


399895
3-14-3
 36
19976
19995
0
GCCAAAGTTGGTCCCCAAAG


 


399896
3-14-3
 38
19997
20016
0
GGCAAAGAGGTCCACACAGC


 


399897
3-14-3
 39
20025
20044
0
TGGAGGCACCAATGATGTCC


 


399899
3-14-3
 45
20691
20710
0
TTGGCAGAGAAGTGGATCAG


 


399900
3-14-3
 50
20726
20745
0
GGTCCTCAGGGAACCAGGCC


 


399901
3-14-3
 87
20733
20752
0
ACCCGCTGGTCCTCAGGGAA


 


399902
3-14-3
119
20762
20781
0
GCAGGGCGGCCACCAGGTTG


 


399904
3-14-3
 54
21121
21140
0
GTAGGCCCCGAGTGTGCTGA


 


399905
3-14-3
 57
22096
22115
0
CGTTGTGGGCCCGGCAGACC


 


399906
3-14-3
 58
22142
22161
0
AGCAGGCAGCACCTGGCAAT


 


399907
3-14-3
 59
22204
22223
0
CACGGGTCCCCATGCTGGCC


 


399908
3-14-3
 60
24095
24114
0
CTTTGCATTCCAGACCTGGG


 


399909
3-14-3
 62
26117
26136
0
GGCAGCAGATGGCAACGGCT


 


399910
3-14-3
154
26217
26236
0
TTTTAAAGCTCAGCCCCAGC


 


399911
3-14-3
 64
26311
26330
0
TCAAGGGCCAGGCCAGCAGC


 


399912
3-14-3
 66
26404
26423
0
ATGCCCCACAGTGAGGGAGG


 


399913
3-14-3
 67
26413
26432
0
AATGGTGAAATGCCCCACAG


 


399914
3-14-3
153
26456
26475
0
TTGGGAGCAGCTGGCAGCAC


 


399915
3-14-3
 68
26557
26576
0
CATGGGAAGAATCCTGCCTC


 


399916
3-14-3
 69
26639
26658
0
GGAGATGAGGGCCATCAGCA


 


399917
3-14-3
 70
26707
26726
0
TAGATGCCATCCAGAAAGCT


 


399918
3-14-3
 72
26790
26809
0
GGCATAGAGCAGAGTAAAGG


 


399919
3-14-3
112
27034
27053
0
GAAGAGGCTTGGCTTCAGAG


 


399920
3-14-3
 75
27244
27263
0
GCTCAAGGAGGGACAGTTGT


 


399921
3-14-3
 76
27279
27298
0
AAAGATAAATGTCTGCTTGC


 


399922
3-14-3
 78
27350
27369
0
TCTTCAAGTTACAAAAGCAA


 


399923
3-14-3
 85
13681
13700
0
ACAAATTCCCAGACTCAGCA


 


399924
3-14-3
129
13816
13835
0
GTGCCATCTGAACAGCACCT


 


399925
3-14-3
110
13926
13945
0
CCTGGAACCCCTGCAGCCAG


 


399926
3-14-3
152
13977
13996
0
TTCAGGCAGGTTGCTGCTAG


 


399927
3-14-3
140
13998
14017
0
TCAGCCAGGCCAAAGGAAGA


 


399928
3-14-3
136
14122
14141
0
TAGGAGAAAGTAGGGAGAGC


 


399929
3-14-3
132
14179
14198
0
TAAAAGCTGCAAGAGACTCA


 


399930
3-14-3
139
14267
14286
0
TCAGAGAAAACAGTCACCGA


 


399931
3-14-3
142
14404
14423
0
TCATTTTAGAGACAGGAAGC


 


399932
3-14-3
113
14441
14460
0
GAATAACAGTGATGTCTGGC


 


399933
3-14-3
138
14494
14513
0
TCACAGCTCACCGAGTCTGC


 


399934
3-14-3
 98
14524
14543
0
AGTGTAAAATAAAGCCCCTA


 


399935
3-14-3
 96
14601
14620
0
AGGACCCAAGTCATCCTGCT


 


399936
3-14-3
124
14631
14650
0
GGCCATCAGCTGGCAATGCT


 


399937
3-14-3
133
14675
14694
0
TAGACAAGGAAAGGGAGGCC


 


399938
3-14-3
103
14681
14700
0
ATTTCATAGACAAGGAAAGG


 


399939
3-14-3
155
14801
14820
0
CTTATAGTTAACACACAGAA


 


399943
3-14-3
102
 5590
 5609
0
ATGTGCAGAGATCAATCACA


 


399944
3-14-3
127
10633
10652
0
GGTGGTAATTTGTCACAGCA


 


399945
3-14-3
 84
11308
11327
0
AAGGTCACACAGTTAAGAGT


 


399946
3-14-3
157
18561
18580
0
AGGAACAAAGCCAAGGTCAC


 


399947
3-14-3
126
22292
22311
0
GGTGCATAAGGAGAAAGAGA


 


399948
3-14-3
147
24858
24877
0
TGAGTTCATTTAAGAGTGGA


 


399949
3-14-3
  8
 2556
 2575
0
CCTCGGAACGCAAGGCTAGC


 


399950
3-14-3
 12
 6543
 6562
0
GGATCTTGGTGAGGTATCCC


 


399951
3-14-3
 13
 6552
 6571
0
AGACATGCAGGATCTTGGTG


 


399952
3-14-3
 16
 6588
 6607
0
TCATCTTCACCAGGAAGCCA


 


399953
3-14-3
 20
14896
14915
0
GTATGCTGGTGTCTAGGAGA


 


399954
3-14-3
 22
14922
14941
0
GCCCTCGATTTCCCGGTGGT


 


399955
3-14-3
 23
14936
14955
0
GTGACCATGACCCTGCCCTC


 


399956
3-14-3
 27
15269
15288
0
TGACCACCCCTGCCAGGTGG


 


399957
3-14-3
 30
15339
15358
0
TTCCCTTGGCAGTTGAGCAC


 


399958
3-14-3
 35
19971
19990
0
AGTTGGTCCCCAAAGTCCCC


 


399959
3-14-3
 37
19988
20007
0
GTCCACACAGCGGCCAAAGT


 


399960
3-14-3
 40
20036
20055
0
GCTGCAGTCGCTGGAGGCAC


 


399961
3-14-3
 41
20047
20066
0
ACAAAGCAGGTGCTGCAGTC


 


399963
3-14-3
 43
20635
20654
0
GGCAGACAGCATCATGGCTG


 


399964
3-14-3
 44
20683
20702
0
GAAGTGGATCAGTCTCTGCC


 


399965
3-14-3
 46
20696
20715
0
CATCTTTGGCAGAGAAGTGG


 


399966
3-14-3
 47
20702
20721
0
TGATGACATCTTTGGCAGAG


 


399967
3-14-3
 48
20709
20728
0
GCCTCATTGATGACATCTTT


 


399968
3-14-3
 49
20721
20740
0
TCAGGGAACCAGGCCTCATT


 


399969
3-14-3
 51
20740
20759
0
GGTCAGTACCCGCTGGTCCT


 


399970
3-14-3
 53
21091
21110
0
CTGCAAAACAGCTGCCAACC


 


399971
3-14-3
 55
21186
21205
0
AGAAACTGGAGCAGCTCAGC


 


399972
3-14-3
 56
21192
21211
0
TCCTGGAGAAACTGGAGCAG


 


399973
3-14-3
 61
24100
24119
0
CTTGACTTTGCATTCCAGAC


 


399974
3-14-3
 63
26222
26241
0
AACCATTTTAAAGCTCAGCC


 


399975
3-14-3
 65
26316
26335
0
CCCACTCAAGGGCCAGGCCA


 


399976
3-14-3
122
26389
26408
0
GGAGGGAGCTTCCTGGCACC


 


399977
3-14-3
 71
26713
26732
0
TCTGGCTAGATGCCATCCAG


 


399978
3-14-3
 73
26795
26814
0
AGCCTGGCATAGAGCAGAGT


 


399979
3-14-3
135
26801
26820
0
TAGCACAGCCTGGCATAGAG


 


399980
3-14-3
 74
27040
27059
0
AAGTAAGAAGAGGCTTGGCT


 


399981
3-14-3
 77
27284
27303
0
ACCCAAAAGATAAATGTCTG


 


399982
3-14-3
 99
27357
27376
0
ATAAATATCTTCAAGTTACA


 


399983
3-14-3
100
13746
13765
0
ATCTCAGGACAGGTGAGCAA


 


399984
3-14-3
116
13760
13779
0
GAGTAGAGATTCTCATCTCA


 


399985
3-14-3
117
13828
13847
0
GAGTCTTCTGAAGTGCCATC


 


399986
3-14-3
 81
13903
13922
0
AAGCAGGGCCTCAGGTGGAA


 


399987
3-14-3
 83
13986
14005
0
AAGGAAGACTTCAGGCAGGT


 


399988
3-14-3
137
14112
14131
0
TAGGGAGAGCTCACAGATGC


 


399989
3-14-3
 92
14397
14416
0
AGAGACAGGAAGCTGCAGCT


 


399990
3-14-3
 82
14670
14689
0
AAGGAAAGGGAGGCCTAGAG


 


399991
3-14-3
156
14809
14828
0
AAGTCAACCTTATAGTTAAC


 


399993
3-14-3
107
 3056
 3075
0
CCCACTATAATGGCAAGCCC


 


399994
3-14-3
 80
 4306
 4325
0
AACCCAGTTCTAATGCACCT


 


399995
3-14-3
106
 5140
 5159
0
CCAGTCAGAGTAGAACAGAG


 


399996
3-14-3
121
 5599
 5618
0
GGAGCCTACATGTGCAGAGA


 


399997
3-14-3
 94
 5667
 5686
0
AGCATGGCACCAGCATCTGC


 


399998
3-14-3
108
 6652
 6671
0
CCCAGCCCTATCAGGAAGTG


 


399999
3-14-3
144
 7099
 7118
0
TGACATCCAGGAGGGAGGAG


 


400000
3-14-3
 91
 7556
 7575
0
AGACTGATGGAAGGCATTGA


 


400001
3-14-3
131
 7565
 7584
0
GTGTTGAGCAGACTGATGGA


 


400002
3-14-3
145
 8836
 8855
0
TGACATCTTGTCTGGGAGCC


 


400003
3-14-3
 90
 8948
 8967
0
AGACTAGGAGCCTGAGTTTT


 


400004
3-14-3
125
 9099
 9118
0
GGCCTGCAGAAGCCAGAGAG


 


400005
3-14-3
148
10252
10271
0
TGGCAGCAACTCAGACATAT


 


400006
3-14-3
 79
11472
11491
0
AAATGCAGGGCTAAAATCAC


 


400007
3-14-3
 88
12715
12734
0
ACTGGATACATTGGCAGACA


 


400008
3-14-3
111
12928
12947
0
CTAGAGGAACCACTAGATAT


 


400009
3-14-3
 86
15471
15490
0
ACAGCATTCTTGGTTAGGAG


 


400010
3-14-3
 97
16134
16153
0
AGTCAAGCTGCTGCCCAGAG


 


400011
3-14-3
120
16668
16687
0
GCTAGTTATTAAGCACCTGC


 


400012
3-14-3
150
17267
17286
0
TGTGAGCTCTGGCCCAGTGG


 


400013
3-14-3
115
18377
18396
0
GAGTAAGGCAGGTTACTCTC


 


400014
3-14-3
134
18408
18427
0
TAGATGTGACTAACATTTAA


 


400015
3-14-3
105
19203
19222
0
CACATTAGCCTTGCTCAAGT


 


400016
3-14-3
151
19913
19932
0
TGTGATGACCTGGAAAGGTG


 


400017
3-14-3
158
20100
20119
0
GTGGTGACTTACCAGCCACG


 


400018
3-14-3
109
20188
20207
0
CCCCTGCACAGAGCCTGGCA


 


400019
3-14-3
141
20624
20643
0
TCATGGCTGCAATGCCTGGT


 


400020
3-14-3
 93
20995
21014
0
AGAGAGGAGGGCTTAAAGAA


 


400021
3-14-3
 95
21082
21101
0
AGCTGCCAACCTGCAAAAAG


 


400022
3-14-3
143
21481
21500
0
TGAAAATCCATCCAGCACTG


 


400023
3-14-3
 89
21589
21608
0
AGAACCATGGAGCACCTGAG


 


400024
3-14-3
123
21696
21715
0
GGCACTGCCCTTCCACCAAA


 


400025
3-14-3
118
24907
24926
0
GCACCATCCAGACCAGAATC


 


400026
3-14-3
114
25413
25432
0
GAGAGGTTCAGATCCAGGCC


 


405604
3-14-3
236
15257
15276
0
CCAGGTGGGTGCCATGACTG


 


405641
3-14-3
373
21183
21202
0
AACTGGAGCAGCTCAGCAGC


 


410574
3-14-3
184
 6534
 6553
0
TGAGGTATCCCCGGCGGGCA


 


410575
3-14-3
185
 6535
 6554
0
GTGAGGTATCCCCGGCGGGC


 


410576
3-14-3
186
 6536
 6555
0
GGTGAGGTATCCCCGGCGGG


 


410577
3-14-3
187
 6538
 6557
0
TTGGTGAGGTATCCCCGGCG


 


410578
3-14-3
188
 6539
 6558
0
CTTGGTGAGGTATCCCCGGC


 


410579
3-14-3
189
 6540
 6559
0
TCTTGGTGAGGTATCCCCGG


 


410580
3-14-3
190
 6541
 6560
0
ATCTTGGTGAGGTATCCCCG


 


410581
3-14-3
191
 6542
 6561
0
GATCTTGGTGAGGTATCCCC


 


410582
3-14-3
192
 6544
 6563
0
AGGATCTTGGTGAGGTATCC


 


410583
3-14-3
243
15291
15310
0
GCCACGCCGGCATCCCGGCC


 


410584
3-14-3
244
15292
15311
0
GGCCACGCCGGCATCCCGGC


 


410585
3-14-3
245
15293
15312
0
TGGCCACGCCGGCATCCCGG


 


410586
3-14-3
246
15294
15313
0
TTGGCCACGCCGGCATCCCG


 


410587
3-14-3
247
15295
15314
0
CTTGGCCACGCCGGCATCCC


 


410588
3-14-3
248
15296
15315
0
CCTTGGCCACGCCGGCATCC


 


410589
3-14-3
249
15297
15316
0
CCCTTGGCCACGCCGGCATC


 


410590
3-14-3
250
15299
15318
0
CACCCTTGGCCACGCCGGCA


 


410591
3-14-3
251
15300
15319
0
GCACCCTTGGCCACGCCGGC


 


410592
3-14-3
252
15301
15320
0
GGCACCCTTGGCCACGCCGG


 


410593
3-14-3
253
15302
15321
0
TGGCACCCTTGGCCACGCCG


 


410594
3-14-3
254
15303
15322
0
CTGGCACCCTTGGCCACGCC


 


410595
3-14-3
255
15304
15323
0
GCTGGCACCCTTGGCCACGC


 


410596
3-14-3
348
20719
20738
0
AGGGAACCAGGCCTCATTGA


 


410597
3-14-3
349
20720
20739
0
CAGGGAACCAGGCCTCATTG


 


410598
3-14-3
350
20722
20741
0
CTCAGGGAACCAGGCCTCAT


 


410599
3-14-3
351
20723
20742
0
CCTCAGGGAACCAGGCCTCA


 


410600
3-14-3
352
20724
20743
0
TCCTCAGGGAACCAGGCCTC


 


410601
3-14-3
353
20725
20744
0
GTCCTCAGGGAACCAGGCCT


 


410602
3-14-3
354
20727
20746
0
TGGTCCTCAGGGAACCAGGC


 


410603
3-14-3
355
20728
20747
0
CTGGTCCTCAGGGAACCAGG


 


410604
3-14-3
356
20729
20748
0
GCTGGTCCTCAGGGAACCAG


 


410605
3-14-3
376
22133
22152
0
CACCTGGCAATGGCGTAGAC


 


410606
3-14-3
377
22134
22153
0
GCACCTGGCAATGGCGTAGA


 


410607
3-14-3
378
22135
22154
0
AGCACCTGGCAATGGCGTAG


 


410608
3-14-3
379
22136
22155
0
CAGCACCTGGCAATGGCGTA


 


410609
3-14-3
380
22137
22156
0
GCAGCACCTGGCAATGGCGT


 


410610
3-14-3
381
22138
22157
0
GGCAGCACCTGGCAATGGCG


 


410611
3-14-3
382
22139
22158
0
AGGCAGCACCTGGCAATGGC


 


410612
3-14-3
383
22140
22159
0
CAGGCAGCACCTGGCAATGG


 


410613
3-14-3
384
22141
22160
0
GCAGGCAGCACCTGGCAATG


 


410614
3-14-3
385
22143
22162
0
TAGCAGGCAGCACCTGGCAA


 


410615
3-14-3
388
22199
22218
0
GTCCCCATGCTGGCCTCAGC


 


410616
3-14-3
389
22200
22219
0
GGTCCCCATGCTGGCCTCAG


 


410617
3-14-3
390
22201
22220
0
GGGTCCCCATGCTGGCCTCA


 


410618
3-14-3
391
22202
22221
0
CGGGTCCCCATGCTGGCCTC


 


410619
3-14-3
392
22203
22222
0
ACGGGTCCCCATGCTGGCCT


 


410620
3-14-3
393
22205
22224
0
ACACGGGTCCCCATGCTGGC


 


410621
3-14-3
394
22206
22225
0
GACACGGGTCCCCATGCTGG


 


410622
3-14-3
395
22207
22226
0
GGACACGGGTCCCCATGCTG


 


410623
3-14-3
396
22208
22227
0
TGGACACGGGTCCCCATGCT


 


410624
3-14-3
397
22209
22228
0
GTGGACACGGGTCCCCATGC


 


410625
3-14-3
398
22210
22229
0
AGTGGACACGGGTCCCCATG


 


410626
3-14-3
399
22211
22230
0
CAGTGGACACGGGTCCCCAT


 


410627
3-14-3
400
22212
22231
0
GCAGTGGACACGGGTCCCCA


 


410628
3-14-3
401
22213
22232
0
GGCAGTGGACACGGGTCCCC


 


410629
3-14-3
402
22214
22233
0
TGGCAGTGGACACGGGTCCC


 


410630
3-14-3
403
22215
22234
0
GTGGCAGTGGACACGGGTCC


 


410631
3-14-3
404
22216
22235
0
GGTGGCAGTGGACACGGGTC


 


410632
3-14-3
405
22217
22236
0
TGGTGGCAGTGGACACGGGT


 


410633
3-14-3
411
24096
24115
0
ACTTTGCATTCCAGACCTGG


 


410634
3-14-3
412
24097
24116
0
GACTTTGCATTCCAGACCTG


 


410635
3-14-3
413
24098
24117
0
TGACTTTGCATTCCAGACCT


 


410636
3-14-3
414
24099
24118
0
TTGACTTTGCATTCCAGACC


 


410637
3-14-3
419
26112
26131
0
CAGATGGCAACGGCTGTCAC


 


410638
3-14-3
420
26113
26132
0
GCAGATGGCAACGGCTGTCA


 


410639
3-14-3
421
26114
26133
0
AGCAGATGGCAACGGCTGTC


 


410640
3-14-3
422
26115
26134
0
CAGCAGATGGCAACGGCTGT


 


410641
3-14-3
423
26116
26135
0
GCAGCAGATGGCAACGGCTG


 


410642
3-14-3
424
26118
26137
0
CGGCAGCAGATGGCAACGGC


 


410643
3-14-3
425
26119
26138
0
CCGGCAGCAGATGGCAACGG


 


410644
3-14-3
426
26120
26139
0
TCCGGCAGCAGATGGCAACG


 


410645
3-14-3
427
26121
26140
0
CTCCGGCAGCAGATGGCAAC


 


410646
3-14-3
428
26122
26141
0
GCTCCGGCAGCAGATGGCAA









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 2-13-5 gap-widened motif. Table 10 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 10







Gapmer antisense compounds having a 2-13-5 motif


targeted to SEQ ID NO: 2
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 2
NO: 2
Mismatches
Sequence (5′-3′)





410647
2-13-5
184
 6534
 6553
0
TGAGGTATCCCCGGCGGGCA


 


410648
2-13-5
185
 6535
 6554
0
GTGAGGTATCCCCGGCGGGC


 


410649
2-13-5
186
 6536
 6555
0
GGTGAGGTATCCCCGGCGGG


 


410650
2-13-5
 11
 6537
 6556
0
TGGTGAGGTATCCCCGGCGG


 


410651
2-13-5
187
 6538
 6557
0
TTGGTGAGGTATCCCCGGCG


 


410652
2-13-5
188
 6539
 6558
0
CTTGGTGAGGTATCCCCGGC


 


410653
2-13-5
189
 6540
 6559
0
TCTTGGTGAGGTATCCCCGG


 


410654
2-13-5
190
 6541
 6560
0
ATCTTGGTGAGGTATCCCCG


 


410655
2-13-5
191
 6542
 6561
0
GATCTTGGTGAGGTATCCCC


 


410656
2-13-5
 12
 6543
 6562
0
GGATCTTGGTGAGGTATCCC


 


410657
2-13-5
192
 6544
 6563
0
AGGATCTTGGTGAGGTATCC


 


410658
2-13-5
243
15291
15310
0
GCCACGCCGGCATCCCGGCC


 


410659
2-13-5
244
15292
15311
0
GGCCACGCCGGCATCCCGGC


 


410660
2-13-5
245
15293
15312
0
TGGCCACGCCGGCATCCCGG


 


410661
2-13-5
246
15294
15313
0
TTGGCCACGCCGGCATCCCG


 


410662
2-13-5
247
15295
15314
0
CTTGGCCACGCCGGCATCCC


 


410663
2-13-5
248
15296
15315
0
CCTTGGCCACGCCGGCATCC


 


410664
2-13-5
249
15297
15316
0
CCCTTGGCCACGCCGGCATC


 


410665
2-13-5
 28
15298
15317
0
ACCCTTGGCCACGCCGGCAT


 


410666
2-13-5
250
15299
15318
0
CACCCTTGGCCACGCCGGCA


 


410667
2-13-5
251
15300
15319
0
GCACCCTTGGCCACGCCGGC


 


410668
2-13-5
252
15301
15320
0
GGCACCCTTGGCCACGCCGG


 


410669
2-13-5
253
15302
15321
0
TGGCACCCTTGGCCACGCCG


 


410670
2-13-5
254
15303
15322
0
CTGGCACCCTTGGCCACGCC


 


410671
2-13-5
255
15304
15323
0
GCTGGCACCCTTGGCCACGC


 


410672
2-13-5
348
20719
20738
0
AGGGAACCAGGCCTCATTGA


 


410673
2-13-5
349
20720
20739
0
CAGGGAACCAGGCCTCATTG


 


410674
2-13-5
 49
20721
20740
0
TCAGGGAACCAGGCCTCATT


 


410675
2-13-5
350
20722
20741
0
CTCAGGGAACCAGGCCTCAT


 


410676
2-13-5
351
20723
20742
0
CCTCAGGGAACCAGGCCTCA


 


410677
2-13-5
352
20724
20743
0
TCCTCAGGGAACCAGGCCTC


 


410678
2-13-5
353
20725
20744
0
GTCCTCAGGGAACCAGGCCT


 


410679
2-13-5
 50
20726
20745
0
GGTCCTCAGGGAACCAGGCC


 


410680
2-13-5
354
20727
20746
0
TGGTCCTCAGGGAACCAGGC


 


410681
2-13-5
355
20728
20747
0
CTGGTCCTCAGGGAACCAGG


 


410682
2-13-5
356
20729
20748
0
GCTGGTCCTCAGGGAACCAG


 


410683
2-13-5
376
22133
22152
0
CACCTGGCAATGGCGTAGAC


 


410684
2-13-5
377
22134
22153
0
GCACCTGGCAATGGCGTAGA


 


410685
2-13-5
378
22135
22154
0
AGCACCTGGCAATGGCGTAG


 


410686
2-13-5
379
22136
22155
0
CAGCACCTGGCAATGGCGTA


 


410687
2-13-5
380
22137
22156
0
GCAGCACCTGGCAATGGCGT


 


410688
2-13-5
381
22138
22157
0
GGCAGCACCTGGCAATGGCG


 


410689
2-13-5
382
22139
22158
0
AGGCAGCACCTGGCAATGGC


 


410690
2-13-5
383
22140
22159
0
CAGGCAGCACCTGGCAATGG


 


410691
2-13-5
384
22141
22160
0
GCAGGCAGCACCTGGCAATG


 


410692
2-13-5
 58
22142
22161
0
AGCAGGCAGCACCTGGCAAT


 


410693
2-13-5
385
22143
22162
0
TAGCAGGCAGCACCTGGCAA


 


410694
2-13-5
388
22199
22218
0
GTCCCCATGCTGGCCTCAGC


 


410695
2-13-5
389
22200
22219
0
GGTCCCCATGCTGGCCTCAG


 


410696
2-13-5
390
22201
22220
0
GGGTCCCCATGCTGGCCTCA


 


410697
2-13-5
391
22202
22221
0
CGGGTCCCCATGCTGGCCTC


 


410698
2-13-5
392
22203
22222
0
ACGGGTCCCCATGCTGGCCT


 


410699
2-13-5
 59
22204
22223
0
CACGGGTCCCCATGCTGGCC


 


410700
2-13-5
393
22205
22224
0
ACACGGGTCCCCATGCTGGC


 


410701
2-13-5
394
22206
22225
0
GACACGGGTCCCCATGCTGG


 


410702
2-13-5
395
22207
22226
0
GGACACGGGTCCCCATGCTG


 


410703
2-13-5
396
22208
22227
0
TGGACACGGGTCCCCATGCT


 


410704
2-13-5
397
22209
22228
0
GTGGACACGGGTCCCCATGC


 


410705
2-13-5
398
22210
22229
0
AGTGGACACGGGTCCCCATG


 


410706
2-13-5
399
22211
22230
0
CAGTGGACACGGGTCCCCAT


 


410707
2-13-5
400
22212
22231
0
GCAGTGGACACGGGTCCCCA


 


410708
2-13-5
401
22213
22232
0
GGCAGTGGACACGGGTCCCC


 


410709
2-13-5
402
22214
22233
0
TGGCAGTGGACACGGGTCCC


 


410710
2-13-5
403
22215
22234
0
GTGGCAGTGGACACGGGTCC


 


410711
2-13-5
404
22216
22235
0
GGTGGCAGTGGACACGGGTC


 


410712
2-13-5
405
22217
22236
0
TGGTGGCAGTGGACACGGGT


 


410713
2-13-5
 60
24095
24114
0
CTTTGCATTCCAGACCTGGG


 


410714
2-13-5
411
24096
24115
0
ACTTTGCATTCCAGACCTGG


 


410715
2-13-5
412
24097
24116
0
GACTTTGCATTCCAGACCTG


 


410716
2-13-5
413
24098
24117
0
TGACTTTGCATTCCAGACCT


 


410717
2-13-5
414
24099
24118
0
TTGACTTTGCATTCCAGACC


 


410718
2-13-5
 61
24100
24119
0
CTTGACTTTGCATTCCAGAC


 


410719
2-13-5
419
26112
26131
0
CAGATGGCAACGGCTGTCAC


 


410720
2-13-5
420
26113
26132
0
GCAGATGGCAACGGCTGTCA


 


410721
2-13-5
421
26114
26133
0
AGCAGATGGCAACGGCTGTC


 


410722
2-13-5
422
26115
26134
0
CAGCAGATGGCAACGGCTGT


 


410723
2-13-5
423
26116
26135
0
GCAGCAGATGGCAACGGCTG


 


410724
2-13-5
 62
26117
26136
0
GGCAGCAGATGGCAACGGCT


 


410725
2-13-5
424 
26118
26137
0
CGGCAGCAGATGGCAACGGC


 


410726
2-13-5
425 
26119
26138
0
CCGGCAGCAGATGGCAACGG


 


410727
2-13-5
426 
26120
26139
0
TCCGGCAGCAGATGGCAACG


 


410728
2-13-5
427 
26121
26140
0
CTCCGGCAGCAGATGGCAAC


 


410729
2-13-5
428 
26122
26141
0
GCTCCGGCAGCAGATGGCAA









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 2. In certain such embodiments, the nucleotide sequences illustrated in Table 7 have a 3-13-4 gap-widened motif. Table 11 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 2, having a 3-13-4 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 11







Gapmer antisense compounds having a 3-13-4 motif


targeted to SEQ ID NO: 2
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 2
NO: 2
Mismatches
Sequence (5′-3′)





405526
3-13-4
236
15257
15276
0
CCAGGTGGGTGCCATGACTG


 


405557
3-13-4
50
20726
20745
0
GGTCCTCAGGGAACCAGGCC


 


405564
3-13-4
373
21183
21202
0
AACTGGAGCAGCTCAGCAGC









The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 2: 2274-2400, 2274-2575, 2433-2570, 2433-2579, 2549-2575, 2552-2579, 2585-2638, 2605-2638, 3056-3075, 4150-5159, 4306-4325, 5590-5618, 5667-5686, 6444-6463, 6482-6518, 6492-6518, 6528-6555, 6528-6623, 6534-6561, 6535-6562, 6536-6563, 6537-6563, 6538-6565, 6539-6565, 6540-6567, 6541-6567, 6542-6569, 6546-6573, 6557-6584, 6575-6602, 6585-6611, 6594-6621, 6596-6623, 6652-6671, 7099-7118, 7556-7584, 8836-8855, 8948-8967, 9099-9118, 9099-9168, 9130-9168, 9207-9233, 9207-9235, 9209-9235, 10252-10271, 10633-10652, 11308-11491, 12715-12734, 12928-12947, 13681-13700, 13746-13779, 13816-13847, 13903-13945, 13977-14141, 14179-14198, 14267-14286, 14397-14423, 14441-14460, 14494-14513, 14494-14543, 14524-14543, 14601-14650, 14670-14700, 14675-14700, 14801-14828, 14877-14912, 14877-14915, 14877-14973, 14916-14943, 14916-14973, 14925-14951, 14934-14963, 14946-14973, 14979-14998, 15254-15280, 15254-15328, 15264-15290, 15279-15305, 15291-15318, 15292-15319, 15293-15320, 15294-15321, 15294-15321, 15295-15322, 15296-15315, 15296-15323, 15297-15323, 15298-15323, 15299-15323, 15300-15323, 15301-15328, 15330-15355, 15330-15490, 15339-15366, 15358-15490, 16134-16153, 16668-16687, 17267-17286, 18377-18427, 18561-18580, 18591-18618, 18591-18646, 18591-18668, 18695-18746, 18705-18730, 18709-18736, 18719-18746, 19203-20080, 19931-19952, 19954-19981, 19964-19990, 19973-19999, 19982-20009, 19992-20016, 20016-20042, 20025-20052, 20036-20062, 20045-20070, 20100-20119, 20188-20207, 20624-20650, 20624-20759, 20629-20804, 20633-20660, 20635-20781, 20643-20662, 20657-20676, 20670-20697, 20680-20706, 20683-20781, 20689-20715, 20698-20725, 20709-20736, 20717-20744, 20718-20745, 20719-20746, 20720-20747, 20721-20748, 20722-20749, 20727-20752, 20735-20759, 20762-21014, 20785-21014, 21082-21107, 21082-21152, 21091-21114, 21118-21144, 21127-21152, 21181-21209, 21181-21211, 21183-21211, 21481-21500, 21589-21608, 21692-21719, 22000-22227, 22096-22115, 22096-22223, 22096-22311, 22133-22160, 22133-22163, 22134-22161, 22135-22162, 22136-22163, 22137-22163, 22138-22163, 22189-22239, 22199-22226, 22199-22227, 22200-22227, 22201-22228, 22202-22229, 22203-22230, 22204-22231, 22205-22232, 22206-22233, 22207-22234, 22208-22235, 22209-22236, 22210-22236, 22210-22239, 22211-22236, 22212-22239, 22292-22311, 23985-24054, 24035-24134, 24095-24121, 24858-24877, 24907-24926, 25413-25432, 25994-26013, 26112-26139, 26112-26161, 26112-27303, 26113-26140, 26114-26141, 26115-26141, 26116-26141, 26117-26141, 26117-26475, 26118-26141, 26120-26141, 26132-26151, 26142-26161, 26217-26241, 26311-26335, 26389-26432, 26456-26576, 26635-26662, 26707-26734, 26707-26736, 26790-26820, 27034-27263, 27279-27303, or 27350-27376.
In certain embodiments, a target region is nucleotides 2274-2400 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2274-2400 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 4 or 5. In certain such embodiments, an antisense compound targeted to nucleotides 2274-2400 of SEQ ID NO: 2 is selected from ISIS NOs: 395149, 399871, 395150 or 399872.
In certain embodiments, a target region is nucleotides 2274-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2274-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 4, 5, 6, 7, 8, 159, 160, 162, 163, 164, 165, 166, 167, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2274-2575 of SEQ ID NO: 2 is selected from ISIS NOs: 395149, 399871, 395150, 399872, 410742, 405999, 395151, 399873, 405861, 405862, 405863, 405864, 395152, 399874, 405865, 405866, 405867, 405868, 399793, or 399949.
In certain embodiments, a target region is nucleotides 2433-2570 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2433-2570 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 6, 7, 159, 160, 162, 163, 164, 165, 166, or 167. In certain such embodiments, an antisense compound targeted to nucleotides 2433-2570 of SEQ ID NO: 2 is selected from ISIS NOs: 395151, 395152, 399873, 399874, 405861, 405862, 405863, 405864, 405865, 405866, 405999, or 410742.
In certain embodiments, a target region is nucleotides 2433-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2433-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 6, 7, 8, 159, 160, 162, 163, 164, 165, 166, 167, 168, 169, or 170. In certain such embodiments, an antisense compound targeted to nucleotides 2433-2579 of SEQ ID NO: 2 is selected from ISIS NOs: 395151, 395152, 399793, 399873, 399874, 399949, 405861, 405862, 405863, 405864, 405865, 405866, 405867, 405868, 405999, 410742, or 410743.
In certain embodiments, a target region is nucleotides 2549-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2549-2575 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 7, 8, 166, 167, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2549-2575 of SEQ ID NO: 2 is selected from ISIS NOs: 395152, 399793, 399874, 399949, 405865, 405866, 405867, or 405868.
In certain embodiments, a target region is nucleotides 2552-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2552-2579 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 8, 168, 169 or 170. In certain such embodiments, an antisense compound targeted to nucleotides 2552-2579 of SEQ ID NO: 2 is selected from ISIS NOs: 399793, 399949, 405867, 405868, or 410743.
In certain embodiments, a target region is nucleotides 2585-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2585-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 9, 171 or 172. In certain such embodiments, an antisense compound targeted to nucleotides 2585-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 395153, 399875, 410744, or 410745.
In certain embodiments, a target region is nucleotides 2605-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 2605-2638 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 8, 168 or 169. In certain such embodiments, an antisense compound targeted to nucleotides 2605-2638 of SEQ ID NO: 2 is selected from ISIS NOs: 410745, 395153, or 399875.
In certain embodiments, a target region is nucleotides 3056-3075 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 3056-3075 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 107. In certain such embodiments, an antisense compound targeted to nucleotides 3056-3075 of SEQ ID NO: 2 is selected from ISIS NOs: 399837 or 399993.
In certain embodiments, a target region is nucleotides 4150-5159 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 4150-5159 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 106. In certain such embodiments, an antisense compound targeted to nucleotides 4150-5159 of SEQ ID NO: 2 is selected from ISIS NOs: 399839 or 399995.
In certain embodiments, a target region is nucleotides 4306-4325 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 4306-4325 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 80. In certain such embodiments, an antisense compound targeted to nucleotides 4306-4325 of SEQ ID NO: 2 is selected from ISIS NOs: 399838 or 399994.
In certain embodiments, a target region is nucleotides 5590-5618 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 5590-5618 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 102 or 121. In certain such embodiments, an antisense compound targeted to nucleotides 5590-5618 of SEQ ID NO: 2 is selected from ISIS NOs: 395221, 399840, 399943, or 399996.
In certain embodiments, a target region is nucleotides 5667-5686 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 5667-5686 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 94. In certain such embodiments, an antisense compound targeted to nucleotides 5667-5686 of SEQ ID NO: 2 is selected from ISIS NOs: 399841 or 399997.
In certain embodiments, a target region is nucleotides 6444-6463 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6444-6463 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 176. In certain such embodiments, an antisense compound targeted to nucleotides 6444-6463 of SEQ ID NO: 2 is selected from ISIS NOs: 410746.
In certain embodiments, a target region is nucleotides 6482-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6482-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 177, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 6482-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154, 399876, 406003, 406004, 406005, 406006 or 410747.
In certain embodiments, a target region is nucleotides 6492-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6492-6518 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 10, 178, 179, 180 or 181. In certain such embodiments, an antisense compound targeted to nucleotides 6492-6518 of SEQ ID NO: 2 is selected from ISIS NOs: 395154, 399876, 406003, 406004, 406005 or 406006.
In certain embodiments, a target region is nucleotides 6528-6555 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6528-6555 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 182, 183, 184, 185 or 186. In certain such embodiments, an antisense compound targeted to nucleotides 6528-6555 of SEQ ID NO: 2 is selected from ISIS NOs: 406007, 410529, 410530, 410531, 410574, 410575, 410576, 410647, 410648, 410649 or 410748.
In certain embodiments, a target region is nucleotides 6528-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6528-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 13, 13, 14, 15, 16, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 192, 193, 194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, or 207. In certain such embodiments, an antisense compound targeted to nucleotides 6528-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 395156, 395157, 399794, 399795, 399796, 399877, 399878, 399879, 399950, 399951, 399952, 406007, 406008, 406009, 406010, 406011, 406012, 406013, 406014, 406015, 406016, 406017, 406018, 406019, 406020, 410529, 410530, 410531, 410532, 410533, 410534, 410535, 410574, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410582, 410647, 410648, 410649, 410650, 410651, 410652, 410653, 410654, 410655, 410656, 410657, 410730, 410731, 410748, 410749, or 410750.
In certain embodiments, a target region is nucleotides 6534-6561 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6534-6561 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 184, 185, 186, 187, 188, 189, 190 or 191. In certain such embodiments, an antisense compound targeted to nucleotides 6534-6561 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399877, 406008, 406009, 410529, 410530, 410531, 410532, 410533, 410534, 410574, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410647, 410648, 410649, 410650, 410651, 410652, 410653, 410654, or 410655.
In certain embodiments, a target region is nucleotides 6535-6562 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6535-6562 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 185, 186, 187, 188, 189, 190 or 191. In certain such embodiments, an antisense compound targeted to nucleotides 6535-6562 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410530, 410531, 410532, 410533, 410534, 410575, 410576, 410577, 410578, 410579, 410580, 410581, 410648, 410649, 410650, 410651, 410652, 410653, 410654, 410655 or 410656.
In certain embodiments, a target region is nucleotides 6536-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6536-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 186, 187, 188, 189, 190, 191 or 192. In certain such embodiments, an antisense compound targeted to nucleotides 6536-6563 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410531, 410532, 410533, 410534, 410535, 410576, 410577, 410578, 410579, 410580, 410581, 410582, 410649, 410650, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6537-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6537-6563 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 11, 12, 187, 188, 189, 190, 191 or 192. In certain such embodiments, an antisense compound targeted to nucleotides 6537-6563 of SEQ ID NO: 2 is selected from ISIS NOs: 395155, 399794, 399877, 399950, 406008, 406009, 410532, 410533, 410534, 410535, 410577, 410578, 410579, 410580, 410581, 410582, 410650, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6538-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6538-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 187, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 6538-6565 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406008, 406009, 406010, 410532, 410533, 410534, 410535, 410577, 410578, 410579, 410580, 410581, 410582, 410651, 410652, 410653, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6539-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6539-6565 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 188, 189, 190, 191, 192 or 193. In certain such embodiments, an antisense compound targeted to nucleotides 6539-6565 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406008, 406009, 406010, 410533, 410534, 410535, 410578, 410579, 410580, 410581, 410582, 410652, 410653, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6540-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6540-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 189, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 6540-6567 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406009, 406010, 406011, 410533, 410534, 410535, 410579, 410580, 410581, 410582, 410653, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6541-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6541-6567 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 190, 191, 192, 193 or 194. In certain such embodiments, an antisense compound targeted to nucleotides 6541-6567 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406009, 406010, 406011, 410534, 410535, 410580, 410581, 410582, 410654, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6542-6569 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6542-6569 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 12, 191, 192, 193, 194 or 195. In certain such embodiments, an antisense compound targeted to nucleotides 6542-6569 of SEQ ID NO: 2 is selected from ISIS NOs: 399794, 399950, 406010, 406011, 406012, 410534, 410535, 410581, 410582, 410655, 410656 or 410657.
In certain embodiments, a target region is nucleotides 6546-6573 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6546-6573 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 13, 193, 194, 195 or 196. In certain such embodiments, an antisense compound targeted to nucleotides 6546-6573 of SEQ ID NO: 2 is selected from ISIS NOs: 399795, 399951, 406010, 406011, 406012 or 406013.
In certain embodiments, a target region is nucleotides 6557-6584 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6557-6584 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 14, 197 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 6557-6584 of SEQ ID NO: 2 is selected from ISIS NOs: 395156, 399878, 410749 or 410750.
In certain embodiments, a target region is nucleotides 6575-6602 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6575-6602 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 15 or 198. In certain such embodiments, an antisense compound targeted to nucleotides 6575-6602 of SEQ ID NO: 2 is selected from ISIS NOs: 395157, 399879 or 410750.
In certain embodiments, a target region is nucleotides 6585-6611 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6585-6611 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 16, 199, 200 or 201. In certain such embodiments, an antisense compound targeted to nucleotides 6585-6611 of SEQ ID NO: 2 is selected from ISIS NOs: 399796, 399952, 406014, 406015 or 406016.
In certain embodiments, a target region is nucleotides 6594-6621 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6594-6621 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 202, 203, 204, 205 or 206. In certain such embodiments, an antisense compound targeted to nucleotides 6594-6621 of SEQ ID NO: 2 is selected from ISIS NOs: 406017, 406018, 406019, 406020 or 410730.
In certain embodiments, a target region is nucleotides 6596-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6596-6623 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 203, 204, 205, 206 or 207. In certain such embodiments, an antisense compound targeted to nucleotides 6596-6623 of SEQ ID NO: 2 is selected from ISIS NOs: 406017, 406018, 406019, 406020 or 410731.
In certain embodiments, a target region is nucleotides 6652-6671 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 6652-6671 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 108. In certain such embodiments, an antisense compound targeted to nucleotides 6652-6671 of SEQ ID NO: 2 is selected from ISIS NOs: 399842 or 399998.
In certain embodiments, a target region is nucleotides 7099-7118 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 7099-7118 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 114. In certain such embodiments, an antisense compound targeted to nucleotides 7099-7118 of SEQ ID NO: 2 is selected from ISIS NOs: 399843 or 399999.
In certain embodiments, a target region is nucleotides 7556-7584 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 7556-7584 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 91 or 131. In certain such embodiments, an antisense compound targeted to nucleotides 7556-7584 of SEQ ID NO: 2 is selected from ISIS NOs: 399844, 399845, 400000 or 400001.
In certain embodiments, a target region is nucleotides 8836-8855 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 8836-8855 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 145. In certain such embodiments, an antisense compound targeted to nucleotides 8836-8855 of SEQ ID NO: 2 is selected from ISIS NOs: 399846 or 400002.
In certain embodiments, a target region is nucleotides 8948-8967 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 8948-8967 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 90. In certain such embodiments, an antisense compound targeted to nucleotides 8948-8967 of SEQ ID NO: 2 is selected from ISIS NOs: 399847 or 400003.
In certain embodiments, a target region is nucleotides 9099-9118 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9099-9118 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 125. In certain such embodiments, an antisense compound targeted to nucleotides 9099-9118 of SEQ ID NO: 2 is selected from ISIS NOs: 399848 or 400004.
In certain embodiments, a target region is nucleotides 9099-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9099-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17, 125 or 209. In certain such embodiments, an antisense compound targeted to nucleotides 9099-9168 of SEQ ID NO: 2 is selected from ISIS NOs: 395158, 399848, 399880, 400004 or 410752.
In certain embodiments, a target region is nucleotides 9130-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9130-9168 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 17. In certain such embodiments, an antisense compound targeted to nucleotides 9130-9168 of SEQ ID NO: 2 is selected from ISIS NOs: 395158 or 399880.
In certain embodiments, a target region is nucleotides 9207-9233 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9207-9233 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212 or 213. In certain such embodiments, an antisense compound targeted to nucleotides 9207-9233 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406021, 406022, 406023 or 406024.
In certain embodiments, a target region is nucleotides 9207-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9207-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 210, 211, 212, 213 or 214. In certain such embodiments, an antisense compound targeted to nucleotides 9207-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406021, 406022, 406023, 406024 or 410732.
In certain embodiments, a target region is nucleotides 9209-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 9209-9235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 18, 211, 212, 213 or 214. In certain such embodiments, an antisense compound targeted to nucleotides 9209-9235 of SEQ ID NO: 2 is selected from ISIS NOs: 395159, 399881, 406022, 406023, 406024 or 410732.
In certain embodiments, a target region is nucleotides 10252-10271 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 10252-10271 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 148. In certain such embodiments, an antisense compound targeted to nucleotides 10252-10271 of SEQ ID NO: 2 is selected from ISIS NOs: 399849 or 400005.
In certain embodiments, a target region is nucleotides 10633-10652 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 10633-10652 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 127. In certain such embodiments, an antisense compound targeted to nucleotides 10633-10652 of SEQ ID NO: 2 is selected from ISIS NOs: 395222 or 399944.
In certain embodiments, a target region is nucleotides 11308-11491 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 11308-11491 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 79 or 84. In certain such embodiments, an antisense compound targeted to nucleotides 11308-11491 of SEQ ID NO: 2 is selected from ISIS NOs: 395223, 399850, 399945 or 400006.
In certain embodiments, a target region is nucleotides 12715-12734 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 12715-12734 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 88. In certain such embodiments, an antisense compound targeted to nucleotides 12715-12734 of SEQ ID NO: 2 is selected from ISIS NOs: 399851 or 400007.
In certain embodiments, a target region is nucleotides 12928-12947 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 12928-12947 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 111. In certain such embodiments, an antisense compound targeted to nucleotides 12928-12947 of SEQ ID NO: 2 is selected from ISIS NOs: 399852 or 400008.
In certain embodiments, a target region is nucleotides 13681-13700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13681-13700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 85. In certain such embodiments, an antisense compound targeted to nucleotides 13681-13700 of SEQ ID NO: 2 is selected from ISIS NOs: 395201 or 399923.
In certain embodiments, a target region is nucleotides 13746-13779 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13746-13779 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 100 or 116. In certain such embodiments, an antisense compound targeted to nucleotides 13746-13779 of SEQ ID NO: 2 is selected from ISIS NOs: 399827, 399828, 399983 or 399984.
In certain embodiments, a target region is nucleotides 13816-13847 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13816-13847 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 117 or 129. In certain such embodiments, an antisense compound targeted to nucleotides 13816-13847 of SEQ ID NO: 2 is selected from ISIS NOs: 395202, 399829, 399924 or 399985.
In certain embodiments, a target region is nucleotides 13903-13945 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13903-13945 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain such embodiments, an antisense compound targeted to nucleotides 13903-13945 of SEQ ID NO: 2 is selected from ISIS NOs: 395203, 399830, 399925 or 399986.
In certain embodiments, a target region is nucleotides 13977-14141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 13977-14141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 83, 136, 137, 140 or 152. In certain such embodiments, an antisense compound targeted to nucleotides 13977-14141 of SEQ ID NO: 2 is selected from ISIS NOs: 395204, 395205, 395206, 399831, 399832, 399926, 399927, 399928, 399987 or 399988.
In certain embodiments, a target region is nucleotides 14179-14198 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14179-14198 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 132. In certain such embodiments, an antisense compound targeted to nucleotides 14179-14198 of SEQ ID NO: 2 is selected from ISIS NOs: 395207 or 399929.
In certain embodiments, a target region is nucleotides 14267-14286 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14267-14286 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 139. In certain such embodiments, an antisense compound targeted to nucleotides 14267-14286 of SEQ ID NO: 2 is selected from ISIS NOs: 395208 or 399930.
In certain embodiments, a target region is nucleotides 14397-14423 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14397-14423 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain such embodiments, an antisense compound targeted to nucleotides 14397-14423 of SEQ ID NO: 2 is selected from ISIS NOs: 395209, 399833, 399931 or 399989.
In certain embodiments, a target region is nucleotides 14441-14460 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14441-14460 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 113. In certain such embodiments, an antisense compound targeted to nucleotides 14441-14460 of SEQ ID NO: 2 is selected from ISIS NOs: 395210 or 399932.
In certain embodiments, a target region is nucleotides 14494-14513 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14494-14513 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 138. In certain such embodiments, an antisense compound targeted to nucleotides 14494-14513 of SEQ ID NO: 2 is selected from ISIS NOs: 395211 or 399933.
In certain embodiments, a target region is nucleotides 14494-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14494-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain such embodiments, an antisense compound targeted to nucleotides 14494-14543 of SEQ ID NO: 2 is selected from ISIS NOs: 395211, 395212, 399933 or 399934.
In certain embodiments, a target region is nucleotides 14524-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14524-14543 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98. In certain such embodiments, an antisense compound targeted to nucleotides 14524-14543 of SEQ ID NO: 2 is selected from ISIS NOs: 395212 or 399934.
In certain embodiments, a target region is nucleotides 14601-14650 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14601-14650 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain such embodiments, an antisense compound targeted to nucleotides 14601-14650 of SEQ ID NO: 2 is selected from ISIS NOs: 395213, 395214, 399935 or 399936.
In certain embodiments, a target region is nucleotides 14670-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14670-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 103 or 133. In certain such embodiments, an antisense compound targeted to nucleotides 14670-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216, 399834, 399937, 399938 or 399990.
In certain embodiments, a target region is nucleotides 14675-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14675-14700 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 103 or 133. In certain such embodiments, an antisense compound targeted to nucleotides 14675-14700 of SEQ ID NO: 2 is selected from ISIS NOs: 395215, 395216, 399937 or 399938.
In certain embodiments, a target region is nucleotides 14801-14828 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14801-14828 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 155 or 156. In certain such embodiments, an antisense compound targeted to nucleotides 14801-14828 of SEQ ID NO: 2 is selected from ISIS NOs: 395217, 399835, 399939 or 399991.
In certain embodiments, a target region is nucleotides 14877-14912 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14912 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14912 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 399882, 406025, 406026 or 410753.
In certain embodiments, a target region is nucleotides 14877-14915 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14915 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 215, 216 or 217. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14915 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 399797, 399882, 399953, 406025, 406026 or 410753.
In certain embodiments, a target region is nucleotides 14877-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14877-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 14877-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395160, 395161, 395162, 399797, 399798, 399799, 399882, 399883, 399884, 399953, 399954, 399955, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 406025, 406026, 406027, 406028, 406029, 406030, 406031, 406032, 410733, 410753, or 410754.
In certain embodiments, a target region is nucleotides 14916-14943 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14916-14943 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 218, 219, 220, 221, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 14916-14943 of SEQ ID NO: 2 is selected from ISIS NOs: 395161, 399798, 399883, 399954, 405869, 405870, 405871, 405872, 405873 or 405874.
In certain embodiments, a target region is nucleotides 14916-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14916-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, or 223. In certain such embodiments, an antisense compound targeted to nucleotides 14916-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395161, 395162, 399798, 399799, 399883, 399884, 399954, 399955, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 406030, 406031, 406032, 410733, or 410754.
In certain embodiments, a target region is nucleotides 14925-14951 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14925-14951 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. In certain such embodiments, an antisense compound targeted to nucleotides 14925-14951 of SEQ ID NO: 2 is selected from ISIS NOs: 405875, 405876, 406027 or 406028.
In certain embodiments, a target region is nucleotides 14934-14963 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14934-14963 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 23, 228, 229, 230, 231 or 232. In certain such embodiments, an antisense compound targeted to nucleotides 14934-14963 of SEQ ID NO: 2 is selected from ISIS NOs: 399799, 399955, 406029, 406030, 406031, 406032 or 410733.
In certain embodiments, a target region is nucleotides 14946-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14946-14973 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 14946-14973 of SEQ ID NO: 2 is selected from ISIS NOs: 395162, 399884 or 410754.
In certain embodiments, a target region is nucleotides 14979-14998 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 14979-14998 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25. In certain such embodiments, an antisense compound targeted to nucleotides 14979-14998 of SEQ ID NO: 2 is selected from ISIS NOs: 395163 or 399885.
In certain embodiments, a target region is nucleotides 15254-15280 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15254-15280 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236 or 237. In certain such embodiments, an antisense compound targeted to nucleotides 15254-15280 of SEQ ID NO: 2 is selected from ISIS NOs: 405526, 405604, 406033 or 410756.
In certain embodiments, a target region is nucleotides 15254-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15254-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 28, 235, 236, 237, 238, 239, 240, 241, 242, 243, 243, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15254-15328 of SEQ ID NO: 2 is selected from ISIS NOs: 395164, 395165, 399800, 399886, 399887, 399956, 405526, 405604, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 406033, 406034, 406035, 406036, 406037, 406038, 409126, 410536, 410537, 410538, 410539, 410583, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410658, 410659, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668, 410669, 410670, 410671, 410756, 410757, or 410758.
In certain embodiments, a target region is nucleotides 15264-15290 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15264-15290 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 27, 238 or 239. In certain such embodiments, an antisense compound targeted to nucleotides 15264-15290 of SEQ ID NO: 2 is selected from ISIS NOs: 395164, 399800, 399886, 399956, 406034, or 406035.
In certain embodiments, a target region is nucleotides 15279-15305 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15279-15305 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241 or 242. In certain such embodiments, an antisense compound targeted to nucleotides 15279-15305 of SEQ ID NO: 2 is selected from ISIS NOs: 406036, 406037, or 410757.
In certain embodiments, a target region is nucleotides 15291-15318 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15291-15318 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 243, 244, 245, 246, 247, 248, 249, 250, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15291-15318 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 406038, 409126, 410536, 410537, 410583, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410658, 410659, 410660, 410661, 410662, 410663, 410664, 410665, or 410666.
In certain embodiments, a target region is nucleotides 15292-15319 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15292-15319 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 244, 245, 246, 247, 248, 249, 250 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15292-15319 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 406038, 409126, 410537, 410584, 410585, 410586, 410587, 410588, 410589, 410590, 410659, 410660, 410661, 410662, 410663, 410664, 410665, or 410666.
In certain embodiments, a target region is nucleotides 15293-15320 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15293-15320 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 245, 246, 247, 248, 249, 250, 251, 252 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15293-15320 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 406038, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410668.
In certain embodiments, a target region is nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668 or 410669.
In certain embodiments, a target region is nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15294-15321 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15294-15321 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668 or 410669.
In certain embodiments, a target region is nucleotides 15295-15322 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15295-15322 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 250, 251, 252, 253, 254 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15295-15322 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410662, 410663, 410664, 410665, 410666, 410667, 410668, 410669 or 410670.
In certain embodiments, a target region is nucleotides 15296-15315 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15296-15315 of SEQ ID NO: 2. In one such embodiment, an antisense compound targeted to nucleotides 15296-15315 of SEQ ID NO: 2 is ISIS NO: 405879.
In certain embodiments, a target region is nucleotides 15296-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15296-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15296-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410588, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410663, 410664, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.
In certain embodiments, a target region is nucleotides 15297-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15297-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15297-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405880, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410589, 410590, 410591, 410592, 410593, 410594, 410595, 410664, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.
In certain embodiments, a target region is nucleotides 15298-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15298-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 15298-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 395165, 399887, 405881, 405882, 405883, 405884, 409126, 410538, 410539, 410590, 410591, 410592, 410593, 410594, 410595, 410665, 410666, 410667, 410668, 410669, 410670 or 410671.
In certain embodiments, a target region is nucleotides 15299-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15299-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254 or 255. In certain such embodiments, an antisense compound targeted to nucleotides 15299-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 405881, 405882, 405883, 405884, 410538, 410539, 410590, 410591, 410592, 410593, 410594, 410595, 410666, 410667, 410668, 410669, 410670 or 410671.
In certain embodiments, a target region is nucleotides 15300-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15300-15323 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254 or 255. In certain such embodiments, an antisense compound targeted to nucleotides 15300-15323 of SEQ ID NO: 2 is selected from ISIS NOs: 405882, 405883, 405884, 410538, 410539, 410591, 410592, 410593, 410594, 410595, 410667, 410668, 410669, 410670 or 410671.
In certain embodiments, a target region is nucleotides 15301-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15301-15328 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 252, 253, 254, 255 or 256. In certain such embodiments, an antisense compound targeted to nucleotides 15301-15328 of SEQ ID NO: 2 is selected from ISIS NOs: 405883, 405884, 410538, 410539, 410592, 410593, 410594, 410595, 410668, 410669, 410670, 410671 or 410758.
In certain embodiments, a target region is nucleotides 15330-15355 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15330-15355 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 257, 258 or 259. In certain such embodiments, an antisense compound targeted to nucleotides 15330-15355 of SEQ ID NO: 2 is selected from ISIS NOs: 395166, 399888, 406039, 406040 or 406041.
In certain embodiments, a target region is nucleotides 15330-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15330-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 86, 257, 258, 259, 260, 261, 262 or 263. In certain such embodiments, an antisense compound targeted to nucleotides 15330-15490 of SEQ ID NO: 2 is selected from ISIS NOs: 395166, 399801, 399853, 399888, 399957, 400009, 406039, 406040, 406041, 406042, 406043, 406044 or 410759.
In certain embodiments, a target region is nucleotides 15339-15366 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15339-15366 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 30, 260, 261 or 262. In certain such embodiments, an antisense compound targeted to nucleotides 15339-15366 of SEQ ID NO: 2 is selected from ISIS NOs: 399801, 399957, 406042, 406043 or 406044.
In certain embodiments, a target region is nucleotides 15358-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 15358-15490 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 86 or 263. In certain such embodiments, an antisense compound targeted to nucleotides 15358-15490 of SEQ ID NO: 2 is selected from ISIS NOs: 399853, 400009 or 410759.
In certain embodiments, a target region is nucleotides 16134-16153 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 16134-16153 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 97. In certain such embodiments, an antisense compound targeted to nucleotides 16134-16153 of SEQ ID NO: 2 is selected from ISIS NOs: 399854 or 400010.
In certain embodiments, a target region is nucleotides 16668-16687 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 16668-16687 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 120. In certain such embodiments, an antisense compound targeted to nucleotides 16668-16687 of SEQ ID NO: 2 is selected from ISIS NOs: 399855 or 400011.
In certain embodiments, a target region is nucleotides 17267-17286 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 17267-17286 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 150. In certain such embodiments, an antisense compound targeted to nucleotides 17267-17286 of SEQ ID NO: 2 is selected from ISIS NOs: 399856 or 400012.
In certain embodiments, a target region is nucleotides 18377-18427 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18377-18427 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 115 or 134. In certain such embodiments, an antisense compound targeted to nucleotides 18377-18427 of SEQ ID NO: 2 is selected from ISIS NOs: 399857, 399858, 400013 or 400014.
In certain embodiments, a target region is nucleotides 18561-18580 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18561-18580 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 157. In certain such embodiments, an antisense compound targeted to nucleotides 18561-18580 of SEQ ID NO: 2 is selected from ISIS NOs: 395224 or 399946.
In certain embodiments, a target region is nucleotides 18591-18618 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18618 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268 or 269. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18618 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911 or 406045.
In certain embodiments, a target region is nucleotides 18591-18646 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18646 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 32, 266, 267, 268, 269, 270, 271 or 272. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18646 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045, 410761 or 410762.
In certain embodiments, a target region is nucleotides 18591-18668 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18591-18668 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, 269, 270, 271, 272 or 273. In certain such embodiments, an antisense compound targeted to nucleotides 18591-18668 of SEQ ID NO: 2 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, 405912, 406045, 410761, 410762 or 410763.
In certain embodiments, a target region is nucleotides 18695-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18695-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284 or 285. In certain such embodiments, an antisense compound targeted to nucleotides 18695-18746 of SEQ ID NO: 2 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, 405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734 or 410764.
In certain embodiments, a target region is nucleotides 18705-18730 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18705-18730 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276 or 277. In certain such embodiments, an antisense compound targeted to nucleotides 18705-18730 of SEQ ID NO: 2 is selected from ISIS NOs: 395169, 399891, 405913, 405914 or 405915.
In certain embodiments, a target region is nucleotides 18709-18736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18709-18736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 276, 277, 278, 279 or 280. In certain such embodiments, an antisense compound targeted to nucleotides 18709-18736 of SEQ ID NO: 2 is selected from ISIS NOs: 405914, 405915, 405916, 405917 or 410734.
In certain embodiments, a target region is nucleotides 18719-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 18719-18746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 281, 282, 283, 284 or 285. In certain such embodiments, an antisense compound targeted to nucleotides 18719-18746 of SEQ ID NO: 2 is selected from ISIS NOs: 405918, 405919, 405920, 405921 or 405922.
In certain embodiments, a target region is nucleotides 19203-20080 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19203-20080 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 35, 36, 37, 38, 39, 40, 41, 105, 128, 149, 151, 288, 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317 or 318. In certain such embodiments, an antisense compound targeted to nucleotides 19203-20080 of SEQ ID NO: 2 is selected from ISIS NOs: 395170, 395171, 395172, 395173, 395174, 395175, 399802, 399803, 399804, 399805, 399859, 399860, 399892, 399893, 399894, 399895, 399896, 399897, 399958, 399959, 399960, 399961, 400015, 400016, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405933, 405934, 405935, 405936, 405937, 405938, 405939, 405940, 405941, 405942, 405943, 405944, 405945, 405946, 405947, 410735, 410736, 410737, 410767, 410768 or 410769.
In certain embodiments, a target region is nucleotides 19931-19952 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19931-19952 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 149 or 188. In certain such embodiments, an antisense compound targeted to nucleotides 19931-19952 of SEQ ID NO: 2 is selected from ISIS NOs: 395170, 399892 or 405923.
In certain embodiments, a target region is nucleotides 19954-19981 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19954-19981 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 34, 290, 291, 292 or 293. In certain such embodiments, an antisense compound targeted to nucleotides 19954-19981 of SEQ ID NO: 2 is selected from ISIS NOs: 395171, 399893, 405924, 405925, 405926 or 410735.
In certain embodiments, a target region is nucleotides 19964-19990 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19964-19990 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 35, 128, 128, 294 or 295. In certain such embodiments, an antisense compound targeted to nucleotides 19964-19990 of SEQ ID NO: 2 is selected from ISIS NOs: 395172, 399802, 399894, 399958, 405927 or 405928.
In certain embodiments, a target region is nucleotides 19973-19999 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19973-19999 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 36, 296, 297 or 298. In certain such embodiments, an antisense compound targeted to nucleotides 19973-19999 of SEQ ID NO: 2 is selected from ISIS NOs: 395173, 399895, 405929, 405930 or 405931.
In certain embodiments, a target region is nucleotides 19982-20009 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19982-20009 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 37, 299, 300, 301 or 302. In certain such embodiments, an antisense compound targeted to nucleotides 19982-20009 of SEQ ID NO: 2 is selected from ISIS NOs: 399803, 399959, 405932, 405933, 405934 or 405935.
In certain embodiments, a target region is nucleotides 19992-20016 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 19992-20016 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 38, 303 or 304. In certain such embodiments, an antisense compound targeted to nucleotides 19992-20016 of SEQ ID NO: 2 is selected from ISIS NOs: 395174, 399896, 405936 or 410736.
In certain embodiments, a target region is nucleotides 20016-20042 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20016-20042 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 305 or 306. In certain such embodiments, an antisense compound targeted to nucleotides 20016-20042 of SEQ ID NO: 2 is selected from ISIS NOs: 405937 or 410768.
In certain embodiments, a target region is nucleotides 20025-20052 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20025-20052 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309 or 310. In certain such embodiments, an antisense compound targeted to nucleotides 20025-20052 of SEQ ID NO: 2 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940 or 405941.
In certain embodiments, a target region is nucleotides 20036-20062 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20036-20062 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 311, 312, 313 or 314. In certain such embodiments, an antisense compound targeted to nucleotides 20036-20062 of SEQ ID NO: 2 is selected from ISIS NOs: 399804, 399960, 405942, 405943, 405944 or 410737.
In certain embodiments, a target region is nucleotides 20045-20070 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20045-20070 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 315, 316 or 317. In certain such embodiments, an antisense compound targeted to nucleotides 20045-20070 of SEQ ID NO: 2 is selected from ISIS NOs: 399805, 399961, 405945, 405946 or 405947.
In certain embodiments, a target region is nucleotides 20100-20119 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20100-20119 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 158. In certain such embodiments, an antisense compound targeted to nucleotides 20100-20119 of SEQ ID NO: 2 is selected from ISIS NOs: 399861 or 400017.
In certain embodiments, a target region is nucleotides 20188-20207 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20188-20207 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 109. In certain such embodiments, an antisense compound targeted to nucleotides 20188-20207 of SEQ ID NO: 2 is selected from ISIS NOs: 399862 or 400018.
In certain embodiments, a target region is nucleotides 20624-20650 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20624-20650 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 141, 320 or 321. In certain such embodiments, an antisense compound targeted to nucleotides 20624-20650 of SEQ ID NO: 2 is selected from ISIS NOs: 399863, 400019, 405949 or 405950.
In certain embodiments, a target region is nucleotides 20624-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20624-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 141, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20624-20759 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399863, 399899, 399900, 399901, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 400019, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740 or 410770.
In certain embodiments, a target region is nucleotides 20629-20804 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20629-20804 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 356, 357, 358, 359 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20629-20804 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740, 410770 or 410771.
In certain embodiments, a target region is nucleotides 20633-20660 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20633-20660 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 322, 323, 324 or 325. In certain such embodiments, an antisense compound targeted to nucleotides 20633-20660 of SEQ ID NO: 2 is selected from ISIS NOs: 399807, 399963, 405951, 405952, 405953 or 410738.
In certain embodiments, a target region is nucleotides 20635-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20635-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20635-20781 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740 or 410770.
In certain embodiments, a target region is nucleotides 20643-20662 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20643-20662 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 326. In certain such embodiments, an antisense compound targeted to nucleotides 20643-20662 of SEQ ID NO: 2 is selected from ISIS NOs: 405954.
In certain embodiments, a target region is nucleotides 20657-20676 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20657-20676 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 327. In certain such embodiments, an antisense compound targeted to nucleotides 20657-20676 of SEQ ID NO: 2 is selected from ISIS NOs: 410770.
In certain embodiments, a target region is nucleotides 20670-20697 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20670-20697 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, 331 or 332. In certain such embodiments, an antisense compound targeted to nucleotides 20670-20697 of SEQ ID NO: 2 is selected from ISIS NOs: 405955, 405956, 405957, 405958 or 405959.
In certain embodiments, a target region is nucleotides 20680-20706 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20680-20706 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 333, 334, 335 or 336. In certain such embodiments, an antisense compound targeted to nucleotides 20680-20706 of SEQ ID NO: 2 is selected from ISIS NOs: 399808, 399964, 405960, 405961, 405962 or 410739.
In certain embodiments, a target region is nucleotides 20683-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20683-20781 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 45, 46, 47, 48, 49, 50, 51, 87, 119, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20683-20781 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 395178, 395179, 395180, 399808, 399809, 399810, 399811, 399812, 399813, 399899, 399900, 399901, 399902, 399964, 399965, 399966, 399967, 399968, 399969, 405557, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410739 or 410740.
In certain embodiments, a target region is nucleotides 20689-20715 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20689-20715 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 46, 337 or 338. In certain such embodiments, an antisense compound targeted to nucleotides 20689-20715 of SEQ ID NO: 2 is selected from ISIS NOs: 395177, 399809, 399899, 399965, 405963 or 405964.
In certain embodiments, a target region is nucleotides 20698-20725 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20698-20725 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 47, 47, 339, 340, 341 or 342. In certain such embodiments, an antisense compound targeted to nucleotides 20698-20725 of SEQ ID NO: 2 is selected from ISIS NOs: 399810, 399966, 405965, 405966, 405967 or 405968.
In certain embodiments, a target region is nucleotides 20709-20736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20709-20736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 48, 343, 344, 345 or 346. In certain such embodiments, an antisense compound targeted to nucleotides 20709-20736 of SEQ ID NO: 2 is selected from ISIS NOs: 399811, 399967, 405885, 405969, 405970 or 410740.
In certain embodiments, a target region is nucleotides 20717-20744 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20717-20744 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, 351, 352 or 353. In certain such embodiments, an antisense compound targeted to nucleotides 20717-20744 of SEQ ID NO: 2 is selected from ISIS NOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 410596, 410597, 410598, 410599, 410600, 410601, 410672, 410673, 410674, 410675, 410676, 410677 or 410678.
In certain embodiments, a target region is nucleotides 20718-20745 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20718-20745 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 347, 348, 349, 350, 351, 352 or 353. In certain such embodiments, an antisense compound targeted to nucleotides 20718-20745 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 410596, 410597, 410598, 410599, 410600, 410601, 410672, 410673, 410674, 410675, 410676, 410677, 410678 or 410679.
In certain embodiments, a target region is nucleotides 20719-20746 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20719-20746 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 348, 349, 350, 351, 352, 353 or 354. In certain such embodiments, an antisense compound targeted to nucleotides 20719-20746 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405887, 405888, 405889, 405890, 405891, 405892, 408653, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679 or 410680.
In certain embodiments, a target region is nucleotides 20720-20747 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20720-20747 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351, 352, 353, 354 or 355. In certain such embodiments, an antisense compound targeted to nucleotides 20720-20747 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405888, 405889, 405890, 405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680 or 410681.
In certain embodiments, a target region is nucleotides 20721-20748 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20721-20748 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355 or 356. In certain such embodiments, an antisense compound targeted to nucleotides 20721-20748 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405557, 405889, 405890, 405891, 405892, 405971, 408653, 410540, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681 or 410682.
In certain embodiments, a target region is nucleotides 20722-20749 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20722-20749 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 20722-20749 of SEQ ID NO: 2 is selected from ISIS NOs: 395178, 399900, 405557, 405889, 405890, 405891, 405892, 405971, 405972, 408653, 410540, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410675, 410676, 410677, 410678, 410679, 410680, 410681 or 410682.
In certain embodiments, a target region is nucleotides 20727-20752 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20727-20752 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 87, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 20727-20752 of SEQ ID NO: 2 is selected from ISIS NOs: 395179, 399901, 405971, 405972, 408653, 410540, 410602, 410603, 410604, 410680, 410681 or 410682.
In certain embodiments, a target region is nucleotides 20735-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20735-20759 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 51, 358 or 359. In certain such embodiments, an antisense compound targeted to nucleotides 20735-20759 of SEQ ID NO: 2 is selected from ISIS NOs: 399813, 399969, 405973 or 405974.
In certain embodiments, a target region is nucleotides 20762-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20762-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 93, 119 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20762-21014 of SEQ ID NO: 2 is selected from ISIS NOs: 395180, 399864, 399902, 400020 or 410771.
In certain embodiments, a target region is nucleotides 20785-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 20785-21014 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 93 or 360. In certain such embodiments, an antisense compound targeted to nucleotides 20785-21014 of SEQ ID NO: 2 is selected from ISIS NOs: 399864, 400020 or 410771.
In certain embodiments, a target region is nucleotides 21082-21107 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21082-21107 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 95 or 361. In certain such embodiments, an antisense compound targeted to nucleotides 21082-21107 of SEQ ID NO: 2 is selected from ISIS NOs: 399865, 400021 or 405975.
In certain embodiments, a target region is nucleotides 21082-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21082-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 54, 95, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370 or 371. In certain such embodiments, an antisense compound targeted to nucleotides 21082-21152 of SEQ ID NO: 2 is selected from ISIS NOs: 395182, 399814, 399865, 399904, 399970, 400021, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741 or 410772.
In certain embodiments, a target region is nucleotides 21091-21114 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21091-21114 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 362 or 363. In certain such embodiments, an antisense compound targeted to nucleotides 21091-21114 of SEQ ID NO: 2 is selected from ISIS NOs: 399814, 399970, 405976 or 405977.
In certain embodiments, a target region is nucleotides 21118-21144 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21118-21144 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366 or 367. In certain such embodiments, an antisense compound targeted to nucleotides 21118-21144 of SEQ ID NO: 2 is selected from ISIS NOs: 395182, 399904, 405978, 405979 or 405980.
In certain embodiments, a target region is nucleotides 21127-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21127-21152 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 368, 369, 370 or 371. In certain such embodiments, an antisense compound targeted to nucleotides 21127-21152 of SEQ ID NO: 2 is selected from ISIS NOs: 405981, 405982, 405983 or 410741.
In certain embodiments, a target region is nucleotides 21181-21209 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21181-21209 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 372, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21181-21209 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399971, 405564, 405641, 405984, 405985, 405986 or 405987.
In certain embodiments, a target region is nucleotides 21181-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21181-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21181-21211 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405564, 405641, 405984, 405985, 405986 or 405987.
In certain embodiments, a target region is nucleotides 21183-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21183-21211 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 373, 374 or 375. In certain such embodiments, an antisense compound targeted to nucleotides 21183-21211 of SEQ ID NO: 2 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405564, 405641, 405985, 405986 or 405987.
In certain embodiments, a target region is nucleotides 21481-21500 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21481-21500 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 143. In certain such embodiments, an antisense compound targeted to nucleotides 21481-21500 of SEQ ID NO: 2 is selected from ISIS NOs: 399866 or 400022.
In certain embodiments, a target region is nucleotides 21589-21608 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21589-21608 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 89. In certain such embodiments, an antisense compound targeted to nucleotides 21589-21608 of SEQ ID NO: 2 is selected from ISIS NOs: 399867 or 400023.
In certain embodiments, a target region is nucleotides 21692-21719 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 21692-21719 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 123, 448, 449, 450, 451, 452, 453, 454 or 455. In certain such embodiments, an antisense compound targeted to nucleotides 21692-21719 of SEQ ID NO: 2 is selected from ISIS NOs: 399868, 400024, 406478, 406479, 406480, 406481, 406482, 406483, 406484 or 406485.
In certain embodiments, a target region is nucleotides 22000-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22000-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22000-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552, 410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.
In certain embodiments, a target region is nucleotides 22096-22115 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22115 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22115 of SEQ ID NO: 2 is selected from ISIS NOs: 395183 or 399905.
In certain embodiments, a target region is nucleotides 22096-22223 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22223 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391 or 392. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22223 of SEQ ID NO: 2 is selected from ISIS NOs: 395183, 395184, 395185, 399905, 399906, 399907, 405988, 405989, 405990, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410549, 410550, 410551, 410552, 410553, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410615, 410616, 410617, 410618, 410619, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699 or 410773.
In certain embodiments, a target region is nucleotides 22096-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22096-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 57, 58, 59, 126, 126, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 401, 401, 402, 402, 402, 403, 403, 403, 404, 404, 404, 405, 405, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22096-22311 of SEQ ID NO: 2 is selected from ISIS NOs: 395183, 395184, 395185, 395225, 399905, 399906, 399907, 399947, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410556, 410557, 410558, 410559, 410560, 410561, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692, 410693, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712, 410773 or 410774.
In certain embodiments, a target region is nucleotides 22133-22160 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22133-22160 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 376, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 22133-22160 of SEQ ID NO: 2 is selected from ISIS NOs: 405988, 405989, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690 or 410691.
In certain embodiments, a target region is nucleotides 22133-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22133-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 376, 377, 378, 379, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22133-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410541, 410542, 410543, 410544, 410545, 410546, 410547, 410548, 410605, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410683, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.
In certain embodiments, a target region is nucleotides 22134-22161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22134-22161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 377, 378, 379, 380, 381, 382, 383 or 384. In certain such embodiments, an antisense compound targeted to nucleotides 22134-22161 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 410542, 410543, 410544, 410545, 410546, 410547, 410606, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410684, 410685, 410686, 410687, 410688, 410689, 410690, 410691 or 410692.
In certain embodiments, a target region is nucleotides 22135-22162 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22135-22162 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 378, 379, 380, 381, 382, 383, 384 or 385. In certain such embodiments, an antisense compound targeted to nucleotides 22135-22162 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 410543, 410544, 410545, 410546, 410547, 410548, 410607, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410685, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.
In certain embodiments, a target region is nucleotides 22136-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22136-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 379, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22136-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410544, 410545, 410546, 410547, 410548, 410608, 410609, 410610, 410611, 410612, 410613, 410614, 410686, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.
In certain embodiments, a target region is nucleotides 22137-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22137-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 380, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22137-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410545, 410546, 410547, 410548, 410609, 410610, 410611, 410612, 410613, 410614, 410687, 410688, 410689, 410690, 410691, 410692 or 410693.
In certain embodiments, a target region is nucleotides 22138-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22138-22163 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 58, 381, 382, 383, 384, 385 or 386. In certain such embodiments, an antisense compound targeted to nucleotides 22138-22163 of SEQ ID NO: 2 is selected from ISIS NOs: 395184, 399906, 405988, 405989, 405990, 410546, 410547, 410548, 410610, 410611, 410612, 410613, 410614, 410688, 410689, 410690, 410691, 410692 or 410693.
In certain embodiments, a target region is nucleotides 22189-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22189-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22189-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 405994, 405995, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410556, 410557, 410558, 410559, 410560, 410561, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712, 410773 or 410774.
In certain embodiments, a target region is nucleotides 22199-22226 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22199-22226 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394 or 395. In certain such embodiments, an antisense compound targeted to nucleotides 22199-22226 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701 or 410702.
In certain embodiments, a target region is nucleotides 22199-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22199-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 388, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22199-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410549, 410550, 410551, 410552, 410553, 410554, 410555, 410615, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410694, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.
In certain embodiments, a target region is nucleotides 22200-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22200-22227 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 389, 390, 391, 392, 393, 394, 395 or 396. In certain such embodiments, an antisense compound targeted to nucleotides 22200-22227 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410550, 410551, 410552, 410553, 410554, 410555, 410616, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410695, 410696, 410697, 410698, 410699, 410700, 410701, 410702 or 410703.
In certain embodiments, a target region is nucleotides 22201-22228 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22201-22228 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 390, 391, 392, 393, 394, 395, 396 or 397. In certain such embodiments, an antisense compound targeted to nucleotides 22201-22228 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 410551, 410552, 410553, 410554, 410555, 410556, 410617, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410696, 410697, 410698, 410699, 410700, 410701, 410702, 410703 or 410704.
In certain embodiments, a target region is nucleotides 22202-22229 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22202-22229 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 391, 392, 393, 394, 395, 396, 397 or 398. In certain such embodiments, an antisense compound targeted to nucleotides 22202-22229 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 410552, 410553, 410554, 410555, 410556, 410618, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410697, 410698, 410699, 410700, 410701, 410702, 410703, 410704 or 410705.
In certain embodiments, a target region is nucleotides 22203-22230 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22203-22230 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 392, 393, 394, 395, 396, 397, 398 or 399. In certain such embodiments, an antisense compound targeted to nucleotides 22203-22230 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 410553, 410554, 410555, 410556, 410557, 410619, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410698, 410699, 410700, 410701, 410702, 410703, 410704, 410705 or 410706.
In certain embodiments, a target region is nucleotides 22204-22231 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22204-22231 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 59, 393, 394, 395, 396, 397, 398, 399 or 400. In certain such embodiments, an antisense compound targeted to nucleotides 22204-22231 of SEQ ID NO: 2 is selected from ISIS NOs: 395185, 399907, 405991, 405992, 405993, 405994, 410554, 410555, 410556, 410557, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410699, 410700, 410701, 410702, 410703, 410704, 410705, 410706 or 410707.
In certain embodiments, a target region is nucleotides 22205-22232 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22205-22232 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 393, 394, 395, 396, 397, 398, 399, 400 or 401. In certain such embodiments, an antisense compound targeted to nucleotides 22205-22232 of SEQ ID NO: 2 is selected from ISIS NOs: 405991, 405992, 405993, 405994, 410554, 410555, 410556, 410557, 410558, 410620, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410700, 410701, 410702, 410703, 410704, 410705, 410706, 410707 or 410708.
In certain embodiments, a target region is nucleotides 22206-22233 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22206-22233 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 394, 395, 396, 397, 398, 399, 400, 401 or 402. In certain such embodiments, an antisense compound targeted to nucleotides 22206-22233 of SEQ ID NO: 2 is selected from ISIS NOs: 405991, 405992, 405993, 405994, 405995, 410555, 410556, 410557, 410558, 410621, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410701, 410702, 410703, 410704, 410705, 410706, 410707, 410708 or 410709.
In certain embodiments, a target region is nucleotides 22207-22234 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22207-22234 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 395, 396, 397, 398, 399, 400, 401, 402 or 403. In certain such embodiments, an antisense compound targeted to nucleotides 22207-22234 of SEQ ID NO: 2 is selected from ISIS NOs: 405992, 405993, 405994, 405995, 410555, 410556, 410557, 410558, 410559, 410622, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410702, 410703, 410704, 410705, 410706, 410707, 410708, 410709 or 410710.
In certain embodiments, a target region is nucleotides 22208-22235 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22208-22235 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 396, 397, 398, 399, 400, 401, 402, 403 or 404. In certain such embodiments, an antisense compound targeted to nucleotides 22208-22235 of SEQ ID NO: 2 is selected from ISIS NOs: 405992, 405993, 405994, 405995, 410556, 410557, 410558, 410559, 410560, 410623, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410703, 410704, 410705, 410706, 410707, 410708, 410709, 410710 or 410711.
In certain embodiments, a target region is nucleotides 22209-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22209-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 397, 398, 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22209-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410556, 410557, 410558, 410559, 410560, 410561, 410624, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410704, 410705, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.
In certain embodiments, a target region is nucleotides 22210-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22210-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22210-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410705, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.
In certain embodiments, a target region is nucleotides 22210-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22210-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 398, 399, 400, 401, 402, 403, 404, 405 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22210-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 405993, 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410625, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410705, 410706, 410707, 410708, 410709, 410710, 410711, 410712 or 410774.
In certain embodiments, a target region is nucleotides 22211-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22211-22236 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 399, 400, 401, 402, 403, 404 or 405. In certain such embodiments, an antisense compound targeted to nucleotides 22211-22236 of SEQ ID NO: 2 is selected from ISIS NOs: 405994, 405995, 410557, 410558, 410559, 410560, 410561, 410626, 410627, 410628, 410629, 410630, 410631, 410632, 410706, 410707, 410708, 410709, 410710, 410711 or 410712.
In certain embodiments, a target region is nucleotides 22212-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22212-22239 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 400, 401, 402, 403, 404, 405 or 406. In certain such embodiments, an antisense compound targeted to nucleotides 22212-22239 of SEQ ID NO: 2 is selected from ISIS NOs: 405994, 405995, 410558, 410559, 410560, 410561, 410627, 410628, 410629, 410630, 410631, 410632, 410707, 410708, 410709, 410710, 410711, 410712 or 410774.
In certain embodiments, a target region is nucleotides 22292-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 22292-22311 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 126. In certain such embodiments, an antisense compound targeted to nucleotides 22292-22311 of SEQ ID NO: 2 is selected from ISIS NOs: 395225 or 399947.
In certain embodiments, a target region is nucleotides 23985-24054 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 23985-24054 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 408, 409 or 410. In certain such embodiments, an antisense compound targeted to nucleotides 23985-24054 of SEQ ID NO: 2 is selected from ISIS NOs: 410776, 410777 or 410778.
In certain embodiments, a target region is nucleotides 24035-24134 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24035-24134 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, 415 or 416. In certain such embodiments, an antisense compound targeted to nucleotides 24035-24134 of SEQ ID NO: 2 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, 410778 or 410779.
In certain embodiments, a target region is nucleotides 24095-24121 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24095-24121 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414 or 415. In certain such embodiments, an antisense compound targeted to nucleotides 24095-24121 of SEQ ID NO: 2 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717 or 410718.
In certain embodiments, a target region is nucleotides 24858-24877 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24858-24877 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 147. In certain such embodiments, an antisense compound targeted to nucleotides 24858-24877 of SEQ ID NO: 2 is selected from ISIS NOs: 395226 or 399948.
In certain embodiments, a target region is nucleotides 24907-24926 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 24907-24926 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 118. In certain such embodiments, an antisense compound targeted to nucleotides 24907-24926 of SEQ ID NO: 2 is selected from ISIS NOs: 399869 or 400025.
In certain embodiments, a target region is nucleotides 25413-25432 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 25413-25432 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 114. In certain such embodiments, an antisense compound targeted to nucleotides 25413-25432 of SEQ ID NO: 2 is selected from ISIS NOs: 399870 or 400026.
In certain embodiments, a target region is nucleotides 25994-26013 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 25994-26013 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 418. In certain such embodiments, an antisense compound targeted to nucleotides 25994-26013 of SEQ ID NO: 2 is selected from ISIS NOs: 410781.
In certain embodiments, a target region is nucleotides 26112-26139 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-26139 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425 or 426. In certain such embodiments, an antisense compound targeted to nucleotides 26112-26139 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726 or 410727.
In certain embodiments, a target region is nucleotides 26112-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429 or 430. In certain such embodiments, an antisense compound targeted to nucleotides 26112-26161 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.
In certain embodiments, a target region is nucleotides 26112-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26112-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 112, 122, 135, 153, 154, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443, 444, 445 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 26112-27303 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188, 395189, 395190, 395191, 395192, 395193, 395194, 395195, 395196, 395197, 395198, 395199, 399818, 399819, 399820, 399821, 399822, 399823, 399824, 399825, 399909, 399910, 399911, 399912, 399913, 399914, 399915, 399916, 399917, 399918, 399919, 399920, 399921, 399974, 399975, 399976, 399977, 399978, 399979, 399980, 399981, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.
In certain embodiments, a target region is nucleotides 26113-26140 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26113-26140 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 420, 421, 422, 423, 424, 425, 426 or 427. In certain such embodiments, an antisense compound targeted to nucleotides 26113-26140 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727 or 410728.
In certain embodiments, a target region is nucleotides 26114-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26114-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427 or 248. In certain such embodiments, an antisense compound targeted to nucleotides 26114-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728 or 410729.
In certain embodiments, a target region is nucleotides 26115-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26115-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 422, 423, 424, 425, 426, 427 or 248. In certain such embodiments, an antisense compound targeted to nucleotides 26115-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410568, 410569, 410570, 410571, 410572, 410573, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410722, 410723, 410724, 410725, 410726, 410727, 410728 or 410729.
In certain embodiments, a target region is nucleotides 26116-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26116-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 423, 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26116-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410569, 410570, 410571, 410572, 410573, 410641, 410642, 410643, 410644, 410645, 410646, 410723, 410724, 410725, 410726, 410727 or 410728.
In certain embodiments, a target region is nucleotides 26117-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26117-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26117-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 399909, 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410724, 410725, 410726, 410727, 410728 or 410729.
In certain embodiments, a target region is nucleotides 26117-26475 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26117-26475 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 63, 64, 65, 66, 67, 122, 153, 154, 424, 425, 426, 427, 428, 429 or 430. In certain such embodiments, an antisense compound targeted to nucleotides 26117-26475 of SEQ ID NO: 2 is selected from ISIS NOs: 395187, 395188, 395189, 395190, 395191, 395192, 399818, 399819, 399820, 399909, 399910, 399911, 399912, 399913, 399914, 399974, 399975, 399976, 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410724, 410725, 410726, 410727, 410728, 410729, 410782 or 410783.
In certain embodiments, a target region is nucleotides 26118-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26118-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26118-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 405998, 410570, 410571, 410572, 410573, 410642, 410643, 410644, 410645, 410646, 410725, 410726, 410727, 410728 or 410729.
In certain embodiments, a target region is nucleotides 26120-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26120-26141 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 26120-26141 of SEQ ID NO: 2 is selected from ISIS NOs: 405998, 410572, 410573, 410644, 410645, 410646, 410727, 410728 or 410729.
In certain embodiments, a target region is nucleotides 26132-26151 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26132-26151 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 429. In certain such embodiments, an antisense compound targeted to nucleotides 26132-26151 of SEQ ID NO: 2 is selected from ISIS NOs: 410782.
In certain embodiments, a target region is nucleotides 26142-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26142-26161 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 430. In certain such embodiments, an antisense compound targeted to nucleotides 26142-26161 of SEQ ID NO: 2 is selected from ISIS NOs: 410783.
In certain embodiments, a target region is nucleotides 26217-26241 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26217-26241 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 26217-26241 of SEQ ID NO: 2 is selected from ISIS NOs: 395188, 399818 or 399910.
In certain embodiments, a target region is nucleotides 26311-26335 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26311-26335 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 26311-26335 of SEQ ID NO: 2 is selected from ISIS NOs: 395189, 399819, 399911 or 399975.
In certain embodiments, a target region is nucleotides 26389-26432 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26389-26432 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67 or 122. In certain such embodiments, an antisense compound targeted to nucleotides 26389-26432 of SEQ ID NO: 2 is selected from ISIS NOs: 395190, 395191, 399820, 399912, 399913 or 399976.
In certain embodiments, a target region is nucleotides 26456-26576 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26456-26576 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68 or 153. In certain such embodiments, an antisense compound targeted to nucleotides 26456-26576 of SEQ ID NO: 2 is selected from ISIS NOs: 395192, 395193, 399914 or 399915.
In certain embodiments, a target region is nucleotides 26635-26662 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26635-26662 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 431, 432, 433, 434, 435, 436, 437 or 438. In certain such embodiments, an antisense compound targeted to nucleotides 26635-26662 of SEQ ID NO: 2 is selected from ISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897, 405898, 405899 or 405900.
In certain embodiments, a target region is nucleotides 26707-26734 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26707-26734 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443 or 444. In certain such embodiments, an antisense compound targeted to nucleotides 26707-26734 of SEQ ID NO: 2 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905 or 405906.
In certain embodiments, a target region is nucleotides 26707-26736 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26707-26736 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 445 or 446. In certain such embodiments, an antisense compound targeted to nucleotides 26707-26736 of SEQ ID NO: 2 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905, 405906, 405907 or 405908.
In certain embodiments, a target region is nucleotides 26790-26820 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 26790-26820 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73 or 135. In certain such embodiments, an antisense compound targeted to nucleotides 26790-26820 of SEQ ID NO: 2 is selected from ISIS NOs: 395196, 399822, 399823, 399918, 399978 or 399979.
In certain embodiments, a target region is nucleotides 27034-27263 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27034-27263 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75 or 112. In certain such embodiments, an antisense compound targeted to nucleotides 27034-27263 of SEQ ID NO: 2 is selected from ISIS NOs: 395197, 395198, 399824, 399919, 399920 or 399980.
In certain embodiments, a target region is nucleotides 27279-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27279-27303 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 27279-27303 of SEQ ID NO: 2 is selected from ISIS NOs: 395199, 399825, 399921 or 399981.
In certain embodiments, a target region is nucleotides 27350-27376 of SEQ ID NO: 2. In certain embodiments, an antisense compound is targeted to nucleotides 27350-27376 of SEQ ID NO: 2. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 27350-27376 of SEQ ID NO: 2 is selected from ISIS NOs: 395200, 399826, 399922 or 399982.
In certain embodiments, antisense compounds target a PCSK9 nucleic acid having the sequence of GENBANK® Accession No. AK124635.1, first deposited with GENBANK® on Sep. 8, 2003, and incorporated herein as SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide is targeted to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is at least 90% complementary to SEQ ID NO: 1. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is at least 95% complementary to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide that is targeted to SEQ ID NO: 3 is 100% complementary to SEQ ID NO: 3. In certain such embodiments, an antisense oligonucleotide comprises a nucleotide sequence selected from a nucleotide sequence set forth in Table 12.







TABLE 12







Nucleotide sequences targeted to 


AK124635.1 (SEQ ID NO: 3)











5′
3′




Target
Target



SEQ
Site on
Site on



ID
SEQ ID
SEQ ID



NO
NO: 3
NO: 3
Sequences (5′-3′)





 85
 155
 174
ACAAATTCCCAGACTCAGCA


 


100
 220
 239
ATCTCAGGACAGGTGAGCAA


 


116
 234
 253
GAGTAGAGATTCTCATCTCA


 


129
 290
 309
GTGCCATCTGAACAGCACCT


 


117
 302
 321
GAGTCTTCTGAAGTGCCATC


 


 81
 377
 396
AAGCAGGGCCTCAGGTGGAA


 


110
 400
 419
CCTGGAACCCCTGCAGCCAG


 


152
 451
 470
TTCAGGCAGGTTGCTGCTAG


 


 83
 460
 479
AAGGAAGACTTCAGGCAGGT


 


140
 472
 491
TCAGCCAGGCCAAAGGAAGA


 


137
 586
 605
TAGGGAGAGCTCACAGATGC


 


136
 596
 615
TAGGAGAAAGTAGGGAGAGC


 


132
 653
 672
TAAAAGCTGCAAGAGACTCA


 


139
 741
 760
TCAGAGAAAACAGTCACCGA


 


 92
 871
 890
AGAGACAGGAAGCTGCAGCT


 


142
 878
 897
TCATTTTAGAGACAGGAAGC


 


113
 915
 934
GAATAACAGTGATGTCTGGC


 


138
 968
 987
TCACAGCTCACCGAGTCTGC


 


 98
 998
1017
AGTGTAAAATAAAGCCCCTA


 


 96
1075
1094
AGGACCCAAGTCATCCTGCT


 


124
1105
1124
GGCCATCAGCTGGCAATGCT


 


 82
1144
1163
AAGGAAAGGGAGGCCTAGAG


 


133
1149
1168
TAGACAAGGAAAGGGAGGCC


 


103
1155
1174
ATTTCATAGACAAGGAAAGG


 


155
1275
1294
CTTATAGTTAACACACAGAA


 


156
1283
1302
AAGTCAACCTTATAGTTAAC


 


146
1315
1334
TGACATTTGTGGGAGAGGAG


 


161
1322
1341
TCCAAGGTGACATTTGTGGG


 


215
1351
1370
ATACACCTCCACCAGGCTGC


 


216
1362
1381
GTGTCTAGGAGATACACCTC


 


 19
1365
1384
CTGGTGTCTAGGAGATACAC


 


217
1367
1386
TGCTGGTGTCTAGGAGATAC


 


 20
1370
1389
GTATGCTGGTGTCTAGGAGA


 


 21
1390
1409
GATTTCCCGGTGGTCACTCT


 


218
1392
1411
TCGATTTCCCGGTGGTCACT


 


219
1393
1412
CTCGATTTCCCGGTGGTCAC


 


220
1394
1413
CCTCGATTTCCCGGTGGTCA


 


221
1395
1414
CCCTCGATTTCCCGGTGGTC


 


 22
1396
1415
GCCCTCGATTTCCCGGTGGT


 


222
1397
1416
TGCCCTCGATTTCCCGGTGG


 


223
1398
1417
CTGCCCTCGATTTCCCGGTG


 


224
1399
1418
CCTGCCCTCGATTTCCCGGT


 


225
1400
1419
CCCTGCCCTCGATTTCCCGG


 


226
1404
1423
ATGACCCTGCCCTCGATTTC


 


227
1406
1425
CCATGACCCTGCCCTCGATT


 


228
1408
1427
GACCATGACCCTGCCCTCGA


 


 23
1410
1429
GTGACCATGACCCTGCCCTC


 


229
1412
1431
CGGTGACCATGACCCTGCCC


 


230
1414
1433
GTCGGTGACCATGACCCTGC


 


231
1416
1435
AAGTCGGTGACCATGACCCT


 


232
1418
1437
CGAAGTCGGTGACCATGACC


 


 24
1420
1439
CTCGAAGTCGGTGACCATGA


 


233
1428
1447
GGCACATTCTCGAAGTCGGT


 


 25
1453
1472
GTGGAAGCGGGTCCCGTCCT


 


234
1463
1482
TGGCCTGTCTGTGGAAGCGG


 


235
1490
1509
GGTGGGTGCCATGACTGTCA


 


236
1493
1512
CCAGGTGGGTGCCATGACTG


 


237
1497
1516
CCTGCCAGGTGGGTGCCATG


 


 26
1500
1519
ACCCCTGCCAGGTGGGTGCC


 


238
1502
1521
CCACCCCTGCCAGGTGGGTG


 


 27
1505
1524
TGACCACCCCTGCCAGGTGG


 


239
1507
1526
GCTGACCACCCCTGCCAGGT


 


240
1515
1534
TCCCGGCCGCTGACCACCCC


 


241
1519
1538
GGCATCCCGGCCGCTGACCA


 


242
1522
1541
GCCGGCATCCCGGCCGCTGA


 


243
1527
1546
GCCACGCCGGCATCCCGGCC


 


244
1528
1547
GGCCACGCCGGCATCCCGGC


 


245
1529
1548
TGGCCACGCCGGCATCCCGG


 


246
1530
1549
TTGGCCACGCCGGCATCCCG


 


247
1531
1550
CTTGGCCACGCCGGCATCCC


 


248
1532
1551
CCTTGGCCACGCCGGCATCC


 


249
1533
1552
CCCTTGGCCACGCCGGCATC


 


 28
1534
1553
ACCCTTGGCCACGCCGGCAT


 


447
1534
1553
ACCCTTGGTCACGCCGGCAT


 


250
1535
1554
CACCCTTGGCCACGCCGGCA


 


251
1536
1555
GCACCCTTGGCCACGCCGGC


 


252
1537
1556
GGCACCCTTGGCCACGCCGG


 


253
1538
1557
TGGCACCCTTGGCCACGCCG


 


254
1539
1558
CTGGCACCCTTGGCCACGCC


 


255
1540
1559
GCTGGCACCCTTGGCCACGC


 


256
1545
1564
CGCATGCTGGCACCCTTGGC


 


257
1566
1585
CAGTTGAGCACGCGCAGGCT


 


258
1568
1587
GGCAGTTGAGCACGCGCAGG


 


 29
1570
1589
TTGGCAGTTGAGCACGCGCA


 


259
1572
1591
CCTTGGCAGTTGAGCACGCG


 


 30
1575
1594
TTCCCTTGGCAGTTGAGCAC


 


260
1577
1596
CCTTCCCTTGGCAGTTGAGC


 


261
1581
1600
GTGCCCTTCCCTTGGCAGTT


 


262
1583
1602
CCGTGCCCTTCCCTTGGCAG


 


263
1594
1613
GGTGCCGCTAACCGTGCCCT


 


264
1606
1625
CAGGCCTATGAGGGTGCCGC


 


 31
1607
1626
CCAGGCCTATGAGGGTGCCG


 


458
1609
1622
GCCTATGAGGGTGC


 


459
1614
1627
TCCAGGCCTATGAG


 


265
1618
1637
CCGAATAAACTCCAGGCCTA


 


266
1626
1645
TGGCTTTTCCGAATAAACTC


 


 32
1628
1647
GCTGGCTTTTCCGAATAAAC


 


267
1630
1649
CAGCTGGCTTTTCCGAATAA


 


268
1632
1651
ACCAGCTGGCTTTTCCGAAT


 


269
1634
1653
GGACCAGCTGGCTTTTCCGA


 


270
1638
1657
GGCTGGACCAGCTGGCTTTT


 


271
1649
1668
GTGGCCCCACAGGCTGGACC


 


272
1662
1681
AGCAGCACCACCAGTGGCCC


 


273
1684
1703
GCTGTACCCACCCGCCAGGG


 


274
1730
1749
CGACCCCAGCCCTCGCCAGG


 


 33
1740
1759
GTGACCAGCACGACCCCAGC


 


275
1742
1761
CGGTGACCAGCACGACCCCA


 


276
1744
1763
AGCGGTGACCAGCACGACCC


 


277
1746
1765
GCAGCGGTGACCAGCACGAC


 


278
1748
1767
CGGCAGCGGTGACCAGCACG


 


279
1749
1768
CCGGCAGCGGTGACCAGCAC


 


280
1752
1771
TTGCCGGCAGCGGTGACCAG


 


281
1754
1773
AGTTGCCGGCAGCGGTGACC


 


282
1756
1775
GAAGTTGCCGGCAGCGGTGA


 


283
1758
1777
CGGAAGTTGCCGGCAGCGGT


 


284
1760
1779
CCCGGAAGTTGCCGGCAGCG


 


285
1762
1781
GTCCCGGAAGTTGCCGGCAG


 


306
1820
1839
GAGGCACCAATGATGTCCTC


 


 39
1822
1841
TGGAGGCACCAATGATGTCC


 


307
1824
1843
GCTGGAGGCACCAATGATGT


 


308
1826
1845
TCGCTGGAGGCACCAATGAT


 


309
1828
1847
AGTCGCTGGAGGCACCAATG


 


310
1830
1849
GCAGTCGCTGGAGGCACCAA


 


 40
1833
1852
GCTGCAGTCGCTGGAGGCAC


 


311
1835
1854
GTGCTGCAGTCGCTGGAGGC


 


312
1837
1856
AGGTGCTGCAGTCGCTGGAG


 


313
1839
1858
GCAGGTGCTGCAGTCGCTGG


 


314
1840
1859
AGCAGGTGCTGCAGTCGCTG


 


315
1842
1861
AAAGCAGGTGCTGCAGTCGC


 


 41
1844
1863
ACAAAGCAGGTGCTGCAGTC


 


316
1846
1865
ACACAAAGCAGGTGCTGCAG


 


317
1848
1867
TGACACAAAGCAGGTGCTGC


 


318
1858
1877
TCCCACTCTGTGACACAAAG


 


101
1898
1917
ATGGCTGCAATGCCAGCCAC


 


319
1900
1919
TCATGGCTGCAATGCCAGCC


 


 42
1903
1922
GCATCATGGCTGCAATGCCA


 


320
1905
1924
CAGCATCATGGCTGCAATGC


 


321
1907
1926
GACAGCATCATGGCTGCAAT


 


322
1909
1928
CAGACAGCATCATGGCTGCA


 


 43
1911
1930
GGCAGACAGCATCATGGCTG


 


323
1913
1932
TCGGCAGACAGCATCATGGC


 


324
1915
1934
GCTCGGCAGACAGCATCATG


 


325
1917
1936
CGGCTCGGCAGACAGCATCA


 


326
1919
1938
TCCGGCTCGGCAGACAGCAT


 


327
1933
1952
CGGCCAGGGTGAGCTCCGGC


 


328
1946
1965
CTCTGCCTCAACTCGGCCAG


 


329
1948
1967
GTCTCTGCCTCAACTCGGCC


 


330
1950
1969
CAGTCTCTGCCTCAACTCGG


 


331
1952
1971
ATCAGTCTCTGCCTCAACTC


 


332
1954
1973
GGATCAGTCTCTGCCTCAAC


 


333
1956
1975
GTGGATCAGTCTCTGCCTCA


 


334
1958
1977
AAGTGGATCAGTCTCTGCCT


 


 44
1959
1978
GAAGTGGATCAGTCTCTGCC


 


335
1961
1980
GAGAAGTGGATCAGTCTCTG


 


336
1963
1982
CAGAGAAGTGGATCAGTCTC


 


337
1965
1984
GGCAGAGAAGTGGATCAGTC


 


 45
1967
1986
TTGGCAGAGAAGTGGATCAG


 


338
1969
1988
CTTTGGCAGAGAAGTGGATC


 


 46
1972
1991
CATCTTTGGCAGAGAAGTGG


 


339
1974
1993
GACATCTTTGGCAGAGAAGT


 


340
1976
1995
ATGACATCTTTGGCAGAGAA


 


 47
1978
1997
TGATGACATCTTTGGCAGAG


 


341
1980
1999
ATTGATGACATCTTTGGCAG


 


342
1982
2001
TCATTGATGACATCTTTGGC


 


 48
1985
2004
GCCTCATTGATGACATCTTT


 


343
1987
2006
AGGCCTCATTGATGACATCT


 


344
1989
2008
CCAGGCCTCATTGATGACAT


 


345
1991
2010
AACCAGGCCTCATTGATGAC


 


346
1993
2012
GGAACCAGGCCTCATTGATG


 


347
1994
2013
GGGAACCAGGCCTCATTGAT


 


348
1995
2014
AGGGAACCAGGCCTCATTGA


 


349
1996
2015
CAGGGAACCAGGCCTCATTG


 


 49
1997
2016
TCAGGGAACCAGGCCTCATT


 


350
1998
2017
CTCAGGGAACCAGGCCTCAT


 


351
1999
2018
CCTCAGGGAACCAGGCCTCA


 


352
2000
2019
TCCTCAGGGAACCAGGCCTC


 


353
2001
2020
GTCCTCAGGGAACCAGGCCT


 


 50
2002
2021
GGTCCTCAGGGAACCAGGCC


 


354
2003
2022
TGGTCCTCAGGGAACCAGGC


 


355
2004
2023
CTGGTCCTCAGGGAACCAGG


 


356
2005
2024
GCTGGTCCTCAGGGAACCAG


 


357
2006
2025
CGCTGGTCCTCAGGGAACCA


 


 87
2009
2028
ACCCGCTGGTCCTCAGGGAA


 


358
2011
2030
GTACCCGCTGGTCCTCAGGG


 


359
2013
2032
CAGTACCCGCTGGTCCTCAG


 


 51
2016
2035
GGTCAGTACCCGCTGGTCCT


 


119
2038
2057
GCAGGGCGGCCACCAGGTTG


 


360
2061
2080
ACCTGCCCCATGGGTGCTGG


 


 52
2073
2092
AAACAGCTGCCAACCTGCCC


 


361
2075
2094
CAAAACAGCTGCCAACCTGC


 


 53
2078
2097
CTGCAAAACAGCTGCCAACC


 


362
2080
2099
TCCTGCAAAACAGCTGCCAA


 


363
2082
2101
AGTCCTGCAAAACAGCTGCC


 


104
2085
2104
CACAGTCCTGCAAAACAGCT


 


130
2095
2114
GTGCTGACCACACAGTCCTG


 


365
2105
2124
GGCCCCGAGTGTGCTGACCA


 


 54
2108
2127
GTAGGCCCCGAGTGTGCTGA


 


366
2110
2129
GTGTAGGCCCCGAGTGTGCT


 


367
2112
2131
CCGTGTAGGCCCCGAGTGTG


 


368
2114
2133
ATCCGTGTAGGCCCCGAGTG


 


369
2116
2135
CCATCCGTGTAGGCCCCGAG


 


370
2118
2137
GGCCATCCGTGTAGGCCCCG


 


371
2120
2139
GTGGCCATCCGTGTAGGCCC


 


372
2168
2187
CTGGAGCAGCTCAGCAGCTC


 


460
2168
2181
CAGCTCAGCAGCTC


 


373
2170
2189
AACTGGAGCAGCTCAGCAGC


 


 55
2173
2192
AGAAACTGGAGCAGCTCAGC


 


374
2175
2194
GGAGAAACTGGAGCAGCTCA


 


375
2177
2196
CTGGAGAAACTGGAGCAGCT


 


 56
2179
2198
TCCTGGAGAAACTGGAGCAG


 


408
2245
2264
GTGCCAAGGTCCTCCACCTC


 


409
2265
2284
TCAGCACAGGCGGCTTGTGG


 


410
2295
2314
CCACGCACTGGTTGGGCTGA


 


 60
2355
2374
CTTTGCATTCCAGACCTGGG


 


411
2356
2375
ACTTTGCATTCCAGACCTGG


 


412
2357
2376
GACTTTGCATTCCAGACCTG


 


413
2358
2377
TGACTTTGCATTCCAGACCT


 


414
2359
2378
TTGACTTTGCATTCCAGACC


 


 61
2360
2379
CTTGACTTTGCATTCCAGAC


 


415
2362
2381
TCCTTGACTTTGCATTCCAG


 


416
2375
2394
CGGGATTCCATGCTCCTTGA


 


417
2405
2424
GCAGGCCACGGTCACCTGCT


 


418
2442
2461
GGAGGGCACTGCAGCCAGTC


 


419
2560
2579
CAGATGGCAACGGCTGTCAC


 


420
2561
2580
GCAGATGGCAACGGCTGTCA


 


421
2562
2581
AGCAGATGGCAACGGCTGTC


 


422
2563
2582
CAGCAGATGGCAACGGCTGT


 


423
2564
2583
GCAGCAGATGGCAACGGCTG


 


 62
2565
2584
GGCAGCAGATGGCAACGGCT


 


424
2566
2585
CGGCAGCAGATGGCAACGGC


 


425
2567
2586
CCGGCAGCAGATGGCAACGG


 


426
2568
2587
TCCGGCAGCAGATGGCAACG


 


427
2569
2588
CTCCGGCAGCAGATGGCAAC


 


428
2570
2589
GCTCCGGCAGCAGATGGCAA


 


429
2580
2599
CCAGGTGCCGGCTCCGGCAG


 


430
2590
2609
GAGGCCTGCGCCAGGTGCCG


 


154
2665
2684
TTTTAAAGCTCAGCCCCAGC


 


 63
2670
2689
AACCATTTTAAAGCTCAGCC


 


 64
2759
2778
TCAAGGGCCAGGCCAGCAGC


 


 65
2764
2783
CCCACTCAAGGGCCAGGCCA


 


122
2837
2856
GGAGGGAGCTTCCTGGCACC


 


 66
2852
2871
ATGCCCCACAGTGAGGGAGG


 


 67
2861
2880
AATGGTGAAATGCCCCACAG


 


153
2904
2923
TTGGGAGCAGCTGGCAGCAC


 


 68
3005
3024
CATGGGAAGAATCCTGCCTC


 


431
3083
3102
ATGAGGGCCATCAGCACCTT


 


432
3084
3103
GATGAGGGCCATCAGCACCT


 


433
3085
3104
AGATGAGGGCCATCAGCACC


 


434
3086
3105
GAGATGAGGGCCATCAGCAC


 


 69
3087
3106
GGAGATGAGGGCCATCAGCA


 


435
3088
3107
TGGAGATGAGGGCCATCAGC


 


436
3089
3108
CTGGAGATGAGGGCCATCAG


 


437
3090
3109
GCTGGAGATGAGGGCCATCA


 


438
3091
3110
AGCTGGAGATGAGGGCCATC


 


461
3132
3145
TTAATCAGGGAGCC


 


 70
3155
3174
TAGATGCCATCCAGAAAGCT


 


439
3157
3176
GCTAGATGCCATCCAGAAAG


 


440
3158
3177
GGCTAGATGCCATCCAGAAA


 


441
3159
3178
TGGCTAGATGCCATCCAGAA


 


442
3160
3179
CTGGCTAGATGCCATCCAGA


 


 71
3161
3180
TCTGGCTAGATGCCATCCAG


 


443
3162
3181
CTCTGGCTAGATGCCATCCA


 


444
3163
3182
CCTCTGGCTAGATGCCATCC


 


445
3164
3183
GCCTCTGGCTAGATGCCATC


 


446
3165
3184
AGCCTCTGGCTAGATGCCAT


 


 72
3238
3257
GGCATAGAGCAGAGTAAAGG


 


 73
3243
3262
AGCCTGGCATAGAGCAGAGT


 


135
3249
3268
TAGCACAGCCTGGCATAGAG


 


112
3482
3501
GAAGAGGCTTGGCTTCAGAG


 


 74
3488
3507
AAGTAAGAAGAGGCTTGGCT


 


 75
3692
3711
GCTCAAGGAGGGACAGTTGT


 


 76
3727
3746
AAAGATAAATGTCTGCTTGC


 


 77
3732
3751
ACCCAAAAGATAAATGTCTG


 


 78
3798
3817
TCTTCAAGTTACAAAAGCAA


 


 99
3805
3824
ATAAATATCTTCAAGTTACA









In certain embodiments, gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 5-10-5 gapmer motif. Table 13 illustrates gapmer antisense compounds targeted to SEQ ID NO: 3, having a 5-10-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 13







Gapmer antisense compounds having a 5-10-5 motif 


targeted to SEQ ID NO: 3
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 3
NO: 3
Mismatches
Sequence (5′-3′)





395201
5-10-5
 85
 155
 174
0
ACAAATTCCCAGACTCAGCA


 


399827
5-10-5
100
 220
 239
0
ATCTCAGGACAGGTGAGCAA


 


399828
5-10-5
116
 234
 253
0
GAGTAGAGATTCTCATCTCA


 


395202
5-10-5
129
 290
 309
0
GTGCCATCTGAACAGCACCT


 


399829
5-10-5
117
 302
 321
0
GAGTCTTCTGAAGTGCCATC


 


399830
5-10-5
 81
 377
 396
0
AAGCAGGGCCTCAGGTGGAA


 


395203
5-10-5
110
 400
 419
0
CCTGGAACCCCTGCAGCCAG


 


395204
5-10-5
152
 451
 470
0
TTCAGGCAGGTTGCTGCTAG


 


399831
5-10-5
 83
 460
 479
0
AAGGAAGACTTCAGGCAGGT


 


395205
5-10-5
140
 472
 491
0
TCAGCCAGGCCAAAGGAAGA


 


399832
5-10-5
137
 586
 605
0
TAGGGAGAGCTCACAGATGC


 


395206
5-10-5
136
 596
 615
0
TAGGAGAAAGTAGGGAGAGC


 


395207
5-10-5
132
 653
 672
0
TAAAAGCTGCAAGAGACTCA


 


395208
5-10-5
139
 741
 760
0
TCAGAGAAAACAGTCACCGA


 


399833
5-10-5
 92
 871
 890
0
AGAGACAGGAAGCTGCAGCT


 


395209
5-10-5
142
 878
 897
0
TCATTTTAGAGACAGGAAGC


 


395210
5-10-5
113
 915
 934
0
GAATAACAGTGATGTCTGGC


 


395211
5-10-5
138
 968
 987
0
TCACAGCTCACCGAGTCTGC


 


395212
5-10-5
 98
 998
1017
0
AGTGTAAAATAAAGCCCCTA


 


395213
5-10-5
 96
1075
1094
0
AGGACCCAAGTCATCCTGCT


 


395214
5-10-5
124
1105
1124
0
GGCCATCAGCTGGCAATGCT


 


399834
5-10-5
 82
1144
1163
0
AAGGAAAGGGAGGCCTAGAG


 


395215
5-10-5
133
1149
1168
0
TAGACAAGGAAAGGGAGGCC


 


395216
5-10-5
103
1155
1174
0
ATTTCATAGACAAGGAAAGG


 


395217
5-10-5
155
1275
1294
0
CTTATAGTTAACACACAGAA


 


399835
5-10-5
156
1283
1302
0
AAGTCAACCTTATAGTTAAC


 


395218
5-10-5
146
1315
1334
0
TGACATTTGTGGGAGAGGAG


 


395219
5-10-5
161
1322
1341
0
TCCAAGGTGACATTTGTGGG


 


395160
5-10-5
 19
1365
1384
0
CTGGTGTCTAGGAGATACAC


 


399797
5-10-5
 20
1370
1389
0
GTATGCTGGTGTCTAGGAGA


 


395161
5-10-5
 21
1390
1409
0
GATTTCCCGGTGGTCACTCT


 


399798
5-10-5
 22
1396
1415
0
GCCCTCGATTTCCCGGTGGT


 


399799
5-10-5
 23
1410
1429
0
GTGACCATGACCCTGCCCTC


 


395162
5-10-5
 24
1420
1439
0
CTCGAAGTCGGTGACCATGA


 


395163
5-10-5
 25
1453
1472
0
GTGGAAGCGGGTCCCGTCCT


 


395164
5-10-5
 26
1500
1519
0
ACCCCTGCCAGGTGGGTGCC


 


399800
5-10-5
 27
1505
1524
0
TGACCACCCCTGCCAGGTGG


 


395165
5-10-5
 28
1534
1553
0
ACCCTTGGCCACGCCGGCAT


 


395166
5-10-5
 29
1570
1589
0
TTGGCAGTTGAGCACGCGCA


 


399801
5-10-5
 30
1575
1594
0
TTCCCTTGGCAGTTGAGCAC


 


395167
5-10-5
 31
1607
1626
0
CCAGGCCTATGAGGGTGCCG


 


395168
5-10-5
 32
1628
1647
0
GCTGGCTTTTCCGAATAAAC


 


395169
5-10-5
 33
1740
1759
0
GTGACCAGCACGACCCCAGC


 


395175
5-10-5
 39
1822
1841
0
TGGAGGCACCAATGATGTCC


 


399804
5-10-5
 40
1833
1852
0
GCTGCAGTCGCTGGAGGCAC


 


399805
5-10-5
 41
1844
1863
0
ACAAAGCAGGTGCTGCAGTC


 


395176
5-10-5
101
1898
1917
0
ATGGCTGCAATGCCAGCCAC


 


399806
5-10-5
 42
1903
1922
0
GCATCATGGCTGCAATGCCA


 


399807
5-10-5
 43
1911
1930
0
GGCAGACAGCATCATGGCTG


 


399808
5-10-5
 44
1959
1978
0
GAAGTGGATCAGTCTCTGCC


 


395177
5-10-5
 45
1967
1986
0
TTGGCAGAGAAGTGGATCAG


 


399809
5-10-5
 46
1972
1991
0
CATCTTTGGCAGAGAAGTGG


 


399810
5-10-5
 47
1978
1997
0
TGATGACATCTTTGGCAGAG


 


399811
5-10-5
 48
1985
2004
0
GCCTCATTGATGACATCTTT


 


399812
5-10-5
 49
1997
2016
0
TCAGGGAACCAGGCCTCATT


 


395178
5-10-5
 50
2002
2021
0
GGTCCTCAGGGAACCAGGCC


 


395179
5-10-5
 87
2009
2028
0
ACCCGCTGGTCCTCAGGGAA


 


399813
5-10-5
 51
2016
2035
0
GGTCAGTACCCGCTGGTCCT


 


395180
5-10-5
119
2038
2057
0
GCAGGGCGGCCACCAGGTTG


 


395181
5-10-5
 52
2073
2092
0
AAACAGCTGCCAACCTGCCC


 


399814
5-10-5
 53
2078
2097
0
CTGCAAAACAGCTGCCAACC


 


395220
5-10-5
104
2085
2104
0
CACAGTCCTGCAAAACAGCT


 


399836
5-10-5
130
2095
2114
0
GTGCTGACCACACAGTCCTG


 


395182
5-10-5
 54
2108
2127
0
GTAGGCCCCGAGTGTGCTGA


 


399815
5-10-5
 55
2173
2192
0
AGAAACTGGAGCAGCTCAGC


 


399816
5-10-5
 56
2179
2198
0
TCCTGGAGAAACTGGAGCAG


 


395186
5-10-5
 60
2355
2374
0
CTTTGCATTCCAGACCTGGG


 


399817
5-10-5
 61
2360
2379
0
CTTGACTTTGCATTCCAGAC


 


395187
5-10-5
 62
2565
2584
0
GGCAGCAGATGGCAACGGCT


 


395188
5-10-5
154
2665
2684
0
TTTTAAAGCTCAGCCCCAGC


 


399818
5-10-5
 63
2670
2689
0
AACCATTTTAAAGCTCAGCC


 


395189
5-10-5
 64
2759
2778
0
TCAAGGGCCAGGCCAGCAGC


 


399819
5-10-5
 65
2764
2783
0
CCCACTCAAGGGCCAGGCCA


 


399820
5-10-5
122
2837
2856
0
GGAGGGAGCTTCCTGGCACC


 


395190
5-10-5
 66
2852
2871
0
ATGCCCCACAGTGAGGGAGG


 


395191
5-10-5
 67
2861
2880
0
AATGGTGAAATGCCCCACAG


 


395192
5-10-5
153
2904
2923
0
TTGGGAGCAGCTGGCAGCAC


 


395193
5-10-5
 68
3005
3024
0
CATGGGAAGAATCCTGCCTC


 


395194
5-10-5
 69
3087
3106
0
GGAGATGAGGGCCATCAGCA


 


395195
5-10-5
 70
3155
3174
0
TAGATGCCATCCAGAAAGCT


 


399821
5-10-5
 71
3161
3180
0
TCTGGCTAGATGCCATCCAG


 


395196
5-10-5
 72
3238
3257
0
GGCATAGAGCAGAGTAAAGG


 


399822
5-10-5
 73
3243
3262
0
AGCCTGGCATAGAGCAGAGT


 


399823
5-10-5
135
3249
3268
0
TAGCACAGCCTGGCATAGAG


 


395197
5-10-5
112
3482
3501
0
GAAGAGGCTTGGCTTCAGAG


 


399824
5-10-5
 74
3488
3507
0
AAGTAAGAAGAGGCTTGGCT


 


395198
5-10-5
 75
3692
3711
0
GCTCAAGGAGGGACAGTTGT


 


395199
5-10-5
 76
3727
3746
0
AAAGATAAATGTCTGCTTGC


 


399825
5-10-5
 77
3732
3751
0
ACCCAAAAGATAAATGTCTG


 


395200
5-10-5
 78
3798
3817
0
TCTTCAAGTTACAAAAGCAA


 


399826
5-10-5
 99
3805
3824
0
ATAAATATCTTCAAGTTACA


 


405869
5-10-5
218
1392
1411
0
TCGATTTCCCGGTGGTCACT


 


405870
5-10-5
219
1393
1412
0
CTCGATTTCCCGGTGGTCAC


 


405871
5-10-5
220
1394
1413
0
CCTCGATTTCCCGGTGGTCA


 


405872
5-10-5
221
1395
1414
0
CCCTCGATTTCCCGGTGGTC


 


405873
5-10-5
222
1397
1416
0
TGCCCTCGATTTCCCGGTGG


 


405874
5-10-5
223
1398
1417
0
CTGCCCTCGATTTCCCGGTG


 


405875
5-10-5
224
1399
1418
0
CCTGCCCTCGATTTCCCGGT


 


405876
5-10-5
225
1400
1419
0
CCCTGCCCTCGATTTCCCGG


 


405877
5-10-5
246
1530
1549
0
TTGGCCACGCCGGCATCCCG


 


405878
5-10-5
247
1531
1550
0
CTTGGCCACGCCGGCATCCC


 


405879
5-10-5
248
1532
1551
0
CCTTGGCCACGCCGGCATCC


 


405880
5-10-5
249
1533
1552
0
CCCTTGGCCACGCCGGCATC


 


405881
5-10-5
250
1535
1554
0
CACCCTTGGCCACGCCGGCA


 


405882
5-10-5
251
1536
1555
0
GCACCCTTGGCCACGCCGGC


 


405883
5-10-5
252
1537
1556
0
GGCACCCTTGGCCACGCCGG


 


405884
5-10-5
253
1538
1557
0
TGGCACCCTTGGCCACGCCG


 


405885
5-10-5
346
1993
2012
0
GGAACCAGGCCTCATTGATG


 


405886
5-10-5
347
1994
2013
0
GGGAACCAGGCCTCATTGAT


 


405887
5-10-5
348
1995
2014
0
AGGGAACCAGGCCTCATTGA


 


405888
5-10-5
349
1996
2015
0
CAGGGAACCAGGCCTCATTG


 


405889
5-10-5
350
1998
2017
0
CTCAGGGAACCAGGCCTCAT


 


405890
5-10-5
351
1999
2018
0
CCTCAGGGAACCAGGCCTCA


 


405891
5-10-5
352
2000
2019
0
TCCTCAGGGAACCAGGCCTC


 


405892
5-10-5
353
2001
2020
0
GTCCTCAGGGAACCAGGCCT


 


405893
5-10-5
431
3083
3102
0
ATGAGGGCCATCAGCACCTT


 


405894
5-10-5
432
3084
3103
0
GATGAGGGCCATCAGCACCT


 


405895
5-10-5
433
3085
3104
0
AGATGAGGGCCATCAGCACC


 


405896
5-10-5
434
3086
3105
0
GAGATGAGGGCCATCAGCAC


 


405897
5-10-5
435
3088
3107
0
TGGAGATGAGGGCCATCAGC


 


405898
5-10-5
436
3089
3108
0
CTGGAGATGAGGGCCATCAG


 


405899
5-10-5
437
3090
3109
0
GCTGGAGATGAGGGCCATCA


 


405900
5-10-5
438
3091
3110
0
AGCTGGAGATGAGGGCCATC


 


405901
5-10-5
439
3157
3176
0
GCTAGATGCCATCCAGAAAG


 


405902
5-10-5
440
3158
3177
0
GGCTAGATGCCATCCAGAAA


 


405903
5-10-5
441
3159
3178
0
TGGCTAGATGCCATCCAGAA


 


405904
5-10-5
442
3160
3179
0
CTGGCTAGATGCCATCCAGA


 


405905
5-10-5
443
3162
3181
0
CTCTGGCTAGATGCCATCCA


 


405906
5-10-5
444
3163
3182
0
CCTCTGGCTAGATGCCATCC


 


405907
5-10-5
445
3164
3183
0
GCCTCTGGCTAGATGCCATC


 


405908
5-10-5
446
3165
3184
0
AGCCTCTGGCTAGATGCCAT


 


405909
5-10-5
267
1630
1649
0
CAGCTGGCTTTTCCGAATAA


 


405910
5-10-5
268
1632
1651
0
ACCAGCTGGCTTTTCCGAAT


 


405911
5-10-5
269
1634
1653
0
GGACCAGCTGGCTTTTCCGA


 


405912
5-10-5
270
1638
1657
0
GGCTGGACCAGCTGGCTTTT


 


405913
5-10-5
275
1742
1761
0
CGGTGACCAGCACGACCCCA


 


405914
5-10-5
276
1744
1763
0
AGCGGTGACCAGCACGACCC


 


405915
5-10-5
277
1746
1765
0
GCAGCGGTGACCAGCACGAC


 


405916
5-10-5
278
1748
1767
0
CGGCAGCGGTGACCAGCACG


 


405917
5-10-5
280
1752
1771
0
TTGCCGGCAGCGGTGACCAG


 


405918
5-10-5
281
1754
1773
0
AGTTGCCGGCAGCGGTGACC


 


405919
5-10-5
282
1756
1775
0
GAAGTTGCCGGCAGCGGTGA


 


405920
5-10-5
283
1758
1777
0
CGGAAGTTGCCGGCAGCGGT


 


405921
5-10-5
284
1760
1779
0
CCCGGAAGTTGCCGGCAGCG


 


405922
5-10-5
285
1762
1781
0
GTCCCGGAAGTTGCCGGCAG


 


405937
5-10-5
306
1820
1839
0
GAGGCACCAATGATGTCCTC


 


405938
5-10-5
307
1824
1843
0
GCTGGAGGCACCAATGATGT


 


405939
5-10-5
308
1826
1845
0
TCGCTGGAGGCACCAATGAT


 


405940
5-10-5
309
1828
1847
0
AGTCGCTGGAGGCACCAATG


 


405941
5-10-5
310
1830
1849
0
GCAGTCGCTGGAGGCACCAA


 


405942
5-10-5
311
1835
1854
0
GTGCTGCAGTCGCTGGAGGC


 


405943
5-10-5
312
1837
1856
0
AGGTGCTGCAGTCGCTGGAG


 


405944
5-10-5
314
1840
1859
0
AGCAGGTGCTGCAGTCGCTG


 


405945
5-10-5
315
1842
1861
0
AAAGCAGGTGCTGCAGTCGC


 


405946
5-10-5
316
1846
1865
0
ACACAAAGCAGGTGCTGCAG


 


405947
5-10-5
317
1848
1867
0
TGACACAAAGCAGGTGCTGC


 


405948
5-10-5
319
1900
1919
0
TCATGGCTGCAATGCCAGCC


 


405949
5-10-5
320
1905
1924
0
CAGCATCATGGCTGCAATGC


 


405950
5-10-5
321
1907
1926
0
GACAGCATCATGGCTGCAAT


 


405951
5-10-5
322
1909
1928
0
CAGACAGCATCATGGCTGCA


 


405952
5-10-5
323
1913
1932
0
TCGGCAGACAGCATCATGGC


 


405953
5-10-5
325
1917
1936
0
CGGCTCGGCAGACAGCATCA


 


405954
5-10-5
326
1919
1938
0
TCCGGCTCGGCAGACAGCAT


 


405955
5-10-5
328
1946
1965
0
CTCTGCCTCAACTCGGCCAG


 


405956
5-10-5
329
1948
1967
0
GTCTCTGCCTCAACTCGGCC


 


405957
5-10-5
330
1950
1969
0
CAGTCTCTGCCTCAACTCGG


 


405958
5-10-5
331
1952
1971
0
ATCAGTCTCTGCCTCAACTC


 


405959
5-10-5
332
1954
1973
0
GGATCAGTCTCTGCCTCAAC


 


405960
5-10-5
333
1956
1975
0
GTGGATCAGTCTCTGCCTCA


 


405961
5-10-5
334
1958
1977
0
AAGTGGATCAGTCTCTGCCT


 


405962
5-10-5
335
1961
1980
0
GAGAAGTGGATCAGTCTCTG


 


405963
5-10-5
337
1965
1984
0
GGCAGAGAAGTGGATCAGTC


 


405964
5-10-5
338
1969
1988
0
CTTTGGCAGAGAAGTGGATC


 


405965
5-10-5
339
1974
1993
0
GACATCTTTGGCAGAGAAGT


 


405966
5-10-5
340
1976
1995
0
ATGACATCTTTGGCAGAGAA


 


405967
5-10-5
341
1980
1999
0
ATTGATGACATCTTTGGCAG


 


405968
5-10-5
342
1982
2001
0
TCATTGATGACATCTTTGGC


 


405969
5-10-5
343
1987
2006
0
AGGCCTCATTGATGACATCT


 


405970
5-10-5
344
1989
2008
0
CCAGGCCTCATTGATGACAT


 


405971
5-10-5
355
2004
2023
0
CTGGTCCTCAGGGAACCAGG


 


405972
5-10-5
357
2006
2025
0
CGCTGGTCCTCAGGGAACCA


 


405973
5-10-5
358
2011
2030
0
GTACCCGCTGGTCCTCAGGG


 


405974
5-10-5
359
2013
2032
0
CAGTACCCGCTGGTCCTCAG


 


405975
5-10-5
361
2075
2094
0
CAAAACAGCTGCCAACCTGC


 


405976
5-10-5
362
2080
2099
0
TCCTGCAAAACAGCTGCCAA


 


405977
5-10-5
363
2082
2101
0
AGTCCTGCAAAACAGCTGCC


 


405978
5-10-5
365
2105
2124
0
GGCCCCGAGTGTGCTGACCA


 


405979
5-10-5
366
2110
2129
0
GTGTAGGCCCCGAGTGTGCT


 


405980
5-10-5
367
2112
2131
0
CCGTGTAGGCCCCGAGTGTG


 


405981
5-10-5
368
2114
2133
0
ATCCGTGTAGGCCCCGAGTG


 


405982
5-10-5
370
2118
2137
0
GGCCATCCGTGTAGGCCCCG


 


405983
5-10-5
371
2120
2139
0
GTGGCCATCCGTGTAGGCCC


 


405984
5-10-5
372
2168
2187
0
CTGGAGCAGCTCAGCAGCTC


 


405985
5-10-5
373
2170
2189
0
AACTGGAGCAGCTCAGCAGC


 


405986
5-10-5
374
2175
2194
0
GGAGAAACTGGAGCAGCTCA


 


405987
5-10-5
375
2177
2196
0
CTGGAGAAACTGGAGCAGCT


 


405996
5-10-5
412
2357
2376
0
GACTTTGCATTCCAGACCTG


 


405997
5-10-5
415
2362
2381
0
TCCTTGACTTTGCATTCCAG


 


405998
5-10-5
426
2568
2587
0
TCCGGCAGCAGATGGCAACG


 


406025
5-10-5
216
1362
1381
0
GTGTCTAGGAGATACACCTC


 


406026
5-10-5
217
1367
1386
0
TGCTGGTGTCTAGGAGATAC


 


406027
5-10-5
226
1404
1423
0
ATGACCCTGCCCTCGATTTC


 


406028
5-10-5
227
1406
1425
0
CCATGACCCTGCCCTCGATT


 


406029
5-10-5
228
1408
1427
0
GACCATGACCCTGCCCTCGA


 


406030
5-10-5
229
1412
1431
0
CGGTGACCATGACCCTGCCC


 


406031
5-10-5
230
1414
1433
0
GTCGGTGACCATGACCCTGC


 


406032
5-10-5
232
1418
1437
0
CGAAGTCGGTGACCATGACC


 


406033
5-10-5
237
1497
1516
0
CCTGCCAGGTGGGTGCCATG


 


406034
5-10-5
238
1502
1521
0
CCACCCCTGCCAGGTGGGTG


 


406035
5-10-5
239
1507
1526
0
GCTGACCACCCCTGCCAGGT


 


406036
5-10-5
241
1519
1538
0
GGCATCCCGGCCGCTGACCA


 


406037
5-10-5
242
1522
1541
0
GCCGGCATCCCGGCCGCTGA


 


406038
5-10-5
245
1529
1548
0
TGGCCACGCCGGCATCCCGG


 


406039
5-10-5
257
1566
1585
0
CAGTTGAGCACGCGCAGGCT


 


406040
5-10-5
258
1568
1587
0
GGCAGTTGAGCACGCGCAGG


 


406041
5-10-5
259
1572
1591
0
CCTTGGCAGTTGAGCACGCG


 


406042
5-10-5
260
1577
1596
0
CCTTCCCTTGGCAGTTGAGC


 


406043
5-10-5
261
1581
1600
0
GTGCCCTTCCCTTGGCAGTT


 


406044
5-10-5
262
1583
1602
0
CCGTGCCCTTCCCTTGGCAG


 


406045
5-10-5
266
1626
1645
0
TGGCTTTTCCGAATAAACTC


 


408642
5-10-5
264
1606
1625
0
CAGGCCTATGAGGGTGCCGC


 


408653
5-10-5
354
2003
2022
0
TGGTCCTCAGGGAACCAGGC


 


409126
5-10-5
447
1534
1553
1
ACCCTTGGTCACGCCGGCAT


 


410536
5-10-5
243
1527
1546
0
GCCACGCCGGCATCCCGGCC


 


410537
5-10-5
244
1528
1547
0
GGCCACGCCGGCATCCCGGC


 


410538
5-10-5
254
1539
1558
0
CTGGCACCCTTGGCCACGCC


 


410539
5-10-5
255
1540
1559
0
GCTGGCACCCTTGGCCACGC


 


410540
5-10-5
356
2005
2024
0
GCTGGTCCTCAGGGAACCAG


 


410562
5-10-5
411
2356
2375
0
ACTTTGCATTCCAGACCTGG


 


410563
5-10-5
413
2358
2377
0
TGACTTTGCATTCCAGACCT


 


410564
5-10-5
414
2359
2378
0
TTGACTTTGCATTCCAGACC


 


410565
5-10-5
419
2560
2579
0
CAGATGGCAACGGCTGTCAC


 


410566
5-10-5
420
2561
2580
0
GCAGATGGCAACGGCTGTCA


 


410567
5-10-5
421
2562
2581
0
AGCAGATGGCAACGGCTGTC


 


410568
5-10-5
422
2563
2582
0
CAGCAGATGGCAACGGCTGT


 


410569
5-10-5
423
2564
2583
0
GCAGCAGATGGCAACGGCTG


 


410570
5-10-5
424
2566
2585
0
CGGCAGCAGATGGCAACGGC


 


410571
5-10-5
425
2567
2586
0
CCGGCAGCAGATGGCAACGG


 


410572
5-10-5
427
2569
2588
0
CTCCGGCAGCAGATGGCAAC


 


410573
5-10-5
428
2570
2589
0
GCTCCGGCAGCAGATGGCAA


 


410733
5-10-5
231
1416
1435
0
AAGTCGGTGACCATGACCCT


 


410734
5-10-5
279
1749
1768
0
CCGGCAGCGGTGACCAGCAC


 


410737
5-10-5
313
1839
1858
0
GCAGGTGCTGCAGTCGCTGG


 


410738
5-10-5
324
1915
1934
0
GCTCGGCAGACAGCATCATG


 


410739
5-10-5
336
1963
1982
0
CAGAGAAGTGGATCAGTCTC


 


410740
5-10-5
345
1991
2010
0
AACCAGGCCTCATTGATGAC


 


410741
5-10-5
369
2116
2135
0
CCATCCGTGTAGGCCCCGAG


 


410753
5-10-5
215
1351
1370
0
ATACACCTCCACCAGGCTGC


 


410754
5-10-5
233
1428
1447
0
GGCACATTCTCGAAGTCGGT


 


410755
5-10-5
234
1463
1482
0
TGGCCTGTCTGTGGAAGCGG


 


410756
5-10-5
235
1490
1509
0
GGTGGGTGCCATGACTGTCA


 


410757
5-10-5
240
1515
1534
0
TCCCGGCCGCTGACCACCCC


 


410758
5-10-5
256
1545
1564
0
CGCATGCTGGCACCCTTGGC


 


410759
5-10-5
263
1594
1613
0
GGTGCCGCTAACCGTGCCCT


 


410760
5-10-5
265
1618
1637
0
CCGAATAAACTCCAGGCCTA


 


410761
5-10-5
271
1649
1668
0
GTGGCCCCACAGGCTGGACC


 


410762
5-10-5
272
1662
1681
0
AGCAGCACCACCAGTGGCCC


 


410763
5-10-5
273
1684
1703
0
GCTGTACCCACCCGCCAGGG


 


410764
5-10-5
274
1730
1749
0
CGACCCCAGCCCTCGCCAGG


 


410769
5-10-5
318
1858
1877
0
TCCCACTCTGTGACACAAAG


 


410770
5-10-5
327
1933
1952
0
CGGCCAGGGTGAGCTCCGGC


 


410771
5-10-5
360
2061
2080
0
ACCTGCCCCATGGGTGCTGG


 


410776
5-10-5
408
2245
2264
0
GTGCCAAGGTCCTCCACCTC


 


410777
5-10-5
409
2265
2284
0
TCAGCACAGGCGGCTTGTGG


 


410778
5-10-5
410
2295
2314
0
CCACGCACTGGTTGGGCTGA


 


410779
5-10-5
416
2375
2394
0
CGGGATTCCATGCTCCTTGA


 


410780
5-10-5
417
2405
2424
0
GCAGGCCACGGTCACCTGCT


 


410781
5-10-5
418
2442
2461
0
GGAGGGCACTGCAGCCAGTC


 


410782
5-10-5
429
2580
2599
0
CCAGGTGCCGGCTCCGGCAG


 


410783
5-10-5
430
2590
2609
0
GAGGCCTGCGCCAGGTGCCG









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 3-14-3 gap-widened motif. Table 14 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 3, having a 3-14-3 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 14







Gapmer antisense compounds having a 3-14-3 motif


targeted to SEQ ID NO: 3
















5′
3′







Target
Target






SEQ
Site on
Site on




Isis

ID
SEQ ID
SEQ ID




No
Motif
NO
NO: 3
NO: 3
Mismatches
Sequence (5′-3′)





399923
3-14-3
85
 155
 174
0
ACAAATTCCCAGACTCAGCA


 


399983
3-14-3
100
 220
 239
0
ATCTCAGGACAGGTGAGCAA


 


399984
3-14-3
116
 234
 253
0
GAGTAGAGATTCTCATCTCA


 


399924
3-14-3
129
 290
 309
0
GTGCCATCTGAACAGCACCT


 


399985
3-14-3
117
 302
 321
0
GAGTCTTCTGAAGTGCCATC


 


399986
3-14-3
81
 377
 396
0
AAGCAGGGCCTCAGGTGGAA


 


399925
3-14-3
110
 400
 419
0
CCTGGAACCCCTGCAGCCAG


 


399926
3-14-3
152
 451
 470
0
TTCAGGCAGGTTGCTGCTAG


 


399987
3-14-3
83
 460
 479
0
AAGGAAGACTTCAGGCAGGT


 


399927
3-14-3
140
 472
 491
0
TCAGCCAGGCCAAAGGAAGA


 


399988
3-14-3
137
 586
 605
0
TAGGGAGAGCTCACAGATGC


 


399928
3-14-3
136
 596
 615
0
TAGGAGAAAGTAGGGAGAGC


 


399929
3-14-3
132
 653
 672
0
TAAAAGCTGCAAGAGACTCA


 


399930
3-14-3
139
 741
 760
0
TCAGAGAAAACAGTCACCGA


 


399989
3-14-3
92
 871
 890
0
AGAGACAGGAAGCTGCAGCT


 


399931
3-14-3
142
 878
 897
0
TCATTTTAGAGACAGGAAGC


 


399932
3-14-3
113
 915
 934
0
GAATAACAGTGATGTCTGGC


 


399933
3-14-3
138
 968
 987
0
TCACAGCTCACCGAGTCTGC


 


399934
3-14-3
98
 998
1017
0
AGTGTAAAATAAAGCCCCTA


 


399935
3-14-3
96
1075
1094
0
AGGACCCAAGTCATCCTGCT


 


399936
3-14-3
124
1105
1124
0
GGCCATCAGCTGGCAATGCT


 


399990
3-14-3
82
1144
1163
0
AAGGAAAGGGAGGCCTAGAG


 


399937
3-14-3
133
1149
1168
0
TAGACAAGGAAAGGGAGGCC


 


399938
3-14-3
103
1155
1174
0
ATTTCATAGACAAGGAAAGG


 


399939
3-14-3
155
1275
1294
0
CTTATAGTTAACACACAGAA


 


399991
3-14-3
156
1283
1302
0
AAGTCAACCTTATAGTTAAC


 


399940
3-14-3
146
1315
1334
0
TGACATTTGTGGGAGAGGAG


 


399941
3-14-3
161
1322
1341
0
TCCAAGGTGACATTTGTGGG


 


399882
3-14-3
19
1365
1384
0
CTGGTGTCTAGGAGATACAC


 


399953
3-14-3
20
1370
1389
0
GTATGCTGGTGTCTAGGAGA


 


399883
3-14-3
21
1390
1409
0
GATTTCCCGGTGGTCACTCT


 


399954
3-14-3
22
1396
1415
0
GCCCTCGATTTCCCGGTGGT


 


399955
3-14-3
23
1410
1429
0
GTGACCATGACCCTGCCCTC


 


399884
3-14-3
24
1420
1439
0
CTCGAAGTCGGTGACCATGA


 


399885
3-14-3
25
1453
1472
0
GTGGAAGCGGGTCCCGTCCT


 


399886
3-14-3
26
1500
1519
0
ACCCCTGCCAGGTGGGTGCC


 


399956
3-14-3
27
1505
1524
0
TGACCACCCCTGCCAGGTGG


 


399887
3-14-3
28
1534
1553
0
ACCCTTGGCCACGCCGGCAT


 


399888
3-14-3
29
1570
1589
0
TTGGCAGTTGAGCACGCGCA


 


399957
3-14-3
30
1575
1594
0
TTCCCTTGGCAGTTGAGCAC


 


399889
3-14-3
31
1607
1626
0
CCAGGCCTATGAGGGTGCCG


 


399890
3-14-3
32
1628
1647
0
GCTGGCTTTTCCGAATAAAC


 


399891
3-14-3
33
1740
1759
0
GTGACCAGCACGACCCCAGC


 


399897
3-14-3
39
1822
1841
0
TGGAGGCACCAATGATGTCC


 


399960
3-14-3
40
1833
1852
0
GCTGCAGTCGCTGGAGGCAC


 


399961
3-14-3
41
1844
1863
0
ACAAAGCAGGTGCTGCAGTC


 


399898
3-14-3
101
1898
1917
0
ATGGCTGCAATGCCAGCCAC


 


399962
3-14-3
42
1903
1922
0
GCATCATGGCTGCAATGCCA


 


399963
3-14-3
43
1911
1930
0
GGCAGACAGCATCATGGCTG


 


399964
3-14-3
44
1959
1978
0
GAAGTGGATCAGTCTCTGCC


 


399899
3-14-3
45
1967
1986
0
TTGGCAGAGAAGTGGATCAG


 


399965
3-14-3
46
1972
1991
0
CATCTTTGGCAGAGAAGTGG


 


399966
3-14-3
47
1978
1997
0
TGATGACATCTTTGGCAGAG


 


399967
3-14-3
48
1985
2004
0
GCCTCATTGATGACATCTTT


 


399968
3-14-3
49
1997
2016
0
TCAGGGAACCAGGCCTCATT


 


399900
3-14-3
50
2002
2021
0
GGTCCTCAGGGAACCAGGCC


 


399901
3-14-3
87
2009
2028
0
ACCCGCTGGTCCTCAGGGAA


 


399969
3-14-3
51
2016
2035
0
GGTCAGTACCCGCTGGTCCT


 


399902
3-14-3
119
2038
2057
0
GCAGGGCGGCCACCAGGTTG


 


399903
3-14-3
52
2073
2092
0
AAACAGCTGCCAACCTGCCC


 


399970
3-14-3
53
2078
2097
0
CTGCAAAACAGCTGCCAACC


 


399942
3-14-3
104
2085
2104
0
CACAGTCCTGCAAAACAGCT


 


399992
3-14-3
130
2095
2114
0
GTGCTGACCACACAGTCCTG


 


399904
3-14-3
54
2108
2127
0
GTAGGCCCCGAGTGTGCTGA


 


399971
3-14-3
55
2173
2192
0
AGAAACTGGAGCAGCTCAGC


 


399972
3-14-3
56
2179
2198
0
TCCTGGAGAAACTGGAGCAG


 


399908
3-14-3
60
2355
2374
0
CTTTGCATTCCAGACCTGGG


 


399973
3-14-3
61
2360
2379
0
CTTGACTTTGCATTCCAGAC


 


399909
3-14-3
62
2565
2584
0
GGCAGCAGATGGCAACGGCT


 


399910
3-14-3
154
2665
2684
0
TTTTAAAGCTCAGCCCCAGC


 


399974
3-14-3
63
2670
2689
0
AACCATTTTAAAGCTCAGCC


 


399911
3-14-3
64
2759
2778
0
TCAAGGGCCAGGCCAGCAGC


 


399975
3-14-3
65
2764
2783
0
CCCACTCAAGGGCCAGGCCA


 


399976
3-14-3
122
2837
2856
0
GGAGGGAGCTTCCTGGCACC


 


399912
3-14-3
66
2852
2871
0
ATGCCCCACAGTGAGGGAGG


 


399913
3-14-3
67
2861
2880
0
AATGGTGAAATGCCCCACAG


 


399914
3-14-3
153
2904
2923
0
TTGGGAGCAGCTGGCAGCAC


 


399915
3-14-3
68
3005
3024
0
CATGGGAAGAATCCTGCCTC


 


399916
3-14-3
69
3087
3106
0
GGAGATGAGGGCCATCAGCA


 


399917
3-14-3
70
3155
3174
0
TAGATGCCATCCAGAAAGCT


 


399977
3-14-3
71
3161
3180
0
TCTGGCTAGATGCCATCCAG


 


399918
3-14-3
72
3238
3257
0
GGCATAGAGCAGAGTAAAGG


 


399978
3-14-3
73
3243
3262
0
AGCCTGGCATAGAGCAGAGT


 


399979
3-14-3
135
3249
3268
0
TAGCACAGCCTGGCATAGAG


 


399919
3-14-3
112
3482
3501
0
GAAGAGGCTTGGCTTCAGAG


 


399980
3-14-3
74
3488
3507
0
AAGTAAGAAGAGGCTTGGCT


 


399920
3-14-3
75
3692
3711
0
GCTCAAGGAGGGACAGTTGT


 


399921
3-14-3
76
3727
3746
0
AAAGATAAATGTCTGCTTGC


 


399981
3-14-3
77
3732
3751
0
ACCCAAAAGATAAATGTCTG


 


399922
3-14-3
78
3798
3817
0
TCTTCAAGTTACAAAAGCAA


 


399982
3-14-3
99
3805
3824
0
ATAAATATCTTCAAGTTACA


 


405604
3-14-3
236
1493
1512
0
CCAGGTGGGTGCCATGACTG


 


405641
3-14-3
373
2170
2189
0
AACTGGAGCAGCTCAGCAGC


 


410583
3-14-3
243
1527
1546
0
GCCACGCCGGCATCCCGGCC


 


410584
3-14-3
244
1528
1547
0
GGCCACGCCGGCATCCCGGC


 


410585
3-14-3
245
1529
1548
0
TGGCCACGCCGGCATCCCGG


 


410586
3-14-3
246
1530
1549
0
TTGGCCACGCCGGCATCCCG


 


410587
3-14-3
247
1531
1550
0
CTTGGCCACGCCGGCATCCC


 


410588
3-14-3
248
1532
1551
0
CCTTGGCCACGCCGGCATCC


 


410589
3-14-3
249
1533
1552
0
CCCTTGGCCACGCCGGCATC


 


410590
3-14-3
250
1535
1554
0
CACCCTTGGCCACGCCGGCA


 


410591
3-14-3
251
1536
1555
0
GCACCCTTGGCCACGCCGGC


 


410592
3-14-3
252
1537
1556
0
GGCACCCTTGGCCACGCCGG


 


410593
3-14-3
253
1538
1557
0
TGGCACCCTTGGCCACGCCG


 


410594
3-14-3
254
1539
1558
0
CTGGCACCCTTGGCCACGCC


 


410595
3-14-3
255
1540
1559
0
GCTGGCACCCTTGGCCACGC


 


410596
3-14-3
348
1995
2014
0
AGGGAACCAGGCCTCATTGA


 


410597
3-14-3
349
1996
2015
0
CAGGGAACCAGGCCTCATTG


 


410598
3-14-3
350
1998
2017
0
CTCAGGGAACCAGGCCTCAT


 


410599
3-14-3
351
1999
2018
0
CCTCAGGGAACCAGGCCTCA


 


410600
3-14-3
352
2000
2019
0
TCCTCAGGGAACCAGGCCTC


 


410601
3-14-3
353
2001
2020
0
GTCCTCAGGGAACCAGGCCT


 


410602
3-14-3
354
2003
2022
0
TGGTCCTCAGGGAACCAGGC


 


410603
3-14-3
355
2004
2023
0
CTGGTCCTCAGGGAACCAGG


 


410604
3-14-3
356
2005
2024
0
GCTGGTCCTCAGGGAACCAG


 


410633
3-14-3
411
2356
2375
0
ACTTTGCATTCCAGACCTGG


 


410634
3-14-3
412
2357
2376
0
GACTTTGCATTCCAGACCTG


 


410635
3-14-3
413
2358
2377
0
TGACTTTGCATTCCAGACCT


 


410636
3-14-3
414
2359
2378
0
TTGACTTTGCATTCCAGACC


 


410637
3-14-3
419
2560
2579
0
CAGATGGCAACGGCTGTCAC


 


410638
3-14-3
420
2561
2580
0
GCAGATGGCAACGGCTGTCA


 


410639
3-14-3
421
2562
2581
0
AGCAGATGGCAACGGCTGTC


 


410640
3-14-3
422
2563
2582
0
CAGCAGATGGCAACGGCTGT


 


410641
3-14-3
423
2564
2583
0
GCAGCAGATGGCAACGGCTG


 


410642
3-14-3
424
2566
2585
0
CGGCAGCAGATGGCAACGGC


 


410643
3-14-3
425
2567
2586
0
CCGGCAGCAGATGGCAACGG


 


410644
3-14-3
426
2568
2587
0
TCCGGCAGCAGATGGCAACG


 


410645
3-14-3
427
2569
2588
0
CTCCGGCAGCAGATGGCAAC


 


410646
3-14-3
428
2570
2589
0
GCTCCGGCAGCAGATGGCAA









In certain embodiments, gap-widened antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gap-widened antisense compounds are targeted to SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 12 have a 2-13-5 gap-widened motif. Table 15 illustrates gap-widened antisense compounds targeted to SEQ ID NO: 3, having a 2-13-5 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 15







Gapmer antisense compounds having a 2-13-5 motif


targeted to SEQ ID NO: 3
















5′
3′







Target
Target






SEQ
Site to
Site to




ISIS

ID
SEQ ID
SEQ ID




No
Motiff
NO
NO: 3
NO: 3
Mismatches
Sequences (5′-3′)





410658
2-13-5
243
1527
1546
0
GCCACGCCGGCATCCCGGCC


 


410659
2-13-5
244
1528
1547
0
GGCCACGCCGGCATCCCGGC


 


410660
2-13-5
245
1529
1548
0
TGGCCACGCCGGCATCCCGG


 


410661
2-13-5
246
1530
1549
0
TTGGCCACGCCGGCATCCCG


 


410662
2-13-5
247
1531
1550
0
CTTGGCCACGCCGGCATCCC


 


410663
2-13-5
248
1532
1551
0
CCTTGGCCACGCCGGCATCC


 


410664
2-13-5
249
1533
1552
0
CCCTTGGCCACGCCGGCATC


 


410665
2-13-5
 28
1534
1553
0
ACCCTTGGCCACGCCGGCAT


 


410666
2-13-5
250
1535
1554
0
CACCCTTGGCCACGCCGGCA


 


410667
2-13-5
251
1536
1555
0
GCACCCTTGGCCACGCCGGC


 


410668
2-13-5
252
1537
1556
0
GGCACCCTTGGCCACGCCGG


 


410669
2-13-5
253
1538
1557
0
TGGCACCCTTGGCCACGCCG


 


410670
2-13-5
254
1539
1558
0
CTGGCACCCTTGGCCACGCC


 


410671
2-13-5
255
1540
1559
0
GCTGGCACCCTTGGCCACGC


 


410672
2-13-5
348
1995
2014
0
AGGGAACCAGGCCTCATTGA


 


410673
2-13-5
349
1996
2015
0
CAGGGAACCAGGCCTCATTG


 


410674
2-13-5
 49
1997
2016
0
TCAGGGAACCAGGCCTCATT


 


410675
2-13-5
350
1998
2017
0
CTCAGGGAACCAGGCCTCAT


 


410676
2-13-5
351
1999
2018
0
CCTCAGGGAACCAGGCCTCA


 


410677
2-13-5
352
2000
2019
0
TCCTCAGGGAACCAGGCCTC


 


410678
2-13-5
353
2001
2020
0
GTCCTCAGGGAACCAGGCCT


 


410679
2-13-5
 50
2002
2021
0
GGTCCTCAGGGAACCAGGCC


 


410680
2-13-5
354
2003
2022
0
TGGTCCTCAGGGAACCAGGC


 


410681
2-13-5
355
2004
2023
0
CTGGTCCTCAGGGAACCAGG


 


410682
2-13-5
356
2005
2024
0
GCTGGTCCTCAGGGAACCAG


 


410713
2-13-5
 60
2355
2374
0
CTTTGCATTCCAGACCTGGG


 


410714
2-13-5
411
2356
2375
0
ACTTTGCATTCCAGACCTGG


 


410715
2-13-5
412
2357
2376
0
GACTTTGCATTCCAGACCTG


 


410716
2-13-5
413
2358
2377
0
TGACTTTGCATTCCAGACCT


 


410717
2-13-5
414
2359
2378
0
TTGACTTTGCATTCCAGACC


 


410718
2-13-5
 61
2360
2379
0
CTTGACTTTGCATTCCAGAC


 


410719
2-13-5
419
2560
2579
0
CAGATGGCAACGGCTGTCAC


 


410720
2-13-5
420
2561
2580
0
GCAGATGGCAACGGCTGTCA


 


410721
2-13-5
421
2562
2581
0
AGCAGATGGCAACGGCTGTC


 


410722
2-13-5
422
2563
2582
0
CAGCAGATGGCAACGGCTGT


 


410723
2-13-5
423
2564
2583
0
GCAGCAGATGGCAACGGCTG


 


410724
2-13-5
 62
2565
2584
0
GGCAGCAGATGGCAACGGCT


 


410725
2-13-5
424
2566
2585
0
CGGCAGCAGATGGCAACGGC


 


410726
2-13-5
425
2567
2586
0
CCGGCAGCAGATGGCAACGG


 


410727
2-13-5
426
2568
2587
0
TCCGGCAGCAGATGGCAACG


 


410728
2-13-5
427
2569
2588
0
CTCCGGCAGCAGATGGCAAC


 


410729
2-13-5
428
2570
2589
0
GCTCCGGCAGCAGATGGCAA









The following embodiments set forth target regions of PCSK9 nucleic acids. Also illustrated are examples of antisense compounds targeted to the target regions. It is understood that the sequence set forth in each SEQ ID NO is independent of any modification to a sugar moiety, an internucleoside linkage, or a nucleobase. As such, antisense compounds defined by a SEQ ID NO may comprise, independently, one or more modifications to a sugar moiety, an internucleoside linkage, or a nucleobase. Antisense compounds described by Isis Number (Isis No) indicate a combination of nucleobase sequence and motif.
In certain embodiments, antisense compounds target a range of a PCSK9 nucleic acid. In certain embodiment, such compounds contain at least an 8 nucleotide core sequence in common. In certain embodiments, such compounds sharing at least an 8 nucleotide core sequence targets the following nucleotide regions of SEQ ID NO: 3: 220-253, 290-321, 377-419, 451-615, 653-672, 741-760, 871-897, 915-934, 968-1017, 1075-1124, 1075-1174, 1144-1174, 1275-1302, 1315-1341, 1351-1389, 1351-1447, 1365-1439, 1390-1417, 1390-1429, 1390-1439, 1399-1425, 1408-1435, 1420-1447, 1453-1482, 1490-1516, 1490-1564, 1500-1526, 1515-1541, 1527-1553, 1527-1554, 1528-1554, 1529-1555, 1529-1556, 1530-1556, 1530-1557, 1531-1557, 1532-1551, 1532-1558, 1533-1559, 1534-1559, 1535-1559, 1536-1559, 1537-1564, 1566-1602, 1566-1681, 1606-1626, 1618-1645, 1626-1653, 1684-1703, 1730-1781, 1740-1767, 1749-1775, 1758-1781, 1820-1847, 1820-1877, 1822-2198, 1830-1856, 1839-1865, 1840-1867, 1898-1924, 1898-2035, 1903-2127, 1907-1934, 1911-1938, 1946-1971, 1954-1980, 1959-2035, 1959-2057, 1963-1988, 1967-2035, 1972-1999, 1982-2008, 1991-2018, 1993-2019, 1995-2022, 1996-2023, 1997-2024, 1998-2025, 1999-2025, 2000-2025, 2009-2035, 2038-2139, 2061-2139, 2073-2099, 2078-2104, 2105-2131, 2112-2139, 2168-2198, 2170-2177, 2245-2284, 2295-2394, 2355-2381, 2355-2394, 2405-2461, 2560-2587, 2560-2609, 2561-2588, 2562-2589, 2563-2589, 2564-2589, 2565-2589, 2566-2589, 2567-2589, 2568-2589, 2665-2689, 2759-2783, 2837-2880, 2904-2923, 3005-3024, 3005-3174, 3083-3110, 3155-3184, 3238-3268, 3482-3711, or 3727-3751, or 3798-3824.
In certain embodiments, a target region is nucleotides 290-321 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 290-321 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 117 or 129. In certain such embodiments, an antisense compound targeted to nucleotides 290-321 of SEQ ID NO: 3 is selected from ISIS NOs: 395202, 399924, 399829 or 399985.
In certain embodiments, a target region is nucleotides 220-253 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 220-253 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 100 or 116. In certain such embodiments, an antisense compound targeted to nucleotides 220-253 of SEQ ID NO: 3 is selected from ISIS NOs: 399827, 399983, 399828 or 399984.
In certain embodiments, a target region is nucleotides 377-419 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 377-419 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 81 or 110. In certain such embodiments, an antisense compound targeted to nucleotides 377-419 of SEQ ID NO: 3 is selected from ISIS NOs: 399830, 399986, 395203 or 399925.
In certain embodiments, a target region is nucleotides 451-615 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 451-615 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 152, 140, 137 or 136. In certain such embodiments, an antisense compound targeted to nucleotides 451-615 of SEQ ID NO: 3 is selected from ISIS NOs: 395204, 399926, 399831, 399987, 395205, 399927, 399832, 399988, 395206 or 399928.
In certain embodiments, a target region is nucleotides 653-672 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 653-672 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 132. In certain such embodiments, an antisense compound targeted to nucleotides 653-672 of SEQ ID NO: 3 is selected from ISIS NOs: 395207 or 399929.
In certain embodiments, a target region is nucleotides 741-760 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 741-760 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 139. In certain such embodiments, an antisense compound targeted to nucleotides 741-760 of SEQ ID NO: 3 is selected from ISIS NOs: 395208 or 399930.
In certain embodiments, a target region is nucleotides 871-897 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 871-897 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 92 or 142. In certain such embodiments, an antisense compound targeted to nucleotides 871-897 of SEQ ID NO: 3 is selected from ISIS NOs: 399833, 399989, 395209 or 399931.
In certain embodiments, a target region is nucleotides 915-934 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 915-934 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 113. In certain such embodiments, an antisense compound targeted to nucleotides 915-934 of SEQ ID NO: 3 is selected from ISIS NOs: 395210 or 399932.
In certain embodiments, a target region is nucleotides 968-1017 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 968-1017 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 98 or 138. In certain such embodiments, an antisense compound targeted to nucleotides 968-1017 of SEQ ID NO: 3 is selected from ISIS NOs: 395211, 399933, 395212, or 399934.
In certain embodiments, a target region is nucleotides 1075-1124 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1075-1124 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 96 or 124. In certain such embodiments, an antisense compound targeted to nucleotides 1075-1124 of SEQ ID NO: 3 is selected from ISIS NOs: 395213, 399935, 395214, or 399936.
In certain embodiments, a target region is nucleotides 1075-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1075-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 96, 103, 124, or 133. In certain such embodiments, an antisense compound targeted to nucleotides 1075-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 395213, 399935, 395214, 399936, 399834, 399990, 395215, 399937, 395216, or 399938.
In certain embodiments, a target region is nucleotides 1144-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1144-1174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 82, 133 or 103. In certain such embodiments, an antisense compound targeted to nucleotides 1144-1174 of SEQ ID NO: 3 is selected from ISIS NOs: 399834, 399990, 395215, 399937, 395216 or 399938.
In certain embodiments, a target region is nucleotides 1275-1302 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1275-1302 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 155 or 156. In certain such embodiments, an antisense compound targeted to nucleotides 1275-1302 of SEQ ID NO: 3 is selected from ISIS NOs: 395217, 399939, 399835 or 399991.
In certain embodiments, a target region is nucleotides 1315-1341 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1315-1341 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 146 or 161. In certain such embodiments, an antisense compound targeted to nucleotides 1315-1341 of SEQ ID NO: 3 is selected from ISIS NOs: 395218, 399940, 395219 or 399941.
In certain embodiments, a target region is nucleotides 1351-1389 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1351-1389 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217 or 20. In certain such embodiments, an antisense compound targeted to nucleotides 1351-1389 of SEQ ID NO: 3 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797 or 399953.
In certain embodiments, a target region is nucleotides 1351-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1351-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 215, 216, 19, 217, 20, 21, 218, 219, 220, 221, 22, 222, 223, 224, 225, 226, 227, 228, 23, 229, 230, 231, 232, 24, or 233. In certain such embodiments, an antisense compound targeted to nucleotides 1351-1447 of SEQ ID NO: 3 is selected from ISIS NOs: 410753, 406025, 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, 399884 or 410754.
In certain embodiments, a target region is nucleotides 1365-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1365-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 19, 20, 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, or 232. In certain such embodiments, an antisense compound targeted to nucleotides 1365-1439 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.
In certain embodiments, a target region is nucleotides 1390-1417 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1417 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 218, 219, 220, 221, 22, 222 or 223. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1417 of SEQ ID NO: 3 is selected from ISIS NOs: 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873 or 405874.
In certain embodiments, a target region is nucleotides 1390-1429 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1429 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, or 228. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1429 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, or 399955.
In certain embodiments, a target region is nucleotides 1390-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1390-1439 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 21, 22, 23, 24, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, or 232. In certain such embodiments, an antisense compound targeted to nucleotides 1390-1439 of SEQ ID NO: 3 is selected from ISIS NOs: 395160, 399882, 406026, 399797, 399953, 395161, 399883, 405869, 405870, 405871, 405872, 399798, 399954, 405873, 405874, 405875, 405876, 406027, 406028, 406029, 399799, 399955, 406030, 406031, 410733, 406032, 395162, or 399884.
In certain embodiments, a target region is nucleotides 1399-1425 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1399-1425 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 224, 225, 226 or 227. In certain such embodiments, an antisense compound targeted to nucleotides 1399-1425 of SEQ ID NO: 3 is selected from ISIS NOs: 405875, 405876, 406027 or 406028.
In certain embodiments, a target region is nucleotides 1408-1435 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1408-1435 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 228, 23, 229, 230 or 231. In certain such embodiments, an antisense compound targeted to nucleotides 1408-1435 of SEQ ID NO: 3 is selected from ISIS NOs: 406029, 399799, 399955, 406030, 406031 or 410733.
In certain embodiments, a target region is nucleotides 1420-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1420-1447 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 24 or 233. In certain such embodiments, an antisense compound targeted to nucleotides 1420-1447 of SEQ ID NO: 3 is selected from ISIS NOs: 395162, 399884 or 410754.
In certain embodiments, a target region is nucleotides 1453-1482 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1453-1482 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 25 or 234. In certain such embodiments, an antisense compound targeted to nucleotides 1453-1482 of SEQ ID NO: 3 is selected from ISIS NOs: 395163, 399885 or 410755.
In certain embodiments, a target region is nucleotides 1490-1516 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1490-1516 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, or 237. In certain such embodiments, an antisense compound targeted to nucleotides 1490-1516 of SEQ ID NO: 3 is selected from ISIS NOs: 410756, 405604, or 406033.
In certain embodiments, a target region is nucleotides 1490-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1490-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 235, 236, 237, 26, 238, 27, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 447, 28, 250, 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1490-1564 of SEQ ID NO: 3 is selected from ISIS NOs: 410756, 405604, 406033, 395164, 399886, 406034, 399800, 399956, 406035, 410757, 406036, 406037, 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, or 410758.
In certain embodiments, a target region is nucleotides 1500-1526 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1500-1526 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 26, 238, 27, or 239. In certain such embodiments, an antisense compound targeted to nucleotides 1500-1526 of SEQ ID NO: 3 is selected from ISIS NOs: 395164, 399886, 406034, 399800, 399956, or 406035.
In certain embodiments, a target region is nucleotides 1515-1541 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1515-1541 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 240, 241, or 242. In certain such embodiments, an antisense compound targeted to nucleotides 1515-1541 of SEQ ID NO: 3 is selected from ISIS NOs: 399886, 406034, 399800, 399956, or 406035.
In certain embodiments, a target region is nucleotides 1527-1553 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1527-1553 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 447, 28 or 249. In certain such embodiments, an antisense compound targeted to nucleotides 1527-1553 of SEQ ID NO: 3 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, or 410665.
In certain embodiments, a target region is nucleotides 1527-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1527-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 243, 244, 245, 246, 247, 248, 249, 447, 28, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 1527-1554 of SEQ ID NO: 3 is selected from ISIS NOs: 410536, 410583, 410658, 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 405881, 410590, 410666, or 410665.
In certain embodiments, a target region is nucleotides 1528-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1528-1554 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 244, 245, 246, 247, 248, 249, 447, 28, or 250. In certain such embodiments, an antisense compound targeted to nucleotides 1528-1554 of SEQ ID NO: 3 is selected from ISIS NOs: 410537, 410584, 410659, 406038, 410585, 410660, 405877, 410586, 410661, 405878, 410587, 410662, 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 409126, 410665, or 405881.
In certain embodiments, a target region is nucleotides 1529-1555 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1529-1555 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249, 447, 28, 250 or 251. In certain such embodiments, an antisense compound targeted to nucleotides 1529-1555 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410667.
In certain embodiments, a target region is nucleotides 1529-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1529-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 245, 246, 247, 248, 249, 447, 28, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 1529-1556 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, or 410668.
In certain embodiments, a target region is nucleotides 1530-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1530-1556 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 246, 247, 248, 249, 447, 28, 250, 251, or 252. In certain such embodiments, an antisense compound targeted to nucleotides 1530-1556 of SEQ ID NO: 3 is selected from ISIS NOs: 406038, 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 409126, 410585, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410660, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 405884, 410593, 410669 or 410668.
In certain embodiments, a target region is nucleotides 1530-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1530-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 246, 247, 248, 249, 250, 251, 252, 253, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1530-1557 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410586, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410661, 410662, 410663, 410664, 410665, 410666, 410667, 410668, or 410669.
In certain embodiments, a target region is nucleotides 1531-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1531-1557 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 247, 248, 249, 250, 251, 252, 253, or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1531-1557 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 409126, 410587, 410588, 410589, 410590, 410591, 410592, 410593, 410662, 410663, 410664, 410665, 410666, 410667, 410668, or 410669.
In certain embodiments, a target region is nucleotides 1532-1551 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1551 of SEQ ID NO: 3. In one such embodiment, an antisense compound targeted to nucleotides 1532-1551 of SEQ ID NO: 3 is ISIS NO: 405879.
In certain embodiments, a target region is nucleotides 1532-1558 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1532-1558 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 248, 249, 250, 251, 252, 253, 254 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1532-1558 of SEQ ID NO: 3 is selected from ISIS NOs: 405879, 410588, 410663, 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, or 410670.
In certain embodiments, a target region is nucleotides 1533-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1533-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 249, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1533-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405880, 410589, 410664, 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1534-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1534-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 28, 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1534-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 395165, 399887, 410665, 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, or 410671.
In certain embodiments, a target region is nucleotides 1535-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1535-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 250, 251, 252, 253, 254, 255 or 447. In certain such embodiments, an antisense compound targeted to nucleotides 1535-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405881, 410590, 410666, 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594 or 410671.
In certain embodiments, a target region is nucleotides 1536-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1536-1559 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, or 255. In certain such embodiments, an antisense compound targeted to nucleotides 1536-1559 of SEQ ID NO: 3 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, or 410671.
In certain embodiments, a target region is nucleotides 1537-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1537-1564 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 251, 252, 253, 254, 255, or 256. In certain such embodiments, an antisense compound targeted to nucleotides 1537-1564 of SEQ ID NO: 3 is selected from ISIS NOs: 405882, 410591, 410667, 405883, 410592, 410668, 405884, 410593, 410669, 410538, 410594, 410670, 410539, 410595, 410671, or 410758.
In certain embodiments, a target region is nucleotides 1566-1602 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1602 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258, 259, 260, 261, or 262. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1602 of SEQ ID NO: 3 is selected from ISIS NOs: 395166, 395167, 395168, 399801, 399888, 399889, 399890, 399957, 405909, 405910, 405911, 405912, 406039, 406040, 406041, 406042, 406043, or 406044.
In certain embodiments, a target region is nucleotides 1566-1681 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1566-1681 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 29, 30, 31, 32, 257, 258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, or 272. In certain such embodiments, an antisense compound targeted to nucleotides 1566-1681 of SEQ ID NO: 3 is selected from ISIS NOs: 395166, 395167, 395168, 399801, 399888, 399889, 399890, 399957, 405909, 405910, 405911, 405912, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 408642, 410759, 410760, 410761, or 410762.
In certain embodiments, a target region is nucleotides 1606-1626 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1606-1626 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 264 or 31. In certain such embodiments, an antisense compound targeted to nucleotides 1606-1626 of SEQ ID NO: 3 is selected from ISIS NOs: 408642, 395167, or 399889.
In certain embodiments, a target region is nucleotides 1618-1645 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1618-1645 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 265 or 266. In certain such embodiments, an antisense compound targeted to nucleotides 1618-1645 of SEQ ID NO: 3 is selected from ISIS NOs: 410760 or 406045.
In certain embodiments, a target region is nucleotides 1626-1653 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1626-1653 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 32, 266, 267, 268, or 269. In certain such embodiments, an antisense compound targeted to nucleotides 1626-1653 of SEQ ID NO: 3 is selected from ISIS NOs: 395168, 399890, 405909, 405910, 405911, or 406045.
In certain embodiments, a target region is nucleotides 1684-1703 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1684-1703 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 273. In certain such embodiments, an antisense compound targeted to nucleotides 1684-1703 of SEQ ID NO: 3 is selected from ISIS NOs: 410763.
In certain embodiments, a target region is nucleotides 1730-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1730-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 33, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1730-1781 of SEQ ID NO: 3 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, 405916, 405917, 405918, 405919, 405920, 405921, 405922, 410734, or 410764.
In certain embodiments, a target region is nucleotides 1740-1767 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1740-1767 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 33, 275, 276, 277, or 278. In certain such embodiments, an antisense compound targeted to nucleotides 1740-1767 of SEQ ID NO: 3 is selected from ISIS NOs: 395169, 399891, 405913, 405914, 405915, or 405916.
In certain embodiments, a target region is nucleotides 1749-1775 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1749-1775 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 279, 280, 281 or 282. In certain such embodiments, an antisense compound targeted to nucleotides 1749-1775 of SEQ ID NO: 3 is selected from ISIS NOs: 410734, 405917, 405918 or 405919.
In certain embodiments, a target region is nucleotides 1758-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1758-1781 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 283, 284, or 285. In certain such embodiments, an antisense compound targeted to nucleotides 1758-1781 of SEQ ID NO: 3 is selected from ISIS NOs: 405920, 405921, or 405922.
In certain embodiments, a target region is nucleotides 1820-1847 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1820-1847 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 306, 307, 308, or 309. In certain such embodiments, an antisense compound targeted to nucleotides 1820-1847 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399897, 405937, 405938, 405939, or 405940.
In certain embodiments, a target region is nucleotides 1820-1877 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1820-1877 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 40, 41, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, or 318. In certain such embodiments, an antisense compound targeted to nucleotides 1820-1877 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399804, 399805, 399897, 399960, 399961, 405937, 405938, 405939, 405940, 405941, 405942, 405943, 405944, 405945, 405946, 405947, 410737, or 410769.
In certain embodiments, a target region is nucleotides 1822-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1822-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 39, 307, 308, 309, 310, 40, 311, 312, 313, 314, 315, 41, 41, 316, 317, 318, 101, 319, 42, 320, 321, 322, 43, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 3, or 366. In certain such embodiments, an antisense compound targeted to nucleotides 1822-2198 of SEQ ID NO: 3 is selected from ISIS NOs: 395175, 399897, 405938, 405939, 405940, 405941, 399804, 399960, 405942, 405943, 410737, 405944, 405945, 399805, 399961, 405946, 405947, 410769, 395176, 399898, 405948, 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, or 405979.
In certain embodiments, a target region is nucleotides 1830-1856 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1830-1856 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 40, 310, 311, or 312. In certain such embodiments, an antisense compound targeted to nucleotides 1830-1856 of SEQ ID NO: 3 is selected from ISIS NOs: 399804, 399960, 405941, 405942, or 405943.
In certain embodiments, a target region is nucleotides 1839-1865 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1839-1865 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 41, 313, 314, 315, or 316. In certain such embodiments, an antisense compound targeted to nucleotides 1839-1865 of SEQ ID NO: 3 is selected from ISIS NOs: 399805, 399961, 405944, 405945, 405946, or 410737.
In certain embodiments, a target region is nucleotides 1840-1867 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1840-1867 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 314, 315, 316, or 317. In certain such embodiments, an antisense compound targeted to nucleotides 1840-1867 of SEQ ID NO: 3 is selected from ISIS NOs: 399961, 405944, 405945, 405946, 410737, or 405947.
In certain embodiments, a target region is nucleotides 1898-1924 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1898-1924 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 101, 319, or 320. In certain such embodiments, an antisense compound targeted to nucleotides 1898-1924 of SEQ ID NO: 3 is selected from ISIS NOs: 395176, 399898, 405948, 399806, 399962, 405949, or 405949.
In certain embodiments, a target region is nucleotides 1898-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1898-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 87, 101, 319, 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 1898-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395176, 395177, 395178, 395179, 399806, 399807, 399808, 399809, 399810, 399811, 399812, 399813, 399898, 399899, 399900, 399901, 399962, 399963, 399964, 399965, 399966, 399967, 399968, 399969, 405885, 405886, 405887, 405888, 405889, 405890, 405891, 405892, 405948, 405949, 405950, 405951, 405952, 405953, 405954, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 405962, 405963, 405964, 405965, 405966, 405967, 405968, 405969, 405970, 405971, 405972, 405973, 405974, 408653, 410540, 410596, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410604, 410672, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, 410681, 410682, 410738, 410739, 410740, or 410770.
In certain embodiments, a target region is nucleotides 1903-2127 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1903-2127 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 42, 320, 321, 322, 43, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, 51, 119, 360, or 54. In certain such embodiments, an antisense compound targeted to nucleotides 1903-2127 of SEQ ID NO: 3 is selected from ISIS NOs: 399806, 399962, 405949, 405950, 405951, 399807, 399963, 405952, 410738, 405953, 405954, 410770, 405955, 405956, 405957, 405958, 405959, 405960, 405961, 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, or 399967.
In certain embodiments, a target region is nucleotides 1907-1934 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1907-1934 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 321, 322, 323, or 324. In certain such embodiments, an antisense compound targeted to nucleotides 1907-1934 of SEQ ID NO: 3 is selected from ISIS NOs: 405950, 405951, 399807, 399963, 405952, or 410738.
In certain embodiments, a target region is nucleotides 1911-1938 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1911-1938 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 43, 323, 324, 325, or 326. In certain such embodiments, an antisense compound targeted to nucleotides 1911-1938 of SEQ ID NO: 3 is selected from ISIS NOs: 399807, 399963, 405952, 410738, 405953, or 405954.
In certain embodiments, a target region is nucleotides 1946-1971 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1946-1971 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 328, 329, 330, or 331. In certain such embodiments, an antisense compound targeted to nucleotides 1946-1971 of SEQ ID NO: 3 is selected from ISIS NOs: 405955, 405956, 405957, or 405958.
In certain embodiments, a target region is nucleotides 1954-1980 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1954-1980 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 332, 333, 334, or 335. In certain such embodiments, an antisense compound targeted to nucleotides 1954-1980 of SEQ ID NO: 3 is selected from ISIS NOs: 405959, 405960, 405961, 399808, 399964, or 405962.
In certain embodiments, a target region is nucleotides 1959-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1959-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, or 51. In certain such embodiments, an antisense compound targeted to nucleotides 1959-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399969.
In certain embodiments, a target region is nucleotides 1959-2057 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1959-2057 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 44, 335, 336, 337, 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359, 51, or 119. In certain such embodiments, an antisense compound targeted to nucleotides 1959-2057 of SEQ ID NO: 3 is selected from ISIS NOs: 399808, 399964, 405962, 410739, 405963, 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, or 399902.
In certain embodiments, a target region is nucleotides 1963-1988 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1963-1988 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 336, 337 or 338. In certain such embodiments, an antisense compound targeted to nucleotides 1963-1988 of SEQ ID NO: 3 is selected from ISIS NOs: 410739, 405963, 395177, 399899, or 405964.
In certain embodiments, a target region is nucleotides 1967-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1967-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 45, 338, 46, 339, 340, 47, 341, 342, 48, 343, 344, 345, 346, 347, 348, 349, 49, 350, 351, 352, 353, 50, 354, 355, 356, 357, 87, 358, 359 or 51. In certain such embodiments, an antisense compound targeted to nucleotides 1967-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395177, 399899, 405964, 399809, 399965, 405965, 405966, 399810, 399966, 405967, 405968, 399811, 399967, 405969, 405970, 410740, 405885, 405886, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, or 399969.
In certain embodiments, a target region is nucleotides 1972-1999 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1972-1999 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 46, 47, 339, 340, or 341. In certain such embodiments, an antisense compound targeted to nucleotides 1972-1999 of SEQ ID NO: 3 is selected from ISIS NOs: 399809, 399965, 405965, 405966, 399810, 399966, or 405967.
In certain embodiments, a target region is nucleotides 1982-2008 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1982-2008 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 48, 342, 343, or 344. In certain such embodiments, an antisense compound targeted to nucleotides 1982-2008 of SEQ ID NO: 3 is selected from ISIS NOs: 405968, 399811, 399967, 405969, or 405970.
In certain embodiments, a target region is nucleotides 1991-2018 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1991-2018 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 345, 346, 347, 348, 349, 350, or 351. In certain such embodiments, an antisense compound targeted to nucleotides 1991-2018 of SEQ ID NO: 3 is selected from ISIS NOs: 399812, 399968, 405885, 405886, 405887, 405888, 405889, 405890, 410596, 410597, 410598, 410599, 410672, 410673, 410674, 410675, 410676, or 410740.
In certain embodiments, a target region is nucleotides 1993-2019 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1993-2019 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 346, 347, 348, 349, 350, or 354. In certain such embodiments, an antisense compound targeted to nucleotides 1993-2019 of SEQ ID NO: 3 is selected from ISIS NOs: 405885, 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, or 410677.
In certain embodiments, a target region is nucleotides 1995-2022 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1995-2022 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 348, 349, 350, 351, 352, 353, 50, or 351 In certain such embodiments, an antisense compound targeted to nucleotides 1995-2022 of SEQ ID NO: 3 is selected from ISIS NOs: 405887, 410596, 410672, 405888, 410597, 410673, 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, or 410680.
In certain embodiments, a target region is nucleotides 1996-2023 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1996-2023 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 349, 350, 351, 352, 353, 354, or 355. In certain such embodiments, an antisense compound targeted to nucleotides 1996-2023 of SEQ ID NO: 3 is selected from ISIS NOs: 395178, 399812, 399900, 399968, 405888, 405889, 405890, 405891, 405892, 405971, 408653, 410597, 410598, 410599, 410600, 410601, 410602, 410603, 410673, 410674, 410675, 410676, 410677, 410678, 410679, 410680, or 410681.
In certain embodiments, a target region is nucleotides 1997-2024 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1997-2024 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 49, 50, 350, 351, 352, 353, 354, 355, or 356. In certain such embodiments, an antisense compound targeted to nucleotides 1997-2024 of SEQ ID NO: 3 is selected from ISIS NOs: 399812, 399968, 410674, 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, or 410682.
In certain embodiments, a target region is nucleotides 1998-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1998-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 350, 351, 352, 353, 354, 355, 356 or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1998-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 1999-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 1999-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 50, 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 1999-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405889, 410598, 410675, 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 2000-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2000-2025 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 351, 352, 353, 354, 355, 356, or 357. In certain such embodiments, an antisense compound targeted to nucleotides 2000-2025 of SEQ ID NO: 3 is selected from ISIS NOs: 405890, 410599, 410676, 405891, 410600, 410677, 405892, 410601, 410678, 395178, 399900, 410679, 408653, 410602, 410680, 405971, 410603, 410681, 410540, 410604, 410682, or 405972.
In certain embodiments, a target region is nucleotides 2009-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2009-2035 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 51, 87, 358, or 359. In certain such embodiments, an antisense compound targeted to nucleotides 2009-2035 of SEQ ID NO: 3 is selected from ISIS NOs: 395179, 399813, 399901, 399969, 405973, or 405974.
In certain embodiments, a target region is nucleotides 2038-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2038-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 119, 130, 360, 361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2038-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 395180, 395181, 395182, 395220, 399814, 399836, 399902, 399903, 399904, 399942, 399970, 399992, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741, or 410771.
In certain embodiments, a target region is nucleotides 2061-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2061-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 54, 104, 130, 360, 361, 362, 363, 365, 366, 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2061-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 395181, 395182, 395220, 399814, 399836, 399903, 399904, 399942, 399970, 399992, 405975, 405976, 405977, 405978, 405979, 405980, 405981, 405982, 405983, 410741, or 410771.
In certain embodiments, a target region is nucleotides 2073-2099 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2073-2099 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 52, 53, 361, or 362. In certain such embodiments, an antisense compound targeted to nucleotides 2073-2099 of SEQ ID NO: 3 is selected from ISIS NOs: 395181, 399814, 399903, 399970, 405975, or 405976.
In certain embodiments, a target region is nucleotides 2078-2104 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2078-2104 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 53, 104, 362, or 363. In certain such embodiments, an antisense compound targeted to nucleotides 2078-2104 of SEQ ID NO: 3 is selected from ISIS NOs: 395220, 399814, 399942, 399970, 405976, or 405977.
In certain embodiments, a target region is nucleotides 2105-2131 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2105-2131 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 54, 365, 366, or 367. In certain such embodiments, an antisense compound targeted to nucleotides 2105-2131 of SEQ ID NO: 3 is selected from ISIS NOs: 395182, 399904, 405978, 405979, or 405980.
In certain embodiments, a target region is nucleotides 2112-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2112-2139 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 367, 368, 369, 370, or 371. In certain such embodiments, an antisense compound targeted to nucleotides 2112-2139 of SEQ ID NO: 3 is selected from ISIS NOs: 405980, 405981, 410741, 405982, or 405983.
In certain embodiments, a target region is nucleotides 2168-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2168-2198 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 56, 372, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 2168-2198 of SEQ ID NO: 3 is selected from ISIS NOs: 399815, 399816, 399971, 399972, 405641, 405984, 405985, 405986, or 405987.
In certain embodiments, a target region is nucleotides 2170-2177 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2170-2177 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 55, 373, 374, or 375. In certain such embodiments, an antisense compound targeted to nucleotides 2170-2177 of SEQ ID NO: 3 is selected from ISIS NOs: 399815, 399971, 405641, 405985, 405986, or 405987.
In certain embodiments, a target region is nucleotides 2245-2284 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2245-2284 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 408 or 409. In certain such embodiments, an antisense compound targeted to nucleotides 2245-2284 of SEQ ID NO: 3 is selected from ISIS NOs: 410776 or 410777.
In certain embodiments, a target region is nucleotides 2295-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2295-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 410, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2295-2394 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, 410778, or 410779.
In certain embodiments, a target region is nucleotides 2355-2381 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2355-2381 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, or 415. In certain such embodiments, an antisense compound targeted to nucleotides 2355-2381 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, or 410718.
In certain embodiments, a target region is nucleotides 2355-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2355-2394 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 60, 61, 411, 412, 413, 414, 415, or 416. In certain such embodiments, an antisense compound targeted to nucleotides 2355-2394 of SEQ ID NO: 3 is selected from ISIS NOs: 395186, 399817, 399908, 399973, 405996, 405997, 410562, 410563, 410564, 410633, 410634, 410635, 410636, 410713, 410714, 410715, 410716, 410717, 410718, or 410779.
In certain embodiments, a target region is nucleotides 2405-2461 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2405-2461 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 417 or 418. In certain such embodiments, an antisense compound targeted to nucleotides 2405-2461 of SEQ ID NO: 3 is selected from ISIS NOs: 410780 or 410781.
In certain embodiments, a target region is nucleotides 2560-2587 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2560-2587 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, or 426. In certain such embodiments, an antisense compound targeted to nucleotides 2560-2587 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, or 410727.
In certain embodiments, a target region is nucleotides 2560-2609 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2560-2609 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, or 430. In certain such embodiments, an antisense compound targeted to nucleotides 2560-2609 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410565, 410566, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410637, 410638, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410719, 410720, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, or 410783.
In certain embodiments, a target region is nucleotides 2561-2588 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2561-2588 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 420, 421, 422, 423, 424, 425, 426, 427, or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2561-2588 of SEQ ID NO: 3 is selected from ISIS NOs: 410566, 410638, 410720, 410567, 410639, 410721, 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, or 410728.
In certain embodiments, a target region is nucleotides 2562-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2562-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 62, 421, 422, 423, 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2562-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 405998, 410567, 410568, 410569, 410570, 410571, 410572, 410573, 410639, 410640, 410641, 410642, 410643, 410644, 410645, 410646, 410721, 410722, 410723, 410724, 410725, 410726, 410727, 410728, or 410729.
In certain embodiments, a target region is nucleotides 2563-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2563-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 422, 423, 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2563-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410568, 410640, 410722, 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2564-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2564-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 423, 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2564-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410569, 410641, 410723, 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2565-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2565-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, 428 or 62. In certain such embodiments, an antisense compound targeted to nucleotides 2565-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 395187, 399909, 410724, 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2566-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2566-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 424, 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2566-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410570, 410642, 410725, 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2567-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2567-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 425, 426, 427, or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2567-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 410571, 410643, 410726, 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2568-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2568-2589 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 426, 427 or 428. In certain such embodiments, an antisense compound targeted to nucleotides 2568-2589 of SEQ ID NO: 3 is selected from ISIS NOs: 405988, 405998, 410644, 410727, 410572, 410645, 410728, 410573, 410646, or 410729.
In certain embodiments, a target region is nucleotides 2665-2689 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2665-2689 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 63 or 154. In certain such embodiments, an antisense compound targeted to nucleotides 2665-2689 of SEQ ID NO: 3 is selected from ISIS NOs: 395188, 399910, 399818, or 399974.
In certain embodiments, a target region is nucleotides 2759-2783 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2759-2783 SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 64 or 65. In certain such embodiments, an antisense compound targeted to nucleotides 2759-2783 of SEQ ID NO: 3 is selected from ISIS NOs: 395189, 399911, 399819, or 399975.
In certain embodiments, a target region is nucleotides 2837-2880 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2837-2880 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 66, 67, or 122. In certain such embodiments, an antisense compound targeted to nucleotides 2837-2880 of SEQ ID NO: 3 is selected from ISIS NOs: 399820, 399976, 395190, 399912, 395191, or 399913.
In certain embodiments, a target region is nucleotides 2904-2923 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 2904-2923 SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 153. In certain such embodiments, an antisense compound targeted to nucleotides 2904-2923 of SEQ ID NO: 3 is selected from ISIS NOs: 395192 or 399914.
In certain embodiments, a target region is nucleotides 3005-3024 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3005-3024 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68. In certain such embodiments, an antisense compound targeted to nucleotides 3005-3024 of SEQ ID NO: 3 is selected from ISIS NOs: 395193 or 399915.
In certain embodiments, a target region is nucleotides 3005-3174 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3005-3174 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 68, 431, 432, 433, 434, 69, 435, 436, 437, 438 or 70. In certain such embodiments, an antisense compound targeted to nucleotides 3005-3174 of SEQ ID NO: 3 is selected from ISIS NOs: 395193, 399915, 405893, 405894, 405895, 405896, 395194, 399916, 405897, 405898, 405899, 405900, 395195, or 399917.
In certain embodiments, a target region is nucleotides 3083-3110 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3083-3110 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 69, 69, 431, 432, 433, 434, 435, 436, 437, or 438. In certain such embodiments, an antisense compound targeted to nucleotides 3083-3110 of SEQ ID NO: 3 is selected from ISIS NOs: 395194, 399916, 405893, 405894, 405895, 405896, 405897, 405898, 405899, or 405900.
In certain embodiments, a target region is nucleotides 3155-3184 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3155-3184 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 70, 71, 439, 440, 441, 442, 443, 444, 445, or 446. In certain such embodiments, an antisense compound targeted to nucleotides 3155-3184 of SEQ ID NO: 3 is selected from ISIS NOs: 395195, 399821, 399917, 399977, 405901, 405902, 405903, 405904, 405905, 405906, 405907, or 405908.
In certain embodiments, a target region is nucleotides 3238-3268 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3238-3268 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 72, 73, or 135. In certain such embodiments, an antisense compound targeted to nucleotides 3238-3268 of SEQ ID NO: 3 is selected from ISIS NOs: 395196, 399822, 399823, 399918, 399978, or 399979.
In certain embodiments, a target region is nucleotides 3482-3711 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3482-3711 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 74, 75, or 112. In certain such embodiments, an antisense compound targeted to nucleotides 3482-3711 of SEQ ID NO: 3 is selected from ISIS NOs: 395197, 395198, 399824, 399919, 399920, or 399980.
In certain embodiments, a target region is nucleotides 3727-3751 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3727-3751 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 76 or 77. In certain such embodiments, an antisense compound targeted to nucleotides 3727-3751 of SEQ ID NO: 3 is selected from ISIS NOs: 395199, 399921, 399825, or 399981.
In certain embodiments, a target region is nucleotides 3798-3824 of SEQ ID NO: 3. In certain embodiments, an antisense compound is targeted to nucleotides 3798-3824 of SEQ ID NO: 3. In certain embodiments, an antisense compound targeted to a PCSK9 nucleic acid comprises a nucleotide sequence selected from SEQ ID NOs: 78 or 99. In certain such embodiments, an antisense compound targeted to nucleotides 3798-3824 of SEQ ID NO: 3 is selected from ISIS NOs: 395200, 399922, 399826, or 399982.
In certain embodiments, short gapmer antisense compounds are targeted to a PCSK9 nucleic acid. In certain such embodiments, gapmer antisense compounds are targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3. In certain such embodiments, the nucleotide sequences illustrated in Table 15.1 have a 2-10-2 gapmer motif. Table 15.1 illustrates gapmer antisense compounds targeted to SEQ ID NO: 1, SEQ ID NO: 2 and/or SEQ ID NO: 3, having a 2-10-2 motif, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises nucleotides comprising a 2′-O-methoxyethyl sugar modification. Internucleoside linkages are phosphorthioate, and cytidines are 5-methylcytidines.







TABLE 15.1







Gapmer antisense compounds having a 2-10-2 


motif targeted to SEQ ID NO: 1, SEQ ID  


NO: 2 and/or SEQ ID NO: 3

















5′
3′







Target
Target



Target



Site
Site



SEQ



on
on
SEQ


ID
ISIS


SEQ ID
SEQ ID
ID


NO
NO
Motif
OligoSeq
NO: 1
NO: 1
NO





1
406539
2-10-2
GCCTATGAGGGTGC
 1079
 1092
458


 


1
406540
2-10-2
TCCAGGCCTATGAG
 1084
 1097
459


 


1
406551
2-10-2
CAGCTCAGCAGCTC
 1735
 1748
460


 


1
406595
2-10-2
TTAATCAGGGAGCC
 2877
 2890
461


 


2
406551
2-10-2
CAGCTCAGCAGCTC
21181
21194
460


 


2
406595
2-10-2
TTAATCAGGGAGCC
26684
26697
461


 


3
406539
2-10-2
GCCTATGAGGGTGC
 1609
 1622
458


 


3
406540
2-10-2
TCCAGGCCTATGAG
 1614
 1627
459


 


3
406551
2-10-2
CAGCTCAGCAGCTC
 2168
 2181
460


 


3
406595
2-10-2
TTAATCAGGGAGCC
 3132
 3145
461









In a certain embodiment, a short antisense compound has increased potency compared to a 20 nucleotide compound. In a certain embodiment such short antisense compound and 20 nucleotide compound target the same region. In a certain embodiment, a short antisense compound has increased potency compared to a 20 nucleotide compound containing the sequence of the short antisense compound.
In certain embodiments, the desired effect is a reduction in mRNA target nucleic acid levels. In other embodiments, the desired effect is reduction of levels of protein encoded by the target nucleic acid or a phenotypic change associated with the target nucleic acid.
In one embodiment, a target region is a structurally defined region of the nucleic acid. For example, a target region may encompass a 3′ UTR, a 5′ UTR, an exon, an intron, a coding region, a translation initiation region, translation termination region, or other defined nucleic acid region. In other embodiments, a target region may encompass the sequence from a 5′ target site of one target segment within the target region to a 3′ target site of another target segment within the target region.
Targeting includes determination of at least one target segment to which an antisense compound hybridizes, such that a desired effect occurs. A target region may contain one or more target segments. Multiple target segments within a target region may be overlapping. Alternatively, they may be non-overlapping. In one embodiment, target segments within a target region are separated by no more than about 10 nucleotides on the target nucleic acid. In another embodiment, target segments within a target region are separated by no more than about 5 nucleotides on the target nucleic acid. In additional embodiments, target segments are contiguous.
Suitable target segments may be found within a 5′ UTR, a coding region, a 3′ UTR, an intron, or an exon. Target segments containing a start codon or a stop codon are also suitable target segments.
The determination of suitable target segments may include a comparison of the sequence of a target nucleic acid to other sequences throughout the genome. For example, the BLAST algorithm may be used to identify regions of similarity amongst different nucleic acids. This comparison can prevent the selection of antisense compound sequences that may hybridize in a non-specific manner to sequences other than a selected target nucleic acid (i.e., non-target or off-target sequences).
There may be variation in activity (e.g., as defined by percent reduction of target nucleic acid levels) of the antisense compounds within an active target region. In one embodiment, reductions in PCSK9 mRNA levels are indicative of inhibition of PCSK9 expression. Reductions in levels of a PCSK9 protein are also indicative of inhibition of target mRNA expression. Further, phenotypic changes are indicative of inhibition of PCSK9 expression. For example, a decrease in LDL-C levels is indicative of inhibition of PCSK9 expression.
Hybridization
For example, hybridization may occur between an antisense compound disclosed herein and a PCSK9 nucleic acid. The most common mechanism of hybridization involves hydrogen bonding (e.g., Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding) between complementary nucleobases of the nucleic acid molecules.
Hybridization can occur under varying conditions. Stringent conditions are sequence-dependent and are determined by the nature and composition of the nucleic acid molecules to be hybridized.
Methods of determining whether a sequence is specifically hybridizable to a target nucleic acid are well known in the art. In one embodiment, the antisense compounds provided herein are specifically hybridizable with a PCSK9 nucleic acid.
The PCSK9 sequences targeted by some of the preferred antisense compounds disclosed in Table 1 are shown in FIG. 1. The antisense compounds are labeled with the corresponding Isis number. The bar in FIG. 1 labeled “NM_174936.2” displays the relative position of the target sequences within GENBANK® Accession No. NM_174936.2, first deposited with GENBANK® on Jun. 1, 2003 (SEQ ID NO: 1). The bar below that shows the corresponding structural elements of the PCSK9 mRNA, where “CDS” is the protein-encoding region of the mRNA and “UTRs” are the 5′ and 3′ untranslated regions. As shown in FIG. 1, the sequences targeted by the present antisense compounds are found in the protein-encoding region of the PCSK9 mRNA.
Complementarity
An antisense compound and a target nucleic acid are complementary to each other when a sufficient number of nucleobases of the antisense compound can hydrogen bond with the corresponding nucleobases of the target nucleic acid, such that a desired effect will occur (e.g., antisense inhibition of a target nucleic acid, such as a PCSK9 nucleic acid).
Non-complementary nucleobases between an antisense compound and a PCSK9 nucleic acid may be tolerated provided that the antisense compound remains able to specifically hybridize to a target nucleic acid. Moreover, an antisense compound may hybridize over one or more segments of a PCSK9 nucleic acid such that intervening or adjacent segments are not involved in the hybridization event (e.g., a loop structure, mismatch or hairpin structure).
In some embodiments, the antisense compounds provided herein are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% complementary to a PCSK9 nucleic acid. Percent complementarity of an antisense compound with a target nucleic acid can be determined using routine methods.
In other embodiments, the antisense compounds provided herein are fully complementary (i.e, 100% complementary) to a target nucleic acid. For example, antisense compound may be fully complementary to a PCSK9 nucleic acid. As used herein, “fully complementary” means each nucleobase of an antisense compound is capable of precise base pairing with the corresponding nucleobases of a target nucleic acid.
The location of a non-complementary nucleobase may be at the 5′ end or 3′ end of the antisense compound. Alternatively, the non-complementary nucleobase or nucleobases may be at an internal position of the antisense compound. When two or more non-complementary nucleobases are present, they may be contiguous (i.e., linked) or non-contiguous. In one embodiment, a non-complementary nucleobase is located in the wing segment of a gapmer antisense oligonucleotide.
In one embodiment, antisense compounds up to 20 nucleobases in length comprise no more than 4, no more than 3, no more than 2 or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a PCSK9 nucleic acid.
In another embodiment, antisense compounds up to 30 nucleobases in length comprise no more than 6, no more than 5, no more than 4, no more than 3, no more than 2 or no more than 1 non-complementary nucleobase(s) relative to a target nucleic acid, such as a PCSK9 nucleic acid.
The antisense compounds provided herein also include those which are complementary to a portion of a target nucleic acid. As used herein, “portion” refers to a defined number of contiguous (i.e., linked) nucleobases within a region or segment of a target nucleic acid. A “portion” can also refer to a defined number of contiguous nucleobases of an antisense compound. In one embodiment, the antisense compounds are complementary to at least an 8 nucleobase portion of a target segment. In another embodiment, the antisense compounds are complementary to at least a 12 nucleobase portion of a target segment. In yet another embodiment, the antisense compounds are complementary to at least a 15 nucleobase portion of a target segment.
Identity
The antisense compounds provided herein may also have a defined percent identity to a particular nucleotide sequence, SEQ ID NO, or compound represented by a specific Isis number. As used herein, an antisense compound is identical to the sequence disclosed herein if it has the same nucleobase pairing ability. For example, a RNA which contains uracil in place of thymidine in a disclosed DNA sequence would be considered identical to the DNA sequence since both uracil and thymidine pair with adenine. Shortened and lengthened versions of the antisense compounds described herein as well as compounds having non-identical bases relative to the antisense compounds provided herein also are contemplated. The non-identical bases may be adjacent to each other or dispersed throughout the antisense compound. Percent identity of an antisense compound is calculated according to the number of bases that have identical base pairing relative to the sequence to which it is being compared.
In one embodiment, the antisense compounds are at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% identical to one or more of the antisense compounds disclosed herein.
Modifications
A nucleoside is a base-sugar combination. The nucleobase (also known as base) portion of the nucleoside is normally a heterocyclic base moiety. Nucleotides are nucleosides that further include a phosphate group covalently linked to the sugar portion of the nucleoside. For those nucleosides that include a pentofuranosyl sugar, the phosphate group can be linked to the 2′, 3′ or 5′ hydroxyl moiety of the sugar. Oligonucleotides are formed through the covalent linkage of adjacent nucleosides to one another, to form a linear polymeric oligonucleotide. Within the oligonucleotide structure, the phosphate groups are commonly referred to as forming the internucleoside linkages of the oligonucleotide.
Modifications to antisense compounds encompass substitutions or changes to internucleoside linkages, sugar moieties, or nucleobases. Modified antisense compounds are often preferred over native forms because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for nucleic acid target, increased stability in the presence of nucleases, or increased inhibitory activity.
Chemically modified nucleosides may also be employed to increase the binding affinity of a shortened or truncated antisense oligonucleotide for its target nucleic acid. Consequently, comparable results can often be obtained with shorter antisense compounds that have such chemically modified nucleosides.
Modified Internucleoside Linkages
The naturally occurring internucleoside linkage of RNA and DNA is a 3′ to 5′ phosphodiester linkage. Antisense compounds having one or more modified, i.e., non-naturally occurring, internucleoside linkages are often selected over antisense compounds having naturally occurring internucleoside linkages because of desirable properties such as, for example, enhanced cellular uptake, enhanced affinity for target nucleic acids, and increased stability in the presence of nucleases.
Oligonucleotides having modified internucleoside linkages include internucleoside linkages that retain a phosphorus atom as well as internucleoside linkages that do not have a phosphorus atom. Representative phosphorus containing internucleoside linkages include, but are not limited to, phosphodiesters, phosphotriesters, methylphosphonates, phosphoramidate, and phosphorothioates. Methods of preparation of phosphorous-containing and non-phosphorous-containing linkages are well known.
In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more modified internucleoside linkages. In some embodiments, the modified internucleoside linkages are phosphorothioate linkages. In other embodiments, each internucleoside linkage of an antisense compound is a phosphorothioate internucleoside linkage.
Modified Sugar Moieties
Antisense compounds targeted to a PCSK9 nucleic acid may contain one or more nucleotides having modified sugar moieties. Sugar modifications may impart nuclease stability, binding affinity or some other beneficial biological property to the antisense compounds. The furanosyl sugar ring of a nucleoside can be modified in a number of ways including, but not limited to: addition of a substituent group, particularly at the 2′ position; bridging of two non-geminal ring atoms to form a bicyclic nucleic acid (BNA); and substitution of an atom or group such as —S—, —N(R)— or —C(R1)(R2) (R═H, C1-C12 alkyl or a protecting group) for the ring oxygen at the 4′-position and combinations of these such as for example a 2′-F-5′-methyl substituted nucleoside (see WO 2008/101157, published Aug. 21, 2008, for other disclosed 5′,2′-bis substituted nucleosides) or replacement of the ribosyl ring oxygen atom with S with further substitution at the 2′-position (see U.S. Published Application No. 2005/0130923, published on Jun. 16, 2005) or alternatively 5′-substitution of a BNA (see WO 2007/134181, published Nov. 22, 2007, wherein LNA is substituted with for example a 5′-methyl or a 5′-vinyl group).
Modified sugars include, but are not limited to: substituted sugars, especially 2′-substituted sugars having a 2′-F, 2′-OCH3 (2′-OMe) or a 2′-O(CH2)2—OCH3 (2′-O-methoxyethyl or 2′-MOE) substituent group; and bicyclic modified sugars (BNAs), having a 4′-(CH2)n—O-2′ bridge, where n=1 or n=2, including α-L-methyleneoxy (4′-CH2-O-2′ or 4′-α-L-(CH2)-O-2′ (α-L-LNA)) BNA, β-D-methyleneoxy (4′-CH2-O-2′ or 4′-β-D-(CH2)-O-2′ (β-D-LNA)) BNA and ethyleneoxy (4′-(CH2)2-O-2′ (ENA)) BNA. BNAs also include 4′-(CH2)-S-2′ and 4′-C(CH3)2-O-2′ (see International Application No. PCT/US2008/068922); 4′-CH(CH3)-O-2′ and 4′-CH(CH2OCH3)-O-2′ (see U.S. Pat. No. 7,399,845, issued Jul. 15, 2008); 4′-CH2-N(OCH3)-2′ (see International Application No. PCT/US2008/064591); 4′-CH2-O—N(CH3)-2′ (see U.S. Published Application No. 2004/0171570, published Sep. 2, 2004); 4′-CH2-N(R)—O-2′ (see U.S. Pat. No. 7,427,672, issued Sep. 23, 2008); 4′-CH2-C(CH3)-2′ and 4′-CH2-C(═CH2)-2′ (see International Application No. PCT/US2008/066154); and wherein R is, independently, H, C1-C12 alkyl, or a protecting group. Bicyclic modified sugars also include (6′S)-6′methyl BNA, aminooxy (4′-CH2-O—N(R)-2′) BNA, oxyamino (4′-CH2-N(R)—O-2′) BNA wherein R is, independently, H, a protecting group or C1-C12 alkyl. Examples of nucleosides having modified sugar moieties also include without limitation nucleosides comprising 5′-vinyl, 5′-methyl (R or S) and 4′-S.
The substituent at the 2′ position can also be selected from allyl, amino, azido, thio, O-allyl, O—C1-C10 alkyl, OCF3, O(CH2)2SCH3, O(CH2)2—O—N(Rm)(Rn), and O—CH2—C(═O)—N(Rm)(Rn), where each Rm and Rn is, independently, H or substituted or unsubstituted C1-C10 alkyl.
In certain embodiments, such BNA modified nucleotides are high-affinity nucleotides and their incorporation into antisense compounds allows for increased potency and improved therapeutic index. Methods for the preparations of modified sugars are well known to those skilled in the art.
In certain embodiments, nucleosides are modified by substitution of the ribosyl ring with a sugar surrogate. Such modification includes without limitation, replacement of the ribosyl ring with a surrogate ring system (sometimes referred to as DNA analogs) such as a morpholino ring, a cyclohexenyl ring, a cyclohexyl ring or a tetrahydropyranyl ring such as one having one of the formula:




Many other bicyclo and tricyclo sugar surrogate ring systems are also know in the art that can be used to modify nucleosides for incorporation into antisense compounds. Such ring systems can undergo various additional substitutions to enhance activity.
Methods for the preparations of modified sugars are well known to those skilled in the art.
In nucleotides having modified sugar moieties (including substitution of a sugar surrogate), the nucleobase moieties (natural, modified or a combination thereof) are maintained for hybridization with an appropriate nucleic acid target.
In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more nucleotides having modified sugar moieties. In a suitable embodiment, the modified sugar moiety is 2′-MOE. In other embodiments, the 2′-MOE modified nucleotides are arranged in a gapmer motif.
Modified Nucleobases
Nucleobase (or base) modifications or substitutions are structurally distinguishable from, yet functionally interchangeable with, naturally occurring or synthetic unmodified nucleobases. Both natural and modified nucleobases are capable of participating in hydrogen bonding. Such nucleobase modifications may impart nuclease stability, binding affinity or some other beneficial biological property to antisense compounds. Modified nucleobases include synthetic and natural nucleobases such as, for example, 5-methylcytosine (5-me-C). Certain nucleobase substitutions, including 5-methylcytosine substitutions, are particularly useful for increasing the binding affinity of an antisense compound for a target nucleic acid. For example, 5-methylcytosine substitutions have been shown to increase nucleic acid duplex stability by 0.6-1.2° C. (Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., eds., Antisense Research and Applications, CRC Press, Boca Raton, 1993, pp. 276-278).
Additional unmodified nucleobases include 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiouracil, 2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine, 5-propynyl (—C≡C—CH3) uracil and cytosine and other alkynyl derivatives of pyrimidine bases, 6-azo uracil, cytosine and thymine, 5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol, 8-thioalkyl, 8-hydroxyl and other 8-substituted adenines and guanines, 5-halo particularly 5-bromo, 5-trifluoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 2-F-adenine, 2-amino-adenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and 7-deazaadenine and 3-deazaguanine and 3-deazaadenine.
Heterocyclic base moieties may also include those in which the purine or pyrimidine base is replaced with other heterocycles, for example 7-deaza-adenine, 7-deazaguanosine, 2-aminopyridine and 2-pyridone. Nucleobases that are particularly useful for increasing the binding affinity of antisense compounds include 5-substituted pyrimidines, 6-azapyrimidines and N-2, N-6 and 0-6 substituted purines, including 2 aminopropyladenine, 5-propynyluracil and 5-propynylcytosine.
In one embodiment, antisense compounds targeted to a PCSK9 nucleic acid comprise one or more modified nucleobases. In an additional embodiment, gap-widened antisense oligonucleotides targeted to a PCSK9 nucleic acid comprise one or more modified nucleobases. In some embodiments, the modified nucleobase is 5-methylcytosine. In further embodiments, each cytosine is a 5-methylcytosine.
Compositions and Methods for Formulating Pharmaceutical Compositions
Antisense oligonucleotides may be admixed with pharmaceutically acceptable active and/or inert substances for the preparation of pharmaceutical compositions or formulations. Compositions and methods for the formulation of pharmaceutical compositions are dependent upon a number of criteria, including, but not limited to, route of administration, extent of disease, or dose to be administered.
Antisense compound targeted to a PCSK9 nucleic acid can be utilized in pharmaceutical compositions by combining the antisense compound with a suitable pharmaceutically acceptable diluent or carrier. A pharmaceutically acceptable diluent includes phosphate-buffered saline (PBS). PBS is a diluent suitable for use in compositions to be delivered parenterally. Accordingly, in one embodiment, employed in the methods described herein is a pharmaceutical composition comprising an antisense compound targeted to a PCSK9 nucleic acid and a pharmaceutically acceptable diluent. In one embodiment, the pharmaceutically acceptable diluent is PBS. In other embodiments, the antisense compound is an antisense oligonucleotide.
Pharmaceutical compositions comprising antisense compounds encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other oligonucleotide which, upon administration to an animal, including a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof. Accordingly, for example, the disclosure is also drawn to pharmaceutically acceptable salts of antisense compounds, prodrugs, pharmaceutically acceptable salts of such prodrugs, and other bioequivalents. Suitable pharmaceutically acceptable salts include, but are not limited to, sodium and potassium salts.
A prodrug can include the incorporation of additional nucleosides at one or both ends of an antisense compound which are cleaved by endogenous nucleases within the body, to form the active antisense compound.
In certain embodiments, pharmaceutical compositions of the present invention comprise one or more oligonucleotides and one or more excipients. In certain such embodiments, excipients are selected from water, salt solutions, alcohol, polyethylene glycols, gelatin, lactose, amylase, magnesium stearate, talc, silicic acid, viscous paraffin, hydroxymethylcellulosem and polyvinylpyrrolidone.
In certain embodiments, a pharmaceutical composition of the present invention is prepared using known techniques, including, but not limited to mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or tabletting processes.
In certain embodiments, a pharmaceutical composition of the present invention is a liquid (e.g., a suspension, elixir and/or solution). In certain of such embodiments, a liquid pharmaceutical composition is prepared using ingredients known in the art, including, but not limited to, water, glycols, oils, alcohols, flavoring agents, preservatives, and coloring agents.
In certain embodiments, a pharmaceutical composition of the present invention is a solid (e.g., a powder, tablet, and/or capsule). In certain of such embodiments, a solid pharmaceutical composition comprising one or more oligonucleotides is prepared using ingredients known in the art, including, but not limited to, starches, sugars, diluents, granulating agents, lubricants, binders, and disintegrating agents.
In certain embodiments, a pharmaceutical composition of the present invention is formulated as a depot preparation. Certain such depot preparations are typically longer acting than non-depot preparations. In certain embodiments, such preparations are administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection. In certain embodiments, depot preparations are prepared using suitable polymeric or hydrophobic materials (for example an emulsion in an acceptable oil) or ion exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
In certain embodiments, a pharmaceutical composition of the present invention comprises a delivery system. Examples of delivery systems include, but are not limited to, liposomes and emulsions. Certain delivery systems are useful for preparing certain pharmaceutical compositions including those comprising hydrophobic compounds. In certain embodiments, certain organic solvents such as dimethylsulfoxide are used.
In certain embodiments, a pharmaceutical composition of the present invention comprises one or more tissue-specific delivery molecules designed to deliver the one or more pharmaceutical agents of the present invention to specific tissues or cell types. For example, in certain embodiments, pharmaceutical compositions include liposomes coated with a tissue-specific antibody.
In certain embodiments, a pharmaceutical composition of the present invention comprises a co-solvent system. Certain of such co-solvent systems comprise, for example, benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. In certain embodiments, such co-solvent systems are used for hydrophobic compounds. A non-limiting example of such a co-solvent system is the VPD co-solvent system, which is a solution of absolute ethanol comprising 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80TM, and 65% w/v polyethylene glycol 300. The proportions of such co-solvent systems may be varied considerably without significantly altering their solubility and toxicity characteristics. Furthermore, the identity of co-solvent components may be varied: for example, other surfactants may be used instead of Polysorbate 80TM; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars or polysaccharides may substitute for dextrose.
In certain embodiments, a pharmaceutical composition of the present invention comprises a sustained-release system. A non-limiting example of such a sustained-release system is a semi-permeable matrix of solid hydrophobic polymers. In certain embodiments, sustained-release systems may, depending on their chemical nature, release pharmaceutical agents over a period of hours, days, weeks or months.
In certain embodiments, a pharmaceutical composition of the present invention is prepared for oral administration. In certain of such embodiments, a pharmaceutical composition is formulated by combining one or more oligonucleotides with one or more pharmaceutically acceptable carriers. Certain of such carriers enable pharmaceutical compositions to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a subject. In certain embodiments, pharmaceutical compositions for oral use are obtained by mixing oligonucleotide and one or more solid excipient. Suitable excipients include, but are not limited to, fillers, such as sugars, including lactose, sucrose, mannitol, or sorbitol; cellulose preparations such as, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum tragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). In certain embodiments, such a mixture is optionally ground and auxiliaries are optionally added. In certain embodiments, pharmaceutical compositions are formed to obtain tablets or dragee cores. In certain embodiments, disintegrating agents (e.g., cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such as sodium alginate) are added.
In certain embodiments, dragee cores are provided with coatings. In certain such embodiments, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to tablets or dragee coatings.
In certain embodiments, pharmaceutical compositions for oral administration are push-fit capsules made of gelatin. Certain of such push-fit capsules comprise one or more pharmaceutical agents of the present invention in admixture with one or more filler such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate and, optionally, stabilizers. In certain embodiments, pharmaceutical compositions for oral administration are soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. In certain soft capsules, one or more pharmaceutical agents of the present invention are be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added.
In certain embodiments, pharmaceutical compositions are prepared for buccal administration. Certain of such pharmaceutical compositions are tablets or lozenges formulated in conventional manner.
In certain embodiments, a pharmaceutical composition is prepared for administration by injection (e.g., intravenous, subcutaneous, intramuscular, etc.). In certain of such embodiments, a pharmaceutical composition comprises a carrier and is formulated in aqueous solution, such as water or physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. In certain embodiments, other ingredients are included (e.g., ingredients that aid in solubility or serve as preservatives). In certain embodiments, injectable suspensions are prepared using appropriate liquid carriers, suspending agents and the like. Certain pharmaceutical compositions for injection are presented in unit dosage form, e.g., in ampoules or in multi-dose containers. Certain pharmaceutical compositions for injection are suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Certain solvents suitable for use in pharmaceutical compositions for injection include, but are not limited to, lipophilic solvents and fatty oils, such as sesame oil, synthetic fatty acid esters, such as ethyl oleate or triglycerides, and liposomes. Aqueous injection suspensions may contain substances that increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, such suspensions may also contain suitable stabilizers or agents that increase the solubility of the pharmaceutical agents to allow for the preparation of highly concentrated solutions.
In certain embodiments, a pharmaceutical composition is prepared for transmucosal administration. In certain of such embodiments penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
In certain embodiments, a pharmaceutical composition is prepared for administration by inhalation. Certain of such pharmaceutical compositions for inhalation are prepared in the form of an aerosol spray in a pressurized pack or a nebulizer. Certain of such pharmaceutical compositions comprise a propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In certain embodiments using a pressurized aerosol, the dosage unit may be determined with a valve that delivers a metered amount. In certain embodiments, capsules and cartridges for use in an inhaler or insufflator may be formulated. Certain of such formulations comprise a powder mixture of a pharmaceutical agent of the invention and a suitable powder base such as lactose or starch.
In certain embodiments, a pharmaceutical composition is prepared for rectal administration, such as a suppositories or retention enema. Certain of such pharmaceutical compositions comprise known ingredients, such as cocoa butter and/or other glycerides.
In certain embodiments, a pharmaceutical composition is prepared for topical administration. Certain of such pharmaceutical compositions comprise bland moisturizing bases, such as ointments or creams. Exemplary suitable ointment bases include, but are not limited to, petrolatum, petrolatum plus volatile silicones, lanolin and water in oil emulsions such as EucerinTM, available from Beiersdorf (Cincinnati, Ohio). Exemplary suitable cream bases include, but are not limited to, NiveaTM Cream, available from Beiersdorf (Cincinnati, Ohio), cold cream (USP), Purpose CreamTM, available from Johnson & Johnson (New Brunswick, N.J.), hydrophilic ointment (USP) and LubridermTM, available from Pfizer (Morris Plains, N.J.).
In certain embodiments, a pharmaceutical composition of the present invention comprises an oligonucleotide in a therapeutically effective amount. In certain embodiments, the therapeutically effective amount is sufficient to prevent, alleviate or ameliorate symptoms of a disease or to prolong the survival of the subject being treated. Determination of a therapeutically effective amount is well within the capability of those skilled in the art.
In certain embodiments, one or more oligonucleotides of the present invention is formulated as a prodrug. In certain embodiments, upon in vivo administration, a prodrug is chemically converted to the biologically, pharmaceutically or therapeutically more active form of the oligonucleotide. In certain embodiments, prodrugs are useful because they are easier to administer than the corresponding active form. For example, in certain instances, a prodrug may be more bio available (e.g., through oral administration) than is the corresponding active form. In certain instances, a prodrug may have improved solubility compared to the corresponding active form. In certain embodiments, prodrugs are less water soluble than the corresponding active form. In certain instances, such prodrugs possess superior transmittal across cell membranes, where water solubility is detrimental to mobility. In certain embodiments, a prodrug is an ester. In certain such embodiments, the ester is metabolically hydrolyzed to carboxylic acid upon administration. In certain instances the carboxylic acid containing compound is the corresponding active form. In certain embodiments, a prodrug comprises a short peptide (polyaminoacid) bound to an acid group. In certain of such embodiments, the peptide is cleaved upon administration to form the corresponding active form.
In certain embodiments, a prodrug is produced by modifying a pharmaceutically active compound such that the active compound will be regenerated upon in vivo administration. The prodrug can be designed to alter the metabolic stability or the transport characteristics of a drug, to mask side effects or toxicity, to improve the flavor of a drug or to alter other characteristics or properties of a drug. By virtue of knowledge of pharmacodynamic processes and drug metabolism in vivo, those of skill in this art, once a pharmaceutically active compound is known, can design prodrugs of the compound (see, e.g., Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392).
In certain embodiments, a pharmaceutical composition comprising one or more pharmaceutical agents of the present invention is useful for treating a conditions or disorders in a mammalian, and particularly in a human, subject. Suitable administration routes include, but are not limited to, oral, rectal, transmucosal, intestinal, enteral, topical, suppository, through inhalation, intrathecal, intraventricular, intraperitoneal, intranasal, intraocular and parenteral (e.g., intravenous, intramuscular, intramedullary, and subcutaneous). In certain embodiments, pharmaceutical intrathecals are administered to achieve local rather than systemic exposures. For example, pharmaceutical compositions may be injected directly in the area of desired effect (e.g., in the renal or cardiac area).
In certain embodiments, a pharmaceutical composition of the present invention is administered in the form of a dosage unit (e.g., tablet, capsule, bolus, etc.). In certain embodiments, such pharmaceutical compositions comprise an oligonucleotide in a dose selected from 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 105 mg, 110 mg, 115 mg, 120 mg, 125 mg, 130 mg, 135 mg, 140 mg, 145 mg, 150 mg, 155 mg, 160 mg, 165 mg, 170 mg, 175 mg, 180 mg, 185 mg, 190 mg, 195 mg, 200 mg, 205 mg, 210 mg, 215 mg, 220 mg, 225 mg, 230 mg, 235 mg, 240 mg, 245 mg, 250 mg, 255 mg, 260 mg, 265 mg, 270 mg, 270 mg, 280 mg, 285 mg, 290 mg, 295 mg, 300 mg, 305 mg, 310 mg, 315 mg, 320 mg, 325 mg, 330 mg, 335 mg, 340 mg, 345 mg, 350 mg, 355 mg, 360 mg, 365 mg, 370 mg, 375 mg, 380 mg, 385 mg, 390 mg, 395 mg, 400 mg, 405 mg, 410 mg, 415 mg, 420 mg, 425 mg, 430 mg, 435 mg, 440 mg, 445 mg, 450 mg, 455 mg, 460 mg, 465 mg, 470 mg, 475 mg, 480 mg, 485 mg, 490 mg, 495 mg, 500 mg, 505 mg, 510 mg, 515 mg, 520 mg, 525 mg, 530 mg, 535 mg, 540 mg, 545 mg, 550 mg, 555 mg, 560 mg, 565 mg, 570 mg, 575 mg, 580 mg, 585 mg, 590 mg, 595 mg, 600 mg, 605 mg, 610 mg, 615 mg, 620 mg, 625 mg, 630 mg, 635 mg, 640 mg, 645 mg, 650 mg, 655 mg, 660 mg, 665 mg, 670 mg, 675 mg, 680 mg, 685 mg, 690 mg, 695 mg, 700 mg, 705 mg, 710 mg, 715 mg, 720 mg, 725 mg, 730 mg, 735 mg, 740 mg, 745 mg, 750 mg, 755 mg, 760 mg, 765 mg, 770 mg, 775 mg, 780 mg, 785 mg, 790 mg, 795 mg, and 800 mg. In certain such embodiments, a pharmaceutical composition of the present invention comprises a dose of oligonucleotide selected from 25 mg, 50 mg, 75 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 500 mg, 600 mg, 700 mg, and 800 mg. In certain embodiments, a pharmaceutical composition is comprises a dose of oligonucleotide selected from 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, and 400 mg. In certain embodiments the dose is administered at intervals ranging from more than once per day, once per day, once per week, twice per week, three times per week, four times per week, five times per week, 6 times per week, once per month to once per three months, for as long as needed to sustain the desired effect.
In a further aspect, a pharmaceutical agent is sterile lyophilized oligonucleotide that is reconstituted with a suitable diluent, e.g., sterile water for injection. The reconstituted product is administered as a subcutaneous injection or as an intravenous infusion after dilution into saline. The lyophilized drug product consists of the oligonucleotide which has been prepared in water for injection, adjusted to pH 7.0-9.0 with acid or base during preparation, and then lyophilized. The lyophilized oligonucleotide may be 25-800 mg of the oligonucleotide. It is understood that this encompasses 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, and 800 mg of lyophilized oligonucleotide. The lyophilized drug product may be packaged in a 2 mL Type I, clear glass vial (ammonium sulfate-treated), stoppered with a bromobutyl rubber closure and sealed with an aluminum FLIP-OFF® overseal. In one embodiment, the lyophilized pharmaceutical agent comprises ISIS 405879.
The compositions of the present invention may additionally contain other adjunct components conventionally found in pharmaceutical compositions, at their art-established usage levels. Thus, for example, the compositions may contain additional, compatible, pharmaceutically-active materials such as, for example, antipruritics, astringents, local anesthetics or anti-inflammatory agents, or may contain additional materials useful in physically formulating various dosage forms of the compositions of the present invention, such as dyes, flavoring agents, preservatives, antioxidants, opacifiers, thickening agents and stabilizers. However, such materials, when added, should not unduly interfere with the biological activities of the components of the compositions of the present invention. The formulations can be sterilized and, if desired, mixed with auxiliary agents, e.g., lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorings, flavorings and/or aromatic substances and the like which do not deleteriously interact with the oligonucleotide(s) of the formulation.
Conjugated Antisense Compounds
Antisense compounds may be covalently linked to one or more moieties or conjugates which enhance the activity, visualization of cellular distribution, or cellular uptake of the resulting antisense oligonucleotides. Typical conjugate groups include cholesterol moieties and lipid moieties. Additional conjugate groups include carbohydrates, phospholipids, biotin, phenazine, folate, phenanthridine, anthraquinone, acridine, fluoresceins, rhodamines, coumarins, and dyes.
Antisense compounds can also be modified to have one or more stabilizing groups that are generally attached to one or both termini of antisense compounds to enhance properties such as, for example, nuclease stability. Included in stabilizing groups are cap structures. These terminal modifications protect the antisense compound having terminal nucleic acid from exonuclease degradation, and can help in delivery and/or localization within a cell. The cap can be present at the 5′-terminus (5′-cap), or at the 3′-terminus (3′-cap), or can be present on both termini. Cap structures are well known in the art and include, for example, inverted deoxy abasic caps. Further 3′ and 5′-stabilizing groups that can be used to cap one or both ends of an antisense compound to impart nuclease stability include those disclosed in WO 03/004602 published on Jan. 16, 2003.
Cell Culture and Antisense Compounds Treatment
The effects of antisense compounds on the level, activity or expression of PCSK9 nucleic acids can be tested in vitro in a variety of cell types. Cell types used for such analyses are available from commerical vendors (e.g., American Type Culture Collection, Manassas, Va.; Zen-Bio, Inc., Research Triangle Park, NC; Clonetics Corporation, Walkersville, Md.) and cells are cultured according to the vendor's instructions using commercially available reagents (e.g., Invitrogen Life Technologies, Carlsbad, Calif.). Illustrative cell types include, but are not limited to, HepG2 cells, Hep3B cells, and primary hepatocytes.
In Vitro Testing of Antisense Oligonucleotides
Described herein are methods for treatment of cells with antisense oligonucleotides, which can be modified appropriately for treatment with other antisense compounds.
In general, cells are treated with antisense oligonucleotides when they cells reach approximately 60-80% confluency in culture.
One reagent commonly used to introduce antisense oligonucleotides into cultured cells includes the cationic lipid transfection reagent LIPOFECTIN® (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotides are mixed with LIPOFECTIN® in OPTI-MEM® 1 (Invitrogen, Carlsbad, Calif.) to achieve the desired final concentration of antisense oligonucleotide and a LIPOFECTIN® concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.
Another reagent used to introduce antisense oligonucleotides into cultured cells includes LIPOFECTAMINE® (Invitrogen, Carlsbad, Calif.). Antisense oligonucleotide is mixed with LIPOFECTAMINE® in OPTI-MEM® 1 reduced serum medium (Invitrogen, Carlsbad, Calif.) to achieve the desired concentration of antisense oligonucleotide and a LIPOFECTAMINE® concentration that typically ranges 2 to 12 ug/mL per 100 nM antisense oligonucleotide.
Cells are treated with antisense oligonucleotides by routine methods. Cells are typically harvested 16-24 hours after antisense oligonucleotide treatment, at which time RNA or protein levels of target nucleic acids are measured by methods known in the art and described herein. In general, when treatments are performed in multiple replicates, the data are presented as the average of the replicate treatments.
The concentration of antisense oligonucleotide used varies from cell line to cell line. Methods to determine the optimal antisense oligonucleotide concentration for a particular cell line are well known in the art. Antisense oligonucleotides are typically used at concentrations ranging from 1 nM to 300 nM.
RNA Isolation
RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. RNA is prepared using methods well known in the art, for example, using the TRIZOL® Reagent (Invitrogen, Carlsbad, Calif.) according to the manufacturer's recommended protocols.
Analysis of Inhibition of Target Levels or Expression
Inhibition of levels or expression of a PCSK9 nucleic acid can be assayed in a variety of ways known in the art. For example, target nucleic acid levels can be quantitated by, e.g., Northern blot analysis, competitive polymerase chain reaction (PCR), or quantitaive real-time PCR. RNA analysis can be performed on total cellular RNA or poly(A)+ mRNA. Methods of RNA isolation are well known in the art. Northern blot analysis is also routine in the art. Quantitative real-time PCR can be conveniently accomplished using the commercially available ABI PRISM® 7600, 7700, or 7900 Sequence Detection System, available from PE-Applied Biosystems, Foster City, Calif. and used according to manufacturer's instructions.
Quantitative Real-Time PCR Analysis of Target RNA Levels
Quantitation of target RNA levels may be accomplished by quantitative real-time PCR using the ABI PRISM® 7600, 7700, or 7900 Sequence Detection System (PE-Applied Biosystems, Foster City, Calif.) according to manufacturer's instructions. Methods of quantitative real-time PCR are well known in the art.
Prior to real-time PCR, the isolated RNA is subjected to a reverse transcriptase (RT) reaction, which produces complementary DNA (cDNA) that is then used as the substrate for the real-time PCR amplification. The RT and real-time PCR reactions are performed sequentially in the same sample well. RT and real-time PCR reagents are obtained from Invitrogen (Carlsbad, Calif.). RT, real-time-PCR reactions are carried out by methods well known to those skilled in the art.
Gene (or RNA) target quantities obtained by real time PCR are normalized using either the expression level of a gene whose expression is constant, such as GAPDH, or by quantifying total RNA using RIBOGREEN® (Invitrogen, Inc. Carlsbad, Calif.). GAPDH expression is quantified by real time PCR, by being run simultaneously with the target, multiplexing, or separately. Total RNA is quantified using RIBOGREEN® RNA quantification reagent (Invitrogen, Inc. Carlsbad, Calif.). Methods of RNA quantification by RIBOGREEN® are taught in Jones, L. J., et al, (Analytical Biochemistry, 1998, 265, 368-374). A CYTOFLUOR® 4000 instrument (PE Applied Biosystems) is used to measure RIBOGREEN® fluorescence.
Probes and primers are designed to hybridize to a PCSK9 nucleic acid. Methods for designing real-time PCR probes and primers are well known in the art, and may include the use of software such as PRIMER EXPRESS® Software (Applied Biosystems, Foster City, Calif.).
Analysis of Protein Levels
Antisense inhibition of PCSK9 nucleic acids can be assessed by measuring PCSK9 protein levels. Protein levels of PCSK9 can be evaluated or quantitated in a variety of ways well known in the art, such as immunoprecipitation, Western blot analysis (immunoblotting), enzyme-linked immunosorbent assay (ELISA), quantitative protein assays, protein activity assays (for example, caspase activity assays), immunohistochemistry, immunocytochemistry or fluorescence-activated cell sorting (FACS). Antibodies directed to a target can be identified and obtained from a variety of sources, such as the MSRS catalog of antibodies (Aerie Corporation, Birmingham, Mich.), or can be prepared via conventional monoclonal or polyclonal antibody generation methods well known in the art. Antibodies useful for the detection of human and rat PCSK9 are commercially available.
Serum Lipid Analysis
Plasma concentrations of total cholesterol, LDL-C, HDL-C, free cholesterol, triglycerides, glucose, ketones, transaminases, and phospholipids can be measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein and cholesterol profiling can be performed as described by Crooke et al., Lipid Res. (2005) 46: 872-884, using a Beckman System Gold 126 HPLC system, a 507e refrigerated autosampler, a 126 photodiode array detector (Beckman Instruments, Fullerton, Calif.), and a Superose 6 HR 10/30 column (Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions are measured at a wavelength of 505 nm and validated with a cholesterol calibration kit. (Sigma).
Liver Triglyceride Analysis
Liver triglyceride content can be measured according to routine experimental procedures, for example, per procedures described by Desai et al., Diabetes (2001) 50: 2287-2295. Liver triglyceride content may be measured after antisense inhibition of PCSK9 for 6 weeks.
In Vivo Testing of Antisense Compounds
Antisense compounds, for example, antisense oligonucleotides, are tested in animals to assess their ability to inhibit expression of PCSK9 and produce phenotypic changes, such as decreases in LDL-C. Testing may be performed in normal animals, or in experimental disease models. For administration to animals, antisense oligonucleotides are formulated in a pharmaceutically acceptable diluent, such as phosphate-buffered saline. Administration includes parenteral routes of administration, such as intraperitoneal, intravenous, and subcutaneous. Calculation of antisense oligonucleotide dosage and dosing frequency is within the abilities of those skilled in the art, and depends upon factors such as route of administration and animal body weight. Following a period of treatment with antisense oligonucleotides, RNA is isolated from various tissues and changes in PCSK9 nucleic acid expression are measured. Changes in PCSK9 protein levels may also be measured.
Nonlimiting Disclosure and Incorporation by Reference
The following examples serve only to illustrate the methods, antisense oligonucleotides, and compositions provided and are not intended to limit the same. Each of the references, GENBANK® accession numbers, and the like recited in the present application is incorporated herein by reference in its entirety.
EXAMPLES
Example 1
Antisense Inhibition of Human PCSK9: A549 Cells
Antisense oligonucleotides targeted to a PCSK9 nucleic acid were tested for their effects on PCSK9 mRNA in vitro. Cultured A549 cells at a density of 4000 cells per well in a 96-well plate were treated with 60 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells. Antisense oligonucleotides that exhibited at least 30% inhibition of PCSK9 expression are shown in Table 16.
The motif column indicates the wing-gap-wing motif of each antisense oligonucleotide. Antisense oligonucleotides were designed as 5-10-5 gapmers, or alternatively as 3-14-3 gapmers, where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises 2′-MOE nucleotides. As illustrated in Table 16, a single nucleobase sequence may be represented by a 5-10-5 motif as well as a 3-14-3 motif. “5′ target site” indicates the 5′-most nucleotide which the antisense oligonucleotide is targeted on the indicated GENBANK Accession No.







TABLE 16







Antisense inhibition of PCSK9 in human cells (Cell type A549)














SEQ
5′
Target






ID
Target
Nucleic


%


IsisNo
NO
Site
Acid
Sequence
Motif
Inhibition





395150
  5
  242
NM_174936.2
GAGGAGACCTAGAGGCCGTG
5-10-5
57


 


395151
  6
  300
NM_174936.2
AGGACCGCCTGGAGCTGACG
5-10-5
77


 


395152
  7
  410
NM_174936.2
ACGCAAGGCTAGCACCAGCT
5-10-5
81


 


395153
  9
  480
NM_174936.2
CCTTGGCGCAGCGGTGGAAG
5-10-5
51


 


395154
 10
  561
NM_174936.2
GGCGGGCAGTGCGCTCTGAC
5-10-5
40


 


395155
 11
  600
NM_174936.2
TGGTGAGGTATCCCCGGCGG
5-10-5
79


 


395156
 14
  620
NM_174936.2
ATGGAAGACATGCAGGATCT
5-10-5
75


 


395157
 15
  646
NM_174936.2
TTCACCAGGAAGCCAGGAAG
5-10-5
72


 


395158
 17
  705
NM_174936.2
CCTCGATGTAGTCGACATGG
5-10-5
61


 


395159
 18
  785
NM_174936.2
GTATTCATCCGCCCGGTACC
5-10-5
35


 


395160
 19
  835
NM_174936.2
CTGGTGTCTAGGAGATACAC
5-10-5
47


 


395161
 21
  860
NM_174936.2
GATTTCCCGGTGGTCACTCT
5-10-5
70


 


395162
 24
  890
NM_174936.2
CTCGAAGTCGGTGACCATGA
5-10-5
52


 


395163
 25
  923
NM_174936.2
GTGGAAGCGGGTCCCGTCCT
5-10-5
73


 


395164
 26
  970
NM_174936.2
ACCCCTGCCAGGTGGGTGCC
5-10-5
69


 


395165
 28
 1004
NM_174936.2
ACCCTTGGCCACGCCGGCAT
5-10-5
81


 


395166
 29
 1040
NM_174936.2
TTGGCAGTTGAGCACGCGCA
5-10-5
65


 


395167
 31
 1077
NM_174936.2
CCAGGCCTATGAGGGTGCCG
5-10-5
34


 


395168
 32
 1098
NM_174936.2
GCTGGCTTTTCCGAATAAAC
5-10-5
69


 


395169
 33
 1210
NM_174936.2
GTGACCAGCACGACCCCAGC
5-10-5
61


 


395171
 34
 1326
NM_174936.2
CCAAAGTCCCCAGGGTCACC
5-10-5
38


 


395173
 36
 1340
NM_174936.2
GCCAAAGTTGGTCCCCAAAG
5-10-5
64


 


395174
 38
 1361
NM_174936.2
GGCAAAGAGGTCCACACAGC
5-10-5
72


 


395175
 39
 1389
NM_174936.2
TGGAGGCACCAATGATGTCC
5-10-5
41


 


395177
 45
 1534
NM_174936.2
TTGGCAGAGAAGTGGATCAG
5-10-5
41


 


395178
 50
 1569
NM_174936.2
GGTCCTCAGGGAACCAGGCC
5-10-5
67


 


395181
 52
 1640
NM_174936.2
AAACAGCTGCCAACCTGCCC
5-10-5
37


 


395182
 54
 1675
NM_174936.2
GTAGGCCCCGAGTGTGCTGA
5-10-5
51


 


395183
 57
 1812
NM_174936.2
CGTTGTGGGCCCGGCAGACC
5-10-5
74


 


395184
 58
 1858
NM_174936.2
AGCAGGCAGCACCTGGCAAT
5-10-5
61


 


395185
 59
 1920
NM_174936.2
CACGGGTCCCCATGCTGGCC
5-10-5
73


 


395186
 60
 2100
NM_174936.2
CTTTGCATTCCAGACCTGGG
5-10-5
83


 


395187
 62
 2310
NM_174936.2
GGCAGCAGATGGCAACGGCT
5-10-5
80


 


395189
 64
 2504
NM_174936.2
TCAAGGGCCAGGCCAGCAGC
5-10-5
65


 


395190
 66
 2597
NM_174936.2
ATGCCCCACAGTGAGGGAGG
5-10-5
60


 


395191
 67
 2606
NM_174936.2
AATGGTGAAATGCCCCACAG
5-10-5
76


 


395193
 68
 2750
NM_174936.2
CATGGGAAGAATCCTGCCTC
5-10-5
48


 


395194
 69
 2832
NM_174936.2
GGAGATGAGGGCCATCAGCA
5-10-5
64


 


395195
 70
 2900
NM_174936.2
TAGATGCCATCCAGAAAGCT
5-10-5
44


 


395196
 72
 2983
NM_174936.2
GGCATAGAGCAGAGTAAAGG
5-10-5
39


 


395198
 75
 3437
NM_174936.2
GCTCAAGGAGGGACAGTTGT
5-10-5
73


 


395199
 76
 3472
NM_174936.2
AAAGATAAATGTCTGCTTGC
5-10-5
83


 


395200
 78
 3543
NM_174936.2
TCTTCAAGTTACAAAAGCAA
5-10-5
40


 


395202
129
13816
NT_032977.8
GTGCCATCTGAACAGCACCT
5-10-5
36


 


395203
110
13926
NT_032977.8
CCTGGAACCCCTGCAGCCAG
5-10-5
64


 


395204
152
13977
NT_032977.8
TTCAGGCAGGTTGCTGCTAG
5-10-5
55


 


395206
136
14122
NT_032977.8
TAGGAGAAAGTAGGGAGAGC
5-10-5
45


 


395207
132
14179
NT_032977.8
TAAAAGCTGCAAGAGACTCA
5-10-5
49


 


395208
139
14267
NT_032977.8
TCAGAGAAAACAGTCACCGA
5-10-5
56


 


395209
142
14404
NT_032977.8
TCATTTTAGAGACAGGAAGC
5-10-5
73


 


395210
113
14441
NT_032977.8
GAATAACAGTGATGTCTGGC
5-10-5
82


 


395211
138
14494
NT_032977.8
TCACAGCTCACCGAGTCTGC
5-10-5
60


 


395212
 98
14524
NT_032977.8
AGTGTAAAATAAAGCCCCTA
5-10-5
50


 


395213
 96
14601
NT_032977.8
AGGACCCAAGTCATCCTGCT
5-10-5
57


 


395214
124
14631
NT_032977.8
GGCCATCAGCTGGCAATGCT
5-10-5
50


 


395215
133
14675
NT_032977.8
TAGACAAGGAAAGGGAGGCC
5-10-5
60


 


395216
103
14681
NT_032977.8
ATTTCATAGACAAGGAAAGG
5-10-5
51


 


395218
146
 1315
AK124635.1
TGACATTTGTGGGAGAGGAG
5-10-5
35


 


395220
104
 2085
AK124635.1
CACAGTCCTGCAAAACAGCT
5-10-5
54


 


395221
102
 5590
NT_032977.8
ATGTGCAGAGATCAATCACA
5-10-5
69


 


395222
127
10633
NT_032977.8
GGTGGTAATTTGTCACAGCA
5-10-5
64


 


395223
 84
11308
NT_032977.8
AAGGTCACACAGTTAAGAGT
5-10-5
55


 


395226
147
24858
NT_032977.8
TGAGTTCATTTAAGAGTGGA
5-10-5
42


 


399793
  8
  417
NM_174936.2
CCTCGGAACGCAAGGCTAGC
5-10-5
71


 


399794
 12
  606
NM_174936.2
GGATCTTGGTGAGGTATCCC
5-10-5
42


 


399795
 13
  615
NM_174936.2
AGACATGCAGGATCTTGGTG
5-10-5
54


 


399796
 16
  651
NM_174936.2
TCATCTTCACCAGGAAGCCA
5-10-5
50


 


399797
 20
  840
NM_174936.2
GTATGCTGGTGTCTAGGAGA
5-10-5
38


 


399798
 22
  866
NM_174936.2
GCCCTCGATTTCCCGGTGGT
5-10-5
54


 


399799
 23
  880
NM_174936.2
GTGACCATGACCCTGCCCTC
5-10-5
75


 


399800
 27
  975
NM_174936.2
TGACCACCCCTGCCAGGTGG
5-10-5
67


 


399801
 30
 1045
NM_174936.2
TTCCCTTGGCAGTTGAGCAC
5-10-5
79


 


399802
 35
 1335
NM_174936.2
AGTTGGTCCCCAAAGTCCCC
5-10-5
71


 


399803
 37
 1352
NM_174936.2
GTCCACACAGCGGCCAAAGT
5-10-5
63


 


399804
 40
 1400
NM_174936.2
GCTGCAGTCGCTGGAGGCAC
5-10-5
46


 


399805
 41
 1411
NM_174936.2
ACAAAGCAGGTGCTGCAGTC
5-10-5
46


 


399806
 42
 1470
NM_174936.2
GCATCATGGCTGCAATGCCA
5-10-5
85


 


399807
 43
 1478
NM_174936.2
GGCAGACAGCATCATGGCTG
5-10-5
71


 


399808
 44
 1526
NM_174936.2
GAAGTGGATCAGTCTCTGCC
5-10-5
38


 


399809
 46
 1539
NM_174936.2
CATCTTTGGCAGAGAAGTGG
5-10-5
53


 


399810
 47
 1545
NM_174936.2
TGATGACATCTTTGGCAGAG
5-10-5
63


 


399811
 48
 1552
NM_174936.2
GCCTCATTGATGACATCTTT
5-10-5
61


 


399812
 49
 1564
NM_174936.2
TCAGGGAACCAGGCCTCATT
5-10-5
46


 


399813
 51
 1583
NM_174936.2
GGTCAGTACCCGCTGGTCCT
5-10-5
72


 


399814
 53
 1645
NM_174936.2
CTGCAAAACAGCTGCCAACC
5-10-5
88


 


399815
 55
 1740
NM_174936.2
AGAAACTGGAGCAGCTCAGC
5-10-5
38


 


399816
 56
 1746
NM_174936.2
TCCTGGAGAAACTGGAGCAG
5-10-5
51


 


399817
 61
 2105
NM_174936.2
CTTGACTTTGCATTCCAGAC
5-10-5
84


 


399818
 63
 2415
NM_174936.2
AACCATTTTAAAGCTCAGCC
5-10-5
68


 


399819
 65
 2509
NM_174936.2
CCCACTCAAGGGCCAGGCCA
5-10-5
87


 


399821
 71
 2906
NM_174936.2
TCTGGCTAGATGCCATCCAG
5-10-5
83


 


399822
 73
 2988
NM_174936.2
AGCCTGGCATAGAGCAGAGT
5-10-5
70


 


399824
 74
 3233
NM_174936.2
AAGTAAGAAGAGGCTTGGCT
5-10-5
43


 


399825
 77
 3477
NM_174936.2
ACCCAAAAGATAAATGTCTG
5-10-5
61


 


399827
100
13746
NT_032977.8
ATCTCAGGACAGGTGAGCAA
5-10-5
81


 


399828
116
13760
NT_032977.8
GAGTAGAGATTCTCATCTCA
5-10-5
73


 


399829
117
13828
NT_032977.8
GAGTCTTCTGAAGTGCCATC
5-10-5
65


 


399831
 83
13986
NT_032977.8
AAGGAAGACTTCAGGCAGGT
5-10-5
52


 


399833
 92
14397
NT_032977.8
AGAGACAGGAAGCTGCAGCT
5-10-5
54


 


399834
 82
14670
NT_032977.8
AAGGAAAGGGAGGCCTAGAG
5-10-5
44


 


399836
130
 2095
AK124635.1
GTGCTGACCACACAGTCCTG
5-10-5
96


 


399837
107
 3056
NT_032977.8
CCCACTATAATGGCAAGCCC
5-10-5
83


 


399838
 80
 4306
NT_032977.8
AACCCAGTTCTAATGCACCT
5-10-5
81


 


399839
106
 5140
NT_032977.8
CCAGTCAGAGTAGAACAGAG
5-10-5
68


 


399840
121
 5599
NT_032977.8
GGAGCCTACATGTGCAGAGA
5-10-5
46


 


399841
 94
 5667
NT_032977.8
AGCATGGCACCAGCATCTGC
5-10-5
59


 


399842
108
 6652
NT_032977.8
CCCAGCCCTATCAGGAAGTG
5-10-5
67


 


399843
144
 7099
NT_032977.8
TGACATCCAGGAGGGAGGAG
5-10-5
33


 


399844
 91
 7556
NT_032977.8
AGACTGATGGAAGGCATTGA
5-10-5
56


 


399845
131
 7565
NT_032977.8
GTGTTGAGCAGACTGATGGA
5-10-5
48


 


399846
145
 8836
NT_032977.8
TGACATCTTGTCTGGGAGCC
5-10-5
58


 


399847
 90
 8948
NT_032977.8
AGACTAGGAGCCTGAGTTTT
5-10-5
38


 


399849
148
10252
NT_032977.8
TGGCAGCAACTCAGACATAT
5-10-5
74


 


399851
 88
12715
NT_032977.8
ACTGGATACATTGGCAGACA
5-10-5
62


 


399852
111
12928
NT_032977.8
CTAGAGGAACCACTAGATAT
5-10-5
66


 


399854
 97
16134
NT_032977.8
AGTCAAGCTGCTGCCCAGAG
5-10-5
82


 


399855
120
16668
NT_032977.8
GCTAGTTATTAAGCACCTGC
5-10-5
71


 


399856
150
17267
NT_032977.8
TGTGAGCTCTGGCCCAGTGG
5-10-5
64


 


399857
115
18377
NT_032977.8
GAGTAAGGCAGGTTACTCTC
5-10-5
79


 


399858
134
18408
NT_032977.8
TAGATGTGACTAACATTTAA
5-10-5
57


 


399859
105
19203
NT_032977.8
CACATTAGCCTTGCTCAAGT
5-10-5
75


 


399860
151
19913
NT_032977.8
TGTGATGACCTGGAAAGGTG
5-10-5
36


 


399862
109
20188
NT_032977.8
CCCCTGCACAGAGCCTGGCA
5-10-5
59


 


399863
141
20624
NT_032977.8
TCATGGCTGCAATGCCTGGT
5-10-5
46


 


399864
 93
20995
NT_032977.8
AGAGAGGAGGGCTTAAAGAA
5-10-5
30


 


399865
 95
21082
NT_032977.8
AGCTGCCAACCTGCAAAAAG
5-10-5
75


 


399866
143
21481
NT_032977.8
TGAAAATCCATCCAGCACTG
5-10-5
74


 


399867
 89
21589
NT_032977.8
AGAACCATGGAGCACCTGAG
5-10-5
84


 


399868
123
21696
NT_032977.8
GGCACTGCCCTTCCACCAAA
5-10-5
53


 


399869
118
24907
NT_032977.8
GCACCATCCAGACCAGAATC
5-10-5
49


 


399870
114
25413
NT_032977.8
GAGAGGTTCAGATCCAGGCC
5-10-5
66


 


399871
  4
  135
NM_174936.2
GCGCGGAATCCTGGCTGGGA
3-14-3
40


 


399872
  5
  242
NM_174936.2
GAGGAGACCTAGAGGCCGTG
3-14-3
64


 


399873
  6
  300
NM_174936.2
AGGACCGCCTGGAGCTGACG
3-14-3
65


 


399874
  7
  410
NM_174936.2
ACGCAAGGCTAGCACCAGCT
3-14-3
80


 


399875
  9
  480
NM_174936.2
CCTTGGCGCAGCGGTGGAAG
3-14-3
67


 


399876
 10
  561
NM_174936.2
GGCGGGCAGTGCGCTCTGAC
3-14-3
62


 


399877
 11
  600
NM_174936.2
TGGTGAGGTATCCCCGGCGG
3-14-3
73


 


399878
 14
  620
NM_174936.2
ATGGAAGACATGCAGGATCT
3-14-3
54


 


399879
 15
  646
NM_174936.2
TTCACCAGGAAGCCAGGAAG
3-14-3
75


 


399880
 17
  705
NM_174936.2
CCTCGATGTAGTCGACATGG
3-14-3
80


 


399881
 18
  785
NM_174936.2
GTATTCATCCGCCCGGTACC
3-14-3
63


 


399882
 19
  835
NM_174936.2
CTGGTGTCTAGGAGATACAC
3-14-3
66


 


399883
 21
  860
NM_174936.2
GATTTCCCGGTGGTCACTCT
3-14-3
59


 


399884
 24
  890
NM_174936.2
CTCGAAGTCGGTGACCATGA
3-14-3
71


 


399885
 25
  923
NM_174936.2
GTGGAAGCGGGTCCCGTCCT
3-14-3
73


 


399886
 26
  970
NM_174936.2
ACCCCTGCCAGGTGGGTGCC
3-14-3
76


 


399887
 28
 1004
NM_174936.2
ACCCTTGGCCACGCCGGCAT
3-14-3
54


 


399888
 29
 1040
NM_174936.2
TTGGCAGTTGAGCACGCGCA
3-14-3
75


 


399889
 31
 1077
NM_174936.2
CCAGGCCTATGAGGGTGCCG
3-14-3
84


 


399890
 32
 1098
NM_174936.2
GCTGGCTTTTCCGAATAAAC
3-14-3
73


 


399891
 33
 1210
NM_174936.2
GTGACCAGCACGACCCCAGC
3-14-3
99


 


399892
149
 1297
NM_174936.2
TGGGCATTGGTGGCCCCAAC
3-14-3
61


 


399893
 34
 1326
NM_174936.2
CCAAAGTCCCCAGGGTCACC
3-14-3
54


 


399894
128
 1330
NM_174936.2
GTCCCCAAAGTCCCCAGGGT
3-14-3
80


 


399895
 36
 1340
NM_174936.2
GCCAAAGTTGGTCCCCAAAG
3-14-3
82


 


399896
 38
 1361
NM_174936.2
GGCAAAGAGGTCCACACAGC
3-14-3
83


 


399897
 39
 1389
NM_174936.2
TGGAGGCACCAATGATGTCC
3-14-3
70


 


399898
101
 1465
NM_174936.2
ATGGCTGCAATGCCAGCCAC
3-14-3
37


 


399899
 45
 1534
NM_174936.2
TTGGCAGAGAAGTGGATCAG
3-14-3
64


 


399900
 50
 1569
NM_174936.2
GGTCCTCAGGGAACCAGGCC
3-14-3
82


 


399901
 87
 1576
NM_174936.2
ACCCGCTGGTCCTCAGGGAA
3-14-3
76


 


399902
119
 1605
NM_174936.2
GCAGGGCGGCCACCAGGTTG
3-14-3
39


 


399903
 52
 1640
NM_174936.2
AAACAGCTGCCAACCTGCCC
3-14-3
73


 


399904
 54
 1675
NM_174936.2
GTAGGCCCCGAGTGTGCTGA
3-14-3
81


 


399905
 57
 1812
NM_174936.2
CGTTGTGGGCCCGGCAGACC
3-14-3
80


 


399906
 58
 1858
NM_174936.2
AGCAGGCAGCACCTGGCAAT
3-14-3
92


 


399907
 59
 1920
NM_174936.2
CACGGGTCCCCATGCTGGCC
3-14-3
95


 


399908
 60
 2100
NM_174936.2
CTTTGCATTCCAGACCTGGG
3-14-3
84


 


399909
 62
 2310
NM_174936.2
GGCAGCAGATGGCAACGGCT
3-14-3
73


 


399910
154
 2410
NM_174936.2
TTTTAAAGCTCAGCCCCAGC
3-14-3
57


 


399911
 64
 2504
NM_174936.2
TCAAGGGCCAGGCCAGCAGC
3-14-3
82


 


399912
 66
 2597
NM_174936.2
ATGCCCCACAGTGAGGGAGG
3-14-3
68


 


399913
 67
 2606
NM_174936.2
AATGGTGAAATGCCCCACAG
3-14-3
92


 


399914
153
 2649
NM_174936.2
TTGGGAGCAGCTGGCAGCAC
3-14-3
59


 


399915
 68
 2750
NM_174936.2
CATGGGAAGAATCCTGCCTC
3-14-3
78


 


399916
 69
 2832
NM_174936.2
GGAGATGAGGGCCATCAGCA
3-14-3
71


 


399917
 70
 2900
NM_174936.2
TAGATGCCATCCAGAAAGCT
3-14-3
81


 


399918
 72
 2983
NM_174936.2
GGCATAGAGCAGAGTAAAGG
3-14-3
82


 


399919
112
 3227
NM_174936.2
GAAGAGGCTTGGCTTCAGAG
3-14-3
57


 


399920
 75
 3437
NM_174936.2
GCTCAAGGAGGGACAGTTGT
3-14-3
84


 


399921
 76
 3472
NM_174936.2
AAAGATAAATGTCTGCTTGC
3-14-3
78


 


399922
 78
 3543
NM_174936.2
TCTTCAAGTTACAAAAGCAA
3-14-3
61


 


399923
 85
13681
NT_032977.8
ACAAATTCCCAGACTCAGCA
3-14-3
82


 


399924
129
13816
NT_032977.8
GTGCCATCTGAACAGCACCT
3-14-3
84


 


399925
110
13926
NT_032977.8
CCTGGAACCCCTGCAGCCAG
3-14-3
73


 


399926
152
13977
NT_032977.8
TTCAGGCAGGTTGCTGCTAG
3-14-3
47


 


399927
140
13998
NT_032977.8
TCAGCCAGGCCAAAGGAAGA
3-14-3
78


 


399928
136
14122
NT_032977.8
TAGGAGAAAGTAGGGAGAGC
3-14-3
47


 


399929
132
14179
NT_032977.8
TAAAAGCTGCAAGAGACTCA
3-14-3
69


 


399930
139
14267
NT_032977.8
TCAGAGAAAACAGTCACCGA
3-14-3
82


 


399931
142
14404
NT_032977.8
TCATTTTAGAGACAGGAAGC
3-14-3
79


 


399932
113
14441
NT_032977.8
GAATAACAGTGATGTCTGGC
3-14-3
83


 


399933
138
14494
NT_032977.8
TCACAGCTCACCGAGTCTGC
3-14-3
78


 


399934
 98
14524
NT_032977.8
AGTGTAAAATAAAGCCCCTA
3-14-3
79


 


399935
 96
14601
NT_032977.8
AGGACCCAAGTCATCCTGCT
3-14-3
89


 


399936
124
14631
NT_032977.8
GGCCATCAGCTGGCAATGCT
3-14-3
70


 


399937
133
14675
NT_032977.8
TAGACAAGGAAAGGGAGGCC
3-14-3
78


 


399938
103
14681
NT_032977.8
ATTTCATAGACAAGGAAAGG
3-14-3
67


 


399940
146
 1315
AK124635.1
TGACATTTGTGGGAGAGGAG
3-14-3
35


 


399942
104
 2085
AK124635.1
CACAGTCCTGCAAAACAGCT
3-14-3
75


 


399943
102
 5590
NT_032977.8
ATGTGCAGAGATCAATCACA
3-14-3
64


 


399944
127
10633
NT_032977.8
GGTGGTAATTTGTCACAGCA
3-14-3
73


 


399945
 84
11308
NT_032977.8
AAGGTCACACAGTTAAGAGT
3-14-3
89


 


399947
126
22292
NT_032977.8
GGTGCATAAGGAGAAAGAGA
3-14-3
70


 


399948
147
24858
NT_032977.8
TGAGTTCATTTAAGAGTGGA
3-14-3
46


 


399949
  8
  417
NM_174936.2
CCTCGGAACGCAAGGCTAGC
3-14-3
74


 


399950
 12
  606
NM_174936.2
GGATCTTGGTGAGGTATCCC
3-14-3
68


 


399951
 13
  615
NM_174936.2
AGACATGCAGGATCTTGGTG
3-14-3
67


 


399952
 16
  651
NM_174936.2
TCATCTTCACCAGGAAGCCA
3-14-3
74


 


399953
 20
  840
NM_174936.2
GTATGCTGGTGTCTAGGAGA
3-14-3
59


 


399954
 22
  866
NM_174936.2
GCCCTCGATTTCCCGGTGGT
3-14-3
83


 


399955
 23
  880
NM_174936.2
GTGACCATGACCCTGCCCTC
3-14-3
62


 


399956
 27
  975
NM_174936.2
TGACCACCCCTGCCAGGTGG
3-14-3
78


 


399957
 30
 1045
NM_174936.2
TTCCCTTGGCAGTTGAGCAC
3-14-3
82


 


399958
 35
 1335
NM_174936.2
AGTTGGTCCCCAAAGTCCCC
3-14-3
78


 


399959
 37
 1352
NM_174936.2
GTCCACACAGCGGCCAAAGT
3-14-3
89


 


399960
 40
 1400
NM_174936.2
GCTGCAGTCGCTGGAGGCAC
3-14-3
64


 


399961
 41
 1411
NM_174936.2
ACAAAGCAGGTGCTGCAGTC
3-14-3
63


 


399962
 42
 1470
NM_174936.2
GCATCATGGCTGCAATGCCA
3-14-3
68


 


399963
 43
 1478
NM_174936.2
GGCAGACAGCATCATGGCTG
3-14-3
70


 


399964
 44
 1526
NM_174936.2
GAAGTGGATCAGTCTCTGCC
3-14-3
70


 


399965
 46
 1539
NM_174936.2
CATCTTTGGCAGAGAAGTGG
3-14-3
49


 


399966
 47
 1545
NM_174936.2
TGATGACATCTTTGGCAGAG
3-14-3
78


 


399967
 48
 1552
NM_174936.2
GCCTCATTGATGACATCTTT
3-14-3
82


 


399968
 49
 1564
NM_174936.2
TCAGGGAACCAGGCCTCATT
3-14-3
71


 


399969
 51
 1583
NM_174936.2
GGTCAGTACCCGCTGGTCCT
3-14-3
82


 


399970
 53
 1645
NM_174936.2
CTGCAAAACAGCTGCCAACC
3-14-3
74


 


399971
 55
 1740
NM_174936.2
AGAAACTGGAGCAGCTCAGC
3-14-3
48


 


399972
 56
 1746
NM_174936.2
TCCTGGAGAAACTGGAGCAG
3-14-3
63


 


399973
 61
 2105
NM_174936.2
CTTGACTTTGCATTCCAGAC
3-14-3
74


 


399974
 63
 2415
NM_174936.2
AACCATTTTAAAGCTCAGCC
3-14-3
71


 


399975
 65
 2509
NM_174936.2
CCCACTCAAGGGCCAGGCCA
3-14-3
91


 


399976
122
 2582
NM_174936.2
GGAGGGAGCTTCCTGGCACC
3-14-3
67


 


399977
 71
 2906
NM_174936.2
TCTGGCTAGATGCCATCCAG
3-14-3
89


 


399978
 73
 2988
NM_174936.2
AGCCTGGCATAGAGCAGAGT
3-14-3
80


 


399979
135
 2994
NM_174936.2
TAGCACAGCCTGGCATAGAG
3-14-3
83


 


399980
 74
 3233
NM_174936.2
AAGTAAGAAGAGGCTTGGCT
3-14-3
50


 


399981
 77
 3477
NM_174936.2
ACCCAAAAGATAAATGTCTG
3-14-3
55


 


399982
 99
 3550
NM_174936.2
ATAAATATCTTCAAGTTACA
3-14-3
36


 


399983
100
13746
NT_032977.8
ATCTCAGGACAGGTGAGCAA
3-14-3
96


 


399984
116
13760
NT_032977.8
GAGTAGAGATTCTCATCTCA
3-14-3
71


 


399985
117
13828
NT_032977.8
GAGTCTTCTGAAGTGCCATC
3-14-3
76


 


399986
 81
13903
NT_032977.8
AAGCAGGGCCTCAGGTGGAA
3-14-3
68


 


399987
 83
13986
NT_032977.8
AAGGAAGACTTCAGGCAGGT
3-14-3
59


 


399988
137
14112
NT_032977.8
TAGGGAGAGCTCACAGATGC
3-14-3
31


 


399989
 92
14397
NT_032977.8
AGAGACAGGAAGCTGCAGCT
3-14-3
65


 


399990
 82
14670
NT_032977.8
AAGGAAAGGGAGGCCTAGAG
3-14-3
59


 


399992
130
 2095
AK124635.1
GTGCTGACCACACAGTCCTG
3-14-3
94


 


399993
107
 3056
NT_032977.8
CCCACTATAATGGCAAGCCC
3-14-3
76


 


399994
 80
 4306
NT_032977.8
AACCCAGTTCTAATGCACCT
3-14-3
82


 


399995
106
 5140
NT_032977.8
CCAGTCAGAGTAGAACAGAG
3-14-3
74


 


399996
121
 5599
NT_032977.8
GGAGCCTACATGTGCAGAGA
3-14-3
65


 


399997
 94
 5667
NT_032977.8
AGCATGGCACCAGCATCTGC
3-14-3
77


 


399998
108
 6652
NT_032977.8
CCCAGCCCTATCAGGAAGTG
3-14-3
78


 


399999
144
 7099
NT_032977.8
TGACATCCAGGAGGGAGGAG
3-14-3
43


 


400000
 91
 7556
NT_032977.8
AGACTGATGGAAGGCATTGA
3-14-3
84


 


400001
131
 7565
NT_032977.8
GTGTTGAGCAGACTGATGGA
3-14-3
73


 


400002
145
 8836
NT_032977.8
TGACATCTTGTCTGGGAGCC
3-14-3
82


 


400003
 90
 8948
NT_032977.8
AGACTAGGAGCCTGAGTTTT
3-14-3
48


 


400004
125
 9099
NT_032977.8
GGCCTGCAGAAGCCAGAGAG
3-14-3
45


 


400005
148
10252
NT_032977.8
TGGCAGCAACTCAGACATAT
3-14-3
73


 


400006
 79
11472
NT_032977.8
AAATGCAGGGCTAAAATCAC
3-14-3
78


 


400007
 88
12715
NT_032977.8
ACTGGATACATTGGCAGACA
3-14-3
77


 


400008
111
12928
NT_032977.8
CTAGAGGAACCACTAGATAT
3-14-3
78


 


400009
 86
15471
NT_032977.8
ACAGCATTCTTGGTTAGGAG
3-14-3
73


 


400010
 97
16134
NT_032977.8
AGTCAAGCTGCTGCCCAGAG
3-14-3
74


 


400011
120
16668
NT_032977.8
GCTAGTTATTAAGCACCTGC
3-14-3
75


 


400012
150
17267
NT_032977.8
TGTGAGCTCTGGCCCAGTGG
3-14-3
64


 


400013
115
18377
NT_032977.8
GAGTAAGGCAGGTTACTCTC
3-14-3
88


 


400014
134
18408
NT_032977.8
TAGATGTGACTAACATTTAA
3-14-3
58


 


400015
105
19203
NT_032977.8
CACATTAGCCTTGCTCAAGT
3-14-3
89


 


400018
109
20188
NT_032977.8
CCCCTGCACAGAGCCTGGCA
3-14-3
62


 


400019
141
20624
NT_032977.8
TCATGGCTGCAATGCCTGGT
3-14-3
47


 


400020
 93
20995
NT_032977.8
AGAGAGGAGGGCTTAAAGAA
3-14-3
66


 


400021
 95
21082
NT_032977.8
AGCTGCCAACCTGCAAAAAG
3-14-3
61


 


400022
143
21481
NT_032977.8
TGAAAATCCATCCAGCACTG
3-14-3
90


 


400023
 89
21589
NT_032977.8
AGAACCATGGAGCACCTGAG
3-14-3
83


 


400024
123
21696
NT_032977.8
GGCACTGCCCTTCCACCAAA
3-14-3
79


 


400025
118
24907
NT_032977.8
GCACCATCCAGACCAGAATC
3-14-3
78


 


400026
114
25413
NT_032977.8
GAGAGGTTCAGATCCAGGCC
3-14-3
81









Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 399806, 399814, 399817, 399819, 399821, 399827, 399836, 399837, 399854, 399867, 399889, 399891, 399906, 399907, 399908, 399913, 399920, 399924, 399935, 399945, 399959, 399975, 399977, 399983, 399992, 400000, 400013, 400015, 400022, and 400023. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.
ISIS Nos 399814, 399819, 399836, 399891, 399906, 399907, 399913, 399935, 399945, 399959, 399975, 399977, 399983, 399992, 400013, 400015, and 400022 each exhibited at least 85% inhibition of PCSK9 mRNA levels.
ISIS Nos 399836, 399891, 399906, 399907, 399913, 399975, 399983, 399992, and 400022 each exhibited at least 90% inhibition of PCSK9 mRNA levels.
Example 2
Antisense Inhibition of Human PCSK9 in HepG2 Cells
Antisense oligonucleotides were tested for their ability to inhibit the expression of PCSK9 mRNA in cultured HepG2 cells. HepG2 cells at a density of 10000 cells per well in a 96-well plate were treated with 1500 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells.
Isis Nos. 399819, 395149, 395150, 395151, 395152, 395153, 395154, 395155, 395156, 395157, 395158, 395161, 395162, 395163, 395164, 395165, 395166, 395167, 395168, 395172, 395173, 395174, 395178, 395179, 395182, 395183, 395185, 395186, 395187, 395189, 395190, 395191, 395193, 395194, 395195, 395198, 395199, 395203, 395207, 395210, 395211, 395213, 395214, 395221, 395222, 399793, 399794, 399795, 399798, 399801, 399803, 399804, 399806, 399807, 399812, 399813, 399814, 399816, 399817, 399820, 399821, 399822, 399823, 399827, 399828, 399829, 399833, 399834, 399836, 399837, 399841, 399844, 399846, 399848, 399849, 399851, 399852, 399854, 399855, 399856, 399857, 399862, 399865, 399867, 399868, 399872, 399873, 399874, 399875, 399876, 399877, 399879, 399880, 399883, 399884, 399885, 399886, 399887, 399888, 399889, 399890, 399891, 399892, 399894, 399895, 399896, 399900, 399901, 399902, 399904, 399905, 399906, 399907, 399908, 399909, 399911, 399912, 399913, 399915, 399916, 399918, 399920, 399921, 399924, 399925, 399929, 399931, 399935, 399936, 399937, 399943, 399944, 399945, 399949, 399950, 399954, 399957, 399959, 399960, 399962, 399963, 399966, 399967, 399968, 399969, 399970, 399972, 399973, 399975, 399976, 399977, 399978, 399979, 399983, 399984, 399985, 399986, 399989, 399990, 399992, 399996, 399997, 399998, 400000, 400001, 400002, 400004, 400005, 400007, 400008, 400009, 400010, 400012, 400013, 400015, 400018, 400021, 400023, 400024, 400025, and 400026 each inhibited PCSK9 by at least 70% in this experiment. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.
Isis Nos 399819, 395150, 395152, 395153, 395154, 395155, 395156, 395158, 395163, 395164, 395165, 395166, 395167, 395168, 395173, 395178, 395183, 395185, 395186, 395187, 395189, 395190, 395193, 395194, 395199, 395203, 395210, 395213, 395214, 399793, 399798, 399801, 399804, 399806, 399813, 399816, 399820, 399821, 399822, 399836, 399849, 399856, 399857, 399862, 399868, 399875, 399877, 399880, 399885, 399886, 399887, 399888, 399889, 399890, 399894, 399896, 399900, 399901, 399904, 399905, 399906, 399907, 399911, 399912, 399916, 399920, 399925, 399935, 399936, 399937, 399945, 399954, 399957, 399959, 399960, 399962, 399968, 399969, 399973, 399976, 399977, 399978, 399989, 399992, 400002, 400004, 400007, 400012, 400013, 400023, and 400024 each inhibited PCSK9 by at least 80% in this experiment.
Isis Nos 399819, 395152, 395153, 395155, 395156, 395158, 395163, 395164, 395165, 395166, 395168, 395178, 395183, 395185, 395187, 395189, 395190, 395194, 395199, 395203, 395214, 399793, 399804, 399813, 399821, 399822, 399836, 399856, 399868, 399877, 399887, 399888, 399890, 399900, 399904, 399905, 399907, 399911, 399912, 399916, 399925, 399936, 399954, 399960, 399962, 399969, 399976, 399977, 399978, 399989, 399992, 400012, 400013, 400023, and 400024 each inhibited PCSK9 by at least 85% in this experiment.
Isis Nos 399819, 395165, 395183, 395185, 399868, 399900, 399907, 399911, 399912, 399916, 399936, 399969, and 400024 each inhibited PCSK9 by at least 90% in this experiment.
Example 3
Antisense Inhibition of Human PCSK9 (Hep3B Cells)
Antisense oligonucleotides targeted to a PCSK9 nucleic acid were tested for their effects on PCSK9 mRNA in vitro. Cultured Hep3B cells at a density of 4000 cells per well in a 96-well plate were treated with 75 nM of antisense oligonucleotide. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Results are presented as percent inhibition of PCSK9, relative to untreated control cells. Antisense oligonucleotides that exhibited at least 30% inhibition of PCSK9 expression are shown in Tables 17, 18, and 19.
The motif column indicates the wing-gap-wing motif of each antisense oligonucleotide. Antisense oligonucleotides were designed as 5-10-5 gapmers, or alternatively as 3-14-3 gapmers, or alternatively as 2-13-5 gapmers where the gap segment comprises 2′-deoxynucleotides and each wing segment comprises 2′-MOE nucleotides. As illustrated in Table 17, 18 and 19, a single nucleobase sequence may be represented by a 5-10-5 motif as well as a 3-14-3 motif as well as a 2-13-5 motif. “5′ target site” indicates the 5′-most nucleotide which the antisense oligonucleotide is targeted on the indicated GENBANK Accession No.







TABLE 17







Antisense inhibition of PCSK9 in human cells (Hep3B)


targeting SEQ ID NO: 1














5′
3′







Target
Target







Site to
Site to



SEQ


Isis
SEQ ID
SEQ ID


%
ID


No.
NO: 1
NO: 1
Sequence 5′-3′
Motif
inhibition
NO





410742
 294
 313
GCCTGGAGCTGACGGTGCCC
5-10-5
75.1
159


 


405999
 298
 317
GACCGCCTGGAGCTGACGGT
5-10-5
70.3
160


 


395151
 300
 319
AGGACCGCCTGGAGCTGACG
5-10-5
57.3
  6


 


405861
 406
 425
AAGGCTAGCACCAGCTCCTC
5-10-5
90.5
162


 


405862
 407
 426
CAAGGCTAGCACCAGCTCCT
5-10-5
92.3
163


 


405863
 408
 427
GCAAGGCTAGCACCAGCTCC
5-10-5
82.6
164


 


405864
 409
 428
CGCAAGGCTAGCACCAGCTC
5-10-5
91.8
165


 


395152
 410
 429
ACGCAAGGCTAGCACCAGCT
5-10-5
93.3
  7


 


405865
 411
 430
AACGCAAGGCTAGCACCAGC
5-10-5
84.6
166


 


405866
 412
 431
GAACGCAAGGCTAGCACCAG
5-10-5
81.8
167


 


405867
 413
 432
GGAACGCAAGGCTAGCACCA
5-10-5
74.0
168


 


405868
 414
 433
CGGAACGCAAGGCTAGCACC
5-10-5
76.8
169


 


399793
 417
 436
CCTCGGAACGCAAGGCTAGC
5-10-5
77.9
  8


 


410743
 421
 440
TCCTCCTCGGAACGCAAGGC
5-10-5
76.7
170


 


410744
 446
 465
GTGCTCGGGTGCTTCGGCCA
5-10-5
76.9
171


 


410745
 466
 485
TGGAAGGTGGCTGTGGTTCC
5-10-5
41.7
172


 


395153
 480
 499
CCTTGGCGCAGCGGTGGAAG
5-10-5
81.1
  9


 


406000
 482
 501
ATCCTTGGCGCAGCGGTGGA
5-10-5
79.1
173


 


406001
 484
 503
GGATCCTTGGCGCAGCGGTG
5-10-5
63.1
174


 


406002
 488
 507
CCACGGATCCTTGGCGCAGC
5-10-5
80.8
175


 


410746
 507
 526
CGTAGGTGCCAGGCAACCTC
5-10-5
44.8
176


 


410747
 545
 564
TGACTGCGAGAGGTGGGTCT
5-10-5
NA
177


 


406003
 555
 574
CAGTGCGCTCTGACTGCGAG
5-10-5
84.2
178


 


406004
 557
 576
GGCAGTGCGCTCTGACTGCG
5-10-5
56.4
179


 


406005
 559
 578
CGGGCAGTGCGCTCTGACTG
5-10-5
83.4
180


 


406006
 562
 581
CGGCGGGCAGTGCGCTCTGA
5-10-5
71.0
181


 


410748
 591
 610
ATCCCCGGCGGGCAGCCTGG
5-10-5
56.0
182


 


406007
 595
 614
AGGTATCCCCGGCGGGCAGC
5-10-5
76.3
183


 


410574
 597
 616
TGAGGTATCCCCGGCGGGCA
3-14-3
69.1
184


 


410529
 597
 616
TGAGGTATCCCCGGCGGGCA
5-10-5
77.4
184


 


410647
 597
 616
TGAGGTATCCCCGGCGGGCA
2-13-5
90.7
184


 


410575
 598
 617
GTGAGGTATCCCCGGCGGGC
3-14-3
71.5
185


 


410648
 598
 617
GTGAGGTATCCCCGGCGGGC
2-13-5
82.8
185


 


410530
 598
 617
GTGAGGTATCCCCGGCGGGC
5-10-5
83.2
185


 


410649
 599
 618
GGTGAGGTATCCCCGGCGGG
2-13-5
53.5
186


 


410576
 599
 618
GGTGAGGTATCCCCGGCGGG
3-14-3
56.2
186


 


410531
 599
 618
GGTGAGGTATCCCCGGCGGG
5-10-5
63.6
186


 


410650
 600
 619
TGGTGAGGTATCCCCGGCGG
2-13-5
86.1
 11


 


399877
 600
 619
TGGTGAGGTATCCCCGGCGG
3-14-3
92.6
 11


 


395155
 600
 619
TGGTGAGGTATCCCCGGCGG
5-10-5
92.7
 11


 


410577
 601
 620
TTGGTGAGGTATCCCCGGCG
3-14-3
62.4
187


 


410532
 601
 620
TTGGTGAGGTATCCCCGGCG
5-10-5
63.5
187


 


410651
 601
 620
TTGGTGAGGTATCCCCGGCG
2-13-5
75.5
187


 


410652
 602
 621
CTTGGTGAGGTATCCCCGGC
2-13-5
81.2
188


 


410578
 602
 621
CTTGGTGAGGTATCCCCGGC
3-14-3
83.3
188


 


406008
 602
 621
CTTGGTGAGGTATCCCCGGC
5-10-5
95.2
188


 


410653
 603
 622
TCTTGGTGAGGTATCCCCGG
2-13-5
68.9
189


 


410579
 603
 622
TCTTGGTGAGGTATCCCCGG
3-14-3
74.0
189


 


410533
 603
 622
TCTTGGTGAGGTATCCCCGG
5-10-5
79.0
189


 


410580
 604
 623
ATCTTGGTGAGGTATCCCCG
3-14-3
62.3
190


 


410654
 604
 623
ATCTTGGTGAGGTATCCCCG
2-13-5
65.2
190


 


406009
 604
 623
ATCTTGGTGAGGTATCCCCG
5-10-5
90.7
190


 


410534
 605
 624
GATCTTGGTGAGGTATCCCC
5-10-5
59.8
191


 


410581
 605
 624
GATCTTGGTGAGGTATCCCC
3-14-3
72.9
191


 


410655
 605
 624
GATCTTGGTGAGGTATCCCC
2-13-5
77.4
191


 


399794
 606
 625
GGATCTTGGTGAGGTATCCC
5-10-5
56.5
 12


 


410656
 606
 625
GGATCTTGGTGAGGTATCCC
2-13-5
68.0
 12


 


399950
 606
 625
GGATCTTGGTGAGGTATCCC
3-14-3
74.4
 12


 


410535
 607
 626
AGGATCTTGGTGAGGTATCC
5-10-5
65.7
192


 


410582
 607
 626
AGGATCTTGGTGAGGTATCC
3-14-3
69.5
192


 


410657
 607
 626
AGGATCTTGGTGAGGTATCC
2-13-5
73.1
192


 


406010
 609
 628
GCAGGATCTTGGTGAGGTAT
5-10-5
56.9
193


 


406011
 611
 630
ATGCAGGATCTTGGTGAGGT
5-10-5
70.7
194


 


406012
 613
 632
ACATGCAGGATCTTGGTGAG
5-10-5
69.1
195


 


406013
 617
 636
GAAGACATGCAGGATCTTGG
5-10-5
77.4
196


 


395156
 620
 639
ATGGAAGACATGCAGGATCT
5-10-5
76.7
 14


 


410749
 628
 647
AGAAGGCCATGGAAGACATG
5-10-5
59.4
197


 


410750
 638
 657
GAAGCCAGGAAGAAGGCCAT
5-10-5
57.5
198


 


399879
 646
 665
TTCACCAGGAAGCCAGGAAG
3-14-3
91.9
 15


 


406014
 648
 667
TCTTCACCAGGAAGCCAGGA
5-10-5
94.0
199


 


406015
 653
 672
ACTCATCTTCACCAGGAAGC
5-10-5
78.8
200


 


406016
 655
 674
CCACTCATCTTCACCAGGAA
5-10-5
87.2
201


 


410730
 657
 676
CGCCACTCATCTTCACCAGG
5-10-5
85.4
202


 


406017
 659
 678
GTCGCCACTCATCTTCACCA
5-10-5
76.3
203


 


406018
 661
 680
AGGTCGCCACTCATCTTCAC
5-10-5
63.2
204


 


406019
 663
 682
GCAGGTCGCCACTCATCTTC
5-10-5
68.1
205


 


406020
 665
 684
CAGCAGGTCGCCACTCATCT
5-10-5
83.7
206


 


410731
 667
 686
TCCAGCAGGTCGCCACTCAT
5-10-5
72.9
207


 


410751
 685
 704
GGCAACTTCAAGGCCAGCTC
5-10-5
84.6
208


 


395158
 705
 724
CCTCGATGTAGTCGACATGG
5-10-5
93.9
 17


 


410752
 724
 743
GCAAAGACAGAGGAGTCCTC
5-10-5
76.3
209


 


406021
 782
 801
TTCATCCGCCCGGTACCGTG
5-10-5
79.6
210


 


406022
 784
 803
TATTCATCCGCCCGGTACCG
5-10-5
78.7
211


 


406023
 787
 806
TGGTATTCATCCGCCCGGTA
5-10-5
92.5
212


 


406024
 789
 808
GCTGGTATTCATCCGCCCGG
5-10-5
69.7
213


 


410732
 791
 810
GGGCTGGTATTCATCCGCCC
5-10-5
30.4
214


 


410753
 821
 840
ATACACCTCCACCAGGCTGC
5-10-5
72.8
215


 


406025
 832
 851
GTGTCTAGGAGATACACCTC
5-10-5
74.2
216


 


406026
 837
 856
TGCTGGTGTCTAGGAGATAC
5-10-5
80.5
217


 


405869
 862
 881
TCGATTTCCCGGTGGTCACT
5-10-5
87.6
218


 


405870
 863
 882
CTCGATTTCCCGGTGGTCAC
5-10-5
83.3
219


 


405871
 864
 883
CCTCGATTTCCCGGTGGTCA
5-10-5
87.0
220


 


405872
 865
 884
CCCTCGATTTCCCGGTGGTC
5-10-5
88.2
221


 


399798
 866
 885
GCCCTCGATTTCCCGGTGGT
5-10-5
84.3
 22


 


399954
 866
 885
GCCCTCGATTTCCCGGTGGT
3-14-3
90.3
 22


 


405873
 867
 886
TGCCCTCGATTTCCCGGTGG
5-10-5
90.0
222


 


405874
 868
 887
CTGCCCTCGATTTCCCGGTG
5-10-5
91.4
223


 


405875
 869
 888
CCTGCCCTCGATTTCCCGGT
5-10-5
93.5
224


 


405876
 870
 889
CCCTGCCCTCGATTTCCCGG
5-10-5
90.1
225


 


406027
 874
 893
ATGACCCTGCCCTCGATTTC
5-10-5
73.9
226


 


406028
 876
 895
CCATGACCCTGCCCTCGATT
5-10-5
92.3
227


 


406029
 878
 897
GACCATGACCCTGCCCTCGA
5-10-5
91.4
228


 


406030
 882
 901
CGGTGACCATGACCCTGCCC
5-10-5
95.6
229


 


406031
 884
 903
GTCGGTGACCATGACCCTGC
5-10-5
89.6
230


 


410733
 886
 905
AAGTCGGTGACCATGACCCT
5-10-5
65.7
231


 


406032
 888
 907
CGAAGTCGGTGACCATGACC
5-10-5
88.5
232


 


410754
 898
 917
GGCACATTCTCGAAGTCGGT
5-10-5
80.1
233


 


395163
 923
 942
GTGGAAGCGGGTCCCGTCCT
5-10-5
83.9
 25


 


410755
 933
 952
TGGCCTGTCTGTGGAAGCGG
5-10-5
NA
234


 


410756
 960
 979
GGTGGGTGCCATGACTGTCA
5-10-5
67.1
235


 


406033
 967
 986
CCTGCCAGGTGGGTGCCATG
5-10-5
91.2
237


 


406034
 972
 991
CCACCCCTGCCAGGTGGGTG
5-10-5
59.3
238


 


406035
 977
 996
GCTGACCACCCCTGCCAGGT
5-10-5
91.7
239


 


410757
 985
1004
TCCCGGCCGCTGACCACCCC
5-10-5
88.9
240


 


406036
 989
1008
GGCATCCCGGCCGCTGACCA
5-10-5
84.0
241


 


406037
 992
1011
GCCGGCATCCCGGCCGCTGA
5-10-5
55.9
242


 


410536
 997
1016
GCCACGCCGGCATCCCGGCC
5-10-5
63.7
243


 


410658
 997
1016
GCCACGCCGGCATCCCGGCC
2-13-5
82.0
243


 


410583
 997
1016
GCCACGCCGGCATCCCGGCC
3-14-3
87.3
243


 


410537
 998
1017
GGCCACGCCGGCATCCCGGC
5-10-5
58.5
244


 


410659
 998
1017
GGCCACGCCGGCATCCCGGC
2-13-5
73.7
244


 


410584
 998
1017
GGCCACGCCGGCATCCCGGC
3-14-3
79.5
244


 


410660
 999
1018
TGGCCACGCCGGCATCCCGG
2-13-5
63.0
245


 


410585
 999
1018
TGGCCACGCCGGCATCCCGG
3-14-3
73.2
245


 


406038
 999
1018
TGGCCACGCCGGCATCCCGG
5-10-5
86.3
245


 


410661
1000
1019
TTGGCCACGCCGGCATCCCG
2-13-5
53.3
246


 


410586
1000
1019
TTGGCCACGCCGGCATCCCG
3-14-3
60.3
246


 


405877
1000
1019
TTGGCCACGCCGGCATCCCG
5-10-5
83.2
246


 


410587
1001
1020
CTTGGCCACGCCGGCATCCC
3-14-3
63.2
247


 


410662
1001
1020
CTTGGCCACGCCGGCATCCC
2-13-5
67.3
247


 


405878
1001
1020
CTTGGCCACGCCGGCATCCC
5-10-5
91.4
247


 


410588
1002
1021
CCTTGGCCACGCCGGCATCC
3-14-3
65.3
248


 


410663
1002
1021
CCTTGGCCACGCCGGCATCC
2-13-5
67.9
248


 


405879
1002
1021
CCTTGGCCACGCCGGCATCC
5-10-5
94.1
248


 


410589
1003
1022
CCCTTGGCCACGCCGGCATC
3-14-3
80.1
249


 


410664
1003
1022
CCCTTGGCCACGCCGGCATC
2-13-5
81.9
249


 


405880
1003
1022
CCCTTGGCCACGCCGGCATC
5-10-5
93.5
249


 


410665
1004
1023
ACCCTTGGCCACGCCGGCAT
2-13-5
78.7
 28


 


399887
1004
1023
ACCCTTGGCCACGCCGGCAT
3-14-3
91.9
 28


 


395165
1004
1023
ACCCTTGGCCACGCCGGCAT
5-10-5
96.7
 28


 


410590
1005
1024
CACCCTTGGCCACGCCGGCA
3-14-3
82.2
250


 


410666
1005
1024
CACCCTTGGCCACGCCGGCA
2-13-5
82.7
250


 


405881
1005
1024
CACCCTTGGCCACGCCGGCA
5-10-5
98.3
250


 


410667
1006
1025
GCACCCTTGGCCACGCCGGC
2-13-5
84.7
251


 


410591
1006
1025
GCACCCTTGGCCACGCCGGC
3-14-3
86.3
251


 


405882
1006
1025
GCACCCTTGGCCACGCCGGC
5-10-5
91.6
251


 


410668
1007
1026
GGCACCCTTGGCCACGCCGG
2-13-5
91.4
252


 


405883
1007
1026
GGCACCCTTGGCCACGCCGG
5-10-5
92.5
252


 


410592
1007
1026
GGCACCCTTGGCCACGCCGG
3-14-3
93.4
252


 


410669
1008
1027
TGGCACCCTTGGCCACGCCG
2-13-5
69.0
253


 


410593
1008
1027
TGGCACCCTTGGCCACGCCG
3-14-3
88.8
253


 


405884
1008
1027
TGGCACCCTTGGCCACGCCG
5-10-5
90.8
253


 


410670
1009
1028
CTGGCACCCTTGGCCACGCC
2-13-5
74.9
254


 


410594
1009
1028
CTGGCACCCTTGGCCACGCC
3-14-3
75.2
254


 


410538
1009
1028
CTGGCACCCTTGGCCACGCC
5-10-5
78.2
254


 


410539
1010
1029
GCTGGCACCCTTGGCCACGC
5-10-5
61.9
255


 


410595
1010
1029
GCTGGCACCCTTGGCCACGC
3-14-3
83.1
255


 


410671
1010
1029
GCTGGCACCCTTGGCCACGC
2-13-5
83.3
255


 


410758
1015
1034
CGCATGCTGGCACCCTTGGC
5-10-5
73.6
256


 


406039
1036
1055
CAGTTGAGCACGCGCAGGCT
5-10-5
91.2
257


 


406040
1038
1057
GGCAGTTGAGCACGCGCAGG
5-10-5
96.5
258


 


399888
1040
1059
TTGGCAGTTGAGCACGCGCA
3-14-3
90.9
 29


 


395166
1040
1059
TTGGCAGTTGAGCACGCGCA
5-10-5
96.2
 29


 


406041
1042
1061
CCTTGGCAGTTGAGCACGCG
5-10-5
96.6
259


 


399801
1045
1064
TTCCCTTGGCAGTTGAGCAC
5-10-5
95.6
 30


 


406042
1047
1066
CCTTCCCTTGGCAGTTGAGC
5-10-5
92.0
260


 


406043
1051
1070
GTGCCCTTCCCTTGGCAGTT
5-10-5
91.9
261


 


406044
1053
1072
CCGTGCCCTTCCCTTGGCAG
5-10-5
95.4
262


 


410759
1064
1083
GGTGCCGCTAACCGTGCCCT
5-10-5
83.7
263


 


410760
1088
1107
CCGAATAAACTCCAGGCCTA
5-10-5
96.7
265


 


406045
1096
1115
TGGCTTTTCCGAATAAACTC
5-10-5
88.4
266


 


395168
1098
1117
GCTGGCTTTTCCGAATAAAC
5-10-5
91.9
 32


 


405909
1100
1119
CAGCTGGCTTTTCCGAATAA
5-10-5
80.3
267


 


405910
1102
1121
ACCAGCTGGCTTTTCCGAAT
5-10-5
90.9
268


 


405911
1104
1123
GGACCAGCTGGCTTTTCCGA
5-10-5
88.0
269


 


405912
1108
1127
GGCTGGACCAGCTGGCTTTT
5-10-5
78.2
270


 


410761
1119
1138
GTGGCCCCACAGGCTGGACC
5-10-5
53.7
271


 


410762
1132
1151
AGCAGCACCACCAGTGGCCC
5-10-5
57.6
272


 


410763
1154
1173
GCTGTACCCACCCGCCAGGG
5-10-5
68.8
273


 


410764
1200
1219
CGACCCCAGCCCTCGCCAGG
5-10-5
66.3
274


 


399891
1210
1229
GTGACCAGCACGACCCCAGC
3-14-3
91.1
 33


 


405913
1212
1231
CGGTGACCAGCACGACCCCA
5-10-5
93.8
275


 


405914
1214
1233
AGCGGTGACCAGCACGACCC
5-10-5
90.4
276


 


405915
1216
1235
GCAGCGGTGACCAGCACGAC
5-10-5
86.8
277


 


405916
1218
1237
CGGCAGCGGTGACCAGCACG
5-10-5
91.3
278


 


410734
1219
1238
CCGGCAGCGGTGACCAGCAC
5-10-5
65.8
279


 


405917
1222
1241
TTGCCGGCAGCGGTGACCAG
5-10-5
62.0
280


 


405918
1224
1243
AGTTGCCGGCAGCGGTGACC
5-10-5
33.5
281


 


405919
1226
1245
GAAGTTGCCGGCAGCGGTGA
5-10-5
68.2
282


 


405920
1228
1247
CGGAAGTTGCCGGCAGCGGT
5-10-5
86.2
283


 


405921
1230
1249
CCCGGAAGTTGCCGGCAGCG
5-10-5
85.5
284


 


405922
1232
1251
GTCCCGGAAGTTGCCGGCAG
5-10-5
86.0
285


 


410765
1273
1292
ATGACCTCGGGAGCTGAGGC
5-10-5
50.4
286


 


410766
1283
1302
CCCAACTGTGATGACCTCGG
5-10-5
70.6
287


 


405923
1295
1314
GGCATTGGTGGCCCCAACTG
5-10-5
94.3
288


 


410767
1305
1324
GCTGGTCTTGGGCATTGGTG
5-10-5
65.1
289


 


405924
1318
1337
CCCAGGGTCACCGGCTGGTC
5-10-5
93.1
290


 


410735
1320
1339
TCCCCAGGGTCACCGGCTGG
5-10-5
78.1
291


 


405925
1322
1341
AGTCCCCAGGGTCACCGGCT
5-10-5
93.5
292


 


405926
1324
1343
AAAGTCCCCAGGGTCACCGG
5-10-5
94.6
293


 


405927
1328
1347
CCCCAAAGTCCCCAGGGTCA
5-10-5
94.5
294


 


405928
1333
1352
TTGGTCCCCAAAGTCCCCAG
5-10-5
90.0
295


 


405929
1337
1356
AAAGTTGGTCCCCAAAGTCC
5-10-5
85.5
296


 


399895
1340
1359
GCCAAAGTTGGTCCCCAAAG
3-14-3
84.6
 36


 


395173
1340
1359
GCCAAAGTTGGTCCCCAAAG
5-10-5
94.8
 36


 


405930
1342
1361
CGGCCAAAGTTGGTCCCCAA
5-10-5
92.9
297


 


405931
1344
1363
AGCGGCCAAAGTTGGTCCCC
5-10-5
88.8
298


 


405932
1346
1365
ACAGCGGCCAAAGTTGGTCC
5-10-5
90.6
299


 


405933
1348
1367
ACACAGCGGCCAAAGTTGGT
5-10-5
NA
300


 


405934
1350
1369
CCACACAGCGGCCAAAGTTG
5-10-5
92.7
301


 


405935
1354
1373
AGGTCCACACAGCGGCCAAA
5-10-5
86.0
302


 


410736
1356
1375
AGAGGTCCACACAGCGGCCA
5-10-5
73.8
303


 


405936
1358
1377
AAAGAGGTCCACACAGCGGC
5-10-5
93.9
304


 


410768
1380
1399
CAATGATGTCCTCCCCTGGG
5-10-5
79.7
305


 


405937
1387
1406
GAGGCACCAATGATGTCCTC
5-10-5
77.7
306


 


405938
1391
1410
GCTGGAGGCACCAATGATGT
5-10-5
75.5
307


 


405939
1393
1412
TCGCTGGAGGCACCAATGAT
5-10-5
70.6
308


 


405940
1395
1414
AGTCGCTGGAGGCACCAATG
5-10-5
84.4
309


 


405941
1397
1416
GCAGTCGCTGGAGGCACCAA
5-10-5
90.1
310


 


399804
1400
1419
GCTGCAGTCGCTGGAGGCAC
5-10-5
80.8
 40


 


399960
1400
1419
GCTGCAGTCGCTGGAGGCAC
3-14-3
82.4
 40


 


405942
1402
1421
GTGCTGCAGTCGCTGGAGGC
5-10-5
69.8
311


 


405943
1404
1423
AGGTGCTGCAGTCGCTGGAG
5-10-5
83.6
312


 


410737
1406
1425
GCAGGTGCTGCAGTCGCTGG
5-10-5
49.8
313


 


405944
1407
1426
AGCAGGTGCTGCAGTCGCTG
5-10-5
80.7
314


 


405945
1409
1428
AAAGCAGGTGCTGCAGTCGC
5-10-5
41.1
315


 


405946
1413
1432
ACACAAAGCAGGTGCTGCAG
5-10-5
70.4
316


 


405947
1415
1434
TGACACAAAGCAGGTGCTGC
5-10-5
72.5
317


 


410769
1425
1444
TCCCACTCTGTGACACAAAG
5-10-5
84.9
318


 


405948
1467
1486
TCATGGCTGCAATGCCAGCC
5-10-5
45.9
319


 


399806
1470
1489
GCATCATGGCTGCAATGCCA
5-10-5
82.8
 42


 


399962
1470
1489
GCATCATGGCTGCAATGCCA
3-14-3
86.1
 42


 


405949
1472
1491
CAGCATCATGGCTGCAATGC
5-10-5
84.9
320


 


405950
1474
1493
GACAGCATCATGGCTGCAAT
5-10-5
76.9
321


 


405951
1476
1495
CAGACAGCATCATGGCTGCA
5-10-5
75.4
322


 


405952
1480
1499
TCGGCAGACAGCATCATGGC
5-10-5
85.1
323


 


410738
1482
1501
GCTCGGCAGACAGCATCATG
5-10-5
68.4
324


 


405953
1484
1503
CGGCTCGGCAGACAGCATCA
5-10-5
93.6
325


 


405954
1486
1505
TCCGGCTCGGCAGACAGCAT
5-10-5
87.5
326


 


410770
1500
1519
CGGCCAGGGTGAGCTCCGGC
5-10-5
68.5
327


 


405955
1513
1532
CTCTGCCTCAACTCGGCCAG
5-10-5
94.2
328


 


405956
1515
1534
GTCTCTGCCTCAACTCGGCC
5-10-5
94.3
329


 


405958
1519
1538
ATCAGTCTCTGCCTCAACTC
5-10-5
80.3
331


 


405959
1521
1540
GGATCAGTCTCTGCCTCAAC
5-10-5
95.4
332


 


405960
1523
1542
GTGGATCAGTCTCTGCCTCA
5-10-5
90.1
333


 


405961
1525
1544
AAGTGGATCAGTCTCTGCCT
5-10-5
88.8
334


 


405962
1528
1547
GAGAAGTGGATCAGTCTCTG
5-10-5
56.1
335


 


410739
1530
1549
CAGAGAAGTGGATCAGTCTC
5-10-5
NA
336


 


405963
1532
1551
GGCAGAGAAGTGGATCAGTC
5-10-5
59.7
337


 


405964
1536
1555
CTTTGGCAGAGAAGTGGATC
5-10-5
45.3
338


 


405965
1541
1560
GACATCTTTGGCAGAGAAGT
5-10-5
55.9
339


 


405966
1543
1562
ATGACATCTTTGGCAGAGAA
5-10-5
51.3
340


 


405967
1547
1566
ATTGATGACATCTTTGGCAG
5-10-5
64.4
341


 


405968
1549
1568
TCATTGATGACATCTTTGGC
5-10-5
74.6
342


 


399967
1552
1571
GCCTCATTGATGACATCTTT
3-14-3
80.1
 48


 


405969
1554
1573
AGGCCTCATTGATGACATCT
5-10-5
63.0
343


 


405970
1556
1575
CCAGGCCTCATTGATGACAT
5-10-5
66.8
344


 


410740
1558
1577
AACCAGGCCTCATTGATGAC
5-10-5
46.8
345


 


405885
1560
1579
GGAACCAGGCCTCATTGATG
5-10-5
73.4
346


 


410596
1562
1581
AGGGAACCAGGCCTCATTGA
3-14-3
NA
348


 


410672
1562
1581
AGGGAACCAGGCCTCATTGA
2-13-5
NA
348


 


405887
1562
1581
AGGGAACCAGGCCTCATTGA
5-10-5
NA
348


 


410597
1563
1582
CAGGGAACCAGGCCTCATTG
3-14-3
NA
349


 


405888
1563
1582
CAGGGAACCAGGCCTCATTG
5-10-5
NA
349


 


410673
1563
1582
CAGGGAACCAGGCCTCATTG
2-13-5
NA
349


 


410674
1564
1583
TCAGGGAACCAGGCCTCATT
2-13-5
60.7
 49


 


399812
1564
1583
TCAGGGAACCAGGCCTCATT
5-10-5
68.7
 49


 


399968
1564
1583
TCAGGGAACCAGGCCTCATT
3-14-3
84.5
 49


 


410598
1565
1584
CTCAGGGAACCAGGCCTCAT
3-14-3
74.5
350


 


410675
1565
1584
CTCAGGGAACCAGGCCTCAT
2-13-5
76.9
350


 


405889
1565
1584
CTCAGGGAACCAGGCCTCAT
5-10-5
80.2
350


 


410599
1566
1585
CCTCAGGGAACCAGGCCTCA
3-14-3
70.6
351


 


410676
1566
1585
CCTCAGGGAACCAGGCCTCA
2-13-5
77.8
351


 


405890
1566
1585
CCTCAGGGAACCAGGCCTCA
5-10-5
87.7
351


 


410600
1567
1586
TCCTCAGGGAACCAGGCCTC
3-14-3
76.5
352


 


410677
1567
1586
TCCTCAGGGAACCAGGCCTC
2-13-5
88.4
352


 


405891
1567
1586
TCCTCAGGGAACCAGGCCTC
5-10-5
96.1
352


 


405892
1568
1587
GTCCTCAGGGAACCAGGCCT
5-10-5
71.4
353


 


410601
1568
1587
GTCCTCAGGGAACCAGGCCT
3-14-3
72.7
353


 


410678
1568
1587
GTCCTCAGGGAACCAGGCCT
2-13-5
75.0
353


 


395178
1569
1588
GGTCCTCAGGGAACCAGGCC
5-10-5
76.7
 50


 


410679
1569
1588
GGTCCTCAGGGAACCAGGCC
2-13-5
77.6
 50


 


399900
1569
1588
GGTCCTCAGGGAACCAGGCC
3-14-3
92.0
 50


 


408653
1570
1589
TGGTCCTCAGGGAACCAGGC
5-10-5
42.5
354


 


410602
1570
1589
TGGTCCTCAGGGAACCAGGC
3-14-3
66.5
354


 


410680
1570
1589
TGGTCCTCAGGGAACCAGGC
2-13-5
71.6
354


 


410603
1571
1590
CTGGTCCTCAGGGAACCAGG
3-14-3
40.8
355


 


410681
1571
1590
CTGGTCCTCAGGGAACCAGG
2-13-5
44.4
355


 


405971
1571
1590
CTGGTCCTCAGGGAACCAGG
5-10-5
65.5
355


 


410540
1572
1591
GCTGGTCCTCAGGGAACCAG
5-10-5
50.9
356


 


410682
1572
1591
GCTGGTCCTCAGGGAACCAG
2-13-5
54.1
356


 


410604
1572
1591
GCTGGTCCTCAGGGAACCAG
3-14-3
62.5
356


 


405972
1573
1592
CGCTGGTCCTCAGGGAACCA
5-10-5
77.6
357


 


405973
1578
1597
GTACCCGCTGGTCCTCAGGG
5-10-5
83.8
358


 


405974
1580
1599
CAGTACCCGCTGGTCCTCAG
5-10-5
89.0
359


 


399813
1583
1602
GGTCAGTACCCGCTGGTCCT
5-10-5
72.4
 51


 


399969
1583
1602
GGTCAGTACCCGCTGGTCCT
3-14-3
93.4
 51


 


410771
1628
1647
ACCTGCCCCATGGGTGCTGG
5-10-5
NA
360


 


395181
1640
1659
AAACAGCTGCCAACCTGCCC
5-10-5
87.5
 52


 


405975
1642
1661
CAAAACAGCTGCCAACCTGC
5-10-5
77.4
361


 


405976
1647
1666
TCCTGCAAAACAGCTGCCAA
5-10-5
81.2
362


 


405977
1649
1668
AGTCCTGCAAAACAGCTGCC
5-10-5
89.5
363


 


410772
1660
1679
GCTGACCATACAGTCCTGCA
5-10-5
91.2
364


 


405978
1672
1691
GGCCCCGAGTGTGCTGACCA
5-10-5
95.3
365


 


399904
1675
1694
GTAGGCCCCGAGTGTGCTGA
3-14-3
92.6
 54


 


405979
1677
1696
GTGTAGGCCCCGAGTGTGCT
5-10-5
85.5
366


 


405980
1679
1698
CCGTGTAGGCCCCGAGTGTG
5-10-5
86.5
367


 


405981
1681
1700
ATCCGTGTAGGCCCCGAGTG
5-10-5
77.8
368


 


410741
1683
1702
CCATCCGTGTAGGCCCCGAG
5-10-5
78.4
369


 


405982
1685
1704
GGCCATCCGTGTAGGCCCCG
5-10-5
86.7
370


 


405983
1687
1706
GTGGCCATCCGTGTAGGCCC
5-10-5
73.1
371


 


405984
1735
1754
CTGGAGCAGCTCAGCAGCTC
5-10-5
83.4
372


 


405985
1737
1756
AACTGGAGCAGCTCAGCAGC
5-10-5
50.7
373


 


405986
1742
1761
GGAGAAACTGGAGCAGCTCA
5-10-5
75.6
374


 


405987
1744
1763
CTGGAGAAACTGGAGCAGCT
5-10-5
88.0
375


 


399905
1812
1831
CGTTGTGGGCCCGGCAGACC
3-14-3
91.2
 57


 


395183
1812
1831
CGTTGTGGGCCCGGCAGACC
5-10-5
93.1
 57


 


410541
1849
1868
CACCTGGCAATGGCGTAGAC
5-10-5
46.1
376


 


410605
1849
1868
CACCTGGCAATGGCGTAGAC
3-14-3
67.4
376


 


410683
1849
1868
CACCTGGCAATGGCGTAGAC
2-13-5
71.8
376


 


410606
1850
1869
GCACCTGGCAATGGCGTAGA
3-14-3
74.3
377


 


410542
1850
1869
GCACCTGGCAATGGCGTAGA
5-10-5
75.5
377


 


410684
1850
1869
GCACCTGGCAATGGCGTAGA
2-13-5
78.7
377


 


410543
1851
1870
AGCACCTGGCAATGGCGTAG
5-10-5
76.4
378


 


410685
1851
1870
AGCACCTGGCAATGGCGTAG
2-13-5
76.9
378


 


410607
1851
1870
AGCACCTGGCAATGGCGTAG
3-14-3
77.1
378


 


410544
1852
1871
CAGCACCTGGCAATGGCGTA
5-10-5
62.7
379


 


410608
1852
1871
CAGCACCTGGCAATGGCGTA
3-14-3
69.6
379


 


410686
1852
1871
CAGCACCTGGCAATGGCGTA
2-13-5
81.0
379


 


410545
1853
1872
GCAGCACCTGGCAATGGCGT
5-10-5
75.5
380


 


410687
1853
1872
GCAGCACCTGGCAATGGCGT
2-13-5
79.2
380


 


410609
1853
1872
GCAGCACCTGGCAATGGCGT
3-14-3
83.2
380


 


410610
1854
1873
GGCAGCACCTGGCAATGGCG
3-14-3
67.5
381


 


410688
1854
1873
GGCAGCACCTGGCAATGGCG
2-13-5
89.3
381


 


405988
1854
1873
GGCAGCACCTGGCAATGGCG
5-10-5
95.9
381


 


410546
1855
1874
AGGCAGCACCTGGCAATGGC
5-10-5
74.8
382


 


410689
1855
1874
AGGCAGCACCTGGCAATGGC
2-13-5
83.9
382


 


410611
1855
1874
AGGCAGCACCTGGCAATGGC
3-14-3
88.2
382


 


410612
1856
1875
CAGGCAGCACCTGGCAATGG
3-14-3
72.8
383


 


410690
1856
1875
CAGGCAGCACCTGGCAATGG
2-13-5
74.8
383


 


405989
1856
1875
CAGGCAGCACCTGGCAATGG
5-10-5
88.1
383


 


410547
1857
1876
GCAGGCAGCACCTGGCAATG
5-10-5
63.0
384


 


410691
1857
1876
GCAGGCAGCACCTGGCAATG
2-13-5
70.2
384


 


410613
1857
1876
GCAGGCAGCACCTGGCAATG
3-14-3
76.7
384


 


395184
1858
1877
AGCAGGCAGCACCTGGCAAT
5-10-5
68.5
 58


 


410692
1858
1877
AGCAGGCAGCACCTGGCAAT
2-13-5
69.1
 58


 


399906
1858
1877
AGCAGGCAGCACCTGGCAAT
3-14-3
94.2
 58


 


410614
1859
1878
TAGCAGGCAGCACCTGGCAA
3-14-3
36.6
385


 


410548
1859
1878
TAGCAGGCAGCACCTGGCAA
5-10-5
75.0
385


 


410693
1859
1878
TAGCAGGCAGCACCTGGCAA
2-13-5
82.0
385


 


405990
1860
1879
GTAGCAGGCAGCACCTGGCA
5-10-5
96.8
386


 


410773
1905
1924
TGGCCTCAGCTGGTGGAGCT
5-10-5
61.3
387


 


410549
1915
1934
GTCCCCATGCTGGCCTCAGC
5-10-5
63.0
388


 


410615
1915
1934
GTCCCCATGCTGGCCTCAGC
3-14-3
72.5
388


 


410694
1915
1934
GTCCCCATGCTGGCCTCAGC
2-13-5
74.8
388


 


410550
1916
1935
GGTCCCCATGCTGGCCTCAG
5-10-5
69.4
389


 


410616
1916
1935
GGTCCCCATGCTGGCCTCAG
3-14-3
80.7
389


 


410695
1916
1935
GGTCCCCATGCTGGCCTCAG
2-13-5
85.5
389


 


410551
1917
1936
GGGTCCCCATGCTGGCCTCA
5-10-5
68.7
390


 


410696
1917
1936
GGGTCCCCATGCTGGCCTCA
2-13-5
86.3
390


 


410617
1917
1936
GGGTCCCCATGCTGGCCTCA
3-14-3
86.5
390


 


410552
1918
1937
CGGGTCCCCATGCTGGCCTC
5-10-5
77.9
391


 


410618
1918
1937
CGGGTCCCCATGCTGGCCTC
3-14-3
86.7
391


 


410697
1918
1937
CGGGTCCCCATGCTGGCCTC
2-13-5
87.0
391


 


410553
1919
1938
ACGGGTCCCCATGCTGGCCT
5-10-5
72.8
392


 


410619
1919
1938
ACGGGTCCCCATGCTGGCCT
3-14-3
84.1
392


 


410698
1919
1938
ACGGGTCCCCATGCTGGCCT
2-13-5
87.0
392


 


410699
1920
1939
CACGGGTCCCCATGCTGGCC
2-13-5
79.5
 59


 


395185
1920
1939
CACGGGTCCCCATGCTGGCC
5-10-5
92.7
 59


 


399907
1920
1939
CACGGGTCCCCATGCTGGCC
3-14-3
93.7
 59


 


410620
1921
1940
ACACGGGTCCCCATGCTGGC
3-14-3
74.4
393


 


410554
1921
1940
ACACGGGTCCCCATGCTGGC
5-10-5
79.7
393


 


410700
1921
1940
ACACGGGTCCCCATGCTGGC
2-13-5
84.4
393


 


410621
1922
1941
GACACGGGTCCCCATGCTGG
3-14-3
76.0
394


 


410701
1922
1941
GACACGGGTCCCCATGCTGG
2-13-5
83.4
394


 


405991
1922
1941
GACACGGGTCCCCATGCTGG
5-10-5
91.9
394


 


410555
1923
1942
GGACACGGGTCCCCATGCTG
5-10-5
77.0
395


 


410622
1923
1942
GGACACGGGTCCCCATGCTG
3-14-3
79.8
395


 


410702
1923
1942
GGACACGGGTCCCCATGCTG
2-13-5
85.0
395


 


410703
1924
1943
TGGACACGGGTCCCCATGCT
2-13-5
70.4
396


 


410623
1924
1943
TGGACACGGGTCCCCATGCT
3-14-3
78.9
396


 


405992
1924
1943
TGGACACGGGTCCCCATGCT
5-10-5
89.3
396


 


410704
1925
1944
GTGGACACGGGTCCCCATGC
2-13-5
78.6
397


 


410556
1925
1944
GTGGACACGGGTCCCCATGC
5-10-5
81.4
397


 


410624
1925
1944
GTGGACACGGGTCCCCATGC
3-14-3
82.8
397


 


410625
1926
1945
AGTGGACACGGGTCCCCATG
3-14-3
64.4
398


 


410705
1926
1945
AGTGGACACGGGTCCCCATG
2-13-5
74.7
398


 


405993
1926
1945
AGTGGACACGGGTCCCCATG
5-10-5
85.5
398


 


410706
1927
1946
CAGTGGACACGGGTCCCCAT
2-13-5
70.8
399


 


410557
1927
1946
CAGTGGACACGGGTCCCCAT
5-10-5
74.2
399


 


410626
1927
1946
CAGTGGACACGGGTCCCCAT
3-14-3
76.8
399


 


410627
1928
1947
GCAGTGGACACGGGTCCCCA
3-14-3
71.4
400


 


410707
1928
1947
GCAGTGGACACGGGTCCCCA
2-13-5
72.8
400


 


405994
1928
1947
GCAGTGGACACGGGTCCCCA
5-10-5
95.2
400


 


410708
1929
1948
GGCAGTGGACACGGGTCCCC
2-13-5
79.0
401


 


410628
1929
1948
GGCAGTGGACACGGGTCCCC
3-14-3
88.1
401


 


410558
1929
1948
GGCAGTGGACACGGGTCCCC
5-10-5
88.9
401


 


410629
1930
1949
TGGCAGTGGACACGGGTCCC
3-14-3
66.5
402


 


410709
1930
1949
TGGCAGTGGACACGGGTCCC
2-13-5
66.9
402


 


405995
1930
1949
TGGCAGTGGACACGGGTCCC
5-10-5
92.4
402


 


410630
1931
1950
GTGGCAGTGGACACGGGTCC
3-14-3
54.1
403


 


410710
1931
1950
GTGGCAGTGGACACGGGTCC
2-13-5
61.4
403


 


410559
1931
1950
GTGGCAGTGGACACGGGTCC
5-10-5
77.5
403


 


410711
1932
1951
GGTGGCAGTGGACACGGGTC
2-13-5
NA
404


 


410631
1932
1951
GGTGGCAGTGGACACGGGTC
3-14-3
NA
404


 


410560
1932
1951
GGTGGCAGTGGACACGGGTC
5-10-5
42.5
404


 


410712
1933
1952
TGGTGGCAGTGGACACGGGT
2-13-5
NA
405


 


410632
1933
1952
TGGTGGCAGTGGACACGGGT
3-14-3
NA
405


 


410561
1933
1952
TGGTGGCAGTGGACACGGGT
5-10-5
NA
405


 


410774
1936
1955
TGTTGGTGGCAGTGGACACG
5-10-5
47.8
406


 


410775
1962
1981
AGCTGCAGCCTGTGAGGACG
5-10-5
36.3
407


 


410776
1990
2009
GTGCCAAGGTCCTCCACCTC
5-10-5
77.3
408


 


410777
2010
2029
TCAGCACAGGCGGCTTGTGG
5-10-5
40.0
409


 


410778
2040
2059
CCACGCACTGGTTGGGCTGA
5-10-5
52.7
410


 


399908
2100
2119
CTTTGCATTCCAGACCTGGG
3-14-3
74.9
 60


 


410713
2100
2119
CTTTGCATTCCAGACCTGGG
2-13-5
84.9
 60


 


395186
2100
2119
CTTTGCATTCCAGACCTGGG
5-10-5
93.7
 60


 


410714
2101
2120
ACTTTGCATTCCAGACCTGG
2-13-5
82.4
411


 


410633
2101
2120
ACTTTGCATTCCAGACCTGG
3-14-3
83.8
411


 


410562
2101
2120
ACTTTGCATTCCAGACCTGG
5-10-5
87.3
411


 


410715
2102
2121
GACTTTGCATTCCAGACCTG
2-13-5
80.5
412


 


410634
2102
2121
GACTTTGCATTCCAGACCTG
3-14-3
81.8
412


 


405996
2102
2121
GACTTTGCATTCCAGACCTG
5-10-5
91.0
412


 


410563
2103
2122
TGACTTTGCATTCCAGACCT
5-10-5
75.4
413


 


410635
2103
2122
TGACTTTGCATTCCAGACCT
3-14-3
75.9
413


 


410716
2103
2122
TGACTTTGCATTCCAGACCT
2-13-5
88.3
413


 


410636
2104
2123
TTGACTTTGCATTCCAGACC
3-14-3
61.3
414


 


410564
2104
2123
TTGACTTTGCATTCCAGACC
5-10-5
71.3
414


 


410717
2104
2123
TTGACTTTGCATTCCAGACC
2-13-5
76.7
414


 


410718
2105
2124
CTTGACTTTGCATTCCAGAC
2-13-5
55.8
 61


 


399973
2105
2124
CTTGACTTTGCATTCCAGAC
3-14-3
82.1
 61


 


399817
2105
2124
CTTGACTTTGCATTCCAGAC
5-10-5
93.6
 61


 


405997
2107
2126
TCCTTGACTTTGCATTCCAG
5-10-5
95.6
415


 


410779
2120
2139
CGGGATTCCATGCTCCTTGA
5-10-5
79.7
416


 


410780
2150
2169
GCAGGCCACGGTCACCTGCT
5-10-5
60.9
417


 


410781
2187
2206
GGAGGGCACTGCAGCCAGTC
5-10-5
66.0
418


 


410719
2305
2324
CAGATGGCAACGGCTGTCAC
2-13-5
61.0
419


 


410637
2305
2324
CAGATGGCAACGGCTGTCAC
3-14-3
66.0
419


 


410565
2305
2324
CAGATGGCAACGGCTGTCAC
5-10-5
71.6
419


 


410638
2306
2325
GCAGATGGCAACGGCTGTCA
3-14-3
71.9
420


 


410720
2306
2325
GCAGATGGCAACGGCTGTCA
2-13-5
75.1
420


 


410566
2306
2325
GCAGATGGCAACGGCTGTCA
5-10-5
77.7
420


 


410639
2307
2326
AGCAGATGGCAACGGCTGTC
3-14-3
67.7
421


 


410721
2307
2326
AGCAGATGGCAACGGCTGTC
2-13-5
68.8
421


 


410567
2307
2326
AGCAGATGGCAACGGCTGTC
5-10-5
72.1
421


 


410568
2308
2327
CAGCAGATGGCAACGGCTGT
5-10-5
54.2
422


 


410640
2308
2327
CAGCAGATGGCAACGGCTGT
3-14-3
57.7
422


 


410722
2308
2327
CAGCAGATGGCAACGGCTGT
2-13-5
59.2
422


 


410569
2309
2328
GCAGCAGATGGCAACGGCTG
5-10-5
57.7
423


 


410641
2309
2328
GCAGCAGATGGCAACGGCTG
3-14-3
66.1
423


 


410723
2309
2328
GCAGCAGATGGCAACGGCTG
2-13-5
81.1
423


 


399909
2310
2329
GGCAGCAGATGGCAACGGCT
3-14-3
69.9
 62


 


410724
2310
2329
GGCAGCAGATGGCAACGGCT
2-13-5
87.4
 62


 


395187
2310
2329
GGCAGCAGATGGCAACGGCT
5-10-5
92.7
 62


 


410642
2311
2330
CGGCAGCAGATGGCAACGGC
3-14-3
70.4
424


 


410725
2311
2330
CGGCAGCAGATGGCAACGGC
2-13-5
75.6
424


 


410570
2311
2330
CGGCAGCAGATGGCAACGGC
5-10-5
78.3
424


 


410571
2312
2331
CCGGCAGCAGATGGCAACGG
5-10-5
52.8
425


 


410643
2312
2331
CCGGCAGCAGATGGCAACGG
3-14-3
59.8
425


 


410726
2312
2331
CCGGCAGCAGATGGCAACGG
2-13-5
69.8
425


 


410644
2313
2332
TCCGGCAGCAGATGGCAACG
3-14-3
59.0
426


 


410727
2313
2332
TCCGGCAGCAGATGGCAACG
2-13-5
68.3
426


 


405998
2313
2332
TCCGGCAGCAGATGGCAACG
5-10-5
75.6
426


 


410572
2314
2333
CTCCGGCAGCAGATGGCAAC
5-10-5
40.9
427


 


410728
2314
2333
CTCCGGCAGCAGATGGCAAC
2-13-5
56.4
427


 


410645
2314
2333
CTCCGGCAGCAGATGGCAAC
3-14-3
70.2
427


 


410729
2315
2334
GCTCCGGCAGCAGATGGCAA
2-13-5
67.7
428


 


410573
2315
2334
GCTCCGGCAGCAGATGGCAA
5-10-5
71.8
428


 


410646
2315
2334
GCTCCGGCAGCAGATGGCAA
3-14-3
72.8
428


 


410782
2325
2344
CCAGGTGCCGGCTCCGGCAG
5-10-5
43.2
429


 


410783
2335
2354
GAGGCCTGCGCCAGGTGCCG
5-10-5
63.7
430


 


399819
2509
2528
CCCACTCAAGGGCCAGGCCA
5-10-5
93.4
 65


 


399912
2597
2616
ATGCCCCACAGTGAGGGAGG
3-14-3
84.3
 66


 


405893
2828
2847
ATGAGGGCCATCAGCACCTT
5-10-5
93.9
431


 


405894
2829
2848
GATGAGGGCCATCAGCACCT
5-10-5
93.9
432


 


405895
2830
2849
AGATGAGGGCCATCAGCACC
5-10-5
91.8
433


 


405896
2831
2850
GAGATGAGGGCCATCAGCAC
5-10-5
88.5
434


 


395194
2832
2851
GGAGATGAGGGCCATCAGCA
5-10-5
90.0
 69


 


399916
2832
2851
GGAGATGAGGGCCATCAGCA
3-14-3
94.5
 69


 


405897
2833
2852
TGGAGATGAGGGCCATCAGC
5-10-5
85.1
435


 


405898
2834
2853
CTGGAGATGAGGGCCATCAG
5-10-5
86.9
436


 


405899
2835
2854
GCTGGAGATGAGGGCCATCA
5-10-5
91.5
437


 


405900
2836
2855
AGCTGGAGATGAGGGCCATC
5-10-5
83.7
438


 


405901
2902
2921
GCTAGATGCCATCCAGAAAG
5-10-5
88.4
439


 


405902
2903
2922
GGCTAGATGCCATCCAGAAA
5-10-5
89.3
440


 


405903
2904
2923
TGGCTAGATGCCATCCAGAA
5-10-5
92.6
441


 


405904
2905
2924
CTGGCTAGATGCCATCCAGA
5-10-5
90.4
442


 


399821
2906
2925
TCTGGCTAGATGCCATCCAG
5-10-5
91.7
 71


 


405905
2907
2926
CTCTGGCTAGATGCCATCCA
5-10-5
90.7
443


 


405906
2908
2927
CCTCTGGCTAGATGCCATCC
5-10-5
93.3
444


 


405907
2909
2928
GCCTCTGGCTAGATGCCATC
5-10-5
90.2
445


 


405908
2910
2929
AGCCTCTGGCTAGATGCCAT
5-10-5
83.3
446


 


399978
2988
3007
AGCCTGGCATAGAGCAGAGT
3-14-3
96.4
 73









Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.
Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155, 395158, 395163, 395165, 395166, 395168, 395173, 395181, 395183, 395185, 395186, 395187, 395194, 399798, 399801, 399804, 399806, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904, 399905, 399906, 399907, 399912, 399916, 399954, 399960, 399962, 399967, 399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865, 405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406002, 406003, 406005, 406008, 406009, 406014, 406016, 406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617, 410618, 410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650, 410652, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686, 410688, 410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701, 410702, 410713, 410714, 410715, 410716, 410723, 410724, 410730, 410751, 410754, 410757, 410759, 410760, 410769, and 410772. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.
ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395181, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399954, 399962, 399969, 399978, 405861, 405862, 405864, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558, 410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628, 410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698, 410702, 410716, 410724, 410730, 410757, 410760, and 410772 each exhibited at least 85% inhibition of PCSK9 mRNA levels.
ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399954, 399969, 399978, 405861, 405862, 405864, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405904, 405905, 405906, 405907, 405910, 405913, 405914, 405916, 405923, 405924, 405925, 405926, 405927, 405928, 405930, 405932, 405934, 405936, 405941, 405953, 405955, 405956, 405959, 405960, 405978, 405988, 405990, 405991, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406023, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410647, 410668, 410760, and 410772 each exhibited at least 90% inhibition of PCSK9 mRNA levels.
ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, 406044, and 410760 each exhibited at least 95% inhibition of PCSK9 mRNA levels.







TABLE 18







Antisense inhibition of PCSK9 in human cells (Hep3B) 


targeting SEQ ID NO: 2














5′ 
3′ 







Target
Target







Site to
Site to



SEQ


Isis
SEQ ID
SEQ ID


%
ID


No.
NO: 2
NO: 2
Sequence 5′-3′
Motif
inhibition
NO:





410742
 2433
 2452
GCCTGGAGCTGACGGTGCCC
5-10-5
75.1
159


 


405999
 2437
 2456
GACCGCCTGGAGCTGACGGT
5-10-5
70.3
160


 


395151
 2439
 2458
AGGACCGCCTGGAGCTGACG
5-10-5
57.3
  6


 


405861
 2545
 2564
AAGGCTAGCACCAGCTCCTC
5-10-5
90.5
162


 


405862
 2546
 2565
CAAGGCTAGCACCAGCTCCT
5-10-5
92.3
163


 


405863
 2547
 2566
GCAAGGCTAGCACCAGCTCC
5-10-5
82.6
164


 


405864
 2548
 2567
CGCAAGGCTAGCACCAGCTC
5-10-5
91.8
165


 


395152
 2549
 2568
ACGCAAGGCTAGCACCAGCT
5-10-5
93.3
  7


 


405865
 2550
 2569
AACGCAAGGCTAGCACCAGC
5-10-5
84.6
166


 


405866
 2551
 2570
GAACGCAAGGCTAGCACCAG
5-10-5
81.8
167


 


405867
 2552
 2571
GGAACGCAAGGCTAGCACCA
5-10-5
74.0
168


 


405868
 2553
 2572
CGGAACGCAAGGCTAGCACC
5-10-5
76.8
169


 


399793
 2556
 2575
CCTCGGAACGCAAGGCTAGC
5-10-5
77.9
  8


 


410743
 2560
 2579
TCCTCCTCGGAACGCAAGGC
5-10-5
76.7
170


 


410744
 2585
 2604
GTGCTCGGGTGCTTCGGCCA
5-10-5
76.9
171


 


410745
 2605
 2624
TGGAAGGTGGCTGTGGTTCC
5-10-5
41.7
172


 


395153
 2619
 2638
CCTTGGCGCAGCGGTGGAAG
5-10-5
81.1
  9


 


410746
 6444
 6463
CGTAGGTGCCAGGCAACCTC
5-10-5
44.8
176


 


410747
 6482
 6501
TGACTGCGAGAGGTGGGTCT
5-10-5
NA
177


 


406003
 6492
 6511
CAGTGCGCTCTGACTGCGAG
5-10-5
84.2
178


 


406004
 6494
 6513
GGCAGTGCGCTCTGACTGCG
5-10-5
56.4
179


 


406005
 6496
 6515
CGGGCAGTGCGCTCTGACTG
5-10-5
83.4
180


 


406006
 6499
 6518
CGGCGGGCAGTGCGCTCTGA
5-10-5
71.0
181


 


410748
 6528
 6547
ATCCCCGGCGGGCAGCCTGG
5-10-5
56.0
182


 


406007
 6532
 6551
AGGTATCCCCGGCGGGCAGC
5-10-5
76.3
183


 


410574
 6534
 6553
TGAGGTATCCCCGGCGGGCA
3-14-3
69.1
184


 


410529
 6534
 6553
TGAGGTATCCCCGGCGGGCA
5-10-5
77.4
184


 


410647
 6534
 6553
TGAGGTATCCCCGGCGGGCA
2-13-5
90.7
184


 


410575
 6535
 6554
GTGAGGTATCCCCGGCGGGC
3-14-3
71.5
185


 


410648
 6535
 6554
GTGAGGTATCCCCGGCGGGC
2-13-5
82.8
185


 


410530
 6535
 6554
GTGAGGTATCCCCGGCGGGC
5-10-5
83.2
185


 


410649
 6536
 6555
GGTGAGGTATCCCCGGCGGG
2-13-5
53.5
186


 


410576
 6536
 6555
GGTGAGGTATCCCCGGCGGG
3-14-3
56.2
186


 


410531
 6536
 6555
GGTGAGGTATCCCCGGCGGG
5-10-5
63.6
186


 


410650
 6537
 6556
TGGTGAGGTATCCCCGGCGG
2-13-5
86.1
 11


 


399877
 6537
 6556
TGGTGAGGTATCCCCGGCGG
3-14-3
92.6
 11


 


395155
 6537
 6556
TGGTGAGGTATCCCCGGCGG
5-10-5
92.7
 11


 


410577
 6538
 6557
TTGGTGAGGTATCCCCGGCG
3-14-3
62.4
187


 


410532
 6538
 6557
TTGGTGAGGTATCCCCGGCG
5-10-5
63.5
187


 


410651
 6538
 6557
TTGGTGAGGTATCCCCGGCG
2-13-5
75.5
187


 


410652
 6539
 6558
CTTGGTGAGGTATCCCCGGC
2-13-5
81.2
188


 


410578
 6539
 6558
CTTGGTGAGGTATCCCCGGC
3-14-3
83.3
188


 


406008
 6539
 6558
CTTGGTGAGGTATCCCCGGC
5-10-5
95.2
188


 


410653
 6540
 6559
TCTTGGTGAGGTATCCCCGG
2-13-5
68.9
189


 


410579
 6540
 6559
TCTTGGTGAGGTATCCCCGG
3-14-3
74.0
189


 


410533
 6540
 6559
TCTTGGTGAGGTATCCCCGG
5-10-5
79.0
189


 


410580
 6541
 6560
ATCTTGGTGAGGTATCCCCG
3-14-3
62.3
190


 


410654
 6541
 6560
ATCTTGGTGAGGTATCCCCG
2-13-5
65.2
190


 


406009
 6541
 6560
ATCTTGGTGAGGTATCCCCG
5-10-5
90.7
190


 


410534
 6542
 6561
GATCTTGGTGAGGTATCCCC
5-10-5
59.8
191


 


410581
 6542
 6561
GATCTTGGTGAGGTATCCCC
3-14-3
72.9
191


 


410655
 6542
 6561
GATCTTGGTGAGGTATCCCC
2-13-5
77.4
191


 


399794
 6543
 6562
GGATCTTGGTGAGGTATCCC
5-10-5
56.5
 12


 


410656
 6543
 6562
GGATCTTGGTGAGGTATCCC
2-13-5
68.0
 12


 


399950
 6543
 6562
GGATCTTGGTGAGGTATCCC
3-14-3
74.4
 12


 


410535
 6544
 6563
AGGATCTTGGTGAGGTATCC
5-10-5
65.7
192


 


410582
 6544
 6563
AGGATCTTGGTGAGGTATCC
3-14-3
69.5
192


 


410657
 6544
 6563
AGGATCTTGGTGAGGTATCC
2-13-5
73.1
192


 


406010
 6546
 6565
GCAGGATCTTGGTGAGGTAT
5-10-5
56.9
193


 


406011
 6548
 6567
ATGCAGGATCTTGGTGAGGT
5-10-5
70.7
194


 


406012
 6550
 6569
ACATGCAGGATCTTGGTGAG
5-10-5
69.1
195


 


406013
 6554
 6573
GAAGACATGCAGGATCTTGG
5-10-5
77.4
196


 


395156
 6557
 6576
ATGGAAGACATGCAGGATCT
5-10-5
76.7
 14


 


410749
 6565
 6584
AGAAGGCCATGGAAGACATG
5-10-5
59.4
197


 


410750
 6575
 6594
GAAGCCAGGAAGAAGGCCAT
5-10-5
57.5
198


 


399879
 6583
 6602
TTCACCAGGAAGCCAGGAAG
3-14-3
91.9
 15


 


406014
 6585
 6604
TCTTCACCAGGAAGCCAGGA
5-10-5
94.0
199


 


406015
 6590
 6609
ACTCATCTTCACCAGGAAGC
5-10-5
78.8
200


 


406016
 6592
 6611
CCACTCATCTTCACCAGGAA
5-10-5
87.2
201


 


410730
 6594
 6613
CGCCACTCATCTTCACCAGG
5-10-5
85.4
202


 


406017
 6596
 6615
GTCGCCACTCATCTTCACCA
5-10-5
76.3
203


 


406018
 6598
 6617
AGGTCGCCACTCATCTTCAC
5-10-5
63.2
204


 


406019
 6600
 6619
GCAGGTCGCCACTCATCTTC
5-10-5
68.1
205


 


406020
 6602
 6621
CAGCAGGTCGCCACTCATCT
5-10-5
83.7
206


 


410731
 6604
 6623
TCCAGCAGGTCGCCACTCAT
5-10-5
72.9
207


 


395158
 9130
 9149
CCTCGATGTAGTCGACATGG
5-10-5
93.9
 17


 


410752
 9149
 9168
GCAAAGACAGAGGAGTCCTC
5-10-5
76.3
209


 


406021
 9207
 9226
TTCATCCGCCCGGTACCGTG
5-10-5
79.6
210


 


406022
 9209
 9228
TATTCATCCGCCCGGTACCG
5-10-5
78.7
211


 


406023
 9212
 9231
TGGTATTCATCCGCCCGGTA
5-10-5
92.5
212


 


406024
 9214
 9233
GCTGGTATTCATCCGCCCGG
5-10-5
69.7
213


 


410732
 9216
 9235
GGGCTGGTATTCATCCGCCC
5-10-5
30.4
214


 


399935
14601
14620
AGGACCCAAGTCATCCTGCT
3-14-3
81.1
 96


 


399936
14631
14650
GGCCATCAGCTGGCAATGCT
3-14-3
89.0
124


 


410753
14877
14896
ATACACCTCCACCAGGCTGC
5-10-5
72.8
315


 


406025
14888
14907
GTGTCTAGGAGATACACCTC
5-10-5
74.2
216


 


406026
14893
14912
TGCTGGTGTCTAGGAGATAC
5-10-5
80.5
217


 


405869
14918
14937
TCGATTTCCCGGTGGTCACT
5-10-5
87.6
218


 


405870
14919
14938
CTCGATTTCCCGGTGGTCAC
5-10-5
83.3
219


 


405871
14920
14939
CCTCGATTTCCCGGTGGTCA
5-10-5
87.0
220


 


405872
14921
14940
CCCTCGATTTCCCGGTGGTC
5-10-5
88.2
221


 


399798
14922
14941
GCCCTCGATTTCCCGGTGGT
5-10-5
84.3
 22


 


399954
14922
14941
GCCCTCGATTTCCCGGTGGT
3-14-3
90.3
 22


 


405873
14923
14942
TGCCCTCGATTTCCCGGTGG
5-10-5
90.0
222


 


405874
14924
14943
CTGCCCTCGATTTCCCGGTG
5-10-5
91.4
223


 


405875
14925
14944
CCTGCCCTCGATTTCCCGGT
5-10-5
93.5
224


 


405876
14926
14945
CCCTGCCCTCGATTTCCCGG
5-10-5
90.1
225


 


406027
14930
14949
ATGACCCTGCCCTCGATTTC
5-10-5
73.9
226


 


406028
14932
14951
CCATGACCCTGCCCTCGATT
5-10-5
92.3
227


 


406029
14934
14953
GACCATGACCCTGCCCTCGA
5-10-5
91.4
228


 


406030
14938
14957
CGGTGACCATGACCCTGCCC
5-10-5
95.6
229


 


406031
14940
14959
GTCGGTGACCATGACCCTGC
5-10-5
89.6
230


 


410733
14942
14961
AAGTCGGTGACCATGACCCT
5-10-5
65.7
231


 


406032
14944
14963
CGAAGTCGGTGACCATGACC
5-10-5
88.5
232


 


410754
14954
14973
GGCACATTCTCGAAGTCGGT
5-10-5
80.1
233


 


395163
14979
14998
GTGGAAGCGGGTCCCGTCCT
5-10-5
83.9
 25


 


410756
15254
15273
GGTGGGTGCCATGACTGTCA
5-10-5
67.1
235


 


406033
15261
15280
CCTGCCAGGTGGGTGCCATG
5-10-5
91.2
237


 


406034
15266
15285
CCACCCCTGCCAGGTGGGTG
5-10-5
59.3
238


 


406035
15271
15290
GCTGACCACCCCTGCCAGGT
5-10-5
91.7
239


 


410757
15279
15298
TCCCGGCCGCTGACCACCCC
5-10-5
88.9
240


 


406036
15283
15302
GGCATCCCGGCCGCTGACCA
5-10-5
84.0
241


 


406037
15286
15305
GCCGGCATCCCGGCCGCTGA
5-10-5
55.9
242


 


410536
15291
15310
GCCACGCCGGCATCCCGGCC
5-10-5
63.7
243


 


410658
15291
15310
GCCACGCCGGCATCCCGGCC
2-13-5
82.0
243


 


410583
15291
15310
GCCACGCCGGCATCCCGGCC
3-14-3
87.3
243


 


410537
15292
15311
GGCCACGCCGGCATCCCGGC
5-10-5
58.5
244


 


410659
15292
15311
GGCCACGCCGGCATCCCGGC
2-13-5
73.7
244


 


410584
15292
15311
GGCCACGCCGGCATCCCGGC
3-14-3
79.5
244


 


410660
15293
15312
TGGCCACGCCGGCATCCCGG
2-13-5
63.0
245


 


410585
15293
15312
TGGCCACGCCGGCATCCCGG
3-14-3
73.2
245


 


406038
15293
15312
TGGCCACGCCGGCATCCCGG
5-10-5
86.3
245


 


410661
15294
15313
TTGGCCACGCCGGCATCCCG
2-13-5
53.3
246


 


410586
15294
15313
TTGGCCACGCCGGCATCCCG
3-14-3
60.3
246


 


405877
15294
15313
TTGGCCACGCCGGCATCCCG
5-10-5
83.2
246


 


410587
15295
15314
CTTGGCCACGCCGGCATCCC
3-14-3
63.2
247


 


410662
15295
15314
CTTGGCCACGCCGGCATCCC
2-13-5
67.3
247


 


405878
15295
15314
CTTGGCCACGCCGGCATCCC
5-10-5
91.4
247


 


410588
15296
15315
CCTTGGCCACGCCGGCATCC
3-14-3
65.3
248


 


410663
15296
15315
CCTTGGCCACGCCGGCATCC
2-13-5
67.9
248


 


405879
15296
15315
CCTTGGCCACGCCGGCATCC
5-10-5
94.1
248


 


410589
15297
15316
CCCTTGGCCACGCCGGCATC
3-14-3
80.1
249


 


410664
15297
15316
CCCTTGGCCACGCCGGCATC
2-13-5
81.9
249


 


405880
15297
15316
CCCTTGGCCACGCCGGCATC
5-10-5
93.5
249


 


410665
15298
15317
ACCCTTGGCCACGCCGGCAT
2-13-5
78.7
 28


 


399887
15298
15317
ACCCTTGGCCACGCCGGCAT
3-14-3
91.9
 28


 


395165
15298
15317
ACCCTTGGCCACGCCGGCAT
5-10-5
96.7
 28


 


410590
15299
15318
CACCCTTGGCCACGCCGGCA
3-14-3
82.2
250


 


410666
15299
15318
CACCCTTGGCCACGCCGGCA
2-13-5
82.7
250


 


405881
15299
15318
CACCCTTGGCCACGCCGGCA
5-10-5
98.3
250


 


410667
15300
15319
GCACCCTTGGCCACGCCGGC
2-13-5
84.7
251


 


410591
15300
15319
GCACCCTTGGCCACGCCGGC
3-14-3
86.3
251


 


405882
15300
15319
GCACCCTTGGCCACGCCGGC
5-10-5
91.6
251


 


410668
15301
15320
GGCACCCTTGGCCACGCCGG
2-13-5
91.4
252


 


405883
15301
15320
GGCACCCTTGGCCACGCCGG
5-10-5
92.5
252


 


410592
15301
15320
GGCACCCTTGGCCACGCCGG
3-14-3
93.4
252


 


410669
15302
15321
TGGCACCCTTGGCCACGCCG
2-13-5
69.0
253


 


410593
15302
15321
TGGCACCCTTGGCCACGCCG
3-14-3
88.8
253


 


405884
15302
15321
TGGCACCCTTGGCCACGCCG
5-10-5
90.8
253


 


410670
15303
15322
CTGGCACCCTTGGCCACGCC
2-13-5
74.9
254


 


410594
15303
15322
CTGGCACCCTTGGCCACGCC
3-14-3
75.2
254


 


410538
15303
15322
CTGGCACCCTTGGCCACGCC
5-10-5
78.2
254


 


410539
15304
15323
GCTGGCACCCTTGGCCACGC
5-10-5
61.9
255


 


410595
15304
15323
GCTGGCACCCTTGGCCACGC
3-14-3
83.1
255


 


410671
15304
15323
GCTGGCACCCTTGGCCACGC
2-13-5
83.3
255


 


410758
15309
15328
CGCATGCTGGCACCCTTGGC
5-10-5
73.6
256


 


406039
15330
15349
CAGTTGAGCACGCGCAGGCT
5-10-5
91.2
257


 


406040
15332
15351
GGCAGTTGAGCACGCGCAGG
5-10-5
96.5
258


 


399888
15334
15353
TTGGCAGTTGAGCACGCGCA
3-14-3
90.9
 29


 


395166
15334
15353
TTGGCAGTTGAGCACGCGCA
5-10-5
96.2
 29


 


406041
15336
15355
CCTTGGCAGTTGAGCACGCG
5-10-5
96.6
259


 


399801
15339
15358
TTCCCTTGGCAGTTGAGCAC
5-10-5
95.6
 30


 


406042
15341
15360
CCTTCCCTTGGCAGTTGAGC
5-10-5
92.0
260


 


406043
15345
15364
GTGCCCTTCCCTTGGCAGTT
5-10-5
91.9
261


 


406044
15347
15366
CCGTGCCCTTCCCTTGGCAG
5-10-5
95.4
262


 


410759
15358
15377
GGTGCCGCTAACCGTGCCCT
5-10-5
83.7
263


 


406045
18591
18610
TGGCTTTTCCGAATAAACTC
5-10-5
88.4
266


 


395168
18593
18612
GCTGGCTTTTCCGAATAAAC
5-10-5
91.9
 32


 


405909
18595
18614
CAGCTGGCTTTTCCGAATAA
5-10-5
80.3
267


 


405910
18597
18616
ACCAGCTGGCTTTTCCGAAT
5-10-5
90.9
268


 


405911
18599
18618
GGACCAGCTGGCTTTTCCGA
5-10-5
88.0
269


 


405912
18603
18622
GGCTGGACCAGCTGGCTTTT
5-10-5
78.2
270


 


410761
18614
18633
GTGGCCCCACAGGCTGGACC
5-10-5
53.7
271


 


410762
18627
18646
AGCAGCACCACCAGTGGCCC
5-10-5
57.6
272


 


410763
18649
18668
GCTGTACCCACCCGCCAGGG
5-10-5
68.8
273


 


410764
18695
18714
CGACCCCAGCCCTCGCCAGG
5-10-5
66.3
274


 


399891
18705
18724
GTGACCAGCACGACCCCAGC
3-14-3
91.1
 33


 


405913
18707
18726
CGGTGACCAGCACGACCCCA
5-10-5
93.8
275


 


405914
18709
18728
AGCGGTGACCAGCACGACCC
5-10-5
90.4
276


 


405915
18711
18730
GCAGCGGTGACCAGCACGAC
5-10-5
86.8
277


 


405916
18713
18732
CGGCAGCGGTGACCAGCACG
5-10-5
91.3
278


 


410734
18714
18733
CCGGCAGCGGTGACCAGCAC
5-10-5
65.8
279


 


405917
18717
18736
TTGCCGGCAGCGGTGACCAG
5-10-5
62.0
280


 


405918
18719
18738
AGTTGCCGGCAGCGGTGACC
5-10-5
33.5
281


 


405919
18721
18740
GAAGTTGCCGGCAGCGGTGA
5-10-5
68.2
282


 


405920
18723
18742
CGGAAGTTGCCGGCAGCGGT
5-10-5
86.2
283


 


405921
18725
18744
CCCGGAAGTTGCCGGCAGCG
5-10-5
85.5
284


 


405922
18727
18746
GTCCCGGAAGTTGCCGGCAG
5-10-5
86.0
285


 


405923
19931
19950
GGCATTGGTGGCCCCAACTG
5-10-5
94.3
288


 


410767
19941
19960
GCTGGTCTTGGGCATTGGTG
5-10-5
65.1
289


 


405924
19954
19973
CCCAGGGTCACCGGCTGGTC
5-10-5
93.1
290


 


410735
19956
19975
TCCCCAGGGTCACCGGCTGG
5-10-5
78.1
291


 


405925
19958
19977
AGTCCCCAGGGTCACCGGCT
5-10-5
93.5
292


 


405926
19960
19979
AAAGTCCCCAGGGTCACCGG
5-10-5
94.6
293


 


405927
19964
19983
CCCCAAAGTCCCCAGGGTCA
5-10-5
94.5
294


 


405928
19969
19988
TTGGTCCCCAAAGTCCCCAG
5-10-5
90.0
295


 


405929
19973
19992
AAAGTTGGTCCCCAAAGTCC
5-10-5
85.5
296


 


399895
19976
19995
GCCAAAGTTGGTCCCCAAAG
3-14-3
84.6
 36


 


395173
19976
19995
GCCAAAGTTGGTCCCCAAAG
5-10-5
94.8
 36


 


405930
19978
19997
CGGCCAAAGTTGGTCCCCAA
5-10-5
92.9
297


 


405931
19980
19999
AGCGGCCAAAGTTGGTCCCC
5-10-5
88.8
298


 


405932
19982
20001
ACAGCGGCCAAAGTTGGTCC
5-10-5
90.6
299


 


405933
19984
20003
ACACAGCGGCCAAAGTTGGT
5-10-5
NA
300


 


405934
19986
20005
CCACACAGCGGCCAAAGTTG
5-10-5
92.7
301


 


405935
19990
20009
AGGTCCACACAGCGGCCAAA
5-10-5
86.0
302


 


410736
19992
20011
AGAGGTCCACACAGCGGCCA
5-10-5
73.8
303


 


405936
19994
20013
AAAGAGGTCCACACAGCGGC
5-10-5
93.9
304


 


410768
20016
20035
CAATGATGTCCTCCCCTGGG
5-10-5
79.7
305


 


405937
20023
20042
GAGGCACCAATGATGTCCTC
5-10-5
77.7
306


 


405938
20027
20046
GCTGGAGGCACCAATGATGT
5-10-5
75.5
307


 


405939
20029
20048
TCGCTGGAGGCACCAATGAT
5-10-5
70.6
308


 


405940
20031
20050
AGTCGCTGGAGGCACCAATG
5-10-5
84.4
309


 


405941
20033
20052
GCAGTCGCTGGAGGCACCAA
5-10-5
90.1
310


 


399804
20036
20055
GCTGCAGTCGCTGGAGGCAC
5-10-5
80.8
 40


 


399960
20036
20055
GCTGCAGTCGCTGGAGGCAC
3-14-3
82.4
 40


 


405942
20038
20057
GTGCTGCAGTCGCTGGAGGC
5-10-5
69.8
311


 


405943
20040
20059
AGGTGCTGCAGTCGCTGGAG
5-10-5
83.6
312


 


410737
20042
20061
GCAGGTGCTGCAGTCGCTGG
5-10-5
49.8
313


 


405944
20043
20062
AGCAGGTGCTGCAGTCGCTG
5-10-5
80.7
314


 


405945
20045
20064
AAAGCAGGTGCTGCAGTCGC
5-10-5
41.1
315


 


405946
20049
20068
ACACAAAGCAGGTGCTGCAG
5-10-5
70.4
316


 


405947
20051
20070
TGACACAAAGCAGGTGCTGC
5-10-5
72.5
317


 


410769
20061
20080
TCCCACTCTGTGACACAAAG
5-10-5
84.9
318


 


405949
20629
20648
CAGCATCATGGCTGCAATGC
5-10-5
84.9
320


 


405950
20631
20650
GACAGCATCATGGCTGCAAT
5-10-5
76.9
321


 


405951
20633
20652
CAGACAGCATCATGGCTGCA
5-10-5
75.4
322


 


405952
20637
20656
TCGGCAGACAGCATCATGGC
5-10-5
85.1
323


 


410738
20639
20658
GCTCGGCAGACAGCATCATG
5-10-5
68.4
324


 


405953
20641
20660
CGGCTCGGCAGACAGCATCA
5-10-5
93.6
325


 


405954
20643
20662
TCCGGCTCGGCAGACAGCAT
5-10-5
87.5
326


 


410770
20657
20676
CGGCCAGGGTGAGCTCCGGC
5-10-5
68.5
327


 


405955
20670
20689
CTCTGCCTCAACTCGGCCAG
5-10-5
94.2
328


 


405956
20672
20691
GTCTCTGCCTCAACTCGGCC
5-10-5
94.3
329


 


405958
20676
20695
ATCAGTCTCTGCCTCAACTC
5-10-5
80.3
331


 


405959
20678
20697
GGATCAGTCTCTGCCTCAAC
5-10-5
95.4
332


 


405960
20680
20699
GTGGATCAGTCTCTGCCTCA
5-10-5
90.1
333


 


405961
20682
20701
AAGTGGATCAGTCTCTGCCT
5-10-5
88.8
334


 


405962
20685
20704
GAGAAGTGGATCAGTCTCTG
5-10-5
56.1
335


 


410739
20687
20706
CAGAGAAGTGGATCAGTCTC
5-10-5
NA
336


 


405963
20689
20708
GGCAGAGAAGTGGATCAGTC
5-10-5
59.7
337


 


405964
20693
20712
CTTTGGCAGAGAAGTGGATC
5-10-5
45.3
338


 


405965
20698
20717
GACATCTTTGGCAGAGAAGT
5-10-5
55.9
339


 


405966
20700
20719
ATGACATCTTTGGCAGAGAA
5-10-5
51.3
340


 


405967
20704
20723
ATTGATGACATCTTTGGCAG
5-10-5
64.4
341


 


405968
20706
20725
TCATTGATGACATCTTTGGC
5-10-5
74.6
342


 


399967
20709
20728
GCCTCATTGATGACATCTTT
3-14-3
80.1
 48


 


405969
20711
20730
AGGCCTCATTGATGACATCT
5-10-5
63.0
343


 


405970
20713
20732
CCAGGCCTCATTGATGACAT
5-10-5
66.8
344


 


410740
20715
20734
AACCAGGCCTCATTGATGAC
5-10-5
46.8
345


 


405885
20717
20736
GGAACCAGGCCTCATTGATG
5-10-5
73.4
346


 


410596
20719
20738
AGGGAACCAGGCCTCATTGA
3-14-3
NA
348


 


410672
20719
20738
AGGGAACCAGGCCTCATTGA
2-13-5
NA
348


 


405887
20719
20738
AGGGAACCAGGCCTCATTGA
5-10-5
NA
348


 


410597
20720
20739
CAGGGAACCAGGCCTCATTG
3-14-3
NA
349


 


405888
20720
20739
CAGGGAACCAGGCCTCATTG
5-10-5
NA
349


 


410673
20720
20739
CAGGGAACCAGGCCTCATTG
2-13-5
NA
349


 


410674
20721
20740
TCAGGGAACCAGGCCTCATT
2-13-5
60.7
 49


 


399812
20721
20740
TCAGGGAACCAGGCCTCATT
5-10-5
68.7
 49


 


399968
20721
20740
TCAGGGAACCAGGCCTCATT
3-14-3
84.5
 49


 


410598
20722
20741
CTCAGGGAACCAGGCCTCAT
3-14-3
74.5
350


 


410675
20722
20741
CTCAGGGAACCAGGCCTCAT
2-13-5
76.9
350


 


405889
20722
20741
CTCAGGGAACCAGGCCTCAT
5-10-5
80.2
350


 


410599
20723
20742
CCTCAGGGAACCAGGCCTCA
3-14-3
70.6
351


 


410676
20723
20742
CCTCAGGGAACCAGGCCTCA
2-13-5
77.8
351


 


405890
20723
20742
CCTCAGGGAACCAGGCCTCA
5-10-5
87.7
351


 


410600
20724
20743
TCCTCAGGGAACCAGGCCTC
3-14-3
76.5
352


 


410677
20724
20743
TCCTCAGGGAACCAGGCCTC
2-13-5
88.4
352


 


405891
20724
20743
TCCTCAGGGAACCAGGCCTC
5-10-5
96.1
352


 


405892
20725
20744
GTCCTCAGGGAACCAGGCCT
5-10-5
71.4
353


 


410601
20725
20744
GTCCTCAGGGAACCAGGCCT
3-14-3
72.7
353


 


410678
20725
20744
GTCCTCAGGGAACCAGGCCT
2-13-5
75.0
353


 


395178
20726
20745
GGTCCTCAGGGAACCAGGCC
5-10-5
76.7
 50


 


410679
20726
20745
GGTCCTCAGGGAACCAGGCC
2-13-5
77.6
 50


 


399900
20726
20745
GGTCCTCAGGGAACCAGGCC
3-14-3
92.0
 50


 


408653
20727
20746
TGGTCCTCAGGGAACCAGGC
5-10-5
42.5
354


 


410602
20727
20746
TGGTCCTCAGGGAACCAGGC
3-14-3
66.5
354


 


410680
20727
20746
TGGTCCTCAGGGAACCAGGC
2-13-5
71.6
354


 


410603
20728
20747
CTGGTCCTCAGGGAACCAGG
3-14-3
40.8
355


 


410681
20728
20747
CTGGTCCTCAGGGAACCAGG
2-13-5
44.4
355


 


405971
20728
20747
CTGGTCCTCAGGGAACCAGG
5-10-5
65.5
355


 


410540
20729
20748
GCTGGTCCTCAGGGAACCAG
5-10-5
50.9
356


 


410682
20729
20748
GCTGGTCCTCAGGGAACCAG
2-13-5
54.1
356


 


410604
20729
20748
GCTGGTCCTCAGGGAACCAG
3-14-3
62.5
356


 


405972
20730
20749
CGCTGGTCCTCAGGGAACCA
5-10-5
77.6
357


 


405973
20735
20754
GTACCCGCTGGTCCTCAGGG
5-10-5
83.8
358


 


405974
20737
20756
CAGTACCCGCTGGTCCTCAG
5-10-5
89.0
359


 


399813
20740
20759
GGTCAGTACCCGCTGGTCCT
5-10-5
72.4
 51


 


399969
20740
20759
GGTCAGTACCCGCTGGTCCT
3-14-3
93.4
 51


 


410771
20785
20804
ACCTGCCCCATGGGTGCTGG
5-10-5
NA
360


 


405975
21088
21107
CAAAACAGCTGCCAACCTGC
5-10-5
77.4
361


 


405976
21093
21112
TCCTGCAAAACAGCTGCCAA
5-10-5
81.2
362


 


405977
21095
21114
AGTCCTGCAAAACAGCTGCC
5-10-5
89.5
363


 


410772
21106
21125
GCTGACCATACAGTCCTGCA
5-10-5
91.2
264


 


405978
21118
21137
GGCCCCGAGTGTGCTGACCA
5-10-5
95.3
365


 


399904
21121
21140
GTAGGCCCCGAGTGTGCTGA
3-14-3
92.6
 54


 


405979
21123
21142
GTGTAGGCCCCGAGTGTGCT
5-10-5
85.5
366


 


405980
21125
21144
CCGTGTAGGCCCCGAGTGTG
5-10-5
86.5
367


 


405981
21127
21146
ATCCGTGTAGGCCCCGAGTG
5-10-5
77.8
368


 


410741
21129
21148
CCATCCGTGTAGGCCCCGAG
5-10-5
78.4
369


 


405982
21131
21150
GGCCATCCGTGTAGGCCCCG
5-10-5
86.7
370


 


405983
21133
21152
GTGGCCATCCGTGTAGGCCC
5-10-5
73.1
371


 


405984
21181
21200
CTGGAGCAGCTCAGCAGCTC
5-10-5
83.4
372


 


405985
21183
21202
AACTGGAGCAGCTCAGCAGC
5-10-5
50.7
373


 


405986
21188
21207
GGAGAAACTGGAGCAGCTCA
5-10-5
75.6
374


 


405987
21190
21209
CTGGAGAAACTGGAGCAGCT
5-10-5
88.0
375


 


399868
21696
21715
GGCACTGCCCTTCCACCAAA
5-10-5
85.8
123


 


399905
22096
22115
CGTTGTGGGCCCGGCAGACC
3-14-3
91.2
 57


 


395183
22096
22115
CGTTGTGGGCCCGGCAGACC
5-10-5
93.1
 57


 


410541
22133
22152
CACCTGGCAATGGCGTAGAC
5-10-5
46.1
376


 


410605
22133
22152
CACCTGGCAATGGCGTAGAC
3-14-3
67.4
376


 


410683
22133
22152
CACCTGGCAATGGCGTAGAC
2-13-5
71.8
376


 


410606
22134
22153
GCACCTGGCAATGGCGTAGA
3-14-3
74.3
377


 


410542
22134
22153
GCACCTGGCAATGGCGTAGA
5-10-5
75.5
377


 


410684
22134
22153
GCACCTGGCAATGGCGTAGA
2-13-5
78.7
377


 


410543
22135
22154
AGCACCTGGCAATGGCGTAG
5-10-5
76.4
378


 


410685
22135
22154
AGCACCTGGCAATGGCGTAG
2-13-5
76.9
378


 


410607
22135
22154
AGCACCTGGCAATGGCGTAG
3-14-3
77.1
378


 


410544
22136
22155
CAGCACCTGGCAATGGCGTA
5-10-5
62.7
379


 


410608
22136
22155
CAGCACCTGGCAATGGCGTA
3-14-3
69.6
379


 


410686
22136
22155
CAGCACCTGGCAATGGCGTA
2-13-5
81.0
379


 


410545
22137
22156
GCAGCACCTGGCAATGGCGT
5-10-5
75.5
380


 


410687
22137
22156
GCAGCACCTGGCAATGGCGT
2-13-5
79.2
380


 


410609
22137
22156
GCAGCACCTGGCAATGGCGT
3-14-3
83.2
380


 


410610
22138
22157
GGCAGCACCTGGCAATGGCG
3-14-3
67.5
381


 


410688
22138
22157
GGCAGCACCTGGCAATGGCG
2-13-5
89.3
381


 


405988
22138
22157
GGCAGCACCTGGCAATGGCG
5-10-5
95.9
381


 


410546
22139
22158
AGGCAGCACCTGGCAATGGC
5-10-5
74.8
382


 


410689
22139
22158
AGGCAGCACCTGGCAATGGC
2-13-5
83.9
382


 


410611
22139
22158
AGGCAGCACCTGGCAATGGC
3-14-3
88.2
382


 


410612
22140
22159
CAGGCAGCACCTGGCAATGG
3-14-3
72.8
383


 


410690
22140
22159
CAGGCAGCACCTGGCAATGG
2-13-5
74.8
383


 


405989
22140
22159
CAGGCAGCACCTGGCAATGG
5-10-5
88.1
383


 


410547
22141
22160
GCAGGCAGCACCTGGCAATG
5-10-5
63.0
384


 


410691
22141
22160
GCAGGCAGCACCTGGCAATG
2-13-5
70.2
384


 


410613
22141
22160
GCAGGCAGCACCTGGCAATG
3-14-3
76.7
384


 


395184
22142
22161
AGCAGGCAGCACCTGGCAAT
5-10-5
68.5
 58


 


410692
22142
22161
AGCAGGCAGCACCTGGCAAT
2-13-5
69.1
 58


 


399906
22142
22161
AGCAGGCAGCACCTGGCAAT
3-14-3
94.2
 58


 


410614
22143
22162
TAGCAGGCAGCACCTGGCAA
3-14-3
36.6
385


 


410548
22143
22162
TAGCAGGCAGCACCTGGCAA
5-10-5
75.0
385


 


410693
22143
22162
TAGCAGGCAGCACCTGGCAA
2-13-5
82.0
385


 


405990
22144
22163
GTAGCAGGCAGCACCTGGCA
5-10-5
96.8
386


 


410773
22189
22208
TGGCCTCAGCTGGTGGAGCT
5-10-5
61.3
387


 


410549
22199
22218
GTCCCCATGCTGGCCTCAGC
5-10-5
63.0
388


 


410615
22199
22218
GTCCCCATGCTGGCCTCAGC
3-14-3
72.5
388


 


410694
22199
22218
GTCCCCATGCTGGCCTCAGC
2-13-5
74.8
388


 


410550
22200
22219
GGTCCCCATGCTGGCCTCAG
5-10-5
69.4
389


 


410616
22200
22219
GGTCCCCATGCTGGCCTCAG
3-14-3
80.7
389


 


410695
22200
22219
GGTCCCCATGCTGGCCTCAG
2-13-5
85.5
389


 


410551
22201
22220
GGGTCCCCATGCTGGCCTCA
5-10-5
68.7
390


 


410696
22201
22220
GGGTCCCCATGCTGGCCTCA
2-13-5
86.3
390


 


410617
22201
22220
GGGTCCCCATGCTGGCCTCA
3-14-3
86.5
390


 


410552
22202
22221
CGGGTCCCCATGCTGGCCTC
5-10-5
77.9
391


 


410618
22202
22221
CGGGTCCCCATGCTGGCCTC
3-14-3
86.7
391


 


410697
22202
22221
CGGGTCCCCATGCTGGCCTC
2-13-5
87.0
391


 


410553
22203
22222
ACGGGTCCCCATGCTGGCCT
5-10-5
72.8
392


 


410619
22203
22222
ACGGGTCCCCATGCTGGCCT
3-14-3
84.1
392


 


410698
22203
22222
ACGGGTCCCCATGCTGGCCT
2-13-5
87.0
392


 


410699
22204
22223
CACGGGTCCCCATGCTGGCC
2-13-5
79.5
 59


 


395185
22204
22223
CACGGGTCCCCATGCTGGCC
5-10-5
92.7
 59


 


399907
22204
22223
CACGGGTCCCCATGCTGGCC
3-14-3
93.7
 59


 


410620
22205
22224
ACACGGGTCCCCATGCTGGC
3-14-3
74.4
393


 


410554
22205
22224
ACACGGGTCCCCATGCTGGC
5-10-5
79.7
393


 


410700
22205
22224
ACACGGGTCCCCATGCTGGC
2-13-5
84.4
393


 


410621
22206
22225
GACACGGGTCCCCATGCTGG
3-14-3
76.0
394


 


410701
22206
22225
GACACGGGTCCCCATGCTGG
2-13-5
83.4
394


 


405991
22206
22225
GACACGGGTCCCCATGCTGG
5-10-5
91.9
394


 


410555
22207
22226
GGACACGGGTCCCCATGCTG
5-10-5
77.0
395


 


410622
22207
22226
GGACACGGGTCCCCATGCTG
3-14-3
79.8
395


 


410702
22207
22226
GGACACGGGTCCCCATGCTG
2-13-5
85.0
395


 


410703
22208
22227
TGGACACGGGTCCCCATGCT
2-13-5
70.4
396


 


410623
22208
22227
TGGACACGGGTCCCCATGCT
3-14-3
78.9
396


 


405992
22208
22227
TGGACACGGGTCCCCATGCT
5-10-5
89.3
396


 


410704
22209
22228
GTGGACACGGGTCCCCATGC
2-13-5
78.6
397


 


410556
22209
22228
GTGGACACGGGTCCCCATGC
5-10-5
81.4
397


 


410624
22209
22228
GTGGACACGGGTCCCCATGC
3-14-3
82.8
397


 


410625
22210
22229
AGTGGACACGGGTCCCCATG
3-14-3
64.4
398


 


410705
22210
22229
AGTGGACACGGGTCCCCATG
2-13-5
74.7
398


 


405993
22210
22229
AGTGGACACGGGTCCCCATG
5-10-5
85.5
398


 


410706
22211
22230
CAGTGGACACGGGTCCCCAT
2-13-5
70.8
399


 


410557
22211
22230
CAGTGGACACGGGTCCCCAT
5-10-5
74.2
399


 


410626
22211
22230
CAGTGGACACGGGTCCCCAT
3-14-3
76.8
399


 


410627
22212
22231
GCAGTGGACACGGGTCCCCA
3-14-3
71.4
400


 


410707
22212
22231
GCAGTGGACACGGGTCCCCA
2-13-5
72.8
400


 


405994
22212
22231
GCAGTGGACACGGGTCCCCA
5-10-5
95.2
400


 


410708
22213
22232
GGCAGTGGACACGGGTCCCC
2-13-5
79.0
401


 


410628
22213
22232
GGCAGTGGACACGGGTCCCC
3-14-3
88.1
401


 


410558
22213
22232
GGCAGTGGACACGGGTCCCC
5-10-5
88.9
401


 


410629
22214
22233
TGGCAGTGGACACGGGTCCC
3-14-3
66.5
402


 


410709
22214
22233
TGGCAGTGGACACGGGTCCC
2-13-5
66.9
402


 


405995
22214
22233
TGGCAGTGGACACGGGTCCC
5-10-5
92.4
402


 


410630
22215
22234
GTGGCAGTGGACACGGGTCC
3-14-3
54.1
403


 


410710
22215
22234
GTGGCAGTGGACACGGGTCC
2-13-5
61.4
403


 


410559
22215
22234
GTGGCAGTGGACACGGGTCC
5-10-5
77.5
403


 


410711
22216
22235
GGTGGCAGTGGACACGGGTC
2-13-5
NA
404


 


410631
22216
22235
GGTGGCAGTGGACACGGGTC
3-14-3
NA
404


 


410560
22216
22235
GGTGGCAGTGGACACGGGTC
5-10-5
42.5
404


 


410712
22217
22236
TGGTGGCAGTGGACACGGGT
2-13-5
NA
405


 


410632
22217
22236
TGGTGGCAGTGGACACGGGT
3-14-3
NA
405


 


410561
22217
22236
TGGTGGCAGTGGACACGGGT
5-10-5
NA
405


 


410774
22220
22239
TGTTGGTGGCAGTGGACACG
5-10-5
47.8
406


 


410776
23985
24004
GTGCCAAGGTCCTCCACCTC
5-10-5
77.3
408


 


410777
24005
24024
TCAGCACAGGCGGCTTGTGG
5-10-5
40.0
409


 


410778
24035
24054
CCACGCACTGGTTGGGCTGA
5-10-5
52.7
410


 


399908
24095
24114
CTTTGCATTCCAGACCTGGG
3-14-3
74.9
 60


 


410713
24095
24114
CTTTGCATTCCAGACCTGGG
2-13-5
84.9
 60


 


395186
24095
24114
CTTTGCATTCCAGACCTGGG
5-10-5
93.7
 60


 


410714
24096
24115
ACTTTGCATTCCAGACCTGG
2-13-5
82.4
411


 


410633
24096
24115
ACTTTGCATTCCAGACCTGG
3-14-3
83.8
411


 


410562
24096
24115
ACTTTGCATTCCAGACCTGG
5-10-5
87.3
411


 


410715
24097
24116
GACTTTGCATTCCAGACCTG
2-13-5
80.5
412


 


410634
24097
24116
GACTTTGCATTCCAGACCTG
3-14-3
81.8
412


 


405996
24097
24116
GACTTTGCATTCCAGACCTG
5-10-5
91.0
412


 


410563
24098
24117
TGACTTTGCATTCCAGACCT
5-10-5
75.4
413


 


410635
24098
24117
TGACTTTGCATTCCAGACCT
3-14-3
75.9
413


 


410716
24098
24117
TGACTTTGCATTCCAGACCT
2-13-5
88.3
413


 


410636
24099
24118
TTGACTTTGCATTCCAGACC
3-14-3
61.3
414


 


410564
24099
24118
TTGACTTTGCATTCCAGACC
5-10-5
71.3
414


 


410717
24099
24118
TTGACTTTGCATTCCAGACC
2-13-5
76.7
414


 


410718
24100
24119
CTTGACTTTGCATTCCAGAC
2-13-5
55.8
 61


 


399973
24100
24119
CTTGACTTTGCATTCCAGAC
3-14-3
82.1
 61


 


399817
24100
24119
CTTGACTTTGCATTCCAGAC
5-10-5
93.6
 61


 


405997
24102
24121
TCCTTGACTTTGCATTCCAG
5-10-5
95.6
415


 


410779
24115
24134
CGGGATTCCATGCTCCTTGA
5-10-5
79.7
416


 


410781
25994
26013
GGAGGGCACTGCAGCCAGTC
5-10-5
66.0
418


 


410719
26112
26131
CAGATGGCAACGGCTGTCAC
2-13-5
61.0
419


 


410637
26112
26131
CAGATGGCAACGGCTGTCAC
3-14-3
66.0
419


 


410565
26112
26131
CAGATGGCAACGGCTGTCAC
5-10-5
71.6
419


 


410638
26113
26132
GCAGATGGCAACGGCTGTCA
3-14-3
71.9
420


 


410720
26113
26132
GCAGATGGCAACGGCTGTCA
2-13-5
75.1
420


 


410566
26113
26132
GCAGATGGCAACGGCTGTCA
5-10-5
77.7
420


 


410639
26114
26133
AGCAGATGGCAACGGCTGTC
3-14-3
67.7
421


 


410721
26114
26133
AGCAGATGGCAACGGCTGTC
2-13-5
68.8
421


 


410567
26114
26133
AGCAGATGGCAACGGCTGTC
5-10-5
72.1
421


 


410568
26115
26134
CAGCAGATGGCAACGGCTGT
5-10-5
54.2
422


 


410640
26115
26134
CAGCAGATGGCAACGGCTGT
3-14-3
57.7
422


 


410722
26115
26134
CAGCAGATGGCAACGGCTGT
2-13-5
59.2
422


 


410569
26116
26135
GCAGCAGATGGCAACGGCTG
5-10-5
57.7
423


 


410641
26116
26135
GCAGCAGATGGCAACGGCTG
3-14-3
66.1
423


 


410723
26116
26135
GCAGCAGATGGCAACGGCTG
2-13-5
81.1
423


 


399909
26117
26136
GGCAGCAGATGGCAACGGCT
3-14-3
69.9
 62


 


410724
26117
26136
GGCAGCAGATGGCAACGGCT
2-13-5
87.4
 62


 


395187
26117
26136
GGCAGCAGATGGCAACGGCT
5-10-5
92.7
 62


 


410642
26118
26137
CGGCAGCAGATGGCAACGGC
3-14-3
70.4
424


 


410725
26118
26137
CGGCAGCAGATGGCAACGGC
2-13-5
75.6
424


 


410570
26118
26137
CGGCAGCAGATGGCAACGGC
5-10-5
78.3
424


 


410571
26119
26138
CCGGCAGCAGATGGCAACGG
5-10-5
52.8
425


 


410643
26119
26138
CCGGCAGCAGATGGCAACGG
3-14-3
59.8
425


 


410726
26119
26138
CCGGCAGCAGATGGCAACGG
2-13-5
69.8
425


 


410644
26120
26139
TCCGGCAGCAGATGGCAACG
3-14-3
59.0
426


 


410727
26120
26139
TCCGGCAGCAGATGGCAACG
2-13-5
68.3
426


 


405998
26120
26139
TCCGGCAGCAGATGGCAACG
5-10-5
75.6
426


 


410572
26121
26140
CTCCGGCAGCAGATGGCAAC
5-10-5
40.9
427


 


410728
26121
26140
CTCCGGCAGCAGATGGCAAC
2-13-5
56.4
427


 


410645
26121
26140
CTCCGGCAGCAGATGGCAAC
3-14-3
70.2
427


 


410729
26122
26141
GCTCCGGCAGCAGATGGCAA
2-13-5
67.7
428


 


410573
26122
26141
GCTCCGGCAGCAGATGGCAA
5-10-5
71.8
428


 


410646
26122
26141
GCTCCGGCAGCAGATGGCAA
3-14-3
72.8
428


 


410782
26132
26151
CCAGGTGCCGGCTCCGGCAG
5-10-5
43.2
429


 


410783
26142
26161
GAGGCCTGCGCCAGGTGCCG
5-10-5
63.7
430


 


399819
26316
26335
CCCACTCAAGGGCCAGGCCA
5-10-5
93.4
 65


 


399912
26404
26423
ATGCCCCACAGTGAGGGAGG
3-14-3
84.3
 66


 


405893
26635
26654
ATGAGGGCCATCAGCACCTT
5-10-5
93.9
431


 


405894
26636
26655
GATGAGGGCCATCAGCACCT
5-10-5
93.9
432


 


405895
26637
26656
AGATGAGGGCCATCAGCACC
5-10-5
91.8
433


 


405896
26638
26657
GAGATGAGGGCCATCAGCAC
5-10-5
88.5
434


 


395194
26639
26658
GGAGATGAGGGCCATCAGCA
5-10-5
90.0
 69


 


399916
26639
26658
GGAGATGAGGGCCATCAGCA
3-14-3
94.5
 69


 


405897
26640
26659
TGGAGATGAGGGCCATCAGC
5-10-5
85.1
435


 


405898
26641
26660
CTGGAGATGAGGGCCATCAG
5-10-5
86.9
436


 


405899
26642
26661
GCTGGAGATGAGGGCCATCA
5-10-5
91.5
437


 


405900
26643
26662
AGCTGGAGATGAGGGCCATC
5-10-5
83.7
438


 


405901
26709
26728
GCTAGATGCCATCCAGAAAG
5-10-5
88.4
439


 


405902
26710
26729
GGCTAGATGCCATCCAGAAA
5-10-5
89.3
440


 


405903
26711
26730
TGGCTAGATGCCATCCAGAA
5-10-5
92.6
441


 


405904
26712
26731
CTGGCTAGATGCCATCCAGA
5-10-5
90.4
442


 


399821
26713
26732
TCTGGCTAGATGCCATCCAG
5-10-5
91.7
 71


 


405905
26714
26733
CTCTGGCTAGATGCCATCCA
5-10-5
90.7
443


 


405906
26715
26734
CCTCTGGCTAGATGCCATCC
5-10-5
93.3
444


 


405907
26716
26735
GCCTCTGGCTAGATGCCATC
5-10-5
90.2
445


 


405908
26717
26736
AGCCTCTGGCTAGATGCCAT
5-10-5
83.3
446


 


399978
26795
26814
AGCCTGGCATAGAGCAGAGT
3-14-3
96.4
 73









Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.
Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395152, 395153, 395155, 395158, 395163, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399798, 399801, 399804, 399817, 399819, 399821, 399868, 399877, 399879, 399887, 399888, 399891, 399895, 399900, 399904, 399905, 399906, 399907, 399912, 399916, 399935, 399936, 399954, 399960, 399967, 399968, 399969, 399973, 399978, 405861, 405862, 405863, 405864, 405865, 405866, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406003, 406005, 406008, 406009, 406014, 406016, 406020, 406023, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410530, 410556, 410558, 410562, 410578, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410609, 410611, 410616, 410617, 410618, 410619, 410624, 410628, 410633, 410634, 410647, 410648, 410650, 410652, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410686, 410688, 410689, 410693, 410695, 410696, 410697, 410698, 410700, 410701, 410702, 410713, 410714, 410715, 410716, 410723, 410724, 410730, 410754, 410757, 410759, 410769, and 410772. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.
ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399868, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399936, 399954, 399969, 399978, 405861, 405862, 405864, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405923, 405924, 405925, 405926, 405927, 405928, 405929, 405930, 405931, 405932, 405934, 405935, 405936, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405988, 405989, 405990, 405991, 405992, 405993, 405994, 405995, 405996, 405997, 406008, 406009, 406014, 406016, 406023, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410558, 410562, 410583, 410591, 410592, 410593, 410611, 410617, 410618, 410628, 410647, 410650, 410668, 410677, 410688, 410695, 410696, 410697, 410698, 410702, 410716, 410724, 410730, 410757, and 410772 each exhibited at least 85% inhibition of PCSK9 mRNA levels.
ISIS Nos 395152, 395155, 395158, 395165, 395166, 395168, 395173, 395183, 395185, 395186, 395187, 399801, 399817, 399819, 399821, 399877, 399879, 399887, 399888, 399891, 399900, 399904, 399905, 399906, 399907, 399916, 399969, 399978, 405862, 405864, 405874, 405875, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405906, 405910, 405913, 405916, 405923, 405924, 405925, 405926, 405927, 405930, 405934, 405936, 405953, 405955, 405956, 405959, 405978, 405988, 405990, 405991, 405994, 405995, 405996, 405997, 406008, 406014, 406023, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410647, 410668, and 410772 each exhibited at least 90% inhibition of PCSK9 mRNA levels.
ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405988, 405990, 405994, 405997, 406008, 406030, 406040, 406041, and 406044 each exhibited at least 95% inhibition of PCSK9 mRNA levels.







TABLE 19







Antisense inhibition of PCSK9 in human cells (Hep3B)


targeting SEQ ID NO: 3














5′
3′







Target
Target







Site to
Site to



SEQ


Isis
SEQ ID
SEQ ID


%
ID


No.
NO: 3
NO: 3
Sequence 5′-3′
Motif
inhibition
NO:





399935
1075
1094
AGGACCCAAGTCATCCTGCT
3-14-3
81.1
 96


 


399936
1105
1124
GGCCATCAGCTGGCAATGCT
3-14-3
89.0
124


 


410753
1351
1370
ATACACCTCCACCAGGCTGC
5-10-5
72.8
215


 


406025
1362
1381
GTGTCTAGGAGATACACCTC
5-10-5
74.2
216


 


406026
1367
1386
TGCTGGTGTCTAGGAGATAC
5-10-5
80.5
217


 


405869
1392
1411
TCGATTTCCCGGTGGTCACT
5-10-5
87.6
218


 


405870
1393
1412
CTCGATTTCCCGGTGGTCAC
5-10-5
83.3
219


 


405871
1394
1413
CCTCGATTTCCCGGTGGTCA
5-10-5
87.0
220


 


405872
1395
1414
CCCTCGATTTCCCGGTGGTC
5-10-5
88.2
221


 


399798
1396
1415
GCCCTCGATTTCCCGGTGGT
5-10-5
84.3
 22


 


399954
1396
1415
GCCCTCGATTTCCCGGTGGT
3-14-3
90.3
 22


 


405873
1397
1416
TGCCCTCGATTTCCCGGTGG
5-10-5
90.0
222


 


405874
1398
1417
CTGCCCTCGATTTCCCGGTG
5-10-5
91.4
223


 


405875
1399
1418
CCTGCCCTCGATTTCCCGGT
5-10-5
93.5
224


 


405876
1400
1419
CCCTGCCCTCGATTTCCCGG
5-10-5
90.1
225


 


406027
1404
1423
ATGACCCTGCCCTCGATTTC
5-10-5
73.9
226


 


406028
1406
1425
CCATGACCCTGCCCTCGATT
5-10-5
92.3
227


 


406029
1408
1427
GACCATGACCCTGCCCTCGA
5-10-5
91.4
228


 


406030
1412
1431
CGGTGACCATGACCCTGCCC
5-10-5
95.6
229


 


406031
1414
1433
GTCGGTGACCATGACCCTGC
5-10-5
89.6
230


 


410733
1416
1435
AAGTCGGTGACCATGACCCT
5-10-5
65.7
231


 


406032
1418
1437
CGAAGTCGGTGACCATGACC
5-10-5
88.5
232


 


410754
1428
1447
GGCACATTCTCGAAGTCGGT
5-10-5
80.1
233


 


395163
1453
1472
GTGGAAGCGGGTCCCGTCCT
5-10-5
83.9
 25


 


410755
1463
1482
TGGCCTGTCTGTGGAAGCGG
5-10-5
NA
234


 


410756
1490
1509
GGTGGGTGCCATGACTGTCA
5-10-5
67.1
235


 


406033
1497
1516
CCTGCCAGGTGGGTGCCATG
5-10-5
91.2
237


 


406034
1502
1521
CCACCCCTGCCAGGTGGGTG
5-10-5
59.3
238


 


406035
1507
1526
GCTGACCACCCCTGCCAGGT
5-10-5
91.7
239


 


410757
1515
1534
TCCCGGCCGCTGACCACCCC
5-10-5
88.9
240


 


406036
1519
1538
GGCATCCCGGCCGCTGACCA
5-10-5
84.0
241


 


406037
1522
1541
GCCGGCATCCCGGCCGCTGA
5-10-5
55.9
242


 


410536
1527
1546
GCCACGCCGGCATCCCGGCC
5-10-5
63.7
243


 


410658
1527
1546
GCCACGCCGGCATCCCGGCC
2-13-5
82.0
243


 


410583
1527
1546
GCCACGCCGGCATCCCGGCC
3-14-3
87.3
243


 


410537
1528
1547
GGCCACGCCGGCATCCCGGC
5-10-5
58.5
244


 


410659
1528
1547
GGCCACGCCGGCATCCCGGC
2-13-5
73.7
244


 


410584
1528
1547
GGCCACGCCGGCATCCCGGC
3-14-3
79.5
244


 


410660
1529
1548
TGGCCACGCCGGCATCCCGG
2-13-5
63.0
245


 


410585
1529
1548
TGGCCACGCCGGCATCCCGG
3-14-3
73.2
245


 


406038
1529
1548
TGGCCACGCCGGCATCCCGG
5-10-5
86.3
245


 


410661
1530
1549
TTGGCCACGCCGGCATCCCG
2-13-5
53.3
246


 


410586
1530
1549
TTGGCCACGCCGGCATCCCG
3-14-3
60.3
246


 


405877
1530
1549
TTGGCCACGCCGGCATCCCG
5-10-5
83.2
246


 


410587
1531
1550
CTTGGCCACGCCGGCATCCC
3-14-3
63.2
247


 


410662
1531
1550
CTTGGCCACGCCGGCATCCC
2-13-5
67.3
247


 


405878
1531
1550
CTTGGCCACGCCGGCATCCC
5-10-5
91.4
247


 


410588
1532
1551
CCTTGGCCACGCCGGCATCC
3-14-3
65.3
248


 


410663
1532
1551
CCTTGGCCACGCCGGCATCC
2-13-5
67.9
248


 


405879
1532
1551
CCTTGGCCACGCCGGCATCC
5-10-5
94.1
248


 


410589
1533
1552
CCCTTGGCCACGCCGGCATC
3-14-3
80.1
249


 


410664
1533
1552
CCCTTGGCCACGCCGGCATC
2-13-5
81.9
249


 


405880
1533
1552
CCCTTGGCCACGCCGGCATC
5-10-5
93.5
249


 


410665
1534
1553
ACCCTTGGCCACGCCGGCAT
2-13-5
78.7
 28


 


399887
1534
1553
ACCCTTGGCCACGCCGGCAT
3-14-3
91.9
 28


 


395165
1534
1553
ACCCTTGGCCACGCCGGCAT
5-10-5
96.7
 28


 


410590
1535
1554
CACCCTTGGCCACGCCGGCA
3-14-3
82.2
250


 


410666
1535
1554
CACCCTTGGCCACGCCGGCA
2-13-5
82.7
250


 


405881
1535
1554
CACCCTTGGCCACGCCGGCA
5-10-5
98.3
250


 


410667
1536
1555
GCACCCTTGGCCACGCCGGC
2-13-5
84.7
251


 


410591
1536
1555
GCACCCTTGGCCACGCCGGC
3-14-3
86.3
251


 


405882
1536
1555
GCACCCTTGGCCACGCCGGC
5-10-5
91.6
251


 


410668
1537
1556
GGCACCCTTGGCCACGCCGG
2-13-5
91.4
252


 


405883
1537
1556
GGCACCCTTGGCCACGCCGG
5-10-5
92.5
252


 


410592
1537
1556
GGCACCCTTGGCCACGCCGG
3-14-3
93.4
252


 


410669
1538
1557
TGGCACCCTTGGCCACGCCG
2-13-5
69.0
253


 


410593
1538
1557
TGGCACCCTTGGCCACGCCG
3-14-3
88.8
253


 


405884
1538
1557
TGGCACCCTTGGCCACGCCG
5-10-5
90.8
253


 


410670
1539
1558
CTGGCACCCTTGGCCACGCC
2-13-5
74.9
254


 


410594
1539
1558
CTGGCACCCTTGGCCACGCC
3-14-3
75.2
254


 


410538
1539
1558
CTGGCACCCTTGGCCACGCC
5-10-5
78.2
254


 


410539
1540
1559
GCTGGCACCCTTGGCCACGC
5-10-5
61.9
255


 


410595
1540
1559
GCTGGCACCCTTGGCCACGC
3-14-3
83.1
255


 


410671
1540
1559
GCTGGCACCCTTGGCCACGC
2-13-5
83.3
255


 


410758
1545
1564
CGCATGCTGGCACCCTTGGC
5-10-5
73.6
256


 


406039
1566
1585
CAGTTGAGCACGCGCAGGCT
5-10-5
91.2
257


 


406040
1568
1587
GGCAGTTGAGCACGCGCAGG
5-10-5
96.5
258


 


399888
1570
1589
TTGGCAGTTGAGCACGCGCA
3-14-3
90.9
 29


 


395166
1570
1589
TTGGCAGTTGAGCACGCGCA
5-10-5
96.2
 29


 


406041
1572
1591
CCTTGGCAGTTGAGCACGCG
5-10-5
96.6
259


 


399801
1575
1594
TTCCCTTGGCAGTTGAGCAC
5-10-5
95.6
 30


 


406042
1577
1596
CCTTCCCTTGGCAGTTGAGC
5-10-5
92.0
260


 


406043
1581
1600
GTGCCCTTCCCTTGGCAGTT
5-10-5
91.9
261


 


406044
1583
1602
CCGTGCCCTTCCCTTGGCAG
5-10-5
95.4
262


 


410759
1594
1613
GGTGCCGCTAACCGTGCCCT
5-10-5
83.7
263


 


410760
1618
1637
CCGAATAAACTCCAGGCCTA
5-10-5
96.7
265


 


406045
1626
1645
TGGCTTTTCCGAATAAACTC
5-10-5
88.4
266


 


395168
1628
1647
GCTGGCTTTTCCGAATAAAC
5-10-5
91.9
 32


 


405909
1630
1649
CAGCTGGCTTTTCCGAATAA
5-10-5
80.3
267


 


405910
1632
1651
ACCAGCTGGCTTTTCCGAAT
5-10-5
90.9
268


 


405911
1634
1653
GGACCAGCTGGCTTTTCCGA
5-10-5
88.0
269


 


405912
1638
1657
GGCTGGACCAGCTGGCTTTT
5-10-5
78.2
270


 


410761
1649
1668
GTGGCCCCACAGGCTGGACC
5-10-5
53.7
271


 


410762
1662
1681
AGCAGCACCACCAGTGGCCC
5-10-5
57.6
272


 


410763
1684
1703
GCTGTACCCACCCGCCAGGG
5-10-5
68.8
273


 


410764
1730
1749
CGACCCCAGCCCTCGCCAGG
5-10-5
66.3
274


 


399891
1740
1759
GTGACCAGCACGACCCCAGC
3-14-3
91.1
 33


 


405913
1742
1761
CGGTGACCAGCACGACCCCA
5-10-5
93.8
275


 


405914
1744
1763
AGCGGTGACCAGCACGACCC
5-10-5
90.4
276


 


405915
1746
1765
GCAGCGGTGACCAGCACGAC
5-10-5
86.8
277


 


405916
1748
1767
CGGCAGCGGTGACCAGCACG
5-10-5
91.3
278


 


410734
1749
1768
CCGGCAGCGGTGACCAGCAC
5-10-5
65.8
249


 


405917
1752
1771
TTGCCGGCAGCGGTGACCAG
5-10-5
62.0
280


 


405918
1754
1773
AGTTGCCGGCAGCGGTGACC
5-10-5
33.5
281


 


405919
1756
1775
GAAGTTGCCGGCAGCGGTGA
5-10-5
68.2
282


 


405920
1758
1777
CGGAAGTTGCCGGCAGCGGT
5-10-5
86.2
283


 


405921
1760
1779
CCCGGAAGTTGCCGGCAGCG
5-10-5
85.5
284


 


405922
1762
1781
GTCCCGGAAGTTGCCGGCAG
5-10-5
86.0
285


 


405937
1820
1839
GAGGCACCAATGATGTCCTC
5-10-5
77.7
306


 


405938
1824
1843
GCTGGAGGCACCAATGATGT
5-10-5
75.5
307


 


405939
1826
1845
TCGCTGGAGGCACCAATGAT
5-10-5
70.6
308


 


405940
1828
1847
AGTCGCTGGAGGCACCAATG
5-10-5
84.4
309


 


405941
1830
1849
GCAGTCGCTGGAGGCACCAA
5-10-5
90.1
310


 


399804
1833
1852
GCTGCAGTCGCTGGAGGCAC
5-10-5
80.8
 40


 


399960
1833
1852
GCTGCAGTCGCTGGAGGCAC
3-14-3
82.4
 40


 


405942
1835
1854
GTGCTGCAGTCGCTGGAGGC
5-10-5
69.8
311


 


405943
1837
1856
AGGTGCTGCAGTCGCTGGAG
5-10-5
83.6
312


 


410737
1839
1858
GCAGGTGCTGCAGTCGCTGG
5-10-5
49.8
313


 


405944
1840
1859
AGCAGGTGCTGCAGTCGCTG
5-10-5
80.7
314


 


405945
1842
1861
AAAGCAGGTGCTGCAGTCGC
5-10-5
41.1
315


 


405946
1846
1865
ACACAAAGCAGGTGCTGCAG
5-10-5
70.4
316


 


405947
1848
1867
TGACACAAAGCAGGTGCTGC
5-10-5
72.5
317


 


410769
1858
1877
TCCCACTCTGTGACACAAAG
5-10-5
84.9
318


 


405948
1900
1919
TCATGGCTGCAATGCCAGCC
5-10-5
45.9
319


 


399806
1903
1922
GCATCATGGCTGCAATGCCA
5-10-5
82.8
 42


 


399962
1903
1922
GCATCATGGCTGCAATGCCA
3-14-3
86.1
 42


 


405949
1905
1924
CAGCATCATGGCTGCAATGC
5-10-5
84.9
320


 


405950
1907
1926
GACAGCATCATGGCTGCAAT
5-10-5
76.9
321


 


405951
1909
1928
CAGACAGCATCATGGCTGCA
5-10-5
75.4
322


 


405952
1913
1932
TCGGCAGACAGCATCATGGC
5-10-5
85.1
323


 


410738
1915
1934
GCTCGGCAGACAGCATCATG
5-10-5
68.4
324


 


405953
1917
1936
CGGCTCGGCAGACAGCATCA
5-10-5
93.6
325


 


405954
1919
1938
TCCGGCTCGGCAGACAGCAT
5-10-5
87.5
326


 


410770
1933
1952
CGGCCAGGGTGAGCTCCGGC
5-10-5
68.5
327


 


405955
1946
1965
CTCTGCCTCAACTCGGCCAG
5-10-5
94.2
328


 


405956
1948
1967
GTCTCTGCCTCAACTCGGCC
5-10-5
94.3
329


 


405958
1952
1971
ATCAGTCTCTGCCTCAACTC
5-10-5
80.3
331


 


405959
1954
1973
GGATCAGTCTCTGCCTCAAC
5-10-5
95.4
332


 


405960
1956
1975
GTGGATCAGTCTCTGCCTCA
5-10-5
90.1
333


 


405961
1958
1977
AAGTGGATCAGTCTCTGCCT
5-10-5
88.8
334


 


405962
1961
1980
GAGAAGTGGATCAGTCTCTG
5-10-5
56.1
335


 


410739
1963
1982
CAGAGAAGTGGATCAGTCTC
5-10-5
NA
336


 


405963
1965
1984
GGCAGAGAAGTGGATCAGTC
5-10-5
59.7
337


 


405964
1969
1988
CTTTGGCAGAGAAGTGGATC
5-10-5
45.3
338


 


405965
1974
1993
GACATCTTTGGCAGAGAAGT
5-10-5
55.9
339


 


405966
1976
1995
ATGACATCTTTGGCAGAGAA
5-10-5
51.3
340


 


405967
1980
1999
ATTGATGACATCTTTGGCAG
5-10-5
64.4
341


 


405968
1982
2001
TCATTGATGACATCTTTGGC
5-10-5
74.6
342


 


399967
1985
2004
GCCTCATTGATGACATCTTT
3-14-3
80.1
 48


 


405969
1987
2006
AGGCCTCATTGATGACATCT
5-10-5
63.0
343


 


405970
1989
2008
CCAGGCCTCATTGATGACAT
5-10-5
66.8
344


 


410740
1991
2010
AACCAGGCCTCATTGATGAC
5-10-5
46.8
345


 


405885
1993
2012
GGAACCAGGCCTCATTGATG
5-10-5
73.4
346


 


410596
1995
2014
AGGGAACCAGGCCTCATTGA
3-14-3
NA
348


 


410672
1995
2014
AGGGAACCAGGCCTCATTGA
2-13-5
NA
348


 


405887
1995
2014
AGGGAACCAGGCCTCATTGA
5-10-5
NA
348


 


410597
1996
2015
CAGGGAACCAGGCCTCATTG
3-14-3
NA
349


 


405888
1996
2015
CAGGGAACCAGGCCTCATTG
5-10-5
NA
349


 


410673
1996
2015
CAGGGAACCAGGCCTCATTG
2-13-5
NA
349


 


410674
1997
2016
TCAGGGAACCAGGCCTCATT
2-13-5
60.7
 49


 


399812
1997
2016
TCAGGGAACCAGGCCTCATT
5-10-5
68.7
 49


 


399968
1997
2016
TCAGGGAACCAGGCCTCATT
3-14-3
84.5
 49


 


410598
1998
2017
CTCAGGGAACCAGGCCTCAT
3-14-3
74.5
350


 


410675
1998
2017
CTCAGGGAACCAGGCCTCAT
2-13-5
76.9
350


 


405889
1998
2017
CTCAGGGAACCAGGCCTCAT
5-10-5
80.2
350


 


410599
1999
2018
CCTCAGGGAACCAGGCCTCA
3-14-3
70.6
351


 


410676
1999
2018
CCTCAGGGAACCAGGCCTCA
2-13-5
77.8
351


 


405890
1999
2018
CCTCAGGGAACCAGGCCTCA
5-10-5
87.7
351


 


410600
2000
2019
TCCTCAGGGAACCAGGCCTC
3-14-3
76.5
352


 


410677
2000
2019
TCCTCAGGGAACCAGGCCTC
2-13-5
88.4
352


 


405891
2000
2019
TCCTCAGGGAACCAGGCCTC
5-10-5
96.1
352


 


405892
2001
2020
GTCCTCAGGGAACCAGGCCT
5-10-5
71.4
353


 


410601
2001
2020
GTCCTCAGGGAACCAGGCCT
3-14-3
72.7
353


 


410678
2001
2020
GTCCTCAGGGAACCAGGCCT
2-13-5
75.0
353


 


395178
2002
2021
GGTCCTCAGGGAACCAGGCC
5-10-5
76.7
 50


 


410679
2002
2021
GGTCCTCAGGGAACCAGGCC
2-13-5
77.6
 50


 


399900
2002
2021
GGTCCTCAGGGAACCAGGCC
3-14-3
92.0
 50


 


408653
2003
2022
TGGTCCTCAGGGAACCAGGC
5-10-5
42.5
354


 


410602
2003
2022
TGGTCCTCAGGGAACCAGGC
3-14-3
66.5
354


 


410680
2003
2022
TGGTCCTCAGGGAACCAGGC
2-13-5
71.6
354


 


410603
2004
2023
CTGGTCCTCAGGGAACCAGG
3-14-3
40.8
355


 


410681
2004
2023
CTGGTCCTCAGGGAACCAGG
2-13-5
44.4
355


 


405971
2004
2023
CTGGTCCTCAGGGAACCAGG
5-10-5
65.5
355


 


410540
2005
2024
GCTGGTCCTCAGGGAACCAG
5-10-5
50.9
356


 


410682
2005
2024
GCTGGTCCTCAGGGAACCAG
2-13-5
54.1
356


 


410604
2005
2024
GCTGGTCCTCAGGGAACCAG
3-14-3
62.5
356


 


405972
2006
2025
CGCTGGTCCTCAGGGAACCA
5-10-5
77.6
357


 


405973
2011
2030
GTACCCGCTGGTCCTCAGGG
5-10-5
83.8
358


 


405974
2013
2032
CAGTACCCGCTGGTCCTCAG
5-10-5
89.0
359


 


399813
2016
2035
GGTCAGTACCCGCTGGTCCT
5-10-5
72.4
 51


 


399969
2016
2035
GGTCAGTACCCGCTGGTCCT
3-14-3
93.4
 51


 


410771
2061
2080
ACCTGCCCCATGGGTGCTGG
5-10-5
NA
360


 


395181
2073
2092
AAACAGCTGCCAACCTGCCC
5-10-5
87.5
 52


 


405975
2075
2094
CAAAACAGCTGCCAACCTGC
5-10-5
77.4
361


 


405976
2080
2099
TCCTGCAAAACAGCTGCCAA
5-10-5
81.2
362


 


405977
2082
2101
AGTCCTGCAAAACAGCTGCC
5-10-5
89.5
363


 


399992
2095
2114
GTGCTGACCACACAGTCCTG
3-14-3
92.6
130


 


405978
2105
2124
GGCCCCGAGTGTGCTGACCA
5-10-5
95.3
365


 


399904
2108
2127
GTAGGCCCCGAGTGTGCTGA
3-14-3
92.6
 54


 


405979
2110
2129
GTGTAGGCCCCGAGTGTGCT
5-10-5
85.5
366


 


405980
2112
2131
CCGTGTAGGCCCCGAGTGTG
5-10-5
86.5
367


 


405981
2114
2133
ATCCGTGTAGGCCCCGAGTG
5-10-5
77.8
368


 


410741
2116
2135
CCATCCGTGTAGGCCCCGAG
5-10-5
78.4
369


 


405982
2118
2137
GGCCATCCGTGTAGGCCCCG
5-10-5
86.7
370


 


405983
2120
2139
GTGGCCATCCGTGTAGGCCC
5-10-5
73.1
371


 


405984
2168
2187
CTGGAGCAGCTCAGCAGCTC
5-10-5
83.4
372


 


405985
2170
2189
AACTGGAGCAGCTCAGCAGC
5-10-5
50.7
373


 


405986
2175
2194
GGAGAAACTGGAGCAGCTCA
5-10-5
75.6
374


 


405987
2177
2196
CTGGAGAAACTGGAGCAGCT
5-10-5
88.0
375


 


410776
2245
2264
GTGCCAAGGTCCTCCACCTC
5-10-5
77.3
408


 


410777
2265
2284
TCAGCACAGGCGGCTTGTGG
5-10-5
40.0
409


 


410778
2295
2314
CCACGCACTGGTTGGGCTGA
5-10-5
52.7
410


 


399908
2355
2374
CTTTGCATTCCAGACCTGGG
3-14-3
74.9
 60


 


410713
2355
2374
CTTTGCATTCCAGACCTGGG
2-13-5
84.9
 60


 


395186
2355
2374
CTTTGCATTCCAGACCTGGG
5-10-5
93.7
 60


 


410714
2356
2375
ACTTTGCATTCCAGACCTGG
2-13-5
82.4
411


 


410633
2356
2375
ACTTTGCATTCCAGACCTGG
3-14-3
83.8
411


 


410562
2356
2375
ACTTTGCATTCCAGACCTGG
5-10-5
87.3
411


 


410715
2357
2376
GACTTTGCATTCCAGACCTG
2-13-5
80.5
412


 


410634
2357
2376
GACTTTGCATTCCAGACCTG
3-14-3
81.8
412


 


405996
2357
2376
GACTTTGCATTCCAGACCTG
5-10-5
91.0
412


 


410563
2358
2377
TGACTTTGCATTCCAGACCT
5-10-5
75.4
413


 


410635
2358
2377
TGACTTTGCATTCCAGACCT
3-14-3
75.9
413


 


410716
2358
2377
TGACTTTGCATTCCAGACCT
2-13-5
88.3
413


 


410636
2359
2378
TTGACTTTGCATTCCAGACC
3-14-3
61.3
414


 


410564
2359
2378
TTGACTTTGCATTCCAGACC
5-10-5
71.3
414


 


410717
2359
2378
TTGACTTTGCATTCCAGACC
2-13-5
76.7
414


 


410718
2360
2379
CTTGACTTTGCATTCCAGAC
2-13-5
55.8
 61


 


399973
2360
2379
CTTGACTTTGCATTCCAGAC
3-14-3
82.1
 61


 


399817
2360
2379
CTTGACTTTGCATTCCAGAC
5-10-5
93.6
 61


 


405997
2362
2381
TCCTTGACTTTGCATTCCAG
5-10-5
95.6
415


 


410779
2375
2394
CGGGATTCCATGCTCCTTGA
5-10-5
79.7
416


 


410780
2405
2424
GCAGGCCACGGTCACCTGCT
5-10-5
60.9
417


 


410781
2442
2461
GGAGGGCACTGCAGCCAGTC
5-10-5
66.0
418


 


410719
2560
2579
CAGATGGCAACGGCTGTCAC
2-13-5
61.0
419


 


410637
2560
2579
CAGATGGCAACGGCTGTCAC
3-14-3
66.0
419


 


410565
2560
2579
CAGATGGCAACGGCTGTCAC
5-10-5
71.6
419


 


410638
2561
2580
GCAGATGGCAACGGCTGTCA
3-14-3
71.9
420


 


410720
2561
2580
GCAGATGGCAACGGCTGTCA
2-13-5
75.1
420


 


410566
2561
2580
GCAGATGGCAACGGCTGTCA
5-10-5
77.7
420


 


410639
2562
2581
AGCAGATGGCAACGGCTGTC
3-14-3
67.7
421


 


410721
2562
2581
AGCAGATGGCAACGGCTGTC
2-13-5
68.8
421


 


410567
2562
2581
AGCAGATGGCAACGGCTGTC
5-10-5
72.1
421


 


410568
2563
2582
CAGCAGATGGCAACGGCTGT
5-10-5
54.2
422


 


410640
2563
2582
CAGCAGATGGCAACGGCTGT
3-14-3
57.7
422


 


410722
2563
2582
CAGCAGATGGCAACGGCTGT
2-13-5
59.2
422


 


410569
2564
2583
GCAGCAGATGGCAACGGCTG
5-10-5
57.7
423


 


410641
2564
2583
GCAGCAGATGGCAACGGCTG
3-14-3
66.1
423


 


410723
2564
2583
GCAGCAGATGGCAACGGCTG
2-13-5
81.1
423


 


399909
2565
2584
GGCAGCAGATGGCAACGGCT
3-14-3
69.9
 62


 


410724
2565
2584
GGCAGCAGATGGCAACGGCT
2-13-5
87.4
 62


 


395187
2565
2584
GGCAGCAGATGGCAACGGCT
5-10-5
92.7
 62


 


410642
2566
2585
CGGCAGCAGATGGCAACGGC
3-14-3
70.4
424


 


410725
2566
2585
CGGCAGCAGATGGCAACGGC
2-13-5
75.6
424


 


410570
2566
2585
CGGCAGCAGATGGCAACGGC
5-10-5
78.3
424


 


410571
2567
2586
CCGGCAGCAGATGGCAACGG
5-10-5
52.8
425


 


410643
2567
2586
CCGGCAGCAGATGGCAACGG
3-14-3
59.8
425


 


410726
2567
2586
CCGGCAGCAGATGGCAACGG
2-13-5
69.8
425


 


410644
2568
2587
TCCGGCAGCAGATGGCAACG
3-14-3
59.0
426


 


410727
2568
2587
TCCGGCAGCAGATGGCAACG
2-13-5
68.3
426


 


405998
2568
2587
TCCGGCAGCAGATGGCAACG
5-10-5
75.6
426


 


405988
2568
2587
TCCGGCAGCAGATGGCAACG
5-10-5
84.5
426


 


410572
2569
2588
CTCCGGCAGCAGATGGCAAC
5-10-5
40.9
427


 


410728
2569
2588
CTCCGGCAGCAGATGGCAAC
2-13-5
56.4
427


 


410645
2569
2588
CTCCGGCAGCAGATGGCAAC
3-14-3
70.2
427


 


410729
2570
2589
GCTCCGGCAGCAGATGGCAA
2-13-5
67.7
428


 


410573
2570
2589
GCTCCGGCAGCAGATGGCAA
5-10-5
71.8
428


 


410646
2570
2589
GCTCCGGCAGCAGATGGCAA
3-14-3
72.8
428


 


410782
2580
2599
CCAGGTGCCGGCTCCGGCAG
5-10-5
43.2
429


 


410783
2590
2609
GAGGCCTGCGCCAGGTGCCG
5-10-5
63.7
430


 


399819
2764
2783
CCCACTCAAGGGCCAGGCCA
5-10-5
93.4
 65


 


399912
2852
2871
ATGCCCCACAGTGAGGGAGG
3-14-3
84.3
 66


 


405893
3083
3102
ATGAGGGCCATCAGCACCTT
5-10-5
93.9
431


 


405894
3084
3103
GATGAGGGCCATCAGCACCT
5-10-5
93.9
432


 


405895
3085
3104
AGATGAGGGCCATCAGCACC
5-10-5
91.8
433


 


405896
3086
3105
GAGATGAGGGCCATCAGCAC
5-10-5
88.5
434


 


395194
3087
3106
GGAGATGAGGGCCATCAGCA
5-10-5
90.0
 69


 


399916
3087
3106
GGAGATGAGGGCCATCAGCA
3-14-3
94.5
 69


 


405897
3088
3107
TGGAGATGAGGGCCATCAGC
5-10-5
85.1
435


 


405898
3089
3108
CTGGAGATGAGGGCCATCAG
5-10-5
86.9
436


 


405899
3090
3109
GCTGGAGATGAGGGCCATCA
5-10-5
91.5
437


 


405900
3091
3110
AGCTGGAGATGAGGGCCATC
5-10-5
83.7
438


 


405901
3157
3176
GCTAGATGCCATCCAGAAAG
5-10-5
88.4
439


 


405902
3158
3177
GGCTAGATGCCATCCAGAAA
5-10-5
89.3
440


 


405903
3159
3178
TGGCTAGATGCCATCCAGAA
5-10-5
92.6
441


 


405904
3160
3179
CTGGCTAGATGCCATCCAGA
5-10-5
90.4
442


 


399821
3161
3180
TCTGGCTAGATGCCATCCAG
5-10-5
91.7
 71


 


405905
3162
3181
CTCTGGCTAGATGCCATCCA
5-10-5
90.7
443


 


405906
3163
3182
CCTCTGGCTAGATGCCATCC
5-10-5
93.3
444


 


405907
3164
3183
GCCTCTGGCTAGATGCCATC
5-10-5
90.2
445


 


405908
3165
3184
AGCCTCTGGCTAGATGCCAT
5-10-5
83.3
446


 


399978
3243
3262
AGCCTGGCATAGAGCAGAGT
3-14-3
96.4
 73









Antisense oligonucleotides that exhibited less than 30% inhibition of PCSK9 mRNA levels were marked with “NA”.
Antisense oligonucleotides with the following ISIS Nos exhibited at least 80% inhibition of PCSK9 mRNA levels: 395163, 395165, 395166, 395168, 395181, 395186, 395187, 395194, 399798, 399801, 399804, 399806, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399912, 399916, 399935, 399936, 399954, 399960, 399962, 399967, 399968, 399969, 399973, 399978, 399992, 405869, 405870, 405871, 405872, 405873, 405874, 405875, 405876, 405877, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405889, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405900, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405908, 405909, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405940, 405941, 405943, 405944, 405949, 405952, 405953, 405954, 405955, 405956, 405958, 405959, 405960, 405961, 405973, 405974, 405976, 405977, 405978, 405979, 405980, 405982, 405984, 405987, 405988, 405996, 405997, 406026, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406036, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410562, 410583, 410589, 410590, 410591, 410592, 410593, 410595, 410633, 410634, 410658, 410664, 410666, 410667, 410668, 410671, 410677, 410713, 410714, 410715, 410716, 410723, 410724, 410754, 410757, 410759, 410760, and 410769. The target segments to which these antisense oligonucleotides are targeted are active target segments. The target regions to which these antisense oligonucleotides are targeted are active target regions.
ISIS Nos 395165, 395166, 395168, 395181, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916, 399936, 399954, 399962, 399969, 399978, 399992, 405869, 405871, 405872, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405890, 405891, 405893, 405894, 405895, 405896, 405897, 405898, 405899, 405901, 405902, 405903, 405904, 405905, 405906, 405907, 405910, 405911, 405913, 405914, 405915, 405916, 405920, 405921, 405922, 405941, 405952, 405953, 405954, 405955, 405956, 405959, 405960, 405961, 405974, 405977, 405978, 405979, 405980, 405982, 405987, 405996, 405997, 406028, 406029, 406030, 406031, 406032, 406033, 406035, 406038, 406039, 406040, 406041, 406042, 406043, 406044, 406045, 410562, 410583, 410591, 410592, 410593, 410668, 410677, 410716, 410724, 410757, and 410760 each exhibited at least 85% inhibition of PCSK9 mRNA levels.
ISIS Nos 395165, 395166, 395168, 395186, 395187, 395194, 399801, 399817, 399819, 399821, 399887, 399888, 399891, 399900, 399904, 399916, 399954, 399969, 399978, 399992, 405873, 405874, 405875, 405876, 405878, 405879, 405880, 405881, 405882, 405883, 405884, 405891, 405893, 405894, 405895, 405899, 405903, 405904, 405905, 405906, 405907, 405910, 405913, 405914, 405916, 405941, 405953, 405955, 405956, 405959, 405960, 405978, 405996, 405997, 406028, 406029, 406030, 406033, 406035, 406039, 406040, 406041, 406042, 406043, 406044, 410592, 410668, and 410760 each exhibited at least 90% inhibition of PCSK9 mRNA levels.
ISIS Nos 395165, 395166, 399801, 399978, 405881, 405891, 405959, 405978, 405997, 406030, 406040, 406041, 406044, and 410760 each exhibited at least 95% inhibition of PCSK9 mRNA levels.
Example 4
Antisense Reduction of Human PCSK9 mRNA in HepG2 Cells: Dose Response Experiment
Antisense oligonucleotides targeted to PCSK9 were tested at various doses in HepG2 cells. Cells were plated at densities of 10,000 cells per well and treated with nM concentrations of antisense oligonucleotide as indicated in Tables 20 and 21. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 20, and mRNA levels measured using primer probe set 2823 are shown in Table 21. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Tables 20 and 21, PCSK9 mRNA levels were reduced in a dose-dependent manner.







TABLE 20







Antisense Reduction of PCSK9 mRNA in HepG2 cells,


Primer Probe Set 2740


















100.0
200.0


Isis No
6.25 nM
12.5 nM
25.0 nM
50.0 nM
nM
nM
















399819
93
113
84
71
26
12


395185
81
64
50
35
24
13


399916
65
56
41
18
20
25


399907
68
66
42
26
30
25


399954
71
63
38
22
16
15


395165
71
67
47
29
21
10


399936
65
51
31
20
30
28


399793
61
57
44
31
39
15


399969
57
54
40
27
25
19


395152
75
58
58
49
24
28
















TABLE 21







Antisense Reduction of PCSK9 mRNA in HepG2 cells,


Primer Probe Set 2823


















100.0
200.0


Isis No
6.25 nM
12.5 nM
25.0 nM
50.0 nM
nM
nM
















399819
80
264
87
85
34
15


395185
86
66
57
45
29
22


399916
75
47
45
20
30
45


399907
67
72
47
29
31
32


399954
59
50
35
17
22
27


395165
74
54
41
37
26
22


399936
62
41
33
32
28
42


399793
62
51
38
36
44
27


399969
73
40
58
28
29
35


395152
78
53
64
49
22
35









Example 5
Antisense Reduction of Human PCSK9 mRNA in Hep3B Cells: Dose Response Experiment
Antisense oligonucleotides targeted to PCSK9 were tested at various doses in Hep3B cells. Cells were plated at densities of 4,500 cells per well and treated with nM concentrations of antisense oligonucleotide as indicated in Table 22. ISIS 141923 is not complementary to any known gene sequence and is used in this and the following experiments as a negative control. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 22. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Table 22, PCSK9 mRNA levels were reduced in a dose-dependent manner.







TABLE 22







Antisense Reduction of PCSK9 mRNA in 


Hep3B cells, Primer Probe Set 2740











Isis No.
2.78 nM
8.33 nM
25.0 nM
75.0 nM














399819
127.5
85.0
36.1
21.9


405891
133.7
111.4
40.2
27.2


406008
144.6
108.3
49.9
17.7


395186
124.4
106.4
42.2
30.7


395185
150.7
106.6
53.6
38.1


405994
141.9
101.2
49.3
30.8


405988
148.1
117.4
47.8
25.2


406033
138.1
109.9
62.4
35.3


395187
133.0
115.6
59.9
31.6


405995
124.1
109.2
57.8
42.9


406023
130.0
103.5
67.1
28.0


399900
131.2
95.7
81.1
30.9


301012
121.2
100.1
63.8
49.6


395165
113.2
86.9
41.6
13.3


405879
124.6
88.8
44.1
13.1


405991
100.6
101.4
95.9
41.0


405923
130.4
115.0
68.9
35.1


395152
143.8
98.6
61.3
59.0


405881
100.7
77.4
50.2
4.5


141923
131.4
131.9
144.0
167.3









Example 6
Antisense Reduction of Human PCSK9 mRNA in HeLa Cells: Dose Response Experiment
Antisense oligonucleotides targeted to PCSK9 were tested at various doses in HeLa cells. Cells were plated at densities of 5,000 cells per well and treated with nM concentrations of antisense oligonucleotide as indicated in Table 23. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 23. Results are presented as percent PCSK9 mRNA levels relative to a control, as adjusted according to total RNA content measured by RIBOGREEN®. As illustrated in Table 23, PCSK9 mRNA levels were reduced in a dose-dependent manner.







TABLE 23







Antisense Reduction of PCSK9 mRNA 


in HeLa cells, Primer Probe Set 2740













Isis No.
2.963 nM 
8.8889 nM 
26.6667 nM 
80.0 nM

















399819
57
32
16
14



405891
57
46
17
18



406008
45
27
11
5.3



395186
40
28
11
6.6



395185
53
32
21
17



405994
48
34
20
13



405988
44
32
12
7.3



406033
61
42
25
12



395187
42
35
12
6.6



405995
54
39
21
15



406023
76
46
25
6.2



399900
85
48
23
6.4



301012
121
109
76
65



395165
61
30
10
2



405879
64
32
9.7
1



405991
64
35
24
10



405923
71
45
22
5



395152
73
34
13
3.1



405881
58
33
19
2.8



141923
122
118
131
140










Example 7
Antisense Reduction of Human PCSK9 mRNA in Primary Hepatocytes
Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in human primary hepatocytes. Human primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.
The antisense oligonucleotides tested in human primary hepatocytes were Isis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954, 399978, and 399992. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes.
Monkey primary hepatocytes were similarly treated with 10, 25, 50, 150, or 300 nM concentrations of antisense oligonucleotide targeted to PCSK9. The antisense oligonucleotides tested in monkey primary hepatocytes were Isis 395152, 395155, 395165, 395185, 399819, 399821, 399916, 399954, 399978, and 399992. Each of these antisense oligonucleotides is fully complementary to a monkey PCSK9 nucleic acid. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in monkey primary hepatocytes.
Example 8
Additional Antisense Reduction of Human PCSK9 mRNA in Primary Hepatocytes
Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in human primary hepatocytes. Human primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.
The antisense oligonucleotides tested in human primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes as shown in Table 24.







TABLE 24







Dose-dependent Reduction in Human PCSK9 


mRNA Expression in Human Primary Hepatocytes












ISIS No.
10 nM
25 nM
50 nM
150 nM
300 nM















395165
76
48.7
22.5
20.2
16.2


395185
74.5
42.9
52.6
25.5
15.7


395186
123.8
58.3
34.4
21
19.9


395187
98.7
43.5
29.7
17.4
21.1


405879
119.2
176.6
64.9
48.4
47.7


405881
222.4
115.3
49.5
22.3
79.3


405891
169.9
138.7
73.9
86.9
48.7


405988
85.2
105.4
67.8
43.9
43.8


405994
95.7
83.3
43.5
24.6
10.5


406008
155
101.7
116
41.7
32.4









Example 9
Antisense Reduction of PCSK9 mRNA in Cyno Primary Hepatocytes
Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in cyno primary hepatocytes. Cyno primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.
The antisense oligonucleotides tested in cyno primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in cyno primary hepatocytes as shown in Table 25.







TABLE 25







Dose-dependent PCSK9 mRNA Inhibition in Cyno Primary Hepatocytes












ISIS







No.
10 nM
25 nM
50 nM
150 nM
300 nM















395165
36.3
29.9
14.9
8.6
3.2


395185
65.0
31.9
35.1
16.6
12.0


395186
65.8
42.3
21.6
23.3
16.7


395187
61.3
32.9
13.2
7.7
9.02


405879
24.3
21.9
7.7
7.1
3.0


405881
75.0
33.5
26.5
11.2
4.9


405891
52.4
24.8
11.2
10.8
6.7


405988
59.4
23.7
22.5
10.7
8.2


405994
64.8
52.7
22.2
7.9
6.5


406008
56.3
28.2
26.5
13.4
13.3









Example 10
Antisense Reduction of Mouse PCSK9 mRNA In Vitro
Antisense oligonucleotides were designed to target murine PCSK9, and were evaluated for their ability to reduce PCSK9 mRNA in primary mouse hepatocytes.
Primary mouse hepatocytes were treated with antisense oligonucleotides at doses of 50, 150, or 300 nM, for a period of 24 hours. RNA was isolated using QIAGEN® RNeasy isolation kits and subjected to quantitative real-time PCR using commercially available reagents (Invitrogen, Carlsbad, Calif.). PCSK9 mRNA levels were measured using a mouse PCSK9 primer probe set, and normalized to G3PDH mRNA levels and/or RIBOGREEN® levels.
ISIS 394814 (GAGCAACTTCGGAGGCAGC, SEQ ID NO: 456) was identified as a potent inhibitor of mouse PCSK9. PCSK9 mRNA levels ISIS 394814 yielded a 50% reduction in PCSK9 mRNA at a concentration of 25 nM (i.e., an IC50 of 25 nM) after a 24 hour incubation with primary mouse hepatocytes. No effect on cultured cell viability was observed.
Example 11
Antisense Reduction of PCSK9 mRNA in an Animal Model of Hyperlipidemia: High Fat Fed Mice
Treatment
C57BL/6 mice fed a high-fat diet are routinely employed as an animal model of hyperlipidemia, as well as an animal model of atherosclerosis. Accordingly, to evaluate the effects of PCSK9 antisense inhibition on serum lipid levels in vivo, ISIS 394814 was evaluated in C57BL/6 mice fed a high-fat diet. C57BL/6 mice 4-5 weeks of age were obtained from the Jackson Laboratory and maintained on a diet consisting of 60% fat. Treatment groups of 5 mice each were as follows: a group treated with ISIS 394814; a control group treated with saline; and control group treated with an oligonucleotide not complementary to any known gene sequence (ISIS 141923). Oligonucleotide or saline was administered intraperitoneally twice weekly, for a period of 6 weeks; oligonucleotide doses were 50 mg/kg. After the treatment period, whole blood was collected for analysis of serum parameters, and whole liver was collected for RNA analysis, protein analysis, and histological evaluation. Statistical analyses included a nonparametric, two-tailed t-test comparison of experimental samples (ISIS 394814) to control samples (saline or control oligonucleotide).
RNA Analysis
Liver RNA was isolated for real-time PCR analysis of PCSK9, LDL-receptor and apolipoprotein B mRNA levels. Treatment with ISIS 394814 resulted in a 92% reduction in PCSK9 mRNA levels, while no significant reduction in PCSK9 mRNA levels was observed in ISIS 141923-treated or saline-treated mice. Antisense inhibition of PCSK9 expression did not significantly affect liver LDL-receptor or liver apolipoprotein B mRNA levels.
Protein Analysis
Immunoblotting was performed to assess the effects of PCSK9 antisense inhibition on LDL-receptor protein and apolipoprotein B levels. Protein isolated from mouse liver was subjected to electrophoresis, transferred to a polyvinylidene-fluoride membrane, and subsequently probed with an anti-mouse LDL-receptor antibody and a scavenger receptor B1 (SR-B1) antibody. Mouse plasma was similarly subjected to electrophoresis and immunoblotting using an anti-mouse apolipoprotein B antibody. Secondary antibodies conjugated to peroxidase were used to detect primary antibodies. Protein bands were visualized using the ECL plus Western blot detection kit (Amersham Biosciences, UK) and quantified using ImageQuantTM analysis software (MolecularDynamics, Santa Clara, Calif.).
While LDL-receptor mRNA levels were not significantly affected, antisense inhibition of PCSK9 resulted in an approximate 2-fold increase in the level of hepatic LDL-receptor protein relative to 141923-treated controls. No changes in SR-B1 protein were observed. Furthermore, serum apoB-100 levels were significantly reduced by 50%, relative to 141923-treated controls. Serum apoB-48 levels were significantly increased by approximately 3-fold relative to 141923-treated controls. No significant changes in apoA-I protein were observed, relative to 141923-treated controls.
The mRNA levels of the RNA editing enzyme apobec-1 were also measured. Apobec-1 is responsible for the RNA editing that produces apoB-48 in murine liver and intestine. Human apobec-1 is expressed only in intestinal cells, thus these cells are the source of apoB-48 in humans. Antisense inhibition of PCSK9 resulted in an increase in murine hepatic apobec-1 mRNA levels by approximately 2.7 fold compared to saline-treated controls
Serum Lipid Analysis
Plasma concentrations of total cholesterol, LDL-C, HDL-C, free cholesterol, triglycerides, glucose, ketones, transaminases, and phospholipids were measured using an automated clinical chemistry analyzer (Hitachi Olympus AU400e, Melville, N.Y.). Serum lipoprotein and cholesterol profiling was performed as described by Crooke et al. (Lipid Res., 2005, 46, 872-884) using a Beckman System Gold 126 HPLC system, a 507e refrigerated autosampler, a 126 photodiode array detector (Beckman Instruments, Fullerton, Calif.), and a Superose 6 HR 10/30 column (Pfizer, Chicago, Ill.). HDL, LDL and VLDL fractions were measured at a wavelength of 505 nm and validated with a cholesterol calibration kit. (Sigma).
Administration of ISIS 394814 resulted in a 52% reduction in total cholesterol (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS 394814, 87 mg/dL±19, p<0.005) and a 36% reduction in LDL-C (saline 22 mg/dL±4; ISIS 141923, 25 mg/dL±2; ISIS 394814, 14 mg/dL±2, p<0.005). Serum free cholesterol was reduced by 25% (saline 41 mg/dL±10; ISIS 141923, 58 mg/dL±3; ISIS 394814, 31 mg/dL±5, p<0.005) and phospholipids were reduced 54% (saline, 354 mg/dL±29; ISIS 141923, 383 mg/dL±21; ISIS 394814, 169 mg/dL±35, p<0.0001). HDL-C was also reduced by approximately 54% (saline, 183 mg/dL±18; ISIS 141923, 194 mg/dL±14; ISIS 394814, 87 mg/dL±19; p<0.0001). HPLC profiling confirmed the reduction of LDL and HDL lipid classes.
Liver Triglyceride Analysis
Liver triglyceride content was measured according to routine experimental procedures, for example, per procedures described by Desai et al., Diabetes, 2001, 50:2287-2295.
Antisense inhibition of PCSK9 for 6 weeks reduced liver triglyceride content by approximately 65% (p=0.01) relative to saline controls. No statistically significant changes in liver triglyceride content following 141923 treatment were observed.
Accordingly, one embodiment is a method of lowering LDL-C levels through the administration of an antisense oligonucleotide targeted to a PCSK9 nucleic acid. An additional embodiment includes a method of lowering total cholesterol through the administration of an antisense oligonucleotide targeted to a PCSK9 nucleic acid. An addition embodiment is lowering liver triglycerides by administering an antisense oligonucleotide targeted to a PCSK9 target nucleic acid.
Example 12
Antisense Reduction of PCSK9 mRNA in LDL-R Deficient/apoB-100 Animals
LDL-receptor (LDL-R)-deficient/apoB-100 animals do not express the LDL-R gene, and express only the apoB-100 form of apoB. PCSK9 antisense oligonucleotide was administered to LDL-R deficient/apoB-100 mice, to evaluate the effects of antisense inhibition of PCSK9 in the absence of the LDL-R. Mice 4-5 weeks of age were maintained on a standard mouse diet. Treatment groups of 5 mice each were as follows: a group treated with ISIS 394814; a control group treated with saline; and control group treated with an oligonucleotide not complementary to any known gene sequence (ISIS 141923). Oligonucleotide or saline was administered intraperitoneally twice weekly, for a period of 6 weeks; oligonucleotide doses were 50 mg/kg (100 mg/kg/wk total). Real-time PCR of liver mRNA and serum analyses were performed as described for the C57Bl/6 study.
Antisense inhibition of PCSK9 reduced liver PCSK9 mRNA by approximately 90%, relative to saline controls. However, no reduction in serum cholesterol was observed. Liver triglyceride levels were unaffected by antisense inhibition of PCKS9 in these mice. An 80% increase in liver apobec1 mRNA levels was observed, relative to saline controls. These data suggest that a functioning LDL-R is required for cholesterol and liver triglyceride reduction via PCSK9 antisense inhibition.
Example 13
Antisense Reduction of PCSK9 mRNA in Mouse Primary Hepatocytes
Antisense oligonucleotides targeted to PCSK9 were tested for antisense inhibition of PCSK9 in mouse primary hepatocytes. Mouse primary hepatocytes were purchased from a commercial supplier, and cultured according to routine culture procedures. Cells were treated with 6.25, 12.5, 25, 50, 100, or 200 nM of antisense oligonucleotide for a period of 24 hours, after which RNA was isolated and PCSK9 mRNA was measured by real-time PCR as described herein.
The antisense oligonucleotides tested in human primary hepatocytes were Isis 395165, 395185, 395186, 395187, 405879, 405881, 405891, 405988, 405994, and 406008. The antisense oligonucleotides inhibited PCSK9 in a dose-dependent manner in human primary hepatocytes as shown in Table 26.







TABLE 26







Dose-Dependent Reduction PCSK9 mRNA in


Mouse Primary Hepatocytes


















100.0
200.0


ISIS No
6.25 nM
12.5 nM
25.0 nM
50.0 nM
nM
nM
















395165
95.2
94.9
81.7
66.3
57.5
29.9


395185
109.2
100
90.5
81.2
64.7
35.8


395186
101.9
91.5
77.8
59.1
39.5
13.7


395187
102.4
96.1
83.9
66.4
40
15.3


405879
107.3
97.5
87.8
76.6
63
43.1


405881
104.8
102.1
98.1
84.9
68.7
48.9


405891
104.9
103.3
101.5
88.9
79.6
49.5


405988
97.3
101.1
91.7
82.3
61.2
34.8


405994
110.2
101
111.6
94.4
80.1
57.7


406008
99.5
91.3
83.9
73.7
61.8
39.4


157700
97.8
98.3
96.2
94.3
83.2
76.4


141923
84.3
80
79.9
77.3
98.8
64.2









Example 14
Antisense Reduction of Human PCSK9 mRNA in Hep3B and HeLa Cells: Dose Response Experiment
A subset of the antisense oligonucleotides that inhibited PCSK9 expression at high levels in the cell culture systems described above was selected for further screening. The group of selected antisense compounds is shown in Table 27. Table 27 shows the Isis number, nucleotide sequence, and percent inhibition obtained in Hep3B cells using the protocol disclosed in Example 3 (see Table 17) for the various antisense compounds.











TABLE 27









SEQ


Isis


%
ID


No.
Sequence 5′-3′
Motif
inhibition
NO







405881
CACCCTTGGCCACGCCGGCA
5-10-5
98.3
250


 


399819
CCCACTCAAGGGCCAGGCCA
5-10-5
93.4
 65


 


395165
ACCCTTGGCCACGCCGGCAT
5-10-5
96.7
 28


 


405879
CCTTGGCCACGCCGGCATCC
5-10-5
94.1
248


 


406008
CTTGGTGAGGTATCCCCGGC
5-10-5
95.2
188


 


405891
TCCTCAGGGAACCAGGCCTC
5-10-5
96.1
352


 


395186
CTTTGCATTCCAGACCTGGG
5-10-5
93.7
 60


 


405988
GGCAGCACCTGGCAATGGCG
5-10-5
95.9
381


 


405994
GCAGTGGACACGGGTCCCCA
5-10-5
95.2
400


 


406023
TGGTATTCATCCGCCCGGTA
5-10-5
92.5
212


 


395187
GGCAGCAGATGGCAACGGCT
5-10-5
92.7
 62


 


395185
CACGGGTCCCCATGCTGGCC
5-10-5
92.7
 59


 


406033
CCTGCCAGGTGGGTGCCATG
5-10-5
91.2
237


 


405923
GGCATTGGTGGCCCCAACTG
5-10-5
94.3
288


 


399900
GGTCCTCAGGGAACCAGGCC
3-14-3
92.0
 50


 


405995
TGGCAGTGGACACGGGTCCC
5-10-5
92.4
402


 


405991
GACACGGGTCCCCATGCTGG
5-10-5
91.9
394


 


406005
CGGGCAGTGCGCTCTGACTG
5-10-5
83.4
180


 


399793
CCTCGGAACGCAAGGCTAGC
5-10-5
77.9
  8


 


395152
ACGCAAGGCTAGCACCAGCT
5-10-5
93.3
  7









The IC50 of the various antisense compounds was determined using different cell culture systems. Antisense oligonucleotides targeted to PCSK9 were tested at various doses in Hep3B or HeLa cells, for example. Cells were plated at densities of 4,500 cells or 5,000 per well, respectively, and treated with of antisense oligonucleotides at various concentrations. After a treatment period of approximately 24 hours, RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein, relative to untreated control cells. Two different human PCSK9 primer probe sets were used to measure mRNA levels. Results with Primer Probe Set 2740 are shown in Table 28. PCSK9 mRNA levels were adjusted according to total RNA content as measured by RIBOGREEN®. Table 28 shows IC50 values (nM) obtained from two experiments with Hep3B cells and from one experiment with HeLa cells.







TABLE 28







IC50 values













Hep3B
Hep3B
HeLa

SEQ


ISIS
IC50
IC50
IC50
PCSK9 Target
ID


#
(nM)
(nM)
(nM)
Sequence
NO





405881
 21
26
13
CACCCTTGGCCACGCCGGCA
250


 


399819
 21
28
12
CCCACTCAAGGGCCAGGCCA
 65


 


395165
 23
28
13
ACCCTTGGCCACGCCGGCAT
 28


 


405879
 24
28
14
CCTTGGCCACGCCGGCATCC
248


 


406008
 31
35
 8
CTTGGTGAGGTATCCCCGGC
188


 


405891
 32
38
14
TCCTCAGGGAACCAGGCCTC
352


 


395186
 33
33
 7
CTTTGCATTCCAGACCTGGG
 60


 


405988
 34
42
 8
GGCAGCACCTGGCAATGGCG
381


 


405994
 35
39
 9
GCAGTGGACACGGGTCCCCA
400


 


406023
 40
50
18
TGGTATTCATCCGCCCGGTA
212


 


395187
 41
44
 8
GGCAGCAGATGGCAACGGCT
 62


 


395185
 42
43
11
CACGGGTCCCCATGCTGGCC
 59


 


406033
 44
61
15
CCTGCCAGGTGGGTGCCATG
237


 


405923
 47
41
20
GGCATTGGTGGCCCCAACTG
288


 


399900
 48
44
21
GGTCCTCAGGGAACCAGGCC
 50


 


405995
 48
63
12
TGGCAGTGGACACGGGTCCC
402


 


405991
 63
52
15
GACACGGGTCCCCATGCTGG
394


 


406005
 63


CGGGCAGTGCGCTCTGACTG
180


 


399793
>75


CCTCGGAACGCAAGGCTAGC
  8


 


395152

46
16
ACGCAAGGCTAGCACCAGCT
  7









Several of the antisense compounds examined above were selected for further study. These antisense compounds exhibited at least about 93% inhibition of human PCSK9 mRNA expression in the Hep3B cell culture system, as shown in Tables 17-19, for example. As shown in Table 28, the selected antisense compounds also displayed IC50 parameters of less than about 45 nM when tested with Hep3B cells and less than about 15 nM when tested with HeLa cells. The selected antisense compounds, listed in Table 29, are Isis 405881, Isis 395165, Isis 405879, Isis 406008, Isis 405891, Isis 395186, Isis 405988, Isis 405994, Isis 395187, and Isis 395185. Isis 405879, for example, showed superior results in both Hep3B and HeLa cells, with IC50 values of about 26 nM and 14 nM, respectively.
Example 15
Dose-Dependent Antisense Reduction of PCSK9 mRNA in Human Primary Hepatocytes; Cynomolgus Monkey (“Cyno”) Primary Hepatocytes; and Human HEK-293 Cells
The selected antisense oligonucleotides were further tested for dose dependent antisense inhibition in human primary hepatocytes, cyno primary hepatocytes, and HEK-293 cells expressing human PCSK9. Cells were purchased from commercial suppliers and cultured according to routine culture procedures. Cells were treated with 10, 25, 50, 150, or 300 nM antisense oligonucleotide (i.e., a five-point dose response test) for a period of 24 hours, after which RNA was isolated. PCSK9 mRNA was measured by real-time PCR, as described herein.
Results of the five-point dose response tests using HEK-293 cells are shown in FIG. 2. Results of the five-point dose response tests using cyno primary hepatocytes are shown in FIG. 3. The IC50 results, expressed in nM units, for the five-point dose response test using human primary hepatocytes, cyno primary hepatocytes, HEK-293 cells, Hep3B cells, and HeLa cells are summarized in Table 29.







TABLE 29







IC50 (nM) for Selected Antisense Compounds









Isis No.


















395165
395185
395186
395187
405879
405881
405891
405988
405994
406008









SEQ ID NO:


















28
59
60
62
248
250
352
381
400
188





















Human Primary
24
33
39
30
ND
50
297
166
56
163


Hepatocytes


Cyno Primary
5
17
18
14
2
19
10
12
21
11


Hepatocytes


HEK-293
40
116
6
11
35
86
21
11
17
27


Hep3B
28
43
33
44
28
26
38
42
39
35


HeLa
13
11
7
8
14
13
14
8
9
8









ISIS 405879 was tested at various doses in human primary hepatocytes. RNA was isolated from the cells and PCSK9 mRNA levels were measured by quantitative real-time PCR, as described herein. Human PCSK9 primer probe set 2740 was used to measure mRNA levels. Results are shown in Table 29.1.







TABLE 29.1







Dose-dependent Reduction in Human PCSK9 mRNA Expression


in Human Primary Hepatocytes















300
200







ISIS NO.
nM
nM
150 nM
100 nM
50 nM
25 nM
10 nM





405879
13
18
26
30
27
54
75









Isis 405879 has an IC50 value of 29 nM in human primary hepatocytes. Overall Isis 405879 displays superior activity, particularly in cyno primary hepatocytes. Isis 405879 was thus further tested in the various cell models, and the results of the additional testing are shown in Table 30. In these experiments, Isis 405879 was administered to the cells using lipofectin transfection reagents. PCSK9 mRNA was assayed by PCR following 24 hr incubation after transfection. These results confirm that Isis 405879 potently inhibits PCSK9 mRNA expression at low concentrations.







TABLE 30







Isis 405879 activity in cell culture models


















Hepatocyte


Transgenic



Hep3B
HepG2
HeLa
Transgenic
Primary
Primary
HEK293



cells
Cells
Cells
Mouse
Hepatocyte
Hepatocyte
cells



(human)
(human)
(human)
(h-PCSK9)
(human)
(cyno)
(h-PCSK9)


















Mean
17
5
14
140
22
7.4
18


IC50


(nM)









These experiments confirm that the presently disclosed antisense oligonucleotides are capable of potently inhibiting PCSK9 mRNA expression in a variety of cell model systems, including a human primary cell line. Exemplary antisense compounds inhibit PCSK9 mRNA expression with IC50 values in the nM range.
The transgenic PCSK9 cDNA construct expressed in mouse hepatocytes is thought to be rapidly exported to the cytoplasm, because it does not require post-transcriptional editing. The antisense oligonucleotides provided herein act by recruiting the RNase H enzyme, which is restricted to the nucleus. Therefore, inhibition by antisense oligonucleotides in this cell type is significantly diminished.
Example 16
In Vivo Testing Procedures
16.1. High Fat (HF)-Fed Hyperlipidemic Mouse Model
Antisense compounds may be tested in a high fat-fed, hyperlipidemic mouse model to test efficacy of the antisense compounds in reducing PCSK9 mRNA expression, increasing LDL-R expression, reducing total cholesterol and LDL, and affecting other metrics of antisense compound efficacy. A representative hyperlipidemic mouse model is described in Graham et al., J. Lipid Res. (2007) 48: 763-767.
All animal experiments were conducted in accordance with Institutional American Association for the Accreditation of Laboratory Animal Care guidelines. C57BL/6 mice were obtained from Jackson Laboratory (http://www.jax.org). A majority of the mice were male, and studies were initiated when animals were 4-5 weeks of age. The mice were maintained on a 12 h light/12 h dark cycle and fed ad libitum. C57BL/6 mice were fed a diet consisting of 60% lard (Research Diets, New Brunswick, N.J.), whereas experimental Ldlr-deficient/apoB-100 mice were fed regular chow.
Antisense compounds were administered twice weekly (50 mg/kg) for 6 weeks by intraperitoneal injection (10 mg/mL dosing solution formulated in saline; saline control). After the treatment period, whole blood was collected for analysis of serum parameters, and whole liver was collected for RNA analysis, protein analysis, and histological evaluation.
16.2. H-ApoB/CETP Transgenic Mouse Model
A transgenic mouse model that expresses both human apolipoprotein (apo) B and human cholesteryl ester transfer protein (CETP) provides another useful animal model system to test the efficacy of the presently disclosed antisense compounds. The lipoprotein cholesterol distribution in the serum of a chow-fed h-apoB/CETP mouse resembles a human profile. Specifically, the percentages of total cholesterol within the HDL, LDL, and VLDL fractions of apoB/CETP animals are approximately 30%, 65%, and 5%, respectively, similar to the distribution of cholesterol in the plasma of normolipidemic humans. The h-apoB/CETP mouse model is described further in Grass et al., J. Lipid Res. (1995) 36: 1082-1091.
16.3. H-PCSK9 Transgenic Mouse Model
A transgenic mouse expressing human PCSK9 provides another useful animal model to study in vivo repression of PCSK9 mRNA by the present antisense compounds. H-PCSK9 transgenic mice overexpress human PCSK9 in liver and secrete large amounts of the protein into plasma, which increases plasma LDL-C concentrations to levels similar to those of LDLR-knockout mice. H-PCSK9 transgenic mice were constructed by inserting a cDNA encoding human PCSK9 into a pLiv-11 vector that contained the constitutive human apoE promoter and its hepatic control region, as described in Simonet et al., J. Biol. Chem. (1993) 268: 8221-8229. Transgenic mice were generated by injecting linearized pLiv-11-hPCSK9 into the fertilized eggs as described in Shimano et al., J. Clin. Invest. (1996) 98: 1575-1584. H-PCSK9 transgenic mice are further described in Lagace et al., J. Clin. Invest. (2006) 116: 2995-3005.
16.4. Mouse and Rat Models for In Vivo Tolerability, Half-Life and Tissue Distribution
Mouse (CD1) and rat (Sprague-Dawley) models were used to test the in vivo tolerability of various antisense compounds. In the mouse CD1 model, antisense compounds were overdosed at 100 mg/kg/wk for six weeks. Possible proinflammatory effects and hepatotoxcity were assayed following administration of the antisense compounds. Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), blood urea nitrogen (BUN), creatinine, albumin, and bilirubin were assayed. Weights of livers, kidneys, and spleens of animals were determined. Histopathologic screening was conducted to detect hepatocellular swelling, multifocal apoptosis, inflammatory cell infiltrates in the liver or lungs, or follicular hyperplasia. Additional studies were conducted in the rat model to screen for renal toxicity or high renal accumulation and proinflammatory effects. In these experiments, antisense compounds were administered at 60 or 100 mg/kg/wk for six weeks. In addition to the markers examined above, the urine protein/creatine ratio and proteinuria were assayed.
The half-life of administered antisense compounds also was examined in the target organ, i.e., liver, in the mouse model. For these experiments, antisense compounds were administered twice a week for two weeks at 50 mg/kg, and target organs were then harvested and assayed for the presence of the antisense compounds, as described below.
The distribution of administered antisense compounds in various organs also was examined in the rat model. Rats in these studies were administered 20 mg/kg antisense compounds twice weekly for three weeks. Livers and kidneys of test animals were then harvested and assayed for the presence of the antisense compounds, as described below.
16.5. Cynomolgus Monkey (M. fascicularis) Model
Potencies and pharmacodynamics of antisense compounds were compared in a cyno model to identify the compounds exhibiting the best combination of high potency and low renal accumulation, when given subcutaneously to animals for a 5-week period. In another study, dosing was conducted over a 13-week period. The cyno monkey is widely used in toxicity testing, and a substantial historic toxicology database is available for this model.
The PCSK9 target sequence for an antisense compound may differ in humans versus cyno monkey. As a result, an antisense compound for use in humans may not be fully complementary to a monkey PCSK9 nucleic acid. Antisense compounds having identical target sequences in the two species are preferred. Isis 405881 and Isis 395165 each have a one nucleotide mismatch compared to the cyno sequence, meaning that these antisense compounds are less preferred in the cyno model.
Cyno monkeys (Charles River BRF, Inc.) were approximately 26 to 32 months old and 2.5 to 4 kg in weight on the first day of dosing. Monkeys were offered water ad libitum and fed a daily ration of 12 biscuits approximately 2 hours post dose. Antisense compounds were administered subcutaneously at 3 doses every other day during the first week, then by weekly injections. A low dose of 15 mg/kg was used to assess the potency of the antisense compounds. A high dose of 8 subcutaneous injections of 30 mg/kg over 5 weeks or 13 weeks was used to assay renal accumulation and pharmacodynamics. Antisense compounds were administered at a concentration of 60 or 120 mg/mL, respectively, in a vehicle of sterile phosphate buffered saline. Plasma concentrations of antisense compounds were determined at each dose level to estimate systemic exposure in monkeys following subcutaneous administration and to assess whether systemic exposure was altered after repeat dosing.
To measure the host reaction to the administered antisense compounds, 2 mL blood samples were collected from the femoral vein of host individuals in tubes of appropriate size containing K2EDTA. Plasma was obtained by centrifugation and samples were stored frozen. The analysis of the plasma samples included a determination of the level of plasma PCSK9 protein. To measure an effect on pharmacodynamic end points, LDL, HDL, and apoB levels also were analyzed. To determine whether the host individuals suffered side effects, blood samples were further analyzed for an effect on the hematological parameters listed in Table 31 and the serum chemistry parameters listed in Table 32:







TABLE 31





Hematology markers
















erythrocyte count
red cell distribution width


hemoglobin
platelet count


hematocrit
mean platelet volume


mean corpuscular volume
absolute total and differential leukocyte



counts


mean corpuscular hemoglobin 
evaluation of cell morphology


concentration



absolute reticulocyte count
















TABLE 32





Serum chemistry markers


















aspartate aminotransferase
triglycerides



alanine aminotransferase
glucose



gamma glutamyltransferase
urea nitrogen



alkaline phosphatase
creatinine



total bilirubin
calcium



total cholesterol
phosphorus



total protein
sodium



albumin
potassium



globulins
chloride



albumin/globulin ratio










Possible additional side effects were determined by screening individuals administered antisense compounds for gross changes in organs or microscopic changes in tissues. A limited necropsy conducted on all test animals included gross examination of various organs and tissues. Terminal body weights were recorded, and kidneys and spleen were weighed for all test animals. For microscopic evaluation, liver, kidney, spleen and injection sites from all animals were fixed in 10% neutral buffered formalin and were processed, embedded in paraffin, sectioned, and stained with hematoxylin and eosin.
Representative tissue samples of liver and kidney cortex from each scheduled-necropsy monkey from the high-dose and control groups were collected for ultrastructural examination using electron microscopy. Thin slices (2-4 mm in thickness) of liver and kidney cortex were cut and transferred to a piece of dental wax containing a small volume of McDowell-Trump fixative. Thin slices were further minced into approximately 1×1-2 mm cubes in McDowell-Trump fixative and analyzed by electron microscopy.
To evaluate PCSK9 mRNA levels, samples from liver (approximately 1 g) were homogenized using a Polytron tissue disruptor in 10 mL of RLT solution (Qiagen) and snap-frozen and stored at −70° C. or colder. Additionally, samples (approximately 500 mg each) from kidney were collected and snap-frozen for mRNA analysis.
Tissue samples from liver and kidney cortex were also analyzed for the presence of the administered antisense compounds. Antisense compounds present in the tissues can be detected and quantified using capillary gel electrophoresis (CGE), as described in Leeds et al, Drug Metab Dispos. (1998) 26: 670-675, for example. In this procedure, tissues are weighed, mixed with an internal standard (e.g., a 27-mer (T) oligonucleotide) in proteinase K digestion buffer (2.0 mg/ml proteinase K in 20 mM Tris-HCl, pH 8.0, 20 mM EDTA, 100 mM NaCl, 0.5% Nonidet P-40), and homogenized. Overnight incubation at 37° C. is followed by phenol/chloroform extraction. Tissue extracts are purified by sequential anion-exchange solid-phase extraction (SPE) followed by reverse-phase SPE, described in Leeds et al., Anal Biochem (1996) 235: 36-43. Samples are further desalted by membrane dialysis before analysis using gel-filled capillaries, as described in Leeds et al. (1996).
Capillary gel electrophoresis separations can be performed as described by Leeds et al. (1998), using a Beckman P/ACE capillary electrophoresis instrument (model 5010) with a 27-cm column (effective length, 20 cm) containing 12% polyacrylamide, with 8.3 M urea in 100 mM Tris-borate, pH 8.5, as the running buffer. Separation is achieved at 50° C. and 550 V/cm. Oligomers eluting from the column are detected by UV absorption at 260 nm. Quantification of antisense oligonucleotides is based on the tissue weight extracted and the initial T27 concentration.
Example 17
In Vivo Testing Results
17.1. Reducing PCSK9 mRNA In Vivo Using a Murine Antisense Compound Correlates with In Vivo Plasma LDL-C and Liver TG Reduction
In vivo pharmacodynamic studies were performed with the murine antisense compound Isis 394816 to validate the pharmacodynamic endpoint of PCSK9 mRNA reduction. Isis 394816 has the sequence 5′ GGTAAGGTGCGGTAAGTCCT 3′ (SEQ ID NO: 462), which targets position 3100 (5′) of the coding sequence of murine PCSK9 mRNA (NCBI Accession No. NM_153565.1; SEQ ID NO: 463). Isis 394816 has a 3-14-3 motif.
Isis 394816 was administered by intraperitoneal injection to an h-apoB/CETP transgenic mouse for 6 weeks at a dosing regimen of 20 mg/kg/wk or 50 mg/kg/wk. See Example 16.2. As shown in FIG. 4, Isis 394816 administered at either dose caused an approximate 80% reduction in PCSK9 mRNA expression in livers of test animals, compared to control animals. As shown in FIG. 5, panel A, Isis 394816 produced a significant (p<0.0001) decrease in the level of plasma LDL-C at the end of six weeks. Additionally, as shown in FIG. 5, panel B, Isis 394816 produced a significant (p<0.02) decrease in the level of liver TG at the end of six weeks.
Additionally, Isis 394816 reduced PCSK9 mRNA expression, plasma LDL-C, and liver TG in a dose dependent manner, as shown in FIG. 6. The potency of Isis 394816 was much higher in a hyperlipidemic mouse model (see Example 16.1.) than in a lean mouse model (data not shown). The reduction of plasma LDL-C positively correlated with the inhibition of PCSK9 mRNA levels in liver, shown in FIG. 7. These studies demonstrate that the reduction of liver PCSK9 mRNA correlates with the desired endpoint of reducing plasma LDL-C and liver TG. Accordingly, the reduction of liver PCSK9 mRNA provides a valid pharmacodynamic endpoint.
17.2. Differences in the Half-Life (t1/2) of Antisense Compounds in Liver
The half-lives of selected antisense compounds were assayed in mouse CD1 livers, according to the procedures set forth in Example 16.4. Half-lives varied among the tested antisense compounds, which again is attributable to differences in structural features of the antisense compounds, such as nucleotide sequences and/or motifs. For example, the half-life of Isis 405879 (t1/2=27.7 days) was significantly higher than the half-life of Isis 395165 (20.4 days), although the target sequences of these two antisense compounds are shifted by only two nucleotides:









(SEQ ID NO: 248)










Isis 405879:
CCTTGGCCACGCCGGCATCC



 









(SEQ ID NO: 28)










Isis 395165:
ACCCTTGGCCACGCCGGCAT






Half-life data for exemplary antisense compounds is summarized in Table 33. Isis 405879 displayed a particularly high half-life in liver.








TABLE 33







Liver t1/2 (days)










Antisense

Antisense



Compound
Liver t1/2 (d)
Compound
Liver t1/2 (d)





395165
20.4
405881
14.1


395185
20.2
405891
17.1


395186
19.9
405988
14.6


395187
13.7
405994
20.2


405879
27.7
406008
19.7









17.3. Differences in the Liver and Kidney Distributions of Antisense Compounds
The antisense compounds listed in Table 29 were administered to rats to determine the distribution of antisense compounds in liver and kidney. Accumulation of high levels of antisense compounds in the kidneys may cause adverse side effects. Antisense compounds were dosed and administered as described in Example 16.2. FIG. 8 shows the distribution of the antisense compounds shown in Table 29 in rat liver and kidneys. Isis 395185, Isis 395186, Isis 395187, Isis 405879, Isis 405881, Isis 405891, Isis 405988, and Isis 405994 show a particularly advantageous ratio of distribution between liver and kidney.
When tissue distribution was tested in a cyno model, further differences among antisense compounds were observed. Accumulation of Isis 395186, Isis 405879, and Isis 405891 in liver and kidney was examined after dosing each antisense compound at two concentrations. See Example 16.5. The results are shown in FIG. 9. Accumulation at each dosing concentration (15 mg/kg and 30 mg/kg) is shown for kidney (left side bar) and liver (right side bar). Isis 405879 advantageously accumulates at a high concentration in liver, even at a dose of 15 mg/kg, and accumulates at a low level in kidney. Specifically, Isis 405879 accumulated in kidneys at a concentration of 1300 μg/g (15 mg/kg dose) to 1800 μg/g (30 mg/kg dose). The average accumulation of antisense compounds in kidneys is 2500 μg/g (15 mg/kg dose) to 4000 μg/g (30 mg/kg dose). Variation in kidney and liver accumulation is attributable to structural differences, such as nucleotide sequences and/or motifs, between the antisense compounds.
17.4. Differences in Host Tolerance of Antisense Compounds
Rodent tolerability studies, conducted as described in Example 16.4, showed that antisense compounds were well tolerated. Following administration of the exemplary antisense compounds listing in Table 29, serum chemistry markers were normal, with some variably high ALT and AST levels observed for Isis 405988 and Isis 405994. Whole body weight and major organ weights in CD-1 mice also were not significantly affected by administration of the exemplary antisense compounds. Histological findings revealed differences between the various antisense compounds, but were consistent with the overall good tolerability for the antisense compounds. The results of the histological screens are shown in Table 34.
17.5. Results of the 5 wk Cyno Toxicology Study
The methods used to conduct the 5 wk cyno toxicology study are described in Example 16.5. Isis 405879 is referred to as BMS-844421 below and in the corresponding figures. The PCSK9 mRNA levels assayed by q-RT-PCR in the cyno liver tissue samples were decreased by BMS-844421 treatment by an average of 65% and 70% (average of males and females) versus saline controls at the low and high doses, respectively (FIG. 25). The effect was seen in both females and male monkeys, despite relatively high variability of mRNA level between animals. No dose-dependency was seen between the 2 doses. The data are mean+/− for male and female animals combined.
The levels of BMS-844421 ASO were measured in liver at the end of the 5 wk study. As shown in FIG. 26, liver full-length BMS-844421 levels reached  ̃1 mg/gram tissue in both dose groups. This level is consistent with values from previous experiences with other liver-targeting ASOs. Most of the compound was detected as intact full-length BMS-844421. No dose-dependency was seen in the ASO liver accumulation data, in agreement with the lack of dose-dependent effects on liver PCSK9 target mRNA.
Pharmacodynamic responses were assessed in serum lipid assays for the cyno 5 wk ASO study. As seen in FIG. 27, a modest trend (not statistically significant) toward lowering LDL-C was observed for BMS-844421 treated cyno monkeys at 2.5 and 5 wk of dosing. This trend at both time points for BMS-844421 relative to baseline was also seen in the scatterplot of individual animal data in FIG. 28A. HDL/LDL ratios were sometimes employed as a sensitive indicator of plasma lipid status. A chart of the average ratio of HDL/LDL in the cyno study showed a trend for increase in the ratio for BMS-844421 treated cynos at 2.5 and 5 wk (FIG. 28B).
Gene expression profiling by Affymetrix microarray was conducted on the RNA samples from this study. The analysis included liver RNA from animals administered vehicle alone, BMS-844421 at 15 mg/kg and 30 mg/kg and BMS-844419 and BMS-844423 at 30 mg/kg only. By microarray, there were significant (p<0.01) transcriptional changes in approximately 1 to 2% of genes with approximately 1% expected to change at this significance level by chance. For drug-dosed groups relative to control there were alterations in inflammation-, immune system-, and lipid homeostasis-related transcripts with statistically significant animal to animal variability. Other drug-related hepatic transcriptional changes common to multiple dose groups were associated with several or a few genes in various biological processes and are of unknown significance.
17.6. Results of the 13 wk Cyno Toxicology Study
An additional opportunity to monitor pharmacodynamic responses to BMS-844421 in monkeys was the 13 wk toxicology study, described in Example 16.5. Lean cyno monkeys with low LDL levels were again the subject for this study, as for the 5 wk cyno monkey study above. At 13 wks subcutaneous dosing at 4 different dose levels, BMS-844421 suppressed PCSK9 expression as measured by both liver PCSK9 mRNA and plasma PCSK9 protein levels. Despite the inter-animal variability in these values, it is apparent that BMS-844421 strongly suppressed both liver mRNA and plasma PCSK9 protein (FIG. 29). 40 mg/kg suppressed PCSK9 protein levels very robustly with less variation between animals (FIG. 29). No statistically significant differences were seen in plasma LDL-C or other lipid parameters in this study.







TABLE 34







Histological Findings in CD-1 Mice

















Antisense





Lymph



Skeletal


compound
Liver
Kidney
Lung
Heart
Spleen
node
Intestine
Thyroid
Sternum
Muscle





Saline
−
−
−
−
−
−
−
−
−
−


ISIS 395165
+/++
−


−/+
−/+
−
−
−
−


ISIS 395185
+
−
−
−
−/+
−
−
−
−
−


ISIS 395186
−
−
−
−
−
−
−
−
−
−


ISIS 395187
−
−
−
−
−/+
−
−
−
−
−


ISIS 405879
−
−
−
−
−
−
−
−
−
−


ISIS 405881
inflam
−
inflam
−
−/+
−/+
−
−
−
−


ISIS 405891
−
−
−
−
−/+
−/+
−
−
−
−


ISIS 405988
−
−
−
−
−
−
−
−
−
−


ISIS 405994
++
−
−
−
−/+
−
−
−
−
−


ISIS 406008
+/++
−
−
−
−
−
−
−
−
−





Normal: −


Minimal changes: −/+


Mild changes: +


Moderate changes: ++


Severe changes: +++






Microscopic evaluation of mouse organs from animals treated with 10 different human PCSK-9 antisense compounds revealed that all compounds generally were well tolerated, but that some antisense compounds were particularly well tolerated. The following results were seen in the various examined organs.

    
    
        Liver: Mild (+) to moderate (++) liver injuries (cytoplasmic swelling to multifocal apoptosis) were seen in Isis 395165 (+/++), Isis 395185 (+), Isis 405988 (+/++), Isis 405994 (++), and Isis 406008 (+/++) treated livers. Mild inflammatory changes were seen in Isis 405881 treated livers.
        Lung: No significant abnormality was seen in treated animals except for mild histiocytosis noticed in Isis 405881 treated animals.
        Spleen: Mild follicular hyperplasia was seen in Isis 395165, Isis 395185, Isis 395187, Isis 405881, Isis 405891, and Isis 405994 treated spleens.
        Lymph node: Minimal histiocytosis was observed in Isis 395165, Isis 405881 and Isis 405891 treated animals.
        Heart: Minimal levels of macrophage infiltration were seen in Isis 395165, Isis 395186, Isis 405879, Isis 405881, Isis 405891, Isis 405988, and Isis 406008 treated hearts.
        Kidney, Thyroid, small intestine, skeletal muscle and sternum: No abnormality was visualized.
    
    


Differences in the histological findings were seen even between Isis 395165 and Isis 405879. As noted above, the target sequences of these two antisense compounds are only shifted by two nucleotide positions. Nevertheless, Isis 405879 advantageously displays no adverse reactions, whereas Isis 395165 elicits a mild inflammatory response in liver, spleen, and lymph nodes. In summary, Isis 405879, Isis 395186, and Isis 405988 display superior tolerance, compared to the other antisense compounds tested in this study.
17.7. Differences in Achieving Pharmacodynamic Endpoints in the Cyno Model
Isis 395186 (“844419”), Isis 405879 (“844421”), and Isis 405891 (“844423”) were selected for further study in the cyno model, using the procedures described in Example 16.6. As set forth above, these antisense compounds displayed high activity in cell culture models, were well tolerated, and displayed advantageous tissue distribution and half-life properties. Further, none of these antisense compounds targeted a PCSK9 sequence known to have differences in monkeys and humans. In addition, the target sequences in human PCSK9 were known not to contain SNPs having a wide distribution in the population.
The antisense compounds were tested for their ability to reduce PCSK9 mRNA in cyno liver. See Example 16.6. The results of this experiment are shown in FIG. 10. Despite variability between the tested animals, Isis 405879 (labeled “844421”) displayed a superior ability to reduce PCSK9 mRNA levels significantly. When administered at a low dose (15 mg/kg, shown in the left side bar), Isis 405879 significantly (p<0.023) reduced PCSK9 mRNA levels. When administered at a high dose (30 mg/kg, shown in the right side bar), Isis 405879 also significantly (p<0.011) inhibited PCSK9 mRNA expression. The results obtained with Isis 405879 were superior to those obtained with Isis 395186 (labeled “844419”) and Isis 405891 (labeled “844423”). As shown in FIG. 11, Northern analysis of liver PCSK9 mRNA confirms that 15 mg/kg and 30 mg/kg Isis 405879 substantially reduces PCSK9 mRNA levels.
As described in Example 17.1, the reduction of PCSK9 mRNA is expected to correlate with a reduction in plasma LDL-C and liver TG. Consistent with this expectation, Isis 405879 produced a significant (p<0.06) decrease in cyno serum LDL-C after five weeks of administration at 30 mg/kg, as shown in FIG. 12. Isis 405879 additionally produced a significant (p<0.05) increase in the ratio of HDL to LDL following five weeks of administration at 30 mg/kg, as shown in FIG. 13.
Generally confirming earlier data obtained in rodents, Isis 405879 elicited no differences in serum chemistry, hematology, or organ weights, except for a slight increase in spleen weight. Histology revealed indicia of only mild inflammation. The advantageous distribution of Isis 405879 in liver and kidney in the cyno model is described above and shown in FIG. 9.
Example 18
Dose Response Study of Isis 405879 (“844421”) in a Human PCSK9 Transgenic Mouse Model
BMS-844421 is a human specific PCSK9 antisense oligonucleotide (ASO) designed to downregulate hepatic PCSK9 expression, increase LDL receptor activity, and promote clearance of LDL and reduction of plasma LDL-C levels in humans. A human genomic PCSK9 transgenic mouse model was recently established. This mouse line expresses hemizygous transgene in the presence of endogenous mouse PCSK9 (i.e., both genes are simultaneously expressed). It was found that the transgenic PCSK9 mice responded to chronic dosing with BMS-844421 and related human PCSK9 ASOs with decreased liver mRNA expression and decreased circulating human PCSK9 protein levels.
The present study examined the dose response to BMS-844421 in the hemizygous genomic transgenic mouse model on a normal chow diet. Experiments were conducted comparing the effects of BMS-844421 (human specific PCSK9 ASO) to those of ISIS 394816 (murine specific PCSK9 ASO) in this model alone and in combination. BMS-844421 exhibited dose-dependent suppression of liver target mRNA, reflected in dose-dependent decreases in plasma human PCSK9 levels. Both liver mRNA and plasma protein gave ED50 values of approximately 15 mg/kg (given twice weekly) for BMS-844421, while the suppression of plasma PCSK9 protein was greater than liver PCSK9 mRNA at each dose in this model. Trends for increased LDLR protein in liver total membrane fraction, and a feedback effect on liver HMGCoA reductase mRNA expression were observed, but no changes in plasma lipids were seen for BMS-844421 treated hemizygous mice. The combination of ISIS 394816+BMS-844421 (each at 15 mg/kg) elicted a greater increase in liver LDLR protein than either ASO alone, consistent with the proposed mechanism of action and the dual expression of both human and murine PCSK9 in this model.
18.1. Materials and Methods
The sequences of BMS-844421 and other ASOs tested in this report are given in Table 35. The negative control ISIS 141923 is not complementary to any known gene. The design of the dose response study is described in the beginning of the Results section (section 18.2. below).







TABLE 35







Nucleotide sequences of ASOs











SEQ




ID


ASO
Sequence
NO





BMS 844421, 
5′- CCTTGGCCACGCCGGCATCC -3′
248


human PCSK9 




specific*




 


ISIS 141923, 
5′- CCTTCCCTGAAGGTTCCTCC -3′
465


non-targeting




 


ISIS 394816, 
5′ -GGGCTCATAGCACATTATCC-3′
466


murine PCSK9 




specific





* BMS-844421 is identical to ISIS 405879






Parameters evaluated at study termination included hepatic mRNA, hepatic LDLR protein levels, plasma concentrations of total cholesterol, LDL, HDL, triglycerides, transaminases, and organ and body weights. Whole liver was processed for RNA, and protein, and held for possible histological examination and hepatic triglyceride levels. Blood and liver samples were obtained 24 hr after the prior dose. Blood samples were obtained from tail vein or retro orbital plexus and EDTA, chilled on ice, and plasma samples were obtained by standard procedures. For mRNA assays, liver samples (30-50 mg each) were placed in eppendorf tubes containing 1000 uL of TRIzol (Invitrogen #15596-018) and homogenized using stainless steel beads on a Tissue Lyser instrument (Qiagen, Valencia, Calif.). Chloroform (200 μL) was added to each tube, the contents were mixed vigorously by hand and tubes were centrifuged at 12,000×g for 20 minutes at 4° C. About 300 μL of each supernatant was removed and placed in 1.5 mL microcentrifuge tubes. One volume of 70% EtOH was added, the mixtures were vortexed thoroughly and transferred to RNeasy Mini Spin Columns (QIAgen Cat. No. 74106). RNA was purified, treated with DNase and eluted into microcentrifuge tubes following QIAgen's manufacturer's instructions.
Plasma human PCSK9 levels were assayed using the human specific ELISA with mAb 4H5 as capture antibody.
18.2. Results
A dose-response study was run in human PCSK9 genomic transgenic mouse line 66 mice on a normal chow diet. Groups of 6 mice (3 males, 3 females) received the following treatments for 6 weeks. BMS-844421 was administered p.o. at 5, 10, 15, and 30 mg/kg per dose given twice weekly (on Tuesdays and Fridays) for 6 weeks. Controls received saline, and an additional group received the non-targeting negative control ASO ISIS 141923 (30 mg/kg twice weekly). The murine PCSK9 ASO ISIS 394816 was administered to another group at 15 mg/kg per dose. The final group received BMS-844421 (15 mg/kg) plus ISIS 394816 (15 mg/kg) combined.
Liver mRNA assayed by qRT-PCR showed that BMS-844421 decreased liver PCSK9 mRNA in a dose dependent manner, with greatest effect at the top dose studied (FIG. 14). The negative control ASO, ISIS 141923, had no significant effect on target mRNA, while the murine targeted ASO, ISIS 394816, decreased human PCSK9 mRNA by 33%, suggesting partial cross-hybridization to human PCSK9 mRNA.
The ASO effects on plasma human PCSK9 followed a similar pattern as mRNA, while the effects of the higher doses of BMS-844421 resulted in greater effects, up to 70% suppression at 30 mg/kg/dose (FIG. 15). ISIS 394816 suppressed approximately 30% (similar to mRNA effects), while ISIS 141923 had no significant effect.
Plasma human PCSK9 levels exhibited time dependent suppression with effects at 6 weeks, greater than at 3 wks for all doses of BMS-844421 (FIG. 16). At 3 wks dosing, only the highest dose of BMS-844421 showed significant suppression. As seen in FIG. 17, data for 6 wks treatment with BMS-844421 showed a sigmoidal dose-response curve, with ED50 approximately 15 mg/kg/dose (given twice weekly). Plotting the mRNA and protein data together showed that the two endpoints generally, though imperfectly, reflect each other, with protein showing greater sensitivity to suppression (FIG. 18).
Male transgenic mice responded more sensitively to BMS-844421 than the female transgenic mice in this study (FIG. 19). The data was not the result of different plasma baseline levels of human PCSK9 between males and females, which is human PCSK9=67+/−31 ng/mL in males and 60+/−27 ng/mL in females in the predosing samples. The difference in PCSK9 protein response for males vs. females was less apparent for liver target mRNA (FIG. 20).
Levels of endogenous mouse PCSK9 mRNA in liver were decreased by ISIS 394816 (and the combination of ISIS 394816 with BMS-844421), but were not decreased by BMS-844421 only (FIG. 21).
A key cholesterol regulatory gene, HMGCoA reductase (HMGR), is known to respond to sterol feedback in the liver. An assay of endogenous mouse liver HMGR mRNA revealed 45% suppression of HMGR with BMS-844421 as well as ISIS 394816 (FIG. 22). These findings are consistent with downregulation of PCSK9 promoting higher levels of LDLR, in turn leading to increased cholesterol in the regulatory pool in liver cells, resulting in downregulation of HMGR. Similar, though quantitatively lesser effects, were also seen for the sterol pathway genes LDLR mRNA and for SREBP2 mRNA.
LDLR protein levels were assayed in liver samples from the same study. Total hepatic membranes were prepared, and western blots of SDS-PAGE gels were assayed with anti-LDLR antibodies. As a control, transferrin receptor (TR) protein levels were measured with anti-TR antibodies. Liver samples from the ASO study were pooled from n=3 for the preparation of the membrane fraction. Pairs of pools for each group were assayed. Treatment with BMS-844421, and especially the combination of BMS-844421 with the murine ASO ISIS 394816, led to increases up to >2-fold in levels of LDLR protein, compared to the level of TR protein (FIG. 23 and FIG. 24).
The ELISA assay uses a monoclonal capture antibody and a polyclonal detector antibody developed against human recombinant PCSK9 produced in baculovirus. The recombinant human PCSK9 protein also serves as standard antigen for the ELISA assay. Cynomolgus monkey plasma PCSK9 was readily detectable using this assay. Human PCSK9 protein in plasma of human PCSK9 transgenic mouse lines was specifically detected with no cross-reactivity to murine PCSK9.
Despite these pharmacodynamic responses to BMS-844421, hemizygous transgenic mice did not exhibit changes in plasma LDL-C or total cholesterol following ASO treatment, including BMS-844421 (Table 36). However, normal chow fed mice have low levels of LDL-C (Table 36). It is plausible that these low LDL-C levels are not readily decreased by increases in LDLR protein. Another possible explanation is that the hemizygous expression of human PCSK9, i.e., the gene specifically targeted by BMS-844421, contributes only about 1⁄3 to 1⁄2 of the total murine+human PCSK9 in this hemizygous transgenic mouse model.







TABLE 36







Plasma lipids at 6 wk treatment with ASOs in transgenic mice













Total




Both Sexes
Triglycerides
Cholesterol
HDL
LDL





Saline
72 ± 14 
108 ± 8.2
57 ± 6.4
14 + 1


Missense
64 ± 9.2
110 ± 8.6
57 ± 7.1
15 ± 1


BMS-844421 (30 mg/kg) 
54 ± 8.6
129 ± 7.3
67 ± 4.6
18 ± 2


BMS-844421 (15 mg/kg)
54 ± 9.0
126 ± 9.2
65 ± 5.5
18 ± 1


BMS-844421 (10 mg/kg) 
53 ± 9.4
118 ± 9.3
61 ± 6.9
17 ± 2


BMS-844421 (5 mg/kg)
56 ± 9.7
117 ± 8.5
61 ± 6.1
17 ± 1


Mouse ASO
50 ± 7.0
84.2 ± 2.9 
45 ± 3.0
11 +2


I-394816 (15 mg/kg)






BMS-844421 + I-394816
55 ± 9.3
 92 ± 5.3
48 ± 4.6
12 ± 2


(15 mg/kg each)






WT Littermates
60 ± 8.1
111.7 ± 7.2  
57 ± 5.3
15 ± 1









There were no changes in ALT or AST (not shown), suggesting that no liver abnormalities were present after 6 wks treatment.

18.3. Conclusions
The recently established human genomic PCSK9 transgenic mouse model responded to human PCSK9 ASO treatment with robust downregulation of human PCSK9 at both the mRNA and protein levels in liver. Plasma human PCSK9 was also decreased with the magnitude of decrease somewhat greater than liver mRNA suppression, suggesting that plasma PCSK9 levels are a good readout for suppression of the target mRNA. The ED50 for both endpoints was approximately 15 mg/kg (per dose given twice weekly). The transgenic mice did not exhibit significant changes in plasma LDL-C or total cholesterol following BMS-844421 or ISIS 394816 treatment, nor for the combination of the two ASOs. A trend for increasing liver LDLR protein was observed following treatment with BMS-844421, suggesting a functional effect consistent with the downregulation of PCSK9 mRNA and protein. Consistent with this was the downregulation of the sterol response gene HMGR in liver.
The limited pharmacodynamic responses in this model appear to be related both to the small LDL window in normal chow diet fed mice, and to the co-expression of the human PCSK9 gene with the endogenous murine PCSK9 gene. In the hemizygous mice, human PCSK9 appears to represent only about 1⁄3 of the total human+murine PCSK9 expressed. Consistent with this, in a previous study the murine specific PCSK9 ASO ISIS 394816 administered to human apoB-CETP double transgenic mice produced up to 90% suppression of liver PCSK9 mRNA and a corresponding decrease of up to 80% in plasma LDL-C levels.

## Claims

1. A method of reducing PCSK9 expression in an animal comprising administering to the animal a compound comprising a modified oligonucleotide 10 to 30 linked nucleosides in length targeted to region 3472-3562 of SEQ ID NO: 1, wherein the compound reduces PCSK9 expression by at least 30% in the animal.

2. The method of claim 1, wherein PCSK9 expression is reduced by at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%.

3. The method of claim 1, wherein reducing PCSK9 expression in the animal treats hypercholesterolemia, polygenic hypercholesterolemia, mixed dyslipidemia, coronary heart disease, acute coronary syndrome, early onset coronary heart disease, type II diabetes, type II diabetes with dyslipidemia, hepatic steatosis, non-alcoholic steatohepatitis, non-alcoholic fatty liver disease, hypertriglyceridemia, hyperfattyacidemia, hyperlipidemia, metabolic syndrome, elevated ApoB, elevated cholesterol, elevated LDL-cholesterol, elevated VLDL-cholesterol, stroke or elevated non-HDL cholesterol in the animal.

4. The method of claim 1, wherein reducing PCSK9 expression in the animal reduces ApoB, LDL-cholesterol, VLDL-cholesterol, Lp(a), small LDL-particle, small VLDL-particle, non-HDL-cholesterol, liver triglyceride level, serum triglycerides, serum phospholipids, or any combination thereof, in the animal.

5. The method of claim 4, wherein at least one of the ApoB reduction, LDL-cholesterol reduction, VLDL-cholesterol reduction, Lp(a) reduction, small LDL-particle reduction, small VLDL-particle reduction, non-HDL-cholesterol reduction, liver triglyceride level reduction, serum triglycerides reduction, or serum phospholipids reduction is at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or 100%.

6. The method of claim 1, wherein the modified oligonucleotide has a nucleobase sequence at least 80%, at least 85%, at least 90%, at least 95%, or 100% complementary to SEQ ID NO: 1 as measured over the entirety of said modified oligonucleotide.

7. The method of claim 1, wherein the animal is a human.

8. The method of claim 1, wherein at least one internucleoside linkage of said modified oligonucleotide is a modified internucleoside linkage.

9. The method of claim 8, wherein at least one internucleoside linkage is a phosphorothioate internucleoside linkage.

10. The method of claim 8, wherein each internucleoside linkage is a phosphorothioate internucleoside linkage.

11. The method of claim 1, wherein at least one nucleoside of said modified oligonucleotide comprises a modified sugar.

12. The method of claim 11, wherein at least one modified sugar is a bicyclic sugar.

13. The method of claim 11, wherein at least one modified sugar comprises a 2′-O-methoxyethyl, a (4′-CH(CH3)—O-2′) BNA, a (4′ CH2-O-2′) BNA or a 4′ (CH2)n-O-2′ bridge, wherein n is 1 or 2.

14. The method of claim 1, wherein at least one nucleoside of said modified oligonucleotide comprises a modified nucleobase.

15. The method of claim 14, wherein the modified nucleobase is a 5-methylcytosine.

16. The method of claim 1, wherein the modified oligonucleotide consists of 19 to 22 linked nucleosides.

17. The method of claim 1, wherein the compound is co-administered with one or more other pharmaceutical agents.

18. The method of claim 1, wherein the modified oligonucleotide is conjugated.

19. The method of claim 1, wherein the modified oligonucleotide comprises an oligonucleotide consisting of 12 to 30 linked nucleosides, wherein the nucleobase sequence of the oligonucleotide is at least 90% complementary to an equal length portion of region 3472-3562 of SEQ ID NO: 1.

20. The method of claim 1, wherein the compound is single-stranded.