Provided herein are compounds, and pharmaceutically acceptable salts thereof, useful as KRAS G12D and/or KRAS G12C inhibitors, methods of making and using the same (singly or in combination with additional agents), and pharmaceutical compositions thereof.

BACKGROUND

The KRAS protein, Kirsten Rat Sarcoma 2 Viral Oncogene Homolog (“KRAS”), is a GTPase. KRAS gene mutations have been observed in a number of conditions including, for instance, pancreatic cancer, endometrial cancer, lung adenocarcinoma, colorectal cancer, rectal carcinoma, gall bladder cancer, thyroid cancer, bile duct cancer, small cell lung cancer, and non-small cell lung cancer (NSCLC). Accordingly, there is a need for compounds, pharmaceutical compositions, and methods for inhibiting KRAS (e.g., KRAS G12C and/or KRAS G12D) and treating associated cancers.

SUMMARY

In one embodiment, the present disclosure provides a compound of Formula I-A:

In one embodiment, the present disclosure provides a compound of Formula II:

In another embodiment, the present disclosure provides a pharmaceutical composition comprising a compound of the present disclosure, and a pharmaceutically acceptable excipient.

In another embodiment, the present disclosure provides a method of inhibiting KRAS G12D protein in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

In another embodiment, the present disclosure provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of the present disclosure.

In another embodiment, the present disclosure provides a method for manufacturing a medicament for treating cancer in a subject in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used.

In another embodiment, the present disclosure provides a method for manufacturing a medicament for inhibiting cancer metastasis in a subject in need thereof, characterized in that a compound of the present invention, or a pharmaceutically acceptable salt thereof, is used.

In another embodiment, the present disclosure provides use of the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer in a subject.

In another embodiment, the present disclosure provides use of the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for inhibiting cancer metastasis in a subject.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer in a subject in need thereof.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in inhibiting cancer metastasis in a subject in need thereof.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in therapy.

DETAILED DESCRIPTION

The disclosure relates generally to methods and compounds, and pharmaceutically acceptable salts thereof, for inhibiting KRASG12Dand/or KRASG12C. The following description sets forth exemplary methods, parameters and the like. It should be recognized, however, that such description is not intended as a limitation on the scope of the present disclosure but is instead provided as a description of exemplary embodiments.

As used in the present specification, the following words, phrases and symbols are generally intended to have the meanings as set forth below, except to the extent that the context in which they are used indicates otherwise.

A dash (“-”) that is not between two letters or symbols is used to indicate a point of attachment for a substituent. For example, —CONH2is attached through the carbon atom. A dash at the front or end of a chemical group is a matter of convenience; chemical groups can be depicted with or without one or more dashes without losing their ordinary meaning. A wavy line drawn through a line in a structure indicates a point of attachment of a group. Unless chemically or structurally required, no directionality is indicated or implied by the order in which a chemical group is written or named.

A squiggly line on a chemical group as shown below, for example,

indicates a point of attachment, i.e., it shows the broken bond by which the group is connected to another described group.

As used herein, “a compound of the disclosure” can mean a compound of any of the Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), and (IIb-2), or a pharmaceutically acceptable salt thereof. Similarly, the phrase “a compound of Formula (number)” means a compound of that formula and pharmaceutically acceptable salts thereof.

The prefix “Cu-Cv” indicates that the following group has from u to v carbon atoms. For example, “C1-C8alkyl” indicates that the alkyl group has from 1 to 8 carbon atoms.

“Alkynyl” refers to an unbranched or branched hydrocarbon chain containing at least one carbon-carbon triple bond. For example, an alkynyl group can have from 2 to 20 carbon atoms (i.e., C2-20alkynyl), 2 to 8 carbon atoms (i.e., C2-8alkynyl), 2 to 6 carbon atoms (i.e., C2-6alkynyl), or 2 to 4 carbon atoms (i.e., C2-4alkynyl). The term “alkynyl” also includes those groups having one triple bond and one double bond. Examples of C2-6alkynyl include, but are not limited to, ethynyl, prop-1-ynyl, but-1-ynyl, pent-1-ynyl, pent-4-ynyl and penta-1,4-diynyl.

“Alkoxy” means a group having the formula —O-alkyl, in which an alkyl group, as defined above, is attached to the parent molecule via an oxygen atom. The alkyl portion of an alkoxy group can have 1 to 20 carbon atoms (i.e., C1-C20alkoxy), 1 to 12 carbon atoms (i.e., C1-C12alkoxy), 1 to 8 carbon atoms (i.e., C1-C8alkoxy), 1 to 6 carbon atoms (i.e., C1-C6alkoxy) or 1 to 3 carbon atoms (i.e., C1-C3alkoxy). Examples of suitable alkoxy groups include, but are not limited to, methoxy (—O—CH3or —OMe), ethoxy (—OCH2CH3or -OEt), isopropoxy (—O—CH(CH3)2), t-butoxy (—O—C(CH3)3or -OtBu) and the like. Other examples of suitable alkoxy groups include, but are not limited to, sec-butoxy, tert-butoxy, pentoxy, hexoxy, and the like.

“Alkoxyalkyl” refers an alkoxy group linked to an alkyl group which is linked to the remainder of the compound. Alkoxyalkyl can have any suitable number of carbon, such as from 2 to 6 (C2-6alkoxyalkyl), 2 to 5 (C2-5alkoxyalkyl), 2 to 4 (C2-4alkoxyalkyl), or 2 to 3 (C2-3alkoxyalkyl). Alkoxy and alkyl are as defined above. Examples of “alkoxyalkyl” include, but are not limited to, methoxymethyl (CH3OCH2—), and methoxyethyl (CH3OCH2CH2).

“Bridged” means a ring system in which non-adjacent atoms on a ring are connected by a divalent substituent, such as an alkylenyl or heteroalkylenyl group or a single heteroatom.

“Hydroxyalkyl” refers to a hydroxy group, —OH, linked to an alkyl group which is linked to the remainder of the compound such that the alkyl group is divalent. Hydroxyalkyl can have any suitable number of carbons, such as from 1 to 8 (C1-8hydroxyalkyl), 1 to 6 (C1-6hydroxyalkyl), 2 to 6 (C2-6hydroxyalkyl), 2 to 4 (C2-4hydroxyalkyl), or 2 to 3 (C2-3hydroxyalkyl). Alkyl is as defined above where the alkyl is divalent.

“Halo” or “halogen” as used herein refers to fluoro (—F), chloro (—Cl), bromo (—Br) and iodo (—I).

“Haloalkyl” is an alkyl group, as defined above, in which one or more hydrogen atoms of the alkyl group is replaced with a halogen atom. The alkyl portion of a haloalkyl group can have 1 to 20 carbon atoms (i.e., C1-C20haloalkyl), 1 to 12 carbon atoms (i.e., C1-C12haloalkyl), 1 to 8 carbon atoms (i.e., C1-C8haloalkyl), 1 to 6 carbon atoms (i.e., C1-C6alkyl) or 1 to 3 carbon atoms (i.e., C1-C3alkyl). The alkyl groups can be substituted with 1, 2, 3, 4, 5, 6, 7 8 9 or more halogens. Examples of suitable haloalkyl groups include, but are not limited to, —CF3, —CHF2, —CFH2, —CH2CF3, fluorochloromethyl, difluorochloromethyl, 1,1,1-trifluoroethyl and pentafluoroethyl.

“Haloalkoxy” refers to an alkoxy group where some or all of the hydrogen atoms are substituted with halogen atoms. As for an alkyl group, haloalkoxy groups can have any suitable number of carbon atoms, such as C1-6. The alkoxy groups can be substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9 or more halogens. When all the hydrogens are replaced with a halogen, for example by fluorine, the compounds are per-substituted, for example, perfluorinated. Haloalkoxy includes, but is not limited to, trifluoromethoxy, 2,2,2,-trifluoroethoxy, perfluoroethoxy, etc.

“Thioalkyl” refers to a thio group, —SH, linked to an alkyl group which is linked to the remainder of the compound such that the alkyl group is divalent. Thioalkyl can have any suitable number of carbons, such as from 1 to 8 (C1-8thioalkyl), 1 to 6 (C1-6thioalkyl), 2 to 6 (C2-6 thioalkyl), 2 to 4 (C2-4thioalkyl), or 2 to 3 (C2-3thioalkyl). Alkyl is as defined above where the alkyl is divalent.

“Haloalkylthio” is an alkylthio group, as defined above, in which one or more hydrogen atoms of the alkyl group is replaced with a halogen atom. The alkyl portion of a haloalkylthio group can have 1 to 20 carbon atoms (i.e., C1-C20haloalkylthio), 1 to 12 carbon atoms (i.e., C1-C12haloalkylthio), 1 to 8 carbon atoms (i.e., C1-C8haloalkylthio), 1 to 6 carbon atoms (i.e., C1-C6alkylthio) or 1 to 3 carbon atoms (i.e., C1-C3alkylthio). The alkylthio groups can be substituted with 1, 2, 3, 4, 5, 6, 7, 8, 9 or more halogens.

“Heteroalkyl” refers to an unbranched or branched saturated hydrocarbon chain containing from 1 to 4 heteroatoms.

“Cyanoalkyl” refers to a cyano group, —CN, linked to an alkyl group which is linked to the remainder of the compound such that the alkyl group is divalent. Cyanoalkyl can have any suitable number of carbons, such as from 1 to 8 (C1-8cyanoalkyl), 1 to 6 (C1-6cyanoalkyl), 2 to 6 (C2-6 cyanoalkyl), 2 to 4 (C2-4cyanoalkyl), or 2 to 3 (C2-3cyanoalkyl). Alkyl is as defined above where the alkyl is divalent.

“Cycloalkyl” refers to a saturated or partially saturated cyclic alkyl group having a single ring or multiple rings, such as 2, 3, 4 or more, wherein the multiple rings can be fused, bridged, spiro, or any combination thereof. As used herein, cycloalkyl has from 3 to 20 ring carbon atoms (i.e., C3-20cycloalkyl), 3 to 12 ring carbon atoms (i.e., C3-12cycloalkyl), 3 to 10 ring carbon atoms (i.e., C3-10cycloalkyl), 3 to 8 ring carbon atoms (i.e., C3-8cycloalkyl), or 3 to 6 ring carbon atoms (i.e., C3-6cycloalkyl). Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Cycloalkyl groups also include partially unsaturated ring systems containing one or more double bonds, including fused ring systems with one aromatic ring and one non-aromatic ring, but not fully aromatic ring systems.

The term “fused” refers to a ring system in which two or more rings in the system share a pair of adjacent ring atoms.

“Spiro” refers to at least two rings are linked together by one common atom. “Spiro” also refers to a ring substituent which is joined by two bonds at the same carbon atom. Examples of spiro groups include, but are not limited to, 1,1-diethylcyclopentane, dimethyl-dioxolane, and 4-benzyl-4-methylpiperidine, wherein the cyclopentane and piperidine, respectively, are the spiro substituents.

“Alkyl-cycloalkyl” refers to a radical having an alkyl component and a cycloalkyl component, where the alkyl component links the cycloalkyl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, an alkylene, to link to the cycloalkyl component and to the point of attachment. In some instances, the alkyl component can be absent. The alkyl component can include any number of carbons, such as C1-6, C1-2, C1-3, C1-4, C1-5, C2-3, C2-4, C2-5, C2-6, C3-4, C3-5, C3-6, C4-5, C4-6and C5-6. The cycloalkyl component is as defined within. Exemplary alkyl-cycloalkyl groups include, but are not limited to, methyl-cyclopropyl, methyl-cyclobutyl, methyl-cyclopentyl and methyl-cyclohexyl.

“Heterocycle” or “heterocyclyl” or “heterocycloalkyl” refer to a saturated or unsaturated cyclic alkyl group, with one or more ring heteroatoms independently selected from nitrogen, oxygen, sulfur and silicon. A heterocyclyl can be a single ring or multiple rings, such as 2, 3, 4 or more, wherein the multiple rings can be fused, bridged, spiro, or any combination thereof. As used herein, heterocyclyl has 3 to 20 ring atoms (i.e., 3 to 20 membered heterocyclyl), 3 to 12 ring atoms (i.e., 3 to 12 membered heterocyclyl), 3 to 10 ring atoms (i.e., 3 to 10 membered heterocyclyl), 3 to 8 ring atoms (i.e., 3 to 8 membered heterocyclyl), 4 to 12 ring carbon atoms (i.e., 4 to 12 membered heterocyclyl), 4 to 8 ring atoms (i.e., 4 to 8 membered heterocyclyl), or 4 to 6 ring atoms (i.e., 4 to 6 membered heterocyclyl). Examples of heterocyclyl groups include pyrrolidinyl, piperidinyl, piperazinyl, oxetanyl, dioxolanyl, azetidinyl, and morpholinyl.

“Alkyl-heterocycloalkyl” refers to a radical having an alkyl component and a heterocycloalkyl component, where the alkyl component links the heterocycloalkyl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, an alkylene, to link to the heterocycloalkyl component and to the point of attachment. The alkyl component can include any number of carbons, such as C0-6, C1-2, C1-3, C1-4, C1-5, C1-6, C2-3, C2-4, C2-5, C2-6, C3-4, C3-5, C3-6, C4-5, C4-6and C5-6. In some instances, the alkyl component can be absent. The heterocycloalkyl component is as defined above.

“Aryl” means an aromatic hydrocarbon radical derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. For example, an aryl group can have 6 to 20 carbon atoms, 6 to 14 carbon atoms, or 6 to 10 carbon atoms. Exemplary aryl groups include, but are not limited to, radicals derived from benzene (e.g., phenyl), naphthalene, anthracene, biphenyl, and the like.

“Alkyl-aryl” refers to a radical having an alkyl component and an aryl component, where the alkyl component links the aryl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, an alkylene, to link to the aryl component and to the point of attachment. The alkyl component can include any number of carbons, such as C0-6, C1-2, C1-3, C1-4, C1-5, C1-6, C2-3, C2-4, C2-5, C2-6, C3-4, C3-5, C3-6, C4-5, C4-6and C5-6. In some instances, the alkyl component can be absent. The aryl component is as defined above. Examples of alkyl-aryl groups include, but are not limited to, benzyl and ethyl-benzene.

“Heteroaryl” refers to an aromatic group, including groups having an aromatic tautomer or resonance structure, having a single ring, multiple rings, or multiple fused rings, with at least one heteroatom in the ring, i.e., one or more ring heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the nitrogen or sulfur can be oxidized. Thus, the term includes rings having one or more annular O, N, S, S(O), S(O)2, and N-oxide groups. The term includes rings having one or more annular C(O) groups. As used herein, heteroaryl include 5 to 20 ring atoms (i.e., 5- to 20-membered heteroaryl), 5 to 12 ring atoms (i.e., 5- to 12-membered heteroaryl), or 5 to 10 ring atoms (i.e., 5- to 10-membered heteroaryl), and 1 to 5 heteroatoms independently selected from nitrogen, oxygen, and sulfur, and oxidized forms of the heteroatoms. Examples of heteroaryl groups include, but are not limited to, pyridin-2(1H)-one, pyridazin-3(2H)-one, pyrimidin-4(3H)-one, quinolin-2(1H)-one, pyrimidinyl, purinyl, pyridyl, pyridazinyl, benzothiazolyl, and pyrazolyl. Heteroaryl does not encompass or overlap with aryl as defined above.

“Alkyl-heteroaryl” refers to a radical having an alkyl component and a heteroaryl component, where the alkyl component links the heteroaryl component to the point of attachment. The alkyl component is as defined above, except that the alkyl component is at least divalent, an alkylene, to link to the heteroaryl component and to the point of attachment. The alkyl component can include any number of carbons, such as C0-6, C1-2, C1-3, C1-4, C1-5, C1-6, C2-3, C2-4, C2-5, C2-6, C3-4, C3-5, C3-6, C4-5, C4-6 and C5-6. In some instances, the alkyl component can be absent. The heteroaryl component is as defined within.

“KRAS G12D” refers to the G12D mutation of the KRAS protein, where aspartic acid replaces glycine at amino acid position 12.

“KRAS G12D-associated disease or disorder” refers to diseases or disorders associated with or mediated by or having a KRAS G12D mutation. Representative diseases or disorders include, but are not limited to, KRAS G12D-associated cancer.

“Oxo” refers to the group (═O) or (O).

Provided are also pharmaceutically acceptable salts, hydrates, solvates, tautomeric forms, polymorphs, and prodrugs of the compounds described herein. “Pharmaceutically acceptable” or “physiologically acceptable” refer to compounds, salts, formulations, dosage forms and other materials which are useful in preparing a pharmaceutical composition that is suitable for veterinary or human pharmaceutical use.

Examples of “pharmaceutically acceptable salts” of the compounds disclosed herein also include salts derived from an appropriate base, such as an alkali metal (for example, sodium, potassium), an alkaline earth metal (for example, magnesium), ammonium and NX4+(wherein X is C1-C4alkyl). Also included are base addition salts, such as sodium or potassium salts.

Provided are also compounds described herein or pharmaceutically acceptable salts, isomers, or a mixture thereof, in which from 1 to n hydrogen atoms attached to a carbon atom can be replaced by a deuterium atom or D, in which n is the number of hydrogen atoms in the molecule. As known in the art, the deuterium atom is a non-radioactive isotope of the hydrogen atom. Such compounds can increase resistance to metabolism, and thus can be useful for increasing the half-life of the compounds described herein or pharmaceutically acceptable salts, isomer, or a mixture thereof when administered to a mammal. See, e.g., Foster, “Deuterium Isotope Effects in Studies of Drug Metabolism,” TRENDSPHARMACOL. SCI.,5(12):524-527 (1984). Such compounds are synthesized by means well known in the art, for example by employing starting materials in which one or more hydrogen atoms have been replaced by deuterium.

Examples of isotopes that can be incorporated into the disclosed compounds also include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, fluorine, chlorine, and iodine, such as2H,3H,11C,13C,14C,13N,15N,15O,17O,18O,31P,32P,35S,18F,36Cl,123I, and125I, respectively. Substitution with positron emitting isotopes, such as11C,18F,15O and13N, can be useful in Positron Emission Topography (PET) studies for examining substrate receptor occupancy. Isotopically-labeled compounds of Formulas I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), and (IIb-2), can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the Examples as set out below using an appropriate isotopically-labeled reagent in place of the non-labeled reagent previously employed.

“Racemates” refers to a mixture of enantiomers. The mixture can comprise equal or unequal amounts of each enantiomer.

“Stereoisomer” and “stereoisomers” refer to compounds that differ in the chirality of one or more stereocenters. Stereoisomers include enantiomers and diastereomers. The compounds can exist in stereoisomeric form if they possess one or more asymmetric centers or a double bond with asymmetric substitution and, therefore, can be produced as individual stereoisomers or as mixtures. Unless otherwise indicated, the description is intended to include individual stereoisomers as well as mixtures. The methods for the determination of stereochemistry and the separation of stereoisomers are well-known in the art (see, e.g., Chapter 4 of ADVANCEDORGANICCHEMISTRY,4th ed., J. March, John Wiley & Sons, New York, 1992).

A “subject” or “patient” is meant to describe a human or vertebrate animal including a dog, cat, pocket pet, marmoset, horse, cow, pig, sheep, goat, elephant, giraffe, chicken, lion, monkey, owl, rat, squirrel, slender loris, and mouse. A “pocket pet” refers to a group of vertebrate animals capable of fitting into a commodious coat pocket such as, for example, hamsters, chinchillas, ferrets, rats, guinea pigs, gerbils, rabbits and sugar gliders.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. A dash at the front or end of a chemical group is a matter of convenience; chemical groups can be depicted with or without one or more dashes without losing their ordinary meaning. A wavy line drawn through a line in a structure indicates a point of attachment of a group. A dashed line indicates an optional bond. Unless chemically or structurally required, no directionality is indicated or implied by the order in which a chemical group is written or the point at which it is attached to the remainder of the molecule. For instance, the group “—SO2CH2—” is equivalent to “—CH2SO2—” and both can be connected in either direction. Similarly, an “arylalkyl” group, for example, can be attached to the remainder of the molecule at either an aryl or an alkyl portion of the group. A prefix such as “Cu-Cv” or “(Cu-Cv)” indicates that the following group has from u to v carbon atoms. For example, “C1-6alkyl” and “C1-C6alkyl” both indicate that the alkyl group has from 1 to 6 carbon atoms.

Unless otherwise specified, the carbon atoms of the compounds of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2), are intended to have a valence of four. If in some chemical structure representations, carbon atoms do not have a sufficient number of variables attached to produce a valence of four, the remaining carbon substituents needed to provide a valence of four should be assumed to be hydrogen.

“Treatment” or “treating” is an approach for obtaining beneficial or desired results including clinical results. Beneficial or desired clinical results may include one or more of the following: (a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and/or diminishing the extent of the disease or condition); (b) slowing or arresting the development of one or more clinical symptoms associated with the disease or condition (e.g., stabilizing the disease or condition, preventing or delaying the worsening or progression of the disease or condition, and/or preventing or delaying the spread (e.g., metastasis) of the disease or condition); and/or (c) relieving the disease, that is, causing the regression of clinical symptoms (e.g., ameliorating the disease state, providing partial or total remission of the disease or condition, enhancing effect of another medication, delaying the progression of the disease, increasing the quality of life, and/or prolonging survival.

The term “therapeutically effective amount,” as used herein, is the amount of compound disclosed herein present in a formulation described herein that is needed to provide a desired level of drug in the secretions and tissues of the airways and lungs, or alternatively, in the bloodstream of a subject to be treated to give an anticipated physiological response or desired biological effect when such a formulation is administered by the chosen route of administration. The precise amount will depend upon numerous factors, for example the particular compound disclosed herein, the specific activity of the formulation, the delivery device employed, the physical characteristics of the formulation, its intended use, as well as subject considerations such as severity of the disease state, subject cooperation, etc., and can readily be determined by one skilled in the art based upon the information provided herein. The term “therapeutically effective amount” or “effective amount” also means amounts that eliminate or reduce the subject's viral burden and/or viral reservoir.

“Administering” refers to oral administration, administration as a suppository, topical contact, parenteral, intravenous, intraperitoneal, intramuscular, intralesional, intranasal or subcutaneous administration, intrathecal administration, or the implantation of a slow-release device e.g., a mini-osmotic pump, to the subject. The administration can be carried out according to a schedule specifying frequency of administration, dose for administration, and other factors.

“Co-administration” as used herein refers to administration of unit dosages of the compounds disclosed herein before or after administration of unit dosages of one or more additional therapeutic agents, for example, administration of the compound disclosed herein within seconds, minutes, or hours of the administration of one or more additional therapeutic agents. For example, in some embodiments, a unit dose of a compound of the present disclosure is administered first, followed within seconds or minutes by administration of a unit dose of one or more additional therapeutic agents. Alternatively, in other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed by administration of a unit dose of a compound of the present disclosure within seconds or minutes. In some embodiments, a unit dose of a compound of the present disclosure is administered first, followed, after a period of hours (e.g., 1-12 hours), by administration of a unit dose of one or more additional therapeutic agents. In other embodiments, a unit dose of one or more additional therapeutic agents is administered first, followed, after a period of hours (e.g., 1-12 hours), by administration of a unit dose of a compound of the present disclosure. Co-administration of a compound disclosed herein with one or more additional therapeutic agents generally refers to simultaneous or sequential administration of a compound disclosed herein and one or more additional therapeutic agents, such that therapeutically effective amounts of each agent are present in the body of the patient.

“Subject” refers to animals such as mammals, including, but not limited to, primates (e.g., humans), cows, sheep, goats, horses, dogs, cats, rabbits, rats, mice and the like. In some embodiments, the subject is a human.

“Disease” or “condition” refer to a state of being or health status of a patient or subject capable of being treated with a compound, pharmaceutical composition, or method provided herein. The disease may be an autoimmune, inflammatory, cancer, infectious (e.g., a viral infection), metabolic, developmental, cardiovascular, liver, intestinal, endocrine, neurological, or other disease. In some embodiments, the disease is cancer (e.g. lung cancer, ovarian cancer, osteosarcoma, bladder cancer, cervical cancer, liver cancer, kidney cancer, skin cancer (e.g., Merkel cell carcinoma), testicular cancer, leukemia, lymphoma, head and neck cancer, colorectal cancer, prostate cancer, pancreatic cancer, melanoma, breast cancer, neuroblastoma).

“Melanoma” is taken to mean a tumor arising from the melanocytic system of the skin and other organs. Melanomas that may be treated with a compound, pharmaceutical composition, or method provided herein include, for example, acral-lentiginous melanoma, amelanotic melanoma, benign juvenile melanoma, Cloudman's melanoma, S91 melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo maligna melanoma, malignant melanoma, nodular melanoma, subungal melanoma, or superficial spreading melanoma.

“Metastasis,” “metastatic,” and “metastatic cancer” can be used interchangeably and refer to the spread of a proliferative disease or disorder, e.g., cancer, from one organ or another non-adjacent organ or body part. Cancer occurs at an originating site, e.g., breast, which site is referred to as a primary tumor, e.g., primary breast cancer. Some cancer cells in the primary tumor or originating site acquire the ability to penetrate and infiltrate surrounding normal tissue in the local area and/or the ability to penetrate the walls of the lymphatic system or vascular system circulating through the system to other sites and tissues in the body. A second clinically detectable tumor formed from cancer cells of a primary tumor is referred to as a metastatic or secondary tumor. When cancer cells metastasize, the metastatic tumor and its cells are presumed to be similar to those of the original tumor. Thus, if lung cancer metastasizes to the breast, the secondary tumor at the site of the breast consists of abnormal lung cells and not abnormal breast cells. The secondary tumor in the breast is referred to a metastatic lung cancer. Thus, the phrase metastatic cancer refers to a disease in which a subject has or had a primary tumor and has one or more secondary tumors. The phrases non-metastatic cancer or subjects with cancer that is not metastatic refers to diseases in which subjects have a primary tumor but not one or more secondary tumors. For example, metastatic lung cancer refers to a disease in a subject with or with a history of a primary lung tumor and with one or more secondary tumors at a second location or multiple locations, e.g., in the breast.

“Associated” or “associated with” in the context of a substance or substance activity or function associated with a disease (e.g., diabetes, cancer (e.g. prostate cancer, renal cancer, metastatic cancer, melanoma, castration-resistant prostate cancer, breast cancer, triple negative breast cancer, glioblastoma, ovarian cancer, lung cancer, squamous cell carcinoma (e.g., head, neck, or esophagus), colorectal cancer, leukemia, acute myeloid leukemia, lymphoma, B cell lymphoma, or multiple myeloma)) means that the disease (e.g. lung cancer, ovarian cancer, osteosarcoma, bladder cancer, cervical cancer, liver cancer, kidney cancer, skin cancer (e.g., Merkel cell carcinoma), testicular cancer, leukemia, lymphoma, head and neck cancer, colorectal cancer, prostate cancer, pancreatic cancer, melanoma, breast cancer, neuroblastoma) is caused by (in whole or in part), or a symptom of the disease is caused by (in whole or in part) the substance or substance activity or function.

The term “adjacent carbons” as used herein refers to consecutive carbons atoms that are directly attached to each other. For example, in

“Solvate” as used herein refers to the result of the interaction of a solvent and a compound. Solvates of salts of the compounds described herein are also provided. Hydrates of the compounds described herein are also provided.

“Prodrug” as used herein refers to a derivative of a drug that upon administration to the human body is converted to the parent drug according to some chemical or enzymatic pathway.

As used herein, “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” includes, but is not limited to, any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and combinations thereof. The use of pharmaceutically acceptable carriers and pharmaceutically acceptable excipients for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic formulations is contemplated. Supplementary active ingredients can also be incorporated into the formulations. The carrier(s) must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and physiologically innocuous to the recipient thereof.

In some embodiments, the present disclosure provides the compound of Formula I, wherein R1, R2, R3, and R4are each independently H or methyl. In some embodiments, the present disclosure provides the compound of Formula I, wherein one of two of R1, R2, R3, and R4are methyl. In some embodiments, the present disclosure provides the compound of Formula I, wherein R1and R2are methyl. In some embodiments, the present disclosure provides the compound of Formula I, wherein R1, R2, R3, and R4are each H.

In some embodiments, the present disclosure provides the compound of Formula I or I-A, or a pharmaceutically acceptable salt thereof, having the structure of Formula (I-1):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, or I-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (I-2):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, or I-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (I-3):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein X is N. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein X is CH. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis (CH2)mCN, and m is 0, 1, 2 or 3. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis CH2CN. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis halo.

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein L1is O. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ia):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein L1is CHR1b. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein L1is CH2. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein L1is CH(CH3). In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein R1aand R1bare each independently H, C1-C3alkyl, or halo. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein R1bis C1-C3alkyl or halo. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein R1bis H or methyl. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein R1bis H. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, wherein R1bis methyl.

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, or I-3, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ib):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R2aand R2bare each independently H, C1-C3alkyl, halo, or C1-C6haloalkyl. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein L2is CHR2b. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R2bis H or C1-C3alkyl. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R2bis H. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R2bis C1-C3alkyl. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R2bis methyl.

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein L3is CR3aR3b. In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, (Ia), or (Ib), or a pharmaceutically acceptable salt thereof, wherein R3aand R3bare H.

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, or (Ia), or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ia-1):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, or (Ib), or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ib-1):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, Ib, or Ib-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ib-2):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, Ib, or Ib-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ib-3):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, I-1, I-2, I-3, Ib, or Ib-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (Ib-4):

Disclosed herein are, among other things, compounds of Formulas II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), and (IIb-2). In some embodiments, the present disclosure provides a compound of Formula II:

In some embodiments, the present disclosure provides the compound of Formula II, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-1):

In some embodiments, the present disclosure provides the compound of Formula II or II-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-2):

In some embodiments, the present disclosure provides the compound of Formula II or II-1, or a pharmaceutically acceptable salt thereof, having the structure of Formula (II-3):

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is N. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is CH. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis (CH2)mCN, and m is 0, 1, 2 or 3. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis CH2CN. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is CRx, wherein Rxis halo. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein X is C—Cl.

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein L1is O. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, having the structure of Formula (IIa):

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein L1is CHR1b. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein L1is CH2. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein L1is CH(CH3). In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein R1aand R1bare each independently H, C1-C3alkyl, or halo. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein R1bis C1-C3alkyl or halo. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, wherein R1bis methyl.

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, or II-3, or a pharmaceutically acceptable salt thereof, having the structure of Formula (IIb):

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R2aand R2bare each independently H, C1-C3alkyl, halo, or C1-C6haloalkyl. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein L2is CHR2b. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R2bis H or C1-C3alkyl. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R2bis H. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R2bis C1-C3alkyl. In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), or (IIb), or a pharmaceutically acceptable salt thereof, wherein R2bis methyl.

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, or (IIa), or a pharmaceutically acceptable salt thereof, having the structure of Formula (IIa-1):

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, or (IIb), or a pharmaceutically acceptable salt thereof, having the structure of Formula (IIb-1):

In some embodiments, the present disclosure provides the compound of Formula II, II-1, II-2, II-3, (IIa), (IIb), or (IIb-1), or a pharmaceutically acceptable salt thereof, having the structure of Formula (IIb-2):

Y is C or Si; n is 0 or 1; q is 0 or 1; RY1is H or C1-C3alkyl; and RY2is H or C1-C3alkyl;alternatively, RY1and RY2combine to form a C3-C6cycloalkyl.

Y is C or Si; n is 0 or 1; q is 0 or 1; RY1is H or Me; and RY2is H or Me; alternatively, RY1and RY2combine to form a cyclopropyl.

In some embodiments, the present disclosure provides the compound of Formula I, I-A, (I-1), (I-2), or (I-3), or a pharmaceutically acceptable salt thereof, whereinX is N;R1, R2, R3, and R4are each H;L1is CH2or CHCH3;L2is CH2;L3is CH2;RAis

In some embodiments, the present disclosure provides the compound of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2), or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of Formula (Ib-5) or Formula (Ib-6):

In some embodiments, the present disclosure provides the compound of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2), or a pharmaceutically acceptable salt thereof, wherein the compound has the structure of Formula (Ib-7) or Formula (Ib-8):

Also falling within the scope herein are the in vivo metabolic products of the compounds described herein. Such products may result for example from the oxidation, reduction, hydrolysis, amidation, esterification and the like of the administered compound, primarily due to enzymatic processes. Accordingly, included are novel and unobvious compounds produced by a process comprising contacting a compound with a mammal for a period of time sufficient to yield a metabolic product thereof. Such products typically are identified by preparing a radiolabelled (e.g.,14C or3H) compound, administering it parenterally in a detectable dose (e.g., greater than about 0.5 mg/kg) to an animal such as rat, mouse, guinea pig, monkey, or to man, allowing sufficient time for metabolism to occur (typically about 30 seconds to 30 hours) and isolating its conversion products from the urine, blood or other biological samples. These products are easily isolated since they are labeled (others are isolated by the use of antibodies capable of binding epitopes surviving in the metabolite). The metabolite structures are determined in conventional fashion, e.g., by MS or NMR analysis. In general, analysis of metabolites is done in the same way as conventional drug metabolism studies. The conversion products, so long as they are not otherwise found in vivo, are useful in diagnostic assays for therapeutic dosing of the compounds even if they possess no HSV antiviral activity of their own.

Recipes and methods for determining stability of compounds in surrogate gastrointestinal secretions are known. Compounds are defined herein as stable in the gastrointestinal tract where less than about 50 mole percent of the protected groups are deprotected in surrogate intestinal or gastric juice upon incubation for 1 hour at 37° C. Simply because the compounds are stable to the gastrointestinal tract does not mean that they cannot be hydrolyzed in vivo. The prodrugs typically will be stable in the digestive system but may be substantially hydrolyzed to the parental drug in the digestive lumen, liver, lung or other metabolic organ, or within cells in general. As used herein, a prodrug is understood to be a compound that is chemically designed to efficiently liberate the parent drug after overcoming biological barriers to oral delivery.

Also disclosed herein are pharmaceutical compositions comprising a pharmaceutically effective amount of a compound of the present disclosure (e.g., a compound of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2),) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient. Also provided herein is a pharmaceutical composition comprising a pharmaceutically effective amount of a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

In some embodiments, the compounds disclosed herein have pharmacokinetic properties (e.g., oral bioavailability) suitable for oral administration of the compounds. Formulations suitable for oral administration can, for instance, be presented as discrete units such as capsules, cachets or tablets, each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil liquid emulsion. The active ingredient can also be administered, for instance, as a bolus, electuary, or paste.

A tablet can be made by compression or molding, optionally with at least accessory ingredients. Compressed tablets can be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as, for instance, a powder or granules, optionally mixed with a binder, lubricant, inert diluent, preservative, surface active, dispersing agent, or a combination thereof. Molded tablets can be made by molding in a suitable machine a mixture of the powdered active ingredient moistened with an inert liquid diluent. The tablets can optionally be coated or scored and optionally are formulated so as to provide slow or controlled release of the active ingredient therefrom.

For infections of the eye or other external tissues (e.g., mouth and skin), the formulations can be applied as a topical ointment or cream containing the active ingredient(s) in an amount of, for example, 0.075 to 20% w/w (including active ingredient(s) in a range from 0.1% to 20% in increments of 0.1% w/w such as 0.6% w/w, 0.7% w/w, etc.), from 0.2% to 15% w/w, or from 0.5% to 10% w/w. When formulated in an ointment, the active ingredients can be employed in some embodiments with either a paraffinic or a water-miscible ointment base. Alternatively, the active ingredients can be formulated in a cream with an oil-in-water cream base.

In some embodiments, the aqueous phase of the cream base can include, for example, from 30% to 90% (e.g., 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%) w/w of a polyhydric alcohol, i.e., an alcohol having two or more hydroxyl groups such as propylene glycol, butane 1,3-diol, mannitol, sorbitol, glycerol and polyethylene glycol (including PEG 400) and mixtures thereof. In some embodiments, the cream base can include, for instance, a compound that enhances absorption or penetration of the active ingredient through the skin or other affected areas. Examples of such dermal penetration enhancers include, but are not limited to, dimethyl sulfoxide and related analogs. In some embodiments, the cream or emulsion does not include water.

The oily phase of the emulsions can be constituted from known ingredients in a known manner. In some embodiments, the phase comprises merely an emulsifier (otherwise known as an emulgent). In some embodiments, the phase comprises a mixture of at least one emulsifier with a fat, an oil, or a combination thereof. In some embodiments, a hydrophilic emulsifier is included together with a lipophilic emulsifier that acts as a stabilizer. Together, the emulsifier(s) with or without stabilizer(s) can make up the so-called emulsifying wax, and the wax together with the oil and fat make up the so-called emulsifying ointment base that can form the oily dispersed phase of the cream formulations.

The choice of suitable oils or fats for the formulation can be based on achieving the desired cosmetic properties. In some embodiments, the cream can be a non-greasy, non-staining, and washable product with suitable consistency to avoid leakage from tubes or other containers. In some embodiments, esters can be included, such as, for example, straight or branched chain, mono- or dibasic alkyl esters such as di-isoadipate, isocetyl stearate, propylene glycol diester of coconut fatty acids, isopropyl myristate, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate, a blend of branched chain esters known as CRODAMOL® CAP, or a combination thereof. In some embodiments, high melting point lipids such as white soft paraffin and/or liquid paraffin or other mineral oils can be included.

In some embodiments, the compounds disclosed herein are administered alone. In some embodiments, the compounds disclosed herein are administered in pharmaceutical compositions. In some embodiments, the pharmaceutical compositions are for veterinary use. In some embodiments, the pharmaceutical compositions are for human use. In some embodiments, the pharmaceutical compositions disclosed herein include at least one additional therapeutic agent. In some embodiments, the pharmaceutical compositions disclosed herein include one or more additional therapeutic agent. In some embodiments, the one or more additional therapeutic agents is independently a chemotherapeutic agent, an immunotherapeutic agent, a hormonal agent, an anti-hormonal agent, a targeted therapy agent, or an anti-angiogenesis agent.

Pharmaceutical compositions disclosed herein can be in any form suitable for the intended method of administration. The pharmaceutical compositions disclosed herein can be presented in unit dosage form and can be prepared by any of the methods well known in the art of pharmacy. Exemplary techniques and formulations can be found, for instance, in REMINGTON'SPHARMACEUTICALSCIENCES(Mack Publishing Co., Easton, PA). Such methods can include the step of bringing into association a compound disclosed herein with the carrier that constitutes at least accessory ingredients. In general, the formulations can be prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.

When used for oral use for example, tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, solutions, syrups or elixirs can be prepared. Formulations intended for oral use can be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such formulations can contain at least agents including sweetening agents, flavoring agents, coloring agents and preserving agents, in order to provide a palatable preparation. Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipient which are suitable for manufacture of tablets are acceptable. These excipients can be, for example, inert diluents, such as calcium or sodium carbonate, lactose, calcium or sodium phosphate; granulating and disintegrating agents, such as maize starch, or alginic acid; binding agents, such as starch, gelatin or acacia; and lubricating agents, such as magnesium stearate, stearic acid or talc. Tablets can be uncoated or can be coated by known techniques including microencapsulation to delay disintegration and adsorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate alone or with a wax can be employed.

Formulations for oral use can be also presented as hard gelatin capsules where the active ingredient is mixed with an inert solid diluent, for example calcium phosphate or kaolin, or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, such as peanut oil, liquid paraffin, or olive oil.

Oil suspensions can be formulated by suspending the active ingredient in a vegetable oil (e.g., arachis oil, olive oil, sesame oil, coconut oil, or a combination thereof), a mineral oil such as liquid paraffin, or a combination thereof. The oral suspensions can contain, for instance, a thickening agent, such as beeswax, hard paraffin, cetyl alcohol, or a combination thereof. In some embodiments, sweetening agents, such as those set forth above, and/or flavoring agents, are added to provide a palatable oral preparation. In some embodiments, the formulations disclosed herein are preserved by the addition of an antioxidant such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water can provide the active ingredient in admixture with a dispersing or wetting agent, a suspending agent, a preservative, and combinations thereof. Suitable dispersing or wetting agents and suspending agents are exemplified by those disclosed above. Additional excipients, for example sweetening, flavoring and coloring agents, can also be present.

The pharmaceutical compositions can also be in the form of oil-in-water emulsions. The oily phase can be a vegetable oil, such as olive oil or arachis oil, a mineral oil, such as liquid paraffin, or a mixture of these. Suitable emulsifying agents include naturally-occurring gums, such as gum acacia and gum tragacanth, naturally-occurring phosphatides, such as soybean lecithin, esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of these partial esters with ethylene oxide, such as polyoxyethylene sorbitan monooleate. The emulsion can also contain sweetening and flavoring agents. Syrups and elixirs can be formulated with sweetening agents, such as for instance, glycerol, sorbitol or sucrose. Such formulations can also contain, for instance, a demulcent, a preservative, a flavoring, a coloring agent, or a combination thereof.

The pharmaceutical compositions can be in the form of a sterile injectable or intravenous preparation, such as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable or intravenous preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, such as a solution in 1,3-butane-diol or prepared as a lyophilized powder. Among the acceptable vehicles and solvents that can be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile fixed oils can be employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can likewise be used in the preparation of injectables. Among the acceptable vehicles and solvents that can be employed include, but are not limited to, water, Ringer's solution isotonic sodium chloride solution, and hypertonic sodium chloride solution.

The amount of active ingredient that can be combined with the carrier material to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. For example, a time-release formulation intended for oral administration to humans can contain approximately 1 mg to 2000 mg of active material compounded with an appropriate and convenient amount of carrier material, which can vary from 5% to 95% of the total formulations (weight:weight). For example, a time-release formulation intended for oral administration to humans can contain approximately 1 mg to 1000 mg of active material compounded with an appropriate and convenient amount of carrier material, which can vary from 5% to 95% of the total formulations (weight:weight). The pharmaceutical composition can be prepared to provide easily measurable amounts for administration. For example, an aqueous solution intended for intravenous infusion can contain from 3 μg to 500 μg of the active ingredient per milliliter of solution in order that infusion of a suitable volume at a rate of 30 mL/hr can occur.

Formulations suitable for topical administration to the eye also include eye drops wherein the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient. In some embodiments, the compounds disclosed herein are included in the pharmaceutical compositions disclosed herein in a concentration of 0.5% to 20% (e.g., 0.5% to 10%, 1.5% w/w).

Formulations suitable for topical administration in the mouth include lozenges can comprise an active ingredient (i.e., a compound disclosed herein and/or additional therapeutic agents) in a flavored basis, usually sucrose and acacia or tragacanth; pastilles comprising the active ingredient in an inert basis such as gelatin and glycerin, or sucrose and acacia; and mouthwashes comprising the active ingredient in a suitable liquid carrier.

Formulations for rectal administration can be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.

Formulations suitable for vaginal administration can be presented as pessaries, tampons, creams, gels, pastes, foams or spray formulations containing in addition to the active ingredient such carriers as are known in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions that can contain anti-oxidants, buffers, bacteriostats and solutes that render the formulation isotonic with the blood of the intended recipient; and aqueous and non-aqueous sterile suspensions that can include suspending agents and thickening agents.

The formulations can be presented in unit-dose or multi-dose containers, for example, sealed ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injection, immediately before use. Extemporaneous injection solutions and suspensions are prepared from sterile powders, granules and tablets of the kind previously described. Preferred unit-dosage formulations are those containing a daily dose or unit daily sub-dose, as herein above recited, or an appropriate fraction thereof, of the active ingredient.

It should be understood that in addition to the ingredients particularly mentioned above the formulations can include other agents conventional in the art having regard to the type of formulation in question, for example those suitable for oral administration can include flavoring agents.

Further provided are veterinary formulations comprising a compound disclosed herein together with a veterinary carrier therefor.

Veterinary carriers are materials useful for the purpose of administering the formulation and can be solid, liquid or gaseous materials which are otherwise inert or acceptable in the veterinary art and are compatible with the active ingredient. These veterinary formulations can be administered orally, parenterally, or by any other desired route.

Compounds herein are used to provide controlled release pharmaceutical compositions containing as active ingredient one or more of the compounds (“controlled release formulations”) in which the release of the active ingredient can be controlled and regulated to allow less frequency dosing or to improve the pharmacokinetic or toxicity profile of a given active ingredient.

Effective dose of active ingredient depends at least on the nature of the condition being treated, toxicity, whether the compound is being used prophylactically (lower doses) or against an active viral infection, the method of delivery, and the pharmaceutical composition, and will be determined by the clinician using conventional dose escalation studies. In some embodiments, the effective dose is from 0.0001 to 100 mg/kg body weight per day; for instance, from 10 to 30 mg/kg body weight per day; from 15 to 25 mg/kg body weight per day; from 10 to 15 mg/kg body weight per day; or from 20 to 30 mg/kg body weight per day. For example, the daily candidate dose for an adult human of approximately 70 kg body weight can range from 1 mg to 2000 mg (e.g., from 5 mg to 500 mg, from 500 mg to 1000 mg, from 1000 mg to 1500 mg, from 1500 mg to 2000 mg), and can take the form of single or multiple doses. For example, the daily candidate dose for an adult human of approximately 70 kg body weight can range from 1 mg to 1000 mg (e.g., from 5 mg to 500 mg), and can take the form of single or multiple doses.

Also provided herein are kits that includes a compound disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments the kits described herein can comprise a label and/or instructions for use of the compound in the treatment of a disease or condition in a subject (e.g., human) in need thereof. In some embodiments, the disease or condition is viral infection.

In some embodiments, the kit can also comprise one or more additional therapeutic agents and/or instructions for use of additional therapeutic agents in combination with the compound disclosed herein in the treatment of the disease or condition in a subject (e.g., human) in need thereof.

In some embodiments, the kits provided herein comprise individual dose units of a compound as described herein, or a pharmaceutically acceptable salt, racemate, enantiomer, diastereomer, tautomer, polymorph, pseudopolymorph, amorphous form, hydrate or solvate thereof. Examples of individual dosage units can include pills, tablets, capsules, prefilled syringes or syringe cartridges, IV bags, inhalers, nebulizers etc., each comprising a therapeutically effective amount of the compound in question, or a pharmaceutically acceptable salt, racemate, enantiomer, diastereomer, tautomer, polymorph, pseudopolymorph, amorphous form, hydrate or solvate thereof. In some embodiments, the kit can contain a single dosage unit and in others multiple dosage units are present, such as the number of dosage units required for a specified regimen or period.

Also provided are articles of manufacture that include a compound disclosed herein, or a pharmaceutically acceptable salt, stereoisomer, mixture of stereoisomers or tautomer thereof; and a container. In some embodiments, the container of the article of manufacture is a vial, jar, ampoule, preloaded syringe, blister package, tin, can, bottle, box, an intravenous bag, an inhaler, or a nebulizer.

One or more of the compounds of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (11-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2), (herein referred to as the active ingredients) are administered by any route appropriate to the condition to be treated. Suitable routes include oral, rectal, nasal, pulmonary, topical (including buccal and sublingual), vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural), and the like. It will be appreciated that the route may vary with for example the condition of the recipient. An advantage of the compounds herein is that they are orally bioavailable and can be dosed orally.

The compounds of the present disclosure (also referred to herein as the active ingredients), can be administered by any route appropriate to the condition to be treated.

Suitable routes include oral, rectal, nasal, topical (including buccal and sublingual), transdermal, vaginal and parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intrathecal and epidural), and the like. It will be appreciated that the route may vary with for example the condition of the recipient. An advantage of certain compounds disclosed herein is that they are orally bioavailable and can be dosed orally.

A compound of the present disclosure may be administered to an individual in accordance with an effective dosing regimen for a desired period of time or duration, such as at least about one month, at least about 2 months, at least about 3 months, at least about 6 months, or at least about 12 months or longer. In some embodiments, the compound is administered on a daily or intermittent schedule for the duration of the individual's life.

The dosage or dosing frequency of a compound of the present disclosure may be adjusted over the course of the treatment, based on the judgment of the administering physician.

The compound may be administered to an individual (e.g., a human) in an effective amount. In some embodiments, the compound is administered once daily.

The compound can be administered by any useful route and means, such as by oral or parenteral (e.g., intravenous) administration. Therapeutically effective amounts of the compound may include from about 0.00001 mg/kg body weight per day to about 10 mg/kg body weight per day, such as from about 0.0001 mg/kg body weight per day to about 10 mg/kg body weight per day, or such as from about 0.001 mg/kg body weight per day to about 1 mg/kg body weight per day, or such as from about 0.01 mg/kg body weight per day to about 1 mg/kg body weight per day, or such as from about 0.05 mg/kg body weight per day to about 0.5 mg/kg body weight per day, or such as from about 0.3 mg to about 30 mg per day, or such as from about 30 mg to about 300 mg per day.

A compound of the present disclosure may be combined with one or more additional therapeutic agents in any dosage amount of the compound of the present disclosure (e.g., from about 1 mg to about 1000 mg of compound). Therapeutically effective amounts may include from about 1 mg per dose to about 1000 mg per dose, such as from about 50 mg per dose to about 500 mg per dose, or such as from about 100 mg per dose to about 400 mg per dose, or such as from about 150 mg per dose to about 350 mg per dose, or such as from about 200 mg per dose to about 300 mg per dose. Other therapeutically effective amounts of the compound of the present disclosure are about 100, about 125, about 150, about 175, about 200, about 225, about 250, about 275, about 300, about 325, about 350, about 375, about 400, about 425, about 450, about 475, or about 500 mg per dose. Other therapeutically effective amounts of the compound of the present disclosure are about 100 mg per dose, or about 125, about 150, about 175, about 200, about 225, about 250, about 275, about 300, about 325, about 350, about 375, about 400, about 425, about 450, or about 500 mg per dose. A single dose can be administered hourly, daily, or weekly. For example, a single dose can be administered once about every 1 hour, about 2, about 3, about 4, about 6, about 8, about 12, about 16 or once about every 24 hours. A single dose can also be administered once about every 1 day, about 2, about 3, about 4, about 5, about 6, or once about every 7 days. A single dose can also be administered once about every 1 week, about 2, about 3, or once about every 4 weeks. In some embodiments, a single dose can be administered once about every week. A single dose can also be administered once about every month.

Other therapeutically effective amounts of the compound of the present disclosure are about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, or about 100 mg per dose.

The frequency of dosage of the compound of the present disclosure can be determined by the needs of the individual patient and can be, for example, once per day or twice, or more times, per day. Administration of the compound continues for as long as necessary to treat the disease or condition. For example, a compound can be administered to a human having cancer for a period of from about 20 days to about 180 days or, for example, for a period of from about 20 days to about 90 days or, for example, for a period of from about 30 days to about 60 days.

Administration can be intermittent, with a period of several or more days during which a patient receives a daily dose of the compound of the present disclosure followed by a period of several or more days during which a patient does not receive a daily dose of the compound. For example, a patient can receive a dose of the compound every other day, or three times per week. Again by way of example, a patient can receive a dose of the compound each day for a period of from about 1 to about 14 days, followed by a period of about 7 to about 21 days during which the patient does not receive a dose of the compound, followed by a subsequent period (e.g., from about 1 to about 14 days) during which the patient again receives a daily dose of the compound. Alternating periods of administration of the compound, followed by non-administration of the compound, can be repeated as clinically required to treat the patient.

In some embodiments, pharmaceutical compositions comprising a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more (e.g., one, two, three, four, one or two, one to three, or one to four) additional therapeutic agents, and a pharmaceutically acceptable excipient are provided.

In some embodiments, kits comprising a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, in combination with one or more (e.g., one, two, three, four, one or two, one to three, or one to four) additional therapeutic agents are provided.

In some embodiments, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is combined with one, two, three, four or more additional therapeutic agents. In some embodiments, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is combined with two additional therapeutic agents. In some embodiments, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is combined with three additional therapeutic agents. In some embodiments, a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is combined with four additional therapeutic agents. The one, two, three, four or more additional therapeutic agents can be different therapeutic agents selected from the same class of therapeutic agents, and/or they can be selected from different classes of therapeutic agents.

In some embodiments, when a compound of the present disclosure is combined with one or more additional therapeutic agents as described herein, the components of the composition are administered as a simultaneous or sequential regimen. When administered sequentially, the combination may be administered in two or more administrations.

In some embodiments, a compound of the present disclosure is combined with one or more additional therapeutic agents in a unitary dosage form for simultaneous administration to a patient, for example as a solid dosage form for oral administration.

In some embodiments, a compound of the present disclosure is co-administered with one or more additional therapeutic agents.

In order to prolong the effect of a compound of the present disclosure, it is often desirable to slow the absorption of a compound from subcutaneous or intramuscular injection. This may be accomplished by the use of a liquid suspension of crystalline or amorphous material with poor water solubility. The rate of absorption of the compound then depends upon its rate of dissolution that, in turn, may depend upon crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form is accomplished by dissolving or suspending a compound in an oil vehicle. Injectable depot forms are made by forming microencapsule matrices of a compound in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of compound to polymer and the nature of the particular polymer employed, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations are also prepared by entrapping a compound in liposomes or microemulsions that are compatible with body tissues.

VII. Methods of Use

The disclosure further relates to the use of compounds disclosed herein for the treatment and/or prophylaxis of diseases and/or conditions through inhibition of KRAS G12D and/or G12C. Further, the present disclosure relates to the use of said compounds for the preparation of a medicament for the treatment and/or prophylaxis of cancer.

Medicaments as referred to herein can be prepared by conventional processes, including the combination of a compound according to the present disclosure and a pharmaceutically acceptable carrier.

In some embodiments, provided herein is a method of treating and/or preventing a cancer.

In some embodiments, provided herein is a method of treating and/or preventing a KRAS G12D-associated cancer.

In some embodiments, the KRAS G12D associated disease or condition includes cancer. In some embodiments, the cancer is a hematological cancer. In some embodiments, the cancer includes a solid tumor. In some embodiments, the cancer includes a malignant tumor. In some embodiments the cancer includes a metastatic cancer. In some embodiments, the cancer is resistant or refractory to one or more anticancer therapies. In some embodiments, greater than about 50% of the cancer cells detectably express one or more cell surface immune checkpoint receptors (e.g., so-called “hot” cancer or tumor). In some embodiments, greater than about 1% and less than about 50% of the cancer cells detectably express one or more cell surface immune checkpoint receptors (e.g., so called “warm” cancer or tumor). In some embodiments, less than about 1% of the cancer cells detectably express one or more cell surface immune checkpoint receptors (e.g., so called “cold” cancer or tumor).

In some embodiments, the KRAS G12D associated disease or condition is an epithelial tumor (e.g., a carcinoma, a squamous cell carcinoma, a basal cell carcinoma, a squamous intraepithelial neoplasia), a glandular tumor (e.g., an adenocarcinoma, an adenoma, an adenomyoma), a mesenchymal or soft tissue tumor (e.g., a sarcoma, a rhabdomyosarcoma, a leiomyosarcoma, a liposarcoma, a fibrosarcoma, a dermatofibrosarcoma, a neurofibrosarcoma, a fibrous histiocytoma, an angiosarcoma, an angiomyxoma, a leiomyoma, a chondroma, a chondrosarcoma, an alveolar soft-part sarcoma, an epithelioid hemangioendothelioma, a Spitz tumor, a synovial sarcoma), or a lymphoma.

In some embodiments, the KRAS G12D associated disease or condition is a cancer selected from a lung cancer, a colorectal cancer, a breast cancer, a prostate cancer, a cervical cancer, a pancreatic cancer and a head and neck cancer. In some embodiments, the cancer is metastatic.

In some embodiments, the KRAS G12D associated disease or condition is a cancer selected from non-small lung cancer (NSCLC), melanoma, triple-negative breast cancer (TNBC), nasopharyngeal cancer (NPC), microsatellite stable colorectal cancer (mssCRC), thymoma, and gastrointestinal stromal tumor (GIST). In some embodiments, the cancer is metastatic.

In some embodiments, the KRAS G12D associated disease or condition is a cancer of pancreatic cancer, colorectal cancer, non-small cell lung cancer, endometrial cancer, uterine endometrical carcinoma, cholangio carcinoma, testicular germ cell cancer, cervical squamous carcinoma, or myelodysplastic syndrome.

In some embodiments, the cancer is or myelodysplastic syndrome. In some embodiments, the cancer is high risk myelodysplastic syndrome or low risk myelodysplastic syndrome. In some embodiments, the cancer is high risk myelodysplastic syndrome. In some embodiments, the cancer is high risk myelodysplastic syndrome.

In some embodiments, the cancer is colorectal cancer. In some embodiments, the cancer is non-small cell lung cancer. In some embodiments, the cancer is pancreatic cancer. In some embodiments, the cancer is endometrial cancer. In some embodiments, the cancer is uterine endometrical carcinoma. In some embodiments, the cancer is testicular germ cell cancer. In some embodiments, the cancer is cervical squamous carcinoma. In some embodiments, the cancer is cholangio carcinoma.

The effective dosage of active ingredient employed may vary depending on the particular compound employed, the mode of administration, the condition being treated and the severity of the condition being treated. Such dosage may be ascertained readily by a person skilled in the art.

When treating or preventing a KRAS G12D associated disease or condition for which compounds of the present disclosure are indicated, generally satisfactory results are obtained when the compounds of the present disclosure are administered at a daily dosage of from about 0.1 milligram to about 300 milligram per kilogram of animal body weight. In some embodiments, the compounds of the present disclosure are given as a single daily dose or in divided doses two to six times a day, or in sustained release form. For most large mammals, the total daily dosage is from about 1 milligram to about 1000 milligrams, or from about 1 milligram to about 50 milligrams. In the case of a 70 kg adult human, the total daily dose will generally be from about 0.1 milligrams to about 200 milligrams. This dosage regimen may be adjusted to provide the optimal therapeutic response. In some embodiments, the total daily dosage is from about 1 milligram to about 900 milligrams, about 1 milligram to about 800 milligrams, about 1 milligram to about 700 milligrams, about 1 milligram to about 600 milligrams, about 1 milligram to about 400 milligrams, about 1 milligram to about 300 milligrams, about 1 milligram to about 200 milligrams, about 1 milligram to about 100 milligrams, about 1 milligram to about 50 milligrams, about 1 milligram to about 20 milligram, or about 1 milligram to about 10 milligrams.

The compounds of the present application or the compositions thereof may be administered once, twice, three, or four times daily, using any suitable mode described above. Also, administration or treatment with the compounds may be continued for a number of days; for example, commonly treatment would continue for at least 7 days, 14 days, or 28 days, for one cycle of treatment. Treatment cycles are frequently alternated with resting periods of about 1 to 28 days, commonly about 7 days or about 14 days, between cycles. The treatment cycles, in other embodiments, may also be continuous.

In some embodiments, the methods provided herein comprise administering to the subject an initial daily dose of about 1 to 800 mg of a compound described herein and increasing the dose by increments until clinical efficacy is achieved. Increments of about 5, 10, 25, 50, or 100 mg can be used to increase the dose. The dosage can be increased daily, every other day, twice per week, or once per week.

In some embodiments, the compound or pharmaceutically acceptable salt thereof of the present disclosure is administered in combination with one or more additional therapeutic agent or therapeutic modality.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the one or more additional therapeutic agent or additional therapeutic modality comprises one, two, three, or four additional therapeutic agents and/or therapeutic modalities.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the additional therapeutic agent or therapeutic modalities are selected from an immune checkpoint modulator, an antibody-drug conjugate (ADC), an antiapoptotic agent, a targeted anticancer therapeutic, a chemotherapeutic agent, surgery, or radiation therapy.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the immune checkpoint modulator is selected from an anti-PD-(L)1 antibody, an anti-TIGIT antibody, an anti-CTLA4 antibody, an anti-CCR8 antibody, an anti-TREM1 antibody, an anti-TREM2 antibody, a CD47 inhibitor, a DGKα inhibitor, an HPK1 inhibitor, a FLT3 agonist, an adenosine receptor antagonist, a CD39 inhibitor, a CD73 inhibitor, an IL-2 variant (IL-2v), and a CAR-T cell therapy.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the anti-TIGIT antibody is selected from tiragolumab, vibostolimab, domvanalimab, AB308, AK127, BMS-986207, and etigilimab.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the anti-CTLA4 antibody is selected from ipilimumab, tremelimumab, and zalifrelimab.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the CD47 inhibitor is selected from magrolimab, letaplimab, lemzoparlimab, AL-008, RRx-001, CTX-5861, FSI-189 (GS-0189), ES-004, BI-765063, ADU1805, CC-95251, and Q-1801.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the adenosine receptor antagonist is etrumadenant (AB928), taminadenant, TT-10, TT-4, or M1069.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the CD39 inhibitor is TTX-030.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the CD73 inhibitor is quemliclustat (AB680), uliledlimab, mupadolimab, ORIC-533, ATG-037, PT-199, AK131, NZV930, BMS-986179, or oleclumab.

In some embodiments, the present disclosure provides the pharmaceutical composition or the method wherein the ADC is selected from sacituzumab govitecan, datopotamab deruxtecan, enfortumab vedotin, and trastuzumab deruxtecan.

In some embodiments, the method includes administering one or more additional therapeutic agents. The one or more additional therapeutic agents can be one or more therapeutic agents as described below. In some embodiments, the one or more additional therapeutic agents is independently a chemotherapeutic agent, an immunotherapeutic agent, a hormonal agent, an anti-hormonal agent, a targeted therapy agent, or an anti-angiogenesis agent.

In some embodiments, the one or more additional therapeutic agents includes therapeutic agents used to treat high risk myelodysplastic syndrome (HR MDS), low risk myelodyplastic syndrome (LR MDS), colorectal cancer, non-small cell lung cancer (NSCLC), pancreatic cancer, or endometrial cancer. In some embodiments, the one or more additional therapeutic agents includes therapeutic agents used to treat high risk myelodysplastic syndrome (HR MDS). In some embodiments, the one or more additional therapeutic agents includes azacitidine (Vidaza®), decitabine (Dacogen®), lenalidomide (Revlimid®), cytarabine, idarubicin, daunorubicin, cytarabine+daunorubicin, cytarabine+idarubicin, pevonedistat, venetoclax, sabatolimab, guadecitabine, rigosertib, ivosidenib, enasidenib, selinexor, BGB324, DSP-7888, or SNS-301.

In some embodiments, the one or more additional therapeutic agents includes therapeutic agents used to treat low risk myelodyplastic syndrome (LR MDS). In some embodiments, the one or more additional therapeutic agents includes lenalidomide, azacytidine, roxadustat, luspatercept, imetelstat, LB-100, or rigosertib.

In some embodiments, the one or more additional therapeutic agents includes therapeutic agents used to treat pancreatic cancer. In some embodiments, the one or more additional therapeutic agents includes 5-FU, leucovorin, oxaliplatin, irinotecan, gemcitabine, nab-paclitaxel (Abraxane®), FOLFIRINOX, 5-FU+leucovorin+oxaliplatin+irinotecan, 5-FU+nanoliposomal irinotecan, leucovorin+nanoliposomal irinotecan, or gemcitabine+nab-paclitaxel.

In some embodiments, the one or more additional therapeutic agents includes therapeutic agents used to treat endometrial cancer. In some embodiments, the one or more additional therapeutic agents includes carboplatin, paclitaxel, cisplatin, doxorubicin, ifosfamide, progesterone, anastrozole (Arimidex®), letrozole (Femara®), exemestane (Aromasin®), pembrolizumab (Keytruda®), lenvatinib (Lenvima®), or dostarlimab (Jemperli®).

In another embodiment, the present disclosure provides a method for manufacturing a medicament for treating cancer in a subject in need thereof, characterized in that a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, is used.

In another embodiment, the present disclosure provides a method for manufacturing a medicament for inhibiting cancer metastasis in a subject in need thereof, characterized in that a compound of the present invention, or a pharmaceutically acceptable salt thereof, is used.

In another embodiment, the present disclosure provides use of the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of cancer in a subject.

In another embodiment, the present disclosure provides use of the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for inhibiting cancer metastasis in a subject.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in the treatment of cancer in a subject in need thereof.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in inhibiting cancer metastasis in a subject in need thereof.

In another embodiment, the present disclosure provides the compound of the present disclosure, or a pharmaceutically acceptable salt thereof, for use in therapy.

VIII. Combination Therapy

In some embodiments, a compound of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (IIb-1), or (IIb-2), provided herein, or pharmaceutically acceptable salt thereof, is administered in combination with one or more additional therapeutic agents to treat or prevent a disease or condition disclosed herein. In some embodiments, the one or more additional therapeutic agents are one, two, three, or four additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are one additional therapeutic agent. In some embodiments, the one or more additional therapeutic agents are two additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are three additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are four additional therapeutic agents.

In some embodiments, the pharmaceutical compositions provided herein have a compound of Formula I, I-A, (I-1), (I-2), (I-3), (Ia), (Ia-1), (Ib), (Ib-1), (Ib-2), (Ib-3), (Ib-4), (Ib-5), (Ib-6), (Ib-7), (Ib-8), II, (II-1), (IIa), (IIa-1), (IIb), (Ib-1), or (IIb-2), provided herein, or pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are one, two, three, or four additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are one additional therapeutic agent. In some embodiments, the one or more additional therapeutic agents are two additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are three additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents are four additional therapeutic agents.

Illustrative Targets

Illustrative Mechanisms of Action

Immune Checkpoint Modulators

In some embodiments a compound provided herein is administered with one or more blockers or inhibitors of inhibitory immune checkpoint proteins or receptors and/or with one or more stimulators, activators or agonists of one or more stimulatory immune checkpoint proteins or receptors. Blockade or inhibition of inhibitory immune checkpoints can positively regulate T-cell or NK cell activation and prevent immune escape of cancer cells within the tumor microenvironment. Activation or stimulation of stimulatory immune check points can augment the effect of immune checkpoint inhibitors in cancer therapeutics. In some embodiments, the immune checkpoint proteins or receptors regulate T cell responses (e.g., reviewed in Xu, et al.,J Exp Clin Cancer Res. (2018) 37:110). In some embodiments, the immune checkpoint proteins or receptors regulate NK cell responses (e.g., reviewed in Davis, et al.,Semin Immunol. (2017) 31:64-75 and Chiossone, et al.,Nat Rev Immunol.(2018) 18(11):671-688). Inhibition of regulatory T-cells (Treg) or Treg depletion can alleviate their suppression of antitumor immune responses and have anticancer effects (e.g., reviewed in Plitas and Rudensky,Annu. Rev. Cancer Biol. (2020) 4:459-77; Tanaka and Sakaguchi,Eur. J. Immunol. (2019) 49:1140-1146).

In some embodiments a compound provided herein is administered with one or more blockers or inhibitors of one or more T-cell inhibitory immune checkpoint proteins or receptors. Illustrative T-cell inhibitory immune checkpoint proteins or receptors include CD274 (CD274, PDL1, PD-L1); programmed cell death 1 ligand 2 (PDCD1LG2, PD-L2, CD273); programmed cell death 1 (PDCD1, PD1, PD-1); cytotoxic T-lymphocyte associated protein 4 (CTLA4, CD152); CD276 (B7H3); V-set domain containing T cell activation inhibitor 1 (VTCN1, B7H4); V-set immunoregulatory receptor (VSIR, B7H5, VISTA); immunoglobulin superfamily member 11 (IGSF11, VSIG3); TNFRSF14 (HVEM, CD270), TNFSF14 (HVEML); CD272 (B and T lymphocyte associated (BTLA)); PVR related immunoglobulin domain containing (PVRIG, CD112R); T cell immunoreceptor with Ig and ITIM domains (TIGIT); lymphocyte activating 3 (LAG3, CD223); hepatitis A virus cellular receptor 2 (HAVCR2, TIMD3, TIM3); galectin 9 (LGALS9); killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); and killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1). In some embodiments, the compound provided herein is administered with one or more agonist or activators of one or more T-cell stimulatory immune checkpoint proteins or receptors. Illustrative T-cell stimulatory immune checkpoint proteins or receptors include without limitation CD27, CD70; CD40, CD40LG; inducible T cell costimulator (ICOS, CD278); inducible T cell costimulator ligand (ICOSLG, B7H2); TNF receptor superfamily member 4 (TNFRSF4, OX40); TNF superfamily member 4 (TNFSF4, OX40L); TNFRSF9 (CD137), TNFSF9 (CD137L); TNFRSF18 (GITR), TNFSF18 (GITRL); CD80 (B7-1), CD28; nectin cell adhesion molecule 2 (NECTIN2, CD112); CD226 (DNAM-1); CD244 (2B4, SLAMF4), Poliovirus receptor (PVR) cell adhesion molecule (PVR, CD155). See, e.g., Xu, et al.,J Exp Clin Cancer Res.(2018) 37:110.

In some embodiments the compound provided herein is administered with one or more blockers or inhibitors of one or more NK-cell inhibitory immune checkpoint proteins or receptors. Illustrative NK-cell inhibitory immune checkpoint proteins or receptors include killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR, CD158E1); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 1 (KIR2DL1); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 2 (KIR2DL2); killer cell immunoglobulin like receptor, two Ig domains and long cytoplasmic tail 3 (KIR2DL3); killer cell immunoglobulin like receptor, three Ig domains and long cytoplasmic tail 1 (KIR3DL1); killer cell lectin like receptor C1(KLRC1, NKG2A, CD159A); killer cell lectin like receptor D1 (KLRD1, CD94), killer cell lectin like receptor G1 (KLRG1; CLEC15A, MAFA, 2F1); sialic acid binding Ig like lectin 7 (SIGLEC7); and sialic acid binding Ig like lectin 9 (SIGLEC9). In some embodiments the compound provided herein is administered with one or more agonist or activators of one or more NK-cell stimulatory immune checkpoint proteins or receptors. Illustrative NK-cell stimulatory immune checkpoint proteins or receptors include CD16, CD226 (DNAM-1); CD244 (2B4, SLAMF4); killer cell lectin like receptor K1 (KLRK1, NKG2D, CD314); SLAM family member 7 (SLAMF7). See, e.g., Davis, et al.,Semin Immunol. (2017) 31:64-75; Fang, et al.,Semin Immunol. (2017) 31:37-54; and Chiossone, et al.,Nat Rev Immunol.(2018) 18(11):671-688.

In some embodiments the one or more immune checkpoint inhibitors comprises a proteinaceous (e.g., antibody or fragment thereof, or antibody mimetic) inhibitor of PD-L1 (CD274), PD-1 (PDCD1), CTLA4, or TIGIT. In some embodiments the one or more immune checkpoint inhibitors comprises a small organic molecule inhibitor of PD-L1 (CD274), PD-1 (PDCD1), CTLA4, or TIGIT. In some embodiments the one or more immune checkpoint inhibitors comprises a proteinaceous (e.g., antibody or fragment thereof, or antibody mimetic) inhibitor of LAG3.

Examples of inhibitors of LAG3 that can be co-administered include leramilimab (LAG525).

Inhibition of regulatory T-cell (Treg) activity or Treg depletion can alleviate their suppression of antitumor immune responses and have anticancer effects. See, e.g., Plitas and Rudensky,Annu. Rev. Cancer Biol. (2020) 4:459-77; Tanaka and Sakaguchi,Eur. J. Immunol.(2019) 49:1140-1146. In some embodiments, an compound provided herein is administered with one or more inhibitors of Treg activity or a Treg depleting agent. Treg inhibition or depletion can augment the effect of immune checkpoint inhibitors in cancer therapeutics.

In some embodiments an compound provided herein is administered with one or more Treg inhibitors. In some embodiments the Treg inhibitor can suppress the migration of Tregs into the tumor microenvironment. In some embodiments Treg inhibitor can reduce the immunosuppressive function of Tregs. In some embodiments, the Treg inhibitor can modulate the cellular phenotype and induce production of proinflammatory cytokines. Exemplary Treg inhibitors include without limitation, CCR4 (NCBI Gene ID: 1233) antagonists and degraders of Ikaros zinc-finger proteins (e.g., Ikaros (IKZF1; NCBI Gene ID: 10320), Helios (IKZF2; NCBI Gene ID: 22807), Aiolos (IKZF3; NCBI Gene ID: 22806), and Eos (IKZF4; NCBI Gene ID: 64375).

Examples of Helios degraders that can be co-administered include without limitation I-57 (Novartis) and compounds disclosed in WO2019038717, WO2020012334, WO20200117759, and WO2021101919.

In some embodiments an compound provided herein is administered with one or more Treg depleting agents. In some embodiments the Treg depleting agent is an antibody. In some embodiments the Treg depleting antibody has antibody-dependent cytotoxic (ADCC) activity. In some embodiments, the Treg depleting antibody is Fc-engineered to possess an enhanced ADCC activity. In some embodiments the Treg depleting antibody is an antibody-drug conjugate (ADC). Illustrative targets for Treg depleting agents include without limitation CD25 (IL2RA; NCBI Gene ID: 3559), CTLA4 (CD152; NCBI Gene ID: 1493); GITR (TNFRSF18; NCBI Gene ID: 8784); 4-1BB (CD137; NCBI Gene ID: 3604), OX-40 (CD134; NCBI Gene ID: 7293), LAG3 (CD223; NCBI Gene ID: 3902), TIGIT (NCBI Gene ID: 201633), CCR4 (NCBI Gene ID: 1233), and CCR8 (NCBI Gene ID: 1237).

Examples of Treg depleting anti-CCR4 antibodies that can be administered include mogamulizumab.

Inhibiting, depleting, or reprogramming of non-stimulatory myeloid cells in the tumor microenvironment can enhance anti-cancer immune responses (see, e.g., Binnewies et al.,Nat. Med. (2018) 24(5): 541-550; WO2016049641). Illustrative targets for depleting or reprogramming non-stimulatory myeloid cells include triggering receptors expressed on myeloid cells, TREM-1 (CD354, NCBI Gene ID: 54210) and TREM-2 (NCBI Gene ID: 54209). In some embodiments an compound provided herein is administered with one or more myeloid cell depleting or reprogramming agents, such as an anti-TREM-1 antibody (e.g. PY159; antibodies disclosed in WO2019032624) or an anti-TREM-2 antibody (e.g., PY314; antibodies disclosed in WO2019118513).

Cluster of Differentiation Agonists or Activators

Cluster of Differentiation 47 (CD47) Inhibitors

SIRPa Targeting Agents

In some embodiments the compound provided herein is administered with a FLT3R agonist. In some embodiments, the compound provided herein is administered with a FLT3 ligand. In some embodiments, the compound provided herein is administered with a FLT3L-Fc fusion protein, e.g., as described in WO2020263830. In some embodiments the compound provided herein is administered with GS-3583 or CDX-301. In some embodiments the compound provided herein is administered with GS-3583.

TNF Receptor Superfamily (TNFRSF) Member Agonists or Activators

Example anti-TNFRSF5 (CD40) antibodies that can be co-administered include RG7876, SEA-CD40, APX-005M, and ABBV-428.

In some embodiments the anti-TNFRSF17 (BCMA) antibody GSK-2857916 is co-administered.

In some embodiments the compound provided herein is administered with an ASK inhibitor, e.g., mitogen-activated protein kinase kinase kinase 5 (MAP3K5; ASK1, MAPKKK5, MEKK5; NCBI Gene ID: 4217). Examples of ASK1 inhibitors include those described in WO2011008709 (Gilead Sciences) and WO 2013112741 (Gilead Sciences).

In some embodiments the compound provided herein is administered with a targeted E3 ligase ligand conjugate. Such conjugates have a target protein binding moiety and an E3 ligase binding moiety (e.g., an inhibitor of apoptosis protein (IAP) (e.g., XIAP, c-IAP1, c-IAP2, NIL-IAP, Bruce, and surviving) E3 ubiquitin ligase binding moiety, Von Hippel-Lindau E3 ubiquitin ligase (VHL) binding moiety, a cereblon E3 ubiquitin ligase binding moiety, mouse double minute 2 homolog (MDM2) E3 ubiquitin ligase binding moiety), and can be used to promote or increase the degradation of targeted proteins, e.g., via the ubiquitin pathway. In some embodiments the targeted E3 ligase ligand conjugates comprise a targeting or binding moiety that targets or binds a protein described herein, and an E3 ligase ligand or binding moiety. In some embodiments the targeted E3 ligase ligand conjugates comprise a targeting or binding moiety that targets or binds a protein selected from Cbl proto-oncogene B (CBLB; Cbl-b, Nbla00127, RNF56; NCBI Gene ID: 868) and hypoxia inducible factor 1 subunit alpha (HIF1A; NCBI Gene ID: 3091). In some embodiments the targeted E3 ligase ligand conjugates comprise a kinase inhibitor (e.g., a small molecule kinase inhibitor, e.g., of BTK and an E3 ligase ligand or binding moiety. See, e.g., WO2018098280. In some embodiments the targeted E3 ligase ligand conjugates comprise a binding moiety targeting or binding to Interleukin-1 (IL-1) Receptor-Associated Kinase-4 (IRAK-4); Rapidly Accelerated Fibrosarcoma (RAF, such as c-RAF, A-RAF and/or B-RAF), c-Met/p38, or a BRD protein; and an E3 ligase ligand or binding moiety. See, e.g., WO2019099926, WO2018226542, WO2018119448, WO2018223909, WO2019079701. Additional targeted E3 ligase ligand conjugates that can be co-administered are described, e.g., in WO2018237026, WO2019084026, WO2019084030, WO2019067733, WO2019043217, WO2019043208, and WO2018144649.

RAS and RAS Pathway Inhibitors

Chemotherapeutic Agents

In some embodiments the compound provided herein is administered with a chemotherapeutic agent or anti-neoplastic agent.

Also included in the definition of “chemotherapeutic agent” are anti-hormonal agents such as anti-estrogens and selective estrogen receptor modulators (SERMs), inhibitors of the enzyme aromatase, anti-androgens, and pharmaceutically acceptable salts, acids or derivatives of any of the above that act to regulate or inhibit hormone action on tumors.

An example progesterone receptor antagonist includes onapristone. Additional progesterone targeting agents include TRI-CYCLEN LO (norethindrone+ethinyl estradiol), norgestimate+ethinylestradiol (Tri-Cyclen) and levonorgestrel.

In some embodiments the compound provided herein is administered with an anti-fibrotic agent. Anti-fibrotic agents that can be co-administered include the compounds such as beta-aminoproprionitrile (BAPN), as well as the compounds disclosed in U.S. Pat. No. 4,965,288 relating to inhibitors of lysyl oxidase and their use in the treatment of diseases and conditions associated with the abnormal deposition of collagen and U.S. Pat. No. 4,997,854 relating to compounds which inhibit LOX for the treatment of various pathological fibrotic states, which are herein incorporated by reference. Further exemplary inhibitors are described in U.S. Pat. No. 4,943,593 relating to compounds such as 2-isobutyl-3-fluoro-, chloro-, or bromo-allylamine, U.S. Pat. Nos. 5,021,456, 5,059,714, 5,120,764, 5,182,297, 5,252,608 relating to 2-(1-naphthyloxymemyl)-3-fluoroallylamine, and US 20040248871, which are herein incorporated by reference.

Exemplary anti-fibrotic agents also include the primary amines reacting with the carbonyl group of the active site of the lysyl oxidases, and more particularly those which produce, after binding with the carbonyl, a product stabilized by resonance, such as the following primary amines: emylenemamine, hydrazine, phenylhydrazine, and their derivatives; semicarbazide and urea derivatives; aminonitriles such as BAPN or 2-nitroethylamine; unsaturated or saturated haloamines such as 2-bromo-ethylamine, 2-chloroethylamine, 2-trifluoroethylamine, 3-bromopropylamine, and p-halobenzylamines; and selenohomocysteine lactone.

Other anti-fibrotic agents are copper chelating agents penetrating or not penetrating the cells. Exemplary compounds include indirect inhibitors which block the aldehyde derivatives originating from the oxidative deamination of the lysyl and hydroxylysyl residues by the lysyl oxidases. Examples include the thiolamines, particularly D-penicillamine, and its analogs such as 2-amino-5-mercapto-5-methylhexanoic acid, D-2-amino-3-methyl-3-((2-acetamidoethyl)dithio)butanoic acid, p-2-amino-3-methyl-3-((2-aminoethyl)dithio)butanoic acid, sodium-4-((p-1-dimethyl-2-amino-2-carboxyethyl)dithio)butane sulphurate, 2-acetamidoethyl-2-acetamidoethanethiol sulphanate, and sodium-4-mercaptobutanesulphinate trihydrate.

Examples of inhibitors of prostaglandin-endoperoxide synthase 1 (PTGS1, COX-1; NCBI Gene ID: 5742) that can be co-administered include mofezolac, GLY-230, and TRK-700.

Examples of inhibitors of arachidonate 5-lipoxygenase (ALOX5, 5-LOX; NCBI Gene ID: 240) that can be co-administered include meclofenamate sodium, zileuton.

Examples of inhibitors of soluble epoxide hydrolase 2 (EPHX2, SEH; NCBI Gene ID: 2053) that can be co-administered include compounds described in WO2015148954. Dual inhibitors of COX-2/SEH that can be co-administered include compounds described in WO2012082647. Dual inhibitors of SEH and fatty acid amide hydrolase (FAAH; NCBI Gene ID: 2166) that can be co-administered include compounds described in WO2017160861.

Tumor Oxygenation Agents

In some embodiments the compound provided herein is administered with an agent that promotes or increases tumor oxygenation or reoxygenation, or prevents or reduces tumor hypoxia. Illustrative agents that can be co-administered include, e.g., Hypoxia inducible factor-1 alpha (HIF-1α) inhibitors, such as PT-2977, PT-2385; VEGF inhibitors, such as bevasizumab, IMC-3C5, GNR-011, tanibirumab, LYN-00101, ABT-165; and/or an oxygen carrier protein (e.g., a heme nitric oxide and/or oxygen binding protein (HNOX)), such as OMX-302 and HNOX proteins described in WO2007137767, WO2007139791, WO2014107171, and WO2016149562.

Immunotherapeutic Agents

The exemplified therapeutic antibodies can be further labeled or combined with a radioisotope particle such as indium-111, yttrium-90 (90Y-clivatuzumab), or iodine-131.

Illustrative therapeutic agents (e.g., anticancer or antineoplastic agents) that can be conjugated to the drug-conjugated antibodies, fragments thereof, or antibody mimetics include without limitation monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), a calicheamicin, ansamitocin, maytansine or an analog thereof (e.g., mertansine/emtansine (DM1), ravtansine/soravtansine (DM4)), an anthracyline (e.g., doxorubicin, daunorubicin, epirubicin, idarubicin), pyrrolobenzodiazepine (PBD) DNA cross-linking agent SC-DR002 (D6.5), duocarmycin, a microtubule inhibitors (MTI) (e.g., a taxane, a vinca alkaloid, an epothilone), a pyrrolobenzodiazepine (PBD) or dimer thereof, a duocarmycin (A, B1, B2, C1, C2, D, SA, CC-1065), and other anticancer or anti-neoplastic agents described herein. In some embodiments, the therapeutic agent conjugated to the drug-conjugated antibody is a topoisomerase I inhibitor (e.g., a camptothecin analog, such as irinotecan or its active metabolite SN38). In some embodiments, the therapeutic agents (e.g., anticancer or antineoplastic agents) that can be conjugated to the drug-conjugated antibodies, fragments thereof, or antibody mimetics include an immune checkpoint inhibitor. In some embodiments the conjugated immune checkpoint inhibitor is a conjugated small molecule inhibitor of CD274 (PDL1, PD-L1), programmed cell death 1 (PDCD1, PD1, PD-1) or CTLA4. In some embodiments the conjugated small molecule inhibitor of CD274 or PDCD1 is selected from the group consisting of GS-4224, GS-4416, INCB086550 and MAX10181. In some embodiments the conjugated small molecule inhibitor of CTLA4 comprises BPI-002.

In some embodiments the ADCs that can be co-administered include an antibody targeting carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1; CD66a; NCBI Gene ID: 634). In some embodiments the CEACAM1 antibody is hMN-14 (e.g., as described in WO1996011013). In some embodiments the CEACAM1-ADC is as described in WO2010093395 (anti-CEACAM-1-CL2A-SN38). In some embodiments the compound provided herein is administered with the CEACAM1-ADC IMMU-130.

In some embodiments the ADCs that can be co-administered include an antibody targeting MHC class II cell surface receptor encoded by the human leukocyte antigen complex (HLA-DR). In some embodiments the HLA-DR antibody is hL243 (e.g., as described in WO2006094192). In some embodiments the HLA-DR-ADC is as described in WO2010093395 (anti-HLA-DR-CL2A-SN38). In some embodiments the compound provided herein is administered with the HLA-DR-ADC IMMU-140.

Cancer Gene Therapy and Cell Therapy

In some embodiments the compound provided herein is administered with a cancer gene therapy and cell therapy. Cancer gene therapies and cell therapies include the insertion of a normal gene into cancer cells to replace a mutated or altered gene; genetic modification to silence a mutated gene; genetic approaches to directly kill the cancer cells; including the infusion of immune cells designed to replace most of the patient's own immune system to enhance the immune response to cancer cells, or activate the patient's own immune system (T cells or Natural Killer cells) to kill cancer cells, or find and kill the cancer cells; genetic approaches to modify cellular activity to further alter endogenous immune responsiveness against cancer.

Cellular Therapies

In some embodiments the compound provided herein is administered with one or more cellular therapies. Illustrative cellular therapies include without limitation co-administration of one or more of a population of natural killer (NK) cells, NK-T cells, T cells, cytokine-induced killer (CIK) cells, macrophage (MAC) cells, tumor infiltrating lymphocytes (TILs) and/or dendritic cells (DCs). In some embodiments, the cellular therapy entails a T cell therapy, e.g., co-administering a population of alpha/beta TCR T cells, gamma/delta TCR T cells, regulatory T (Treg) cells and/or TRuC™ T cells. In some embodiments, the cellular therapy entails a NK cell therapy, e.g., co-administering NK-92 cells. As appropriate, a cellular therapy can entail the co-administration of cells that are autologous, syngeneic or allogeneic to the subject.

In some embodiments the cellular therapy entails co-administering cells comprising chimeric antigen receptors (CARs). In such therapies, a population of immune effector cells engineered to express a CAR, wherein the CAR comprises a tumor antigen-binding domain. In T cell therapies, the T cell receptors (TCRs) are engineered to target tumor derived peptides presented on the surface of tumor cells.

With respect to the structure of a CAR, in some embodiments, the CAR comprises an antigen binding domain, a transmembrane domain, and an intracellular signaling domain. In some embodiments, the intracellular domain comprises a primary signaling domain, a costimulatory domain, or both of a primary signaling domain and a costimulatory domain. In some embodiments, the primary signaling domain comprises a functional signaling domain of one or more proteins selected from the group consisting of CD3 zeta, CD3 gamma, CD3 delta, CD3 epsilon, common FcR gamma (FCERIG), FcR beta (Fc Epsilon Rlb), CD79a, CD79b, Fcgamma RIIa, DAP10, and DAP12.

Exemplified Combination Therapies

Lymphoma or Leukemia Combination Therapy

One modified approach is radioimmunotherapy, wherein a monoclonal antibody is combined with a radioisotope particle, such as indium-111, yttrium-90, and iodine-131. Examples of combination therapies include, but are not limited to, iodine-131 tositumomab (BEXXAR®), yttrium-90 ibritumomab tiuxetan (ZEVALIN®), and BEXXAR® with CHOP.

The abovementioned therapies can be supplemented or combined with stem cell transplantation or treatment. Therapeutic procedures include peripheral blood stem cell transplantation, autologous hematopoietic stem cell transplantation, autologous bone marrow transplantation, antibody therapy, biological therapy, enzyme inhibitor therapy, total body irradiation, infusion of stem cells, bone marrow ablation with stem cell support, in vitro-treated peripheral blood stem cell transplantation, umbilical cord blood transplantation, immunoenzyme technique, low-LET cobalt-60 gamma ray therapy, bleomycin, conventional surgery, radiation therapy, and nonmyeloablative allogeneic hematopoietic stem cell transplantation.

Non-Hodgkin's Lymphomas Combination Therapy

Treatment of non-Hodgkin's lymphomas (NHL), especially those of B cell origin, includes using monoclonal antibodies, standard chemotherapy approaches (e.g., CHOP (cyclophosphamide, doxorubicin, vincristine, and prednisone), CVP (cyclophosphamide, vincristine, and prednisone), FCM (fludarabine, cyclophosphamide, and mitoxantrone), MCP (Mitoxantrone, Chlorambucil, Prednisolone), all optionally including rituximab (R) and the like), radioimmunotherapy, and combinations thereof, especially integration of an antibody therapy with chemotherapy.

Mantle Cell Lymphoma Combination Therapy

Therapeutic treatments for mantle cell lymphoma (MCL) include combination chemotherapies such as CHOP, hyperCVAD, and FCM. These regimens can also be supplemented with the monoclonal antibody rituximab to form combination therapies R-CHOP, hyperCVAD-R, and R-FCM. Any of the abovementioned therapies may be combined with stem cell transplantation or ICE in order to treat MCL.

An alternative approach to treating MCL is immunotherapy. One immunotherapy uses monoclonal antibodies like rituximab. Another uses cancer vaccines, such as GTOP-99, which are based on the genetic makeup of an individual patient's tumor.

A modified approach to treat MCL is radioimmunotherapy, wherein a monoclonal antibody is combined with a radioisotope particle, such as iodine-131 tositumomab (BEXXAR®) and yttrium-90 ibritumomab tiuxetan (ZEVALIN®). In another example, BEXXAR® is used in sequential treatment with CHOP.

Other approaches to treating MCL include autologous stem cell transplantation coupled with high-dose chemotherapy, administering proteasome inhibitors such as bortezomib (VELCADE® or PS-341), or administering antiangiogenesis agents such as thalidomide, especially in combination with rituximab.

Another treatment approach is administering drugs that lead to the degradation of Bcl-2 protein and increase cancer cell sensitivity to chemotherapy, such as oblimersen, in combination with other chemotherapeutic agents.

A further treatment approach includes administering mTOR inhibitors, which can lead to inhibition of cell growth and even cell death. Non-limiting examples are sirolimus, temsirolimus (TORISEL®, CCI-779), CC-115, CC-223, SF-1126, PQR-309 (bimiralisib), voxtalisib, GSK-2126458, and temsirolimus in combination with RITUXAN®, VELCADE®, or other chemotherapeutic agents.

Examples of therapeutic procedures used to treat WM include peripheral blood stem cell transplantation, autologous hematopoietic stem cell transplantation, autologous bone marrow transplantation, antibody therapy, biological therapy, enzyme inhibitor therapy, total body irradiation, infusion of stem cells, bone marrow ablation with stem cell support, in vitro-treated peripheral blood stem cell transplantation, umbilical cord blood transplantation, immunoenzyme techniques, low-LET cobalt-60 gamma ray therapy, bleomycin, conventional surgery, radiation therapy, and nonmyeloablative allogeneic hematopoietic stem cell transplantation.

Diffuse Large B-cell Lymphoma (DLBCL) Combination Therapy

Chronic Lymphocytic Leukemia Combination Therapy

Therapeutic agents used to treat chronic lymphocytic leukemia (CLL) include chlorambucil, cyclophosphamide, fludarabine, pentostatin, cladribine, doxorubicin, vincristine, prednisone, prednisolone, alemtuzumab, many of the agents listed for WM, and combination chemotherapy and chemoimmunotherapy, including the following common combination regimens: CVP, R-CVP, ICE, R-ICE, FCR, and FR.

High Risk Myelodysplastic Syndrome (HR MDS) Combination Therapy

Low Risk Myelodysplastic Syndrome (LR MDS) Combination Therapy

Therapeutic agents used to treat LR MDS include lenalidomide, azacytidine, and combinations thereof. In some embodiments therapeutic agents used to treat LR MDS include roxadustat, luspatercept, imetelstat, LB-100, or rigosertib.

High Risk Myelodysplastic Syndrome (HR MDS) Combination Therapy

Low Risk Myelodysplastic Syndrome (LR MDS) Combination Therapy

Therapeutic agents used to treat LR MDS include lenalidomide, azacytidine, and combinations thereof. In some embodiments therapeutic agents used to treat LR MDS include roxadustat, luspatercept, imetelstat, LB-100, or rigosertib.

Acute Myeloid Leukemia (AML) Combination Therapy

Multiple Myeloma (MM) Combination Therapy

Breast Cancer Combination Therapy

Triple Negative Breast Cancer (TNBC) Combination Therapy

Bladder Cancer Combination Therapy

Colorectal Cancer (CRC) Combination Therapy

Esophageal and Esophagogastric Junction Cancer Combination Therapy

Therapeutic agents used to treat esophageal and esophagogastric junction cancer include capecitabine, carboplatin, cisplatin, docetaxel, epirubicin, fluoropyrimidine, fluorouracil, irinotecan, leucovorin, oxaliplatin, paclitaxel, ramucirumab, trastuzumab, and any combinations thereof. In some embodiments therapeutic agents used to treat gastroesophageal junction cancer (GEJ) include herceptin, cisplatin, 5-FU, ramicurimab, or paclitaxel. In some embodiments therapeutic agents used to treat GEJ cancer include ALX-148, AO-176, or IBI-188.

Gastric Cancer Combination Therapy

Head and Neck Cancer Combination Therapy

Non-Small Cell Lung Cancer Combination Therapy

Small Cell Lung Cancer Combination Therapy

Ovarian Cancer Combination Therapy

Pancreatic Cancer Combination Therapies

Endometrial Cancer Combination Therapies

Therapeutic treatments used to treat endometrial cancer include surgery, chemotherapy, radiation therapy, hormone therapy, targeted therapy, and immunotherapy. In some embodiments, the therapeutic treatments include Anti-angiogenesis therapy, Mammalian target of rapamycin (mTOR) inhibitors, Targeted therapy to treat a rare type of uterine cancer.

Prostate Cancer Combination Therapies

Additional Exemplified Combination Therapies

In some embodiments the compound provided herein is administered with one or more therapeutic agents selected from a PI3K inhibitor, a Trop-2 binding agent, CD47 antagonist, a SIRPα antagonist, a FLT3R agonist, a PD-1 antagonist, a PD-L1 antagonist, an MCL1 inhibitor, a CCR8 binding agent, an HPK1 antagonist, a DGKa inhibitor, a CISH inhibitor, a PARP-7 inhibitor, a Cbl-b inhibitor, a KRAS inhibitor (e.g., a KRAS G12C or G12D inhibitor), a KRAS degrader, a beta-catenin degrader, a helios degrader, a CD73 inhibitor, an adenosine receptor antagonist, a TIGIT antagonist, a TREM1 binding agent, a TREM2 binding agent, a CD137 agonist, a GITR binding agent, an OX40 binding agent, and a CAR-T cell therapy.

In some embodiments, the present disclosure provides processes and intermediates useful for preparing the compounds disclosed herein or pharmaceutically acceptable salts thereof.

Compounds disclosed herein can be purified by any of the means known in the art, including chromatographic means, including but not limited to high-performance liquid chromatography (HPLC), preparative thin layer chromatography, flash column chromatography, and ion exchange chromatography. Any suitable stationary phase can be used, including but not limited to, normal and reversed phases as well as ionic resins. In some embodiments, the disclosed compounds are purified via silica gel and/or alumina chromatography.

During any of the processes for preparation of the compounds provided herein, it can be necessary and/or desirable to protect sensitive or reactive groups on any of the molecules concerned. This can be achieved by means of conventional protecting groups as described in standard works, such as T. W. Greene and P. G. M. Wuts, PROTECTIVEGROUPS INORGANICSYNTHESIS,4thed., Wiley, New York 2006. The protecting groups can be removed at a convenient subsequent stage using methods known from the art.

Exemplary chemical entities useful in methods of the embodiments will now be described by reference to illustrative synthetic schemes for their general preparation herein and the specific examples that follow. Skilled artisans will recognize that, to obtain the various compounds herein, starting materials can be suitably selected so that the ultimately desired substituents will be carried through the reaction scheme with or without protection as appropriate to yield the desired product. Alternatively, it can be necessary or desirable to employ, in the place of the ultimately desired substituent, a suitable group that can be carried through the reaction scheme and replaced as appropriate with the desired substituent. Furthermore, one of skill in the art will recognize that the transformations shown in the schemes below can be performed in any order that is compatible with the functionality of the particular pendant groups.

The methods of the present disclosure generally provide a specific enantiomer or diastereomer as the desired product, although the stereochemistry of the enantiomer or diastereomer was not determined in all cases. When the stereochemistry of the specific stereocenter in the enantiomer or diastereomer is not determined, the compound is drawn without showing any stereochemistry at that specific stereocenter even though the compound can be substantially enantiomerically or diastereomerically pure.

Compounds disclosed herein can be prepared from commercially available reagents using the synthetic methods and reaction schemes described herein, or using other reagents and conventional methods known to persons of ordinary skill in the art. For instance, representative syntheses of compounds of the present disclosure are described in the schemes below, and the particular examples that follow.

EXAMPLES

Certain abbreviations and acronyms are used in describing the experimental details. Although most of these would be understood by one skilled in the art, Table 1 contains a list of many of these abbreviations and acronyms.

A vigorously stirred mixture of Intermediate 17-9 (50.0 mg, 86.4 μmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (prepared according to WO 2021/041671) (66.4 mg, 130 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (9.5 mg, 13 μmol), cesium carbonate (141 mg, 432 μmol), water (0.4 mL), and 1,4-dioxane (1.0 mL) was heated to 115° C. After 90 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified on C18 reverse phase silica gel (20% to 87% acetonitrile in water) to give Intermediate 2-1. LCMS: 929.5.

Intermediate 3-1 was synthesized in a manner similar to Intermediate 5-6 using 1-bromo-3-(trifluoromethoxy)benzene instead of 4-bromo-2-chloro-1-fluorobenzene. LCMS: 367.4 [M−CH3O]+.

A vigorously stirred mixture of Intermediate 17-9 (50.0 mg, 86.4 μmol), ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (prepared according to WO 2021/041671) (58.6 mg, 130 μmol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (12.6 mg, 17.3 μmol), aqueous potassium phosphate solution (1.5 M, 288 μL, 432 μmol), and tetrahydrofuran (0.5 mL) was heated to 70° C. After 80 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified on C18 reverse phase silica gel (20% to 87% acetonitrile in water) to give Intermediate 4-1. LCMS: 869.4.

To a stirred solution of 4-bromo-2-chloro-1-fluorobenzene (10 g, 47.746 mmol) in THF (140 mL), was added LDA (2M in THF, 28.65 mL, 57.296 mmol) at −78° C. and stirred at same temp for 45 min. Reaction mixture was purged with CO2gas for 30 min at −78° C. The resulting mixture was warmed to room temperature over 1 hour. After completion of the reaction, the reaction mixture was quenched with 2M aq. HCl (30 mL) and extracted with ethyl acetate (2×100 mL). The combined organic layer was washed with brine (100 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to yield Intermediate 5-1.1H NMR (400 MHz, DMSO-d6) δ 14.44 (br s, 1H), 7.66-7.57 (m, 2H).

To a stirred solution of Intermediate 5-1 (8 g, 31.564 mmol) in toluene (200 mL) and MeOH (58.4 mL), was added TMS diazomethane (23.67 mL, 47.346 mmol) at room temperature. The resulting mixture was stirred at ambient temperature for 2 h. After completion of the reaction, reaction mass was quenched with acetic acid (1.44 mL, 25.251 mmol) and extracted with ethyl acetate (2×90 mL). The combined organic layer was washed with brine (70 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to get the crude. The crude material was subjected to silica gel (100-200 mesh) column chromatography eluting with 8%-10% ethyl acetate in petroleum ether to yield Intermediate 5-2.1H NMR (400 MHz, DMSO-d6) δ 7.73 (t, J=8.8 Hz, 1H), 7.64 (dd, J=1.2, 8.8 Hz, 1H), 3.94 (s, 3H).

A solution of methyl Intermediate 5-3 (2.5 g, 8.659 mmol) in 2-Me-THF (25 mL) was heated under stirring at 70° C. and lithium bis(trimethylsilyl)amide (1.0 M in THF, 21.6 mL, 21.648 mmol) was added at same temperature. The resulting mixture was stirred at 70° C. for 45 min. After completion of the reaction, reaction mass was quenched with citric acid (4.12 g, 21.647 mmol), diethyl ether (50 mL) and water (25 mL) were added sequentially. The organic layer was separated, dried over sodium sulfate, filtered and concentrated under reduced pressure to get the crude. The crude material obtained was purified by silica gel (100-200 mesh) column chromatography eluting with 15% ethyl acetate in petroleum ether to yield Intermediate 5-4. LCMS: 257.2.

To a solution of Intermediate 5-4 (1.6 g, 6.233 mmol) in 2-Me-THF (44.8 mL), was added sodium bi(trimethylsilyl)amide (2 M in THF, 3.42 mL, 6.857 mmol) at 0° C. and stirred for 10 min. N-phenyltrifluoromethanesulfonimide (2.45 g, 6.857 mmol) was added at 0° C. and stirred at same temperature for 1 h. After completion of the reaction, reaction mass was quenched with saturated aqueous sodium bicarbonate (20 mL) and added diethyl ether (75 mL) and ethyl acetate (25 mL) sequentially. The combined organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give Intermediate 5-5. LCMS: 387.1 [M−H]−.

To a stirred solution of Intermediate 5-5 (2.42 g, 6.233 mmol) in 1,4-dioxane (25 mL), were added 4,4,4′,4′,5,5,5′,5′-Octamethyl-2,2′-bi(1,3,2-dioxaborolane) (2.37 g, 9.349 mmol) and potassium acetate (3.05 g, 31.165 mmol). Reaction mixture was degassed with argon gas for 10 min. [1,1′-Bis(diphenylphosphino) ferrocene]dichloropalladium(II) (456 mg, 0.623 mmol) was added and resulting mixture was heated to 100° C. under stirring for 1.5 h. After completion of the reaction, reaction mass was filtered through a pad of celite and concentrated under reduced pressure to get the crude material. The crude material was purified by silica gel (100-200 mesh) column chromatography eluting with 10˜12% ethyl acetate in petroleum ether to yield Intermediate 5-6. LCMS: 367.3.

Isopropylmagnesium chloride lithium chloride complex solution (1.3 M in tetrahydrofuran, 6.50 mL, 8.45 mmol) was added over 5 min via syringe pump to a stirred solution of Intermediate 7-1 (2.55 g, 8.05 mmol) in tetrahydrofuran (4.0 mL) at −40° C. After 35 min, copper(I) cyanide di(lithium chloride) complex solution (1.0 M in tetrahydrofuran, 403 μL, 400 μmol) was added via syringe. After 2 min, 3-chloro-2-(methoxymethoxy)prop-1-ene (1.15 mL, 8.86 mmol) was added over 1 min via syringe, and the resulting mixture was warmed to 10° C. over 120 min. Saturated aqueous ammonium chloride solution (5 mL), aqueous ammonia solution (28% wt, 15 mL), diethyl ether (100 mL), and ethyl acetate (25 mL) were added sequentially. The organic layer was washed with water (2×75 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 7% ethyl acetate in hexanes) to give Intermediate 7-2. LCMS: 339.1.

A solution of Intermediate 7-2 (2.37 g, 7.01 mmol) in 2-methyltetrahydrofuran (30 mL) was added over 60 min via syringe pump to a vigorously stirred mixture of lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 17.5 mL, 18 mmol) and 2-methyltetrahydrofuran (110 mL) at 70° C. After 130 min, the resulting mixture was cooled to room temperature, acetic acid (1.20 mL, 21.0 mmol) was added via syringe, and the resulting mixture was concentrated under reduced pressure. Methanol (50 mL) and acetic acid (4.01 mL, 70.1 mmol) were added sequentially. After 20 min, the resulting mixture was concentrated under reduced pressure, and diethyl ether (200 mL), ethyl acetate (25 mL), and saturated aqueous sodium bicarbonate solution (50 mL) were added sequentially. The organic layer was washed with water (150 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 15% ethyl acetate in hexanes) to give Intermediate 7-3. LCMS: 307.1.

Sodium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 6.33 mL, 6.33 mmol) was added over 1 min via syringe to a stirred solution of Intermediate 7-3 (1.76 g, 5.75 mmol) in 2-methyltetrahydrofuran (15 mL) at 0° C. After 5 min, N-phenyl-bis(trifluoromethanesulfonimide) (2.26 g, 6.33 mmol) was added. After 40 min, diethyl ether (200 mL), ethyl acetate (25 mL), and saturated aqueous sodium bicarbonate solution (25 mL) were added sequentially. The organic layer was washed sequentially with water (200 mL) and a mixture of water and saturated aqueous sodium bicarbonate solution (4:1 v:v, 200 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure give Intermediate 7-4. LCMS: 439.0.

A vigorously stirred mixture of Intermediate 7-4 (2.52 g, 5.75 mmol), bis(pinacolato)diboron (2.19 g, 8.63 mmol), potassium acetate (2.82 g, 28.8 mmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (421 mg, 575 μmol), and 1,4-dioxane (12 mL) was heated to 100° C. After 90 min, the resulting mixture was cooled to room temperature and was filtered through celite. The filter cake was extracted with ethyl acetate (60 mL), and the combined filtrates were concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 10% ethyl acetate in hexanes) to give Intermediate 7-5. LCMS: 417.1.

Intermediate 8-1 was synthesized in a manner similar to Intermediate 5-6 using 6-bromo-2-chloro-3-fluorobenzoic acid instead of Intermediate 5-1. LCMS: 367.3.

Intermediate 11-1 was synthesized in a manner similar to Intermediate 7-5 using 6-bromo-2,3,4-trifluoro-benzoic acid instead of 6-bromo-3-fluoro-2-(trifluoromethoxy)benzoic acid. LCMS: 369.1.

2,2,6,6-Tetramethylpiperidinylmagnesium chloride lithium chloride complex solution (1.0 M in tetrahydrofuran/toluene, 6.88 mL, 6.9 mmol) was added over 10 min via syringe pump to a vigorously stirred solution of ethyl 2,3,4,5-tetrafluorobenzoate (1.00 mL, 6.26 mmol) in tetrahydrofuran (3.5 mL) at −40° C., and the resulting mixture was warmed to −20° C. After 103 min, copper(I) cyanide di(lithium chloride) complex solution (1.0 M in tetrahydrofuran, 313 μL, 310 μmol) was added via syringe. After 1 min, 3-chloro-2-(methoxymethoxy)prop-1-ene (895 μL, 6.88 mmol) was added over 1 min via syringe, and the resulting mixture was warmed to 10° C. over 95 min. The resulting mixture was warmed to room temperature. After 60 min, saturated aqueous ammonium chloride solution (10 mL), aqueous ammonia solution (28% wt, 10 mL), diethyl ether (200 mL), and ethyl acetate (25 mL) were added sequentially. The organic layer was washed with water (2×150 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 5% ethyl acetate in hexanes) to give Intermediate 12-1. LCMS: 323.1.

Sodium methoxide solution (25% wt in methanol, 4.54 mL, 20 mmol) was added over 15 min via syringe pump to a vigorously stirred solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (5.01 g, 19.8 mmol) in 2-methyltetrahydrofuran (70 mL) at −20° C. After 11 min, ethanethiol (4.41 mL, 59.5 mmol) was added over 1 min via syringe. After 1 min, N,N-diisopropylethylamine (11.1 mL, 63.5 mmol) was added over 2 min via syringe. After 11 min, the resulting mixture was warmed to room temperature. After 20 min, the resulting mixture was heated to 70° C. After 22 h, the resulting mixture was cooled to room temperature, and citric acid (3.0 g), diethyl ether (200 mL), and ethyl acetate (25 mL) were added sequentially. The organic layer was washed with water (200 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 11% ethyl acetate in hexanes) to give Intermediate 13-1. LCMS: 274.0.

2,2,6,6-Tetramethylpiperidinylmagnesium chloride lithium chloride complex solution (1.0 M in tetrahydrofuran, 14.4 mL, 14 mmol) was added over 20 min via syringe pump to a vigorously stirred solution of Intermediate 13-1 (1.00 g, 3.65 mmol) in tetrahydrofuran (3.0 mL) at 0° C. After 60 min, a solution of 1,2-dibromo-1,1,2,2-tetrachloroethane (4.76 g, 14.6 mmol) in tetrahydrofuran (8.0 mL) was added via syringe. After 120 min, citric acid (5.0 g), diethyl ether (200 mL), and ethyl acetate (25 mL) were added sequentially. The organic layer was washed with water (2×150 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 5% ethyl acetate in hexanes) to give Intermediate 13-2. LCMS: 351.9.

Sodium iodide (1.90 g, 12.7 mmol) was added to a vigorously stirred solution of Intermediate 13-2 (895 mg, 2.54 mmol) in acetic acid (12.0 mL) at room temperature, and the resulting mixture was heated to 80° C. After 2.5 h, the resulting mixture was cooled to room temperature, and ethyl acetate (100 mL) and aqueous sodium thiosulfate solution (1.0 M, 2.0 mL) were added sequentially. The organic layer was washed with water (100 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 13-3. LCMS: 337.9.

tert-Butyldimethylsilyl chloride (2.44 g, 16.2 mmol) was added to a stirred mixture of Intermediate 13-4 (2.61 g, 10.8 mmol), 4-(dimethylamino)pyridine (132 mg, 1.08 mmol), triethylamine (3.00 mL, 21.6 mmol), and dichloromethane (70 mL) at room temperature. After 15 h, diethyl ether (200 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (150 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 70% ethyl acetate in hexanes) to give Intermediate 13-5. LCMS: 357.2.

N,N-Diisopropylethylamine (884 μL, 5.08 mmol) was added via syringe to a mixture of Intermediate 13-3 (839 mg, 2.48 mmol) and phosphorous (V) oxychloride (10 mL) at room temperature, and the resulting mixture was heated to 80° C. After 10 min, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. Dichloromethane (20 mL) was added, the resulting mixture was cooled to 0° C., and N,N-diisopropylethylamine (1.33 mL, 7.61 mmol) and a solution of Intermediate 13-5 (905 mg, 2.54 mmol) in dichloromethane (4.0 mL) were added sequentially. After 50 min, citric acid (2.0 g), diethyl ether (100 mL), and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (2×75 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 13% ethyl acetate in hexanes) to give Intermediate 13-6. LCMS: 676.1.

Tetrabutylammonium fluoride solution (1.0 M in tetrahydrofuran, 8.18 mL, 8.2 mmol) was added over 2 min via syringe to a stirred solution of Intermediate 13-6 (1.39 g, 2.05 mmol) in tetrahydrofuran (110 mL) at 0° C., and the resulting mixture was warmed to room temperature. After 23 h, saturated aqueous ammonium chloride solution (20 mL) and diethyl ether (400 mL) were added sequentially. The organic layer was washed with water (2×400 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 35% ethyl acetate in hexanes) to give Intermediate 13-7. LCMS: 482.1.

A vigorously stirred mixture of Intermediate 13-7 (722 mg, 1.50 mmol), ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (prepared according to, for instance, WO 2021/041671) (768 mg, 1.50 mmol), aqueous potassium phosphate solution (1.5 M, 4.99 mL, 7.5 mmol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (218 mg, 300 μmol), and tetrahydrofuran (8.0 mL) was heated to 70° C. After 105 min, the resulting mixture was cooled to room temperature and diethyl ether (100 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (60 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 45% ethyl acetate in hexanes) to give Intermediate 13-8. LCMS: 832.4.

3-Chloroperoxybenzoic acid (77% wt, 524 mg, 2.3 mmol) was added in two equal portions over 5 min to a vigorously stirred solution of Intermediate 13-8 (884 mg, 1.06 mmol) in dichloromethane (8.0 mL) at 0° C. After 25 min, the resulting mixture was warmed to room temperature. After 60 min, diethyl ether (100 mL), ethyl acetate (20 mL), and aqueous sodium thiosulfate solution (1.0 M, 8.0 mL) were added sequentially. The organic layer was washed sequentially with water (60 mL), a mixture of water and saturated aqueous sodium bicarbonate solution (7:1 v:v, 80 mL), and water (80 mL); was dried over anhydrous magnesium sulfate; was filtered; and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 75% ethyl acetate in hexanes) to give Intermediate 13-9. LCMS: 864.4.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 46.3 μL, 46 μmol) was added over 1 min via syringe to a stirred mixture of Intermediate 13-9 (20.0 mg, 23.1 μmol), Intermediate 13-14 (7.9 mg, 46 μmol), and tetrahydrofuran (0.5 mL) at room temperature. After 10 min, diethyl ether (40 mL), ethyl acetate (20 mL), and saturated aqueous sodium bicarbonate solution (5 mL) were added sequentially. The organic layer was washed with water (30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 13-10. LCMS: 941.4.

Cesium fluoride (84.4 mg, 556 μmol) was added to a vigorously stirred solution of Intermediate 13-10 (21.8 mg, 23.1 μmol) in N,N-dimethylformamide (0.5 mL) at room temperature. After 30 min, diethyl ether (40 mL), ethyl acetate (20 mL), and saturated aqueous sodium bicarbonate solution (10 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 13-11. LCMS: 785.3.

O1-tert-butyl O2-methyl (2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylate (25 g, 100 mmol) and 3-chloro-2-chloromethyl-1-propene (17 mL, 160 mmol) were dissolved in tetrahydrofuran (100 mL) in an oven dried flask under nitrogen atmosphere. The solution was cooled to −78° C. and lithium bis(trimethylsilyl)amide (1 M in tetrahydrofuran, 120 mL, 120 mmol) was added dropwise via syringe over 20 minutes. The resulting solution was allowed to warm slowly to room temperature and was stirred for an additional 72 hours. The pH of the solution was adjusted to 2 with 2N hydrochloric acid. Brine (100 mL) was added and the mixture was extracted with ethyl acetate (3×100 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0% to 50%) to afford Intermediate 13-12. LCMS [M+Na]+: 358.2.

To a flask charged with Intermediate 13-12 (22 g, 66 mmol) was added hydrogen chloride (4 M in 1,4-dioxane, 90 mL, 361 mmol) and the resulting solution was stirred at room temperature for 2 h. The solution was concentrated in vacuo and taken up into N,N-dimethylformamide (330 mL). Potassium iodide (1.1 g, 6.6 mmol) and potassium carbonate (27 g, 200 mmol) were added and the resulting mixture was stirred for 1 h at room temperature. Brine (300 mL) was added and the pH was adjusted to 10 with saturated aqueous sodium carbonate. The mixture was extracted with dichloromethane (3×300 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel using a gradient of methanol in dichloromethane (0 to 30%) to afford Intermediate 13-13. LCMS: 200.2.

A solution of Intermediate 13-13 (11 g, 54 mmol) in tetrahydrofuran (40 mL) was cooled to 0° C. under nitrogen atmosphere and lithium aluminum hydride (1 M in tetrahydrofuran, 108 mL, 108 mmol) was added dropwise via syringe. The resulting solution was allowed to warm to room temperature and stirred overnight. The solution was diluted with diethyl ether (40 mL) and cooled to 0° C. Water (4.1 mL) was added dropwise followed by sodium hydroxide (15% aqueous solution, 4.1 mL) and additional water (12.3 mL). The mixture was stirred vigorously at room temperature for 15 minutes. Magnesium sulfate was added and the mixture was stirred vigorously for an additional 15 minutes. The mixture was filtered, rinsing with diethyl ether, and concentrated in vacuo to give Intermediate 13-14. LCMS: 172.2.

Intermediate 14-1 was synthesized in a manner similar to Intermediate 5-6 using 2-bromo-3-fluoro-6-iodobenzoic acid instead of Intermediate 5-1. LCMS: 299.1 [M−H]−.

(2-(Chloromethoxy)ethyl)trimethylsilane (586 μL, 3.31 mmol) was added over 3 min via syringe to a vigorously stirred mixture of Intermediate 14-1 (949 mg, 3.15 mmol), N,N-diisopropylethylamine (1.10 mL, 6.30 mmol), and dichloromethane (3.5 mL) at 0° C. After 60 min, diethyl ether (100 ml) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (50 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 7% ethyl acetate in hexanes) to give Intermediate 14-2. LCMS: 431.1.

Tetrabutylammonium fluoride solution (1.0 M in tetrahydrofuran, 2.58 mL, 2.6 mmol) was added via syringe to a stirred mixture of Intermediate 14-3 (105 mg, 258 μmol) and N-(2-aminoethyl)acetamide (49.5 μL, 517 μmol) at room temperature, and the resulting mixture was heated to 60° C. After 60 min, the resulting mixture was cooled to room temperature, and diethyl ether (60 mL), citric acid (25 mg), and saturated aqueous ammonium chloride solution (10 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 25% ethyl acetate in hexanes) to give Intermediate 14-4. LCMS: 275.1 [M−H]−.

Intermediate 14-5 was synthesized in a manner similar to Intermediate 7-5 using Intermediate 14-4 instead of Intermediate 7-3. LCMS: 387.2.

The solution of NaOMe (3.51 g, 25% in MeOH) was added dropwise to the solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (4.1 g, 16.2 mmol) in 2-methyltetrahydrofuran (30 mL) at −40° C. The reaction mixture was stirred at −40° C. for 0.5 h. Saturated aqueous ammonium chloride solution (100 mL) was added. The mixture was extracted with EtOAc (3×100 mL). The combined organic phase was washed with brine, dried over Na2SO4and concentrated to give Intermediate 17-1. LCMS: 248.1.

Sodium bis(trimethylsilyl)amide (1.0 M in THF, 8.47 mL) was added dropwise to the solution of [(2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-8-yl]methanol (1.35 g, 8.47 mmol) in 2-methyltetrahydrofuran (10 mL) at 0 C. The resulting solution was stirred at 0° C. for 10 min before it was transferred via a syringe to the solution of Intermediate 17-1 (2.00 g, 8.06 mmol) in 2-methyltetrahydrofuran (20 mL) at 0° C. The mixture was stirred at 0° C. for 20 min. Saturated aqueous solution of NH4Cl (100 mL) was added. The mixture was extracted with dichloromethane (3×200 mL). The combined organic phase was washed with brine (200 mL), dried over Na2SO4and concentrated in vacuo. The resulting crude product was crystallized in EtOAc to give the Intermediate 17-2. LCMS: 371.0

2,2,6,6-Tetramethylpiperidinylmagnesium chloride lithium chloride complex solution (1.0 M in THF/toluene, 17.9 mL) was added dropwise to the solution of Intermediate 17-2 (1.66 g, 4.48 mmol) in 2-methyltetrahydrofuran (15 mL) at 0° C. The resulting solution was stirred at 0° C. for 0.5 h before a solution of 1,2-dibromotetrachloroethane (5.83 g, 17.9 mmol) was added dropwise at 0° C. The resulting solution was stirred at 0° C. for 20 min. Saturated aqueous NH4Cl solution (100 mL) was added to quench the reaction. The mixture was extracted with EtOAc (2×100 mL). The combined organic phase was washed with brine (200 mL), dried over Na2SO4, and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0% to 10% methanol in dichloromethane) to give Intermediate 17-3. LCMS: 450.9.

The suspension of Intermediate 17-3 (1.58 g, 3.51 mmol) and sodium iodide (2.63 g, 17.6 mmol) in acetic acid (15 mL) was stirred at 70° C. for 2 h (LCMS control) before the reaction mixture was cooled to rt. Saturated aqueous solution of sodium thiosulfate (50 mL) was added. The resulting mixture was extracted with dichloromethane (2×150 mL). The combined organic phase was dried over MgSO4and concentrated. The residue was purified by flash column chromatography on silica gel (0% to 10% methanol in dichloromethane) to give Intermediate 17-4. LCMS: 434.9.

DIPEA (0.72 mL, 4.13 mmol) was added dropwise to the solution of Intermediate 17-4 (600 mg, 1.38 mmol) in phosphoryl chloride (8 mL) at rt and the reaction mixture was stirred at 70° C. for 10 min before it was cooled to rt. The mixture was concentrated in vacuo to give the crude product of Intermediate 17-5, which was used for the next step without purification. LCMS: 454.9.

Lithium aluminum hydride (1.0 M in THF, 2.64 mL) was added dropwise to the solution of 8-(tert-butyl) 2-ethyl (1S,2S,5R)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate in dry THF at 0° C. The reaction mixture was stirred at 0° C. for 2 h before it was quenched with 2 M NaOH solution (250 mL). The mixture was extracted with DCM (2×200 mL). The combined organic phase was washed with brine (200 mL), dried with Na2SO4, and concentrated in vacuo to give the Intermediate 17-6. LCMS: 243.2.

Triethylamine (0.185 mL, 1.28 mmol) was added dropwise to the mixture of Intermediate 17-6 (155 mg, 0.640 mmol) and TBSCl (115 mg, 0.763 mmol) in DCM at rt. The reaction mixture was stirred at rt for 12 h before concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0% to 10% methanol in dichloromethane) to give Intermediate 17-7. LCMS: 357.3.

DIPEA (1.8 mL, 10.3 mmol) was added dropwise to the solution of Intermediate 17-5 (600 mg, 1.32 mmol) and Intermediate 17-7 (471 mg, 1.32 mmol) in dichloromethane (8 mL) at 0° C. and the reaction mixture was stirred at 0° C. for 10 min. The reaction mixture was purified by flash column chromatography on silica gel (0% to 10% methanol in dichloromethane) to give Intermediate 17-8. LCMS: 775.0.

TBAF (1.0 M in THF, 3.62 mL) was added dropwise to the solution of Intermediate 17-8 (700 mg, 0.904 mmol) in THF (10 mL) at 0° C. The reaction mixture was warmed to rt and stirred for 12 h. Saturated aqueous solution of NH4Cl (50 mL) was added to the mixture. It was extracted with EtOAc (2×100 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (50% to 100% EtOAc in hexanes) to give Intermediate 17-9. LCMS: 579.0.

n-BuLi (2.70 M in hexanes, 0.051 mL) was added dropwise to the solution of 1-bromo-8-chloro-3-(methoxymethoxy)naphthalene (41.7 mg, 0.138 mmol) in 2-methyltetrahydrofuran (1.5 mL) at −78° C. The resulting solution was stirred at −78° C. for 5 min before the solution of zinc chloride (0.073 mL, 1.9 M in 2-methyltetrahydrofuran) was added dropwise at −78° C. The mixture was warmed to rt and stirred at rt for 10 min. The solution was transferred to a reaction vial containing Intermediate 17-9 (40 mg, 0.069 mmol) and palladium-tetrakis(triphenylphosphine) (8.0 mg, 0.0069 mmol) at rt under N2atmosphere. The reaction mixture was stirred at 90° C. for 90 min before it was cooled to rt. Saturated aqueous solution of NH4Cl (20 mL) was added. The mixture was extracted with EtOAc (2×20 mL). The combined organic phase was washed with brine (30 mL), dried with Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0% to 10% methanol in dichloromethane) to give Intermediate 17-10. LCMS: 765.0.

Intermediate 18-1 was synthesized in a manner similar to Intermediate 17-7 using 8-(tert-butyl) 2-ethyl (1R,2S,5S)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate instead of 8-(tert-butyl) 2-ethyl (1S,2S,5R)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate. LCMS: 357.3

Intermediate 18-2 was synthesized in a manner similar to Intermediate 20-7 using Intermediate 18-1 instead of Intermediate 17-7. LCMS: 921.3.

TBAF (1.0 M in THF, 0.40 mL) was added to the solution of Intermediate 18-2 (50 mg, 0.054 mmol) in THF (1 mL) at 0° C. The mixture was stirred at rt for 2 h. Saturated aqueous solution of NH4Cl (2 mL) was added. The mixture was extracted with EtOAc (2×5 mL). The combined organic phase was washed with brine (10 mL), dried over Na2SO4and concentrated in vacuo. The residue was purified by reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give Intermediate 18-3. LCMS: 806.9.

Sodium hydride (60%, 7.12 mg, 0.186 mmol) was added to the solution of Intermediate 18-3 (15.0 mg, 0.0186 mmol) in dry THF (0.5 mL) at rt. The resulting reaction mixture was stirred at 70° C. for 15 min before it was cooled to rt. Saturated aqueous solution of NH4Cl (1 mL) was added to quench the reaction. It was extracted with EtOAc (2×2 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo to give Intermediate 18-4. LCMS: 687.0.

Intermediate 19-1 was synthesized in a manner similar to Intermediate 18-1 using 8-(tert-butyl) 2-ethyl (1S,2R,5R)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate instead of 8-(tert-butyl) 2-ethyl (1R,2S,5S)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate. LCMS: 357.2.

The solution of NaOMe (1.50 g, 25% in MeOH) was added dropwise to the solution of 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (700 mg, 2.77 mmol) in 2-methyltetrahydrofuran (30 mL) at 0° C. The reaction mixture was stirred at 0° C. for 0.5 h. Saturated aqueous ammonium chloride solution (50 mL) was added. The mixture was extracted with EtOAc (3×50 mL). The combined organic phase was washed with brine, dried over Na2SO4and concentrated to give Intermediate 20-1. LCMS: 244.3.

The reaction mixture of [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-naphthyl] 2,2-dimethylpropanoate (833 mg, 2.35 mmol), Intermediate 20-1 (500 mg, 2.05 mmol), potassium carbonate (567 mg, 4.10 mmol) and Palladium-tetrakis(triphenylphosphine) (237 mg, 0.205 mmol) in dioxane (10 mL) and water (2 mL) was stirred at 90° C. under N2atmosphere for 2 h. After cooling to rt, water (10 mL) was added and the mixture was extracted with EtOAc (2×50 mL). The combined organic phase was washed with brine (100 mL), dried with Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0% to 30% EtOAc in hexanes) to give Intermediate 20-2. LCMS: 436.4.

Intermediate 20-3 was synthesized in a manner similar to Intermediate 17-3 using Intermediate 20-2 instead of Intermediate 17-2. LCMS: 514.1, 516.2.

The reaction mixture of Intermediate 20-3 (80.0 mg, 0.156 mmol) and sodium iodide (117 mg, 0.778 mmol) in acetic acid (2 mL) was stirred at 80° C. for 2 h before it was cooled to rt. The volatile components were evaporated in vacuo. Saturated aqueous solution of sodium bicarbonate (30 mL) was added. The mixture was extracted with EtOAc (2×30 mL) and the combined organic phase was dried with Na2SO4. Evaporation of the solvent gave the mixture of Intermediate 20-4a and Intermediate 20-4b with a ratio of 1:1. LCMS: 486.6 and 488.2 for Intermediate 20-4a; 534.4 for Intermediate 20-4b.

Intermediate 20-5 was synthesized in a manner similar to Intermediate 17-5 using Intermediate 20-4a and Intermediate 20-4b instead of Intermediate 17-4. LCMS: 478.0, 480.0.

Intermediate 20-6 was synthesized in a manner similar to Intermediate 17-8 using Intermediate 20-5 instead of Intermediate 17-5. LCMS: 798.5.

Intermediate 20-7 was synthesized in a manner similar to Intermediate 17-2 using Intermediate 20-6 instead of Intermediate 17-1. LCMS: 921.7.

Intermediate 20-8 was synthesized in a manner similar to Intermediate 17-9 using Intermediate 20-7 instead of Intermediate 17-8. LCMS: 771.1.

TFA was added drop wise to the solution of Intermediate 20-8 (25.0 mg, 0.0324 mmol) in dichloromethane at rt. The mixture was stirred at rt for 1 h. The volatile components were evaporated in vacuo to give Intermediate 20-9. LCMS: 671.1.

Intermediate 21-1 was synthesized in a manner similar to Intermediate 17-7 using 8-(tert-butyl) 2-ethyl (1R,2R,5S)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate instead of 8-(tert-butyl) 2-ethyl (1S,2S,5R)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate. LCMS: 357.3

Intermediate 22-1 was synthesized in a manner similar to Intermediate 27-7 using ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane instead of 2-(7,8-difluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane. LCMS: 927.2.

Intermediate 23-1 was synthesized in a manner similar to Intermediate 24-1 using Intermediate 27-6 instead of Intermediate 25-4. LCMS: 577.2.

The solution of 9-borabicyclo[3.3.1]nonane (0.50 M in THF, 0.067 mL) was added to the solution of Intermediate 25-4 (10.0 mg, 0.0112 mmol) in THF (0.5 mL) at rt. The solution was stirred at 60° C. for 90 min before it was cooled to rt. Water (0.25 mL) was added and the mixture was transferred to a reaction vial containing Pd(dppf)Cl2and cesium carbonate (10.9 mg, 0.0335 mmol). The resulting mixture was stirred at 90° C. for 30 min. It was cooled to rt. Water (2 mL) was added, and the mixture was extracted with EtOAc (2×2 mL). The combined organic phase was washed with brine (5 mL), dried with Na2SO4, and concentrated. The residue was purified by reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give Intermediate 24-1. LCMS: 769.1.

The reaction mixture of [4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-naphthyl] 2,2-dimethylpropanoate (1.18 g, 3.33 mmol), Intermediate 17-2 (1.08 g, 2.90 mmol), Pd(PPh3)4(335 mg, 0.29 mmol), and K2CO3(802 mg, 5.80 mmol) in dioxane (5 mL) and water (1 mL) was stirred at 90° C. under N2atmosphere for 2 h. After cooling to rt, water was added, and the mixture was extracted with EtOAc (2×50 mL). The combined organic phase was washed with brine (100 mL) and dried with Na2SO4. The residue was purified by flash column chromatography on silica gel (0% to 10% MeOH in dichloromethane) to give Intermediate 25-1. LCMS: 563.4.

Intermediate 25-1 was synthesized in a manner similar to Intermediate 17-3 using Intermediate 25-1 instead of Intermediate 17-2. LCMS: 641.1, 643.0.

Intermediate 25-3 was synthesized in a manner similar to Intermediate 17-4 using Intermediate 25-2 instead of Intermediate 17-3. LCMS: 674.9.

HATU (41.9 mg, 0.178 mmol) was added to the reaction mixture of Intermediate 25-3 (40.0 mg, 0.0593 mmol) and Intermediate 27-5 (21.2 mg, 0.0890 mmol) in MeCN followed by the addition of DIPEA (23.0 mg, 0.178 mmol) at rt. The reaction mixture was stirred at rt for 12 h before it was filtered. The filtrate was purified by reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give Intermediate 25-4. LCMS: 895.0.

LAH (1.0 M in tetrahydrofuran, 20.0 mL, 20.0 mmol) was added dropwise to a vigorously stirred solution of 8-(tert-butyl) 2-ethyl (1R,2S,5S)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate (4.00 g, 14.1 mmol) in tetrahydrofuran (40.0 mL) at 0° C. The reaction mixture was stirred at 0° C. for 2 hours before it was quenched with solid sodium sulfate decahydrate. The mixture was filtered and concentrated under reduced pressure to give the crude Intermediate 27-1, which was used for the next step without purification. LCMS: 243.0.

To a vigorously stirred solution of Intermediate 27-1 (1.50 g, 6.19 mmol) in ethyl acetate (15 mL) and water (15 mL) was added sodium bicarbonate (1.56 g, 18.6 mmol) in one portion, then benzyl chloroformate (1.32 mL, 9.28 mmol) was added to the solution slowly with stirring at 0° C. The resulted solution was stirred at room temperature for 12 hours. The organic layer was separated from the reaction mixture. The aqueous phase was extracted with ethyl acetate (3×30 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 27-2. LCMS: 376.8 [M+H]+, 399.1 [M+Na]+.

To a vigorously stirred solution of Intermediate 27-2 (2.01 g, 5.30 mmol) in dichloromethane (25 mL) under nitrogen was added Dess Martin periodinane (2.49 g, 5.80 mmol) at room temperature. The mixture was stirred at room temperature for 12 hours before saturated aqueous sodium bicarbonate (50 mL) was added. The mixture was extracted with ethyl acetate (3×50 mL) and the combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 50% ethyl acetate in hexanes) to give Intermediate 27-3. LCMS: 375.0.

To a vigorously stirred solution of methyltriphenylphosphonium bromide (4.99 g, 14.0 mmol) in tetrahydrofuran (22 mL) at room temperature was added KHMDS solution (1.0 M in tetrahydrofuran, 14.0 mL, 14.0 mmol) dropwise to afford a solution. The mixture was stirred for 1 hour at room temperature and was cooled to −78° C. whereupon a solution of Intermediate 27-3 (1.74 g, 4.65 mmol) in tetrahydrofuran (22 mL) was added dropwise over 20 minutes. The resulting solution was allowed to gradually warm to room temperature and stir for 3 hours. The mixture was quenched with methanol (40 mL) and stirred for 15 min. Saturated aqueous ammonium chloride solution (50 mL) was added and the mixture was extracted with ethyl acetate (3×50 mL). The combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 40% ethyl acetate in hexanes) to give Intermediate 27-4. LCMS: 395.1 [M+Na]+.

To a vigorously stirred solution of palladium(II) acetate (9.5 mg, 0.0426 mmol) in dichloromethane (5.3 mL), triethyl silane (0.27 mL, 1.70 mmol) was added dropwise, followed by the addition of triethylamine (0.12 μL, 0.851 μmol). The resulting mixture was stirred for 15 minutes at room temperature before the solution of Intermediate 27-4 (317 mg, 0.851 mmol) in dichloromethane (2 mL) was added dropwise. The reaction mixture was stirred for 90 hours at room temperature. The mixture was diluted with saturated aqueous sodium bicarbonate (20 mL). The organic layer was separated, and the aqueous layer was extracted with dichloromethane (3×20 mL). The combined organic layer was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 100% ethyl acetate in hexanes then 0% to 20% methanol in dichloromethane) to give Intermediate 27-5. LCMS: 238.9.

To a vigorously stirred solution of Intermediate 17-4 (371.0 mg, 0.851 mmol) in phosphoryl chloride (4.93 mL), N,N-diisopropylethylamine (0.45 mL, 2.55 mmol) was added dropwise at room temperature and the reaction mixture was stirred at 55° C. for 5 min before it was cooled to rt. The mixture was concentrated under reduced pressure. To the solution of the residue and Intermediate 27-5 (203 mg, 0.851 mmol) in dichloromethane (6.4 mL), DIPEA (1.80 mL, 10.3 mmol) was added dropwise at 0° C. The reaction mixture was stirred at 0° C. for 10 min before it was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 100% ethyl acetate in hexanes) to give Intermediate 27-6. LCMS: 656.9.

To a vigorously stirred solution of Intermediate 27-6 (50.0 mg, 0.0112 mmol) in tetrahydrofuran (3 mL), the solution of 9-borabicyclo[3.3.1]nonane (0.50 M in tetrahydrofuran, 0.45 mL, 0.23 mmol) was added at room temperature. The resulting solution was stirred at 60° C. for 90 minutes before it was cooled to room temperature. Degassed water (1.5 mL) was added and the mixture was transferred to a reaction vial charged with chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)] palladium(II) (5.7 mg, 0.00762 mmol), 2-(7,8-difluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (53.4 mg, 0.152 mmol), and sodium carbonate (10.9 mg, 0.0335 mmol). The resulting mixture was stirred at 80° C. for 5 hours then cool to room temperature. The solution was filtered and purified with reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give Intermediate 27-7. LCMS: 765.0.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 35 μL, 35 μmol) was added over 1 min via syringe to a stirred mixture of Intermediate 13-9 (20.0 mg, 23.1 μmol), (1-(morpholinomethyl)cyclopropyl)methanol (7.9 mg, 44 μmol), and tetrahydrofuran (0.5 mL) at room temperature. After 10 min ethyl acetate and saturated aqueous sodium bicarbonate solution were added sequentially. The organic layer was washed with water, was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 28-1. LCMS: 941.2.

Cesium fluoride (80.2 mg, 531 μmol) was added to a vigorously stirred solution of Intermediate 28-1 (20 mg, 21.2 μmol) in N,N-dimethylformamide (0.5 mL) at room temperature. After 30 min, ethyl acetate and saturated aqueous sodium bicarbonate solution were added sequentially. The organic layer was washed with water, was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 28-2. LCMS: 784.9.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 87 μL, 87 μmol) was added over 1 min via syringe to a stirred mixture of Intermediate 13-9 (50.0 mg, 58 μmol), (dimethylsilanediyl)dimethanol (13.9 mg), and tetrahydrofuran (1 mL) at room temperature. After 10 min ethyl acetate, and saturated aqueous sodium bicarbonate solution were added sequentially. The organic layer was washed with water, was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 29-1. LCMS: 890.4.

To a solution of Intermediate 29-1 (50 mg, 0.058 mmol), N,N-diisopropylethylamine (0.06 ml, 0.35 mmol) in DMF (1 mL) was added methane sulfonyl chloride (0.018 ml, 0.232 mmol) at 0° C., the mixture was stirred at the same temperature for 30 min. the reaction mixture was partitioned between EtOAc and water, the organic phase was washed with brine, dried MgSO4, filtered and concentrated. The residue was dissolved in a mixture of acetonitrile/acetone 1:1 (2 ml) then DIPEA (0.05 ml, 0.28 mmol), morpholine (0.029 ml, 0.34 mmol), and NaI (42 mg, 0.284 mmol) were added. Reaction mixture was stirred at 80° C. for 2 h. Ethyl acetate, and saturated aqueous sodium bicarbonate solution were added sequentially. The organic layer was washed with water, was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 29-2. LCMS: 958.9.

Cesium fluoride (118 mg, 0.78 mmol) was added to a vigorously stirred solution of Intermediate 29-2 (30 mg, 31.3 μmol) in N,N-dimethylformamide (1 mL) at room temperature. After 30 min, ethyl acetate and saturated aqueous sodium bicarbonate solution were added sequentially. The organic layer was washed with water, was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 29-3. LCMS: 802.8.

To a solution of Intermediate 30-1 (0.870 g, 3.625 mmol) in 1,4-dioxane (6.090 mL) were added bromoethynyltriisopropylsilane (0.993 g, 3.806 mmol), KOAc (0.712 g, 7.250 mmol) and dichlororuthenium-1-isopropyl-4-methyl-benzene dimer (0.222 g, 0.363 mmol). The resulting mixture was stirred at 110° C. for 2 h. After completion of the reaction, reaction mass was quenched with water (20 mL) and extracted with ethyl acetate (50 mL). The organic layer was washed with brine (20 mL), dried over sodium sulfate, filtered and concentrated under reduced pressure to get the crude material which was subjected to column chromatographic purification using silica (100-200 mesh) and product was eluted at 2%-3% ethyl acetate in pet ether to yield Intermediate 30-2. LCMS: 421.6.

To a stirred solution of Intermediate 30-3 (1.2 g, 2.174 mmol) in 1,4-dioxane (18 mL), were added 4,4,4′,4′,5,5,5′,5′-Octamethyl-2,2′-bi(1,3,2-dioxaborolane) (1.1 g, 4.348 mmol) and KOAc (1.06 g, 10.870 mmol). Reaction mixture was degassed with argon gas for 10 min. PdCl2(dppf) (159 mg, 0.217 mmol) was added and again degassed the reaction mixture for 5 min. Resulting mixture was heated to 130° C. under stirring for 6 h. After completion of the reaction, reaction mass was filtered, diluted with water (30 ml) and extracted with ethyl acetate (50 mL), concentrated under reduced pressure to get the crude material. The crude material was purified by column chromatography using silica (100-200 mesh) and product was eluted at 10%-12% ethyl acetate in pet ether to yield Intermediate 30-4. LCMS: 531.5.

A mixture of Intermediate 13-9 (179.1 mg, 0.21 mmol) and [1-(hydroxymethyl)cyclopropyl]methanol (55.1 mg, 0.54 mmol) was co-evaporated with toluene (×2) and dissolved in THF (2 mL). The solution was stirred at rt as LiHMDS solution (1.0 M in THF, 0.22 mL) was added over 1 min. After 10 min, the reaction mixture was diluted with ether (˜15 mL), ethyl acetate (˜10 mL), and saturated NaHCO3(˜5 mL) and the layers were separated. The organic layer was washed with water (×1), dried (MgSO4), and concentrated. The residue was purified by flash column chromatography on silica gel (30 to 100% ethyl acetate in hexanes) to give Intermediate 35-1. LCMS: 872.5.

A suspension of Intermediate 35-1 (128.1 mg, 147 umol) in CH2Cl2(4 mL) was stirred at rt as Dess-Martin periodinane (178.6 mg, 421 umol) was added. After 20 min, the reaction mixture was diluted with ethyl acetate (˜25 mL) and washed with saturated NaHCO3solution (˜10 mL) with water (˜15 mL), the separated organic layer was washed with water (˜25 mL×1). After the aq. layer was extracted with ethyl acetate (25 mL×1), the two organic layers were combined, dried (MgSO4), and concentrated. The residue was purified by flash column chromatography on silica gel (0 to 100% ethyl acetate in hexanes) to give Intermediate 35-2. LCMS: 870.5.

A mixture of Intermediate 35-2 (8.91 mg, 10.2 umol), 1,4-oxazepane (34.9 mg, 34.9 umol), and sodium triacetoxyborohydride (10.2 mg, 47.9 umol) in THF (1 mL) was stirred at rt. After 16 h, the reaction mixture was diluted with ethyl acetate (25 mL) and washed with saturated NaHCO3(˜25 mL×1) and then water (˜25 mL×1). After the combined aqueous layers were extracted with ethyl acetate (˜20 mL×1), the resulting organic layers were combined, dried (MgSO4), filtered, and concentrated to give crude Intermediate 35-3. LCMS: 955.6.

To a mixture of crude Intermediate 35-3 (10.2 umol) and CsF (40.99 mg, 270 umol) was added DMF (0.5 mL) and the resulting solution was stirred for 1 h at rt. After the reaction mixture was diluted with saturated NaHCO3solution (˜10 mL), water (˜5 mL), and ethyl acetate (˜15 mL), two layers were separated. The organic layer was washed with water (˜15 mL×2) and the combined aqueous layers were extracted with ethyl acetate (˜15 mL×1). The organic layers were combined, dried (MgSO4), and concentrated to give crude Intermediate 35-4. LCMS: 799.4.

A mixture of Intermediate 13-9 (300 mg, 330 μmol) and [(3S,8S)-3-[[tert-butyl(dimethyl)silyl]oxymethyl]-1,2,3,5,6,7-hexahydropyrrolizin-8-yl]methanol (141 mg, 495 mol) was azeotroped from toluene (500 uL) and dissolved in anhydrous N,N-dimethylformamide (6.0 mL) under argon atmosphere. Sodium hydride (60% dispersion in mineral oil, 19 mg, 495 mol) was added and the mixture was stirred overnight at room temperature. Additional sodium hydride (60% dispersion in mineral oil, 19 mg, 495 μmol) was added and the mixture was stirred for a further 2 h. Brine (6 mL) was added and the mixture was extracted with ethyl acetate (3×10 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo to give Intermediate 36-1. LCMS: 1055.6.

Intermediate 36-2 was prepared in a manner analogous to Intermediate 13-11 using Intermediate 36-1 as the starting material. The product was purified by flash column chromatography on basic alumina (0% to 75% ethyl acetate in hexanes) to give Intermediate 36-2. LCMS: 899.5.

Intermediate 36-2 (221 mg, 246 μmol) was dissolved in anhydrous tetrahydrofuran (2 mL) and tetra-n-butyl ammonium fluoride (1 M in tetrahydrofuran, 320 uL, 320 μmol) was added via syringe. The resulting solution was stirred at room temperature overnight. Additional tetra-n-butyl ammonium fluoride (1 M in THF, 64 uL, 64 μmol) was added via syringe and stirring was continued at room temperature for a further 72 hours. The reaction mixture was concentrated in vacuo and purified by flash column chromatography on basic alumina (0% to 100% ethyl acetate in hexanes then 0% to 40% methanol in ethyl acetate) to give Intermediate 36-3. LCMS: 785.4.

Intermediate 36-3 (30 mg, 38 μmol) was dissolve in tetrahydrofuran (1 mL) and cooled to 0° C. Methanesulfonyl chloride (8.9 μL, 115 μmol) was added followed by triethylamine (16 μL, 115 μmol). The mixture was stirred at 0° C. for 30 min. Brine (1 mL) and saturated aqueous sodium carbonate (1 mL) were added and the mixture was extracted with ethyl acetate (2×1 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified via flash column chromatography on basic alumina using a gradient of ethyl acetate in hexanes (0% to 100%) followed by a gradient of methanol in ethyl acetate (0-20%) to afford Intermediate 36-4. LCMS: 863.4.

To a solution of Intermediate 36-4 (28 mg, 32 μmol) in N,N-dimethylformamide (1 mL) was added sodium azide (11 mg, 162 μmol) and the resulting mixture was stirred at 70° C. overnight. Brine (1 mL) and saturated aqueous sodium carbonate (1 mL) were added and the mixture was extracted with ethyl acetate (2×1 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The resulting residue was purified via flash column chromatography on basic alumina using a gradient of ethyl acetate in hexanes (0% to 100%) to afford Intermediate 36-5. LCMS: 810.4.

To a suspension of NaH (7.55 g, 179.26 mmol) and cyclopropanol (4.54 mL, 71.70 mmol) in THF (360 mL), was added methyl 6-bromo-2,3-difluorobenzoate (18 g, 71.70 mmol) at −40° C. The reaction mixture was allowed to warm up to room temperature and stirred for 4 h. After completion of the reaction, reaction mass was carefully quenched with water (70 mL) and extracted with ethyl acetate (2×90 mL). The combined organic layer was washed with brine (70 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to get the crude adduct. The crude material was subjected to silica gel (100-200 mesh) column chromatography eluting with 8-10% ethyl acetate in petroleum ether to yield Intermediate 42-1.1H NMR (400 MHz, CDCl3) δ 7.26-7.20 (m, 1H), 7.06-7.01 (m, 1H), 4.31-4.29 (m, 1H), 3.99 (s, 3H), 0.82 (s, 2H), 0.61-0.57 (m, 2H).

Intermediate 42-2 was synthesized in a manner similar to Intermediate 7-5 using Intermediate 42-1 instead of Intermediate 7-1. LCMS: 389.3.

A vial was charged with Intermediate 36-5 (10 mg, 12 μmol), copper (II) sulfate (0.6 mg, 2.5 μmol), potassium carbonate (8.5 mg, 62 μmol) and sodium ascorbate (0.8 mg, 4.9 μmol). Water (250 μL) and methanol (250 μL) were added followed by trimethylsilylacetylene (44 μL, 310 μmol). The resulting mixture was stirred vigorously at room temperature overnight. Brine (1 mL) was added and the mixture was extracted with ethyl acetate (2×1 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on basic alumina using a gradient of ethyl acetate in hexanes (0% to 100%) followed by methanol in ethyl acetate (0% to 20%) to give Intermediate 49-1. LCMS: 836.4.

Ammonia solution (0.4 M in 1,4-dioxane, 9.57 mL, 4 mmol) was added via syringe to a stirred mixture of methyl 6-bromo-4-chloro-2,3-difluorobenzoate (600 μL, 3.48 mmol), N,N-diisopropylethylamine (666 μL, 3.83 mmol), and acetonitrile (1.5 mL) at room temperature, and the resulting mixture was heated to 80° C. After 91 min, ammonia solution (0.4 M in 1,4-dioxane, 6.00 mL, 2 mmol) was added via syringe, and the resulting mixture was heated to 105° C. After 5.5 h, the resulting mixture was heated to 115° C. After 64 h, the resulting mixture was cooled to room temperature, and diethyl ether (100 mL) and ethyl acetate (25 mL) were added sequentially. The organic layer was washed with a mixture of water and brine (3:1 v:v, 50 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 12% ethyl acetate in hexanes) to give Intermediate 52-1. LCMS: 281.9.

2,2,2-Trichloroacetyl isocyanate (138 μL, 1.16 mmol) was added over 2 min via syringe to a stirred solution of Intermediate 52-1 (298 mg, 1.05 mmol) in tetrahydrofuran (1.5 mL) at 0° C., and the resulting mixture was warmed to room temperature. After 38 min, the resulting mixture was concentrated under reduced pressure. Ammonia solution (20% wt in methanol, 5.50 mL, 39 mmol) was added via syringe, and the resulting mixture was stirred vigorously. After 20 min, the resulting mixture was concentrated under reduced pressure to give Intermediate 52-2. LCMS: 292.9.

N,N-Diisopropylethylamine (802 μL, 4.60 mmol) was added via syringe to a mixture of Intermediate 52-2 (310 mg, 1.06 mmol) and phosphorous (V) oxychloride (10 mL) at 100° C. After 20 min, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 80% dichloromethane in hexanes) to give Intermediate 52-3.1H NMR (400 MHz, Chloroform-d) δ 8.09 (d, J=6.6 Hz, 1H).

Intermediate 27-5 (72.1 mg, 303 μmol) and N,N-diisopropylethylamine (55.4 μL, 318 mol) were added sequentially to a vigorously stirred solution of Intermediate 52-3 (100 mg, 303 μmol) in dichloromethane (1.4 mL) at 0° C. After 30 min, citric acid (0.3 g), diethyl ether (40 mL), and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (2×30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. 2-Methyltetrahydrofuran (0.5 mL) and [(2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-8-yl]methanol (50.6 mg, 318 μmol) were added sequentially, and the resulting mixture was vigorously stirred at room temperature. Potassium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 318 μL, 320 μmol) was added over 1 min via syringe, and the resulting mixture was heated to 90° C. After 120 min, the resulting mixture was cooled to room temperature, and saturated aqueous sodium bicarbonate solution (5 mL), diethyl ether (40 mL), and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (20 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrate under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 5% methanol in dichloromethane) to give Intermediate 52-4. LCMS: 654.2.

9-Borabicyclo[3.3.1]nonane solution (0.50 M in tetrahydrofuran, 759 μL, 380 μmol) was added via syringe to a vigorously stirred solution of Intermediate 52-4 (82.8 mg, 126 μmol) in tetrahydrofuran (0.3 mL) at room temperature, and the resulting mixture was heated to 60° C. After 80 min, the resulting mixture was cooled to room temperature, and water (0.3 mL) was added via syringe. After 60 min, the resulting mixture was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give Intermediate 52-5. LCMS: 700.2.

Aqueous potassium phosphate solution (1.5 M, 126 μL, 190 μmol) was added via syringe to a vigorously stirred mixture of Intermediate 52-5 (40.7 mg, 58.1 μmol), [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (4.6 mg, 6.3 μmol), and 1,4-dioxane (0.8 mL) at room temperature, and the resulting mixture was heated to 90° C. After 15 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (20 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by reverse phase preparative HPLC (0.1% acetic acid in acetonitrile/water) to give Intermediate 52-6. LCMS: 576.2.

Intermediate 52-7 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 52-6 instead of Intermediate 17-9 and using ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane instead of ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane. LCMS: 926.5.

N,N-Diisopropylethylamine (884 μL, 5.08 mmol) was added via syringe to a mixture of Intermediate 13-3 (839 mg, 2.48 mmol) and phosphorous (V) oxychloride (10 mL) at room temperature, and the resulting mixture was stirred at rt for 15 min before it was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 13% ethyl acetate in hexanes) to give Intermediate 53-1. LCMS: 357.9.

A solution of methyl magnesium bromide in 2-methyltetrahydrofuran (3.4 M, 424 μL, 1.44 mmol) was added via syringe to a mixture of Intermediate 27-3 (180 mg, 0.481 mmol) in 2-methyltetrahydrofuran (4 mL) at −78° C. The resulting mixture was stirred at −78° C. for 15 min before it was quenched with 5 mL of saturated aqueous solution of NH4Cl. The mixture was extracted with EtOAc (2×10 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo to give Intermediate 53-2. LCMS: 391.1.

Chlorotrimethylsilane (309 mg, 2.05 mmol) was added dropwise to the solution of 53-2 (320 mg, 0.820 mmol), imidazole (167 mg, 2.46 mmol) and DMAP (20 mg, 0.164 mmol) in DCM (4 mL) at 0° C. 10 mL of saturated aqueous solution of Na2CO3was added. The mixture was extracted with EtOAc (2×15 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0 to 50% EtOAc in hexanes) to give Intermediate 53-3. LCMS: 505.5.

Intermediate 53-4 was synthesized in a manner similar to Intermediate 27-5 using Intermediate 53-3 instead of Intermediate 27-4. LCMS: 371.3.

The solution of Intermediate 53-4 (34.4 mg, 0.0924 mmol), Intermediate 53-1 (30 mg, 0.0840 mmol) and DIPEA (35.4 mg, 0.274 mmol) in DCM (1 mL) was stirred at 60° C. for 3 before it was cooled to rt. The mixture was purified by flash column chromatography on silica gel (0 to 80% EtOAc in hexanes) to give Intermediate 53-5. LCMS: 690.3, 692.2.

TBAF (1.0 M in THF, 0.579 mL) was added dropwise to the solution of Intermediate 53-5 (100 mg, 0.145 mmol) in THF (2 mL) at 0° C. The reaction mixture was warmed to rt and stirred for 1 h. Saturated aqueous solution of NH4Cl (20 mL) was added to the mixture. It was extracted with EtOAc (2×20 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (50% to 100% EtOAc in hexanes) to give Intermediate 53-6. LCMS: 576.2, 578.0.

TBAF (1.0 M in THF, 0.217 mL) was added dropwise to the solution of the solution of Intermediate 53-7 (50 mg, 0.0867) in THF at rt. The resulting solution was stirred at 75° C. for 3 h before it was cooled to rt. Saturated aqueous solution of NH4Cl (20 mL) was added to the mixture. It was extracted with EtOAc (2×20 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (30% to 100% EtOAc in hexanes) to give Intermediate 53-7. LCMS: 496.3.

Intermediate 53-8 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 53-7 instead of Intermediate 13-7. LCMS: 846.8.

Intermediate 53-9 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 53-8 instead of Intermediate 13-8. LCMS: 878.4.

Intermediate 53-10 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 53-9 and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol instead of Intermediate 13-9 and Intermediate 13-14. LCMS: 943.6.

Intermediate 53-11 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 53-10 instead of Intermediate 13-10. LCMS: 787.0.

Tetrabutylammonium fluoride solution (1.0 M in tetrahydrofuran, 2.56 mL, 2.6 mmol) was added via syringe to a stirred mixture of Intermediate 54-1 (116 mg, 256 μmol) and 3-methylpentane-2,4-dione (29.8 μL, 256 μmol) at room temperature, and the resulting mixture was heated to 60° C. After 13 min, the resulting mixture was heated to 80° C. After 104 min, the resulting mixture was cooled to room temperature, and diethyl ether (60 mL), citric acid (25 mg), and saturated aqueous ammonium chloride solution (10 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 15% ethyl acetate in hexanes) to give Intermediate 54-2. LCMS: 321.0 [M−H]−.

Intermediate 54-3 was synthesized in a manner similar to Intermediate 7-5 using Intermediate 54-2 instead of Intermediate 7-3. LCMS: 401.1 [M−CH3O]+.

Intermediate 57-1 was synthesized in a manner similar to Intermediate 52-4 using Intermediate 17-7 instead of Intermediate 27-5. LCMS: 772.2.

Tetrabutylammonium fluoride solution (1.0 M in tetrahydrofuran, 500 μL, 500 μmol) was added via syringe to a stirred solution of Intermediate 57-1 (48.8 mg, 63.1 μmol) in tetrahydrofuran (0.5 mL) at room temperature. After 3 h, diethyl ether (40 mL), ethyl acetate (20 mL), saturated aqueous ammonium chloride solution (3 mL), and saturated aqueous sodium carbonate solution (10 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. Cesium carbonate (61.7 mg, 189 μmol), rac-BINAP-Pd-G3 (3.1 mg, 3.2 μmol), and toluene (1.0 mL) were added sequentially, and the resulting mixture was heated to 90° C. After 25 min, the resulting mixture was heated to 115° C. After 150 min, the resulting mixture was cooled to room temperature and was filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by reverse phase preparative HPLC (0.1% acetic acid in acetonitrile/water) to give Intermediate 57-2. LCMS: 578.2.

Intermediate 61-1 was synthesized in a manner similar to 27-3 using intermediate 53-2 instead of intermediate 27-2. LCMS: 389.1.

Sodium borohydride (69.6 mg, 1.84 mmol) was added in small portions to the solution of intermediate 61-1 (650 mg, 1.67 mmol) in THF (3 mL) and MeOH (3 mL) at 0° C. The resulting reaction mixture was stirred at 0° C. for 30 min before it was quenched with saturated aqueous solution of NH4Cl (20 mL). The mixture was extracted with EtOAc (3×40 mL). The combined organic phase was washed with brine, dried with MgSO4and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel (0 to 40% ethyl acetate in hexanes) to give intermediate 61-2. LCMS: 413.2 [M+Na]+.

Intermediate 62-1 was synthesized in a manner similar to intermediate 7-2 using methyl 2-bromo-4,5-difluorobenzoate instead of intermediate 7-1 and using allyl bromide instead of 3-chloro-2-(methoxymethoxy)prop-1-ene. LCMS: 213.0.

A solution of intermediate 62-2 (200 mg, 943 μmol) in 2-methyltetrahydrofuran (4 mL) was added over 60 min via syringe pump to a vigorously stirred mixture of potassium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 2.36 mL, 2.4 mmol) and 2-methyltetrahydrofuran (16 mL) at 70° C. After 10 min, the resulting mixture was cooled to room temperature, acetic acid (162 μL, 2.83 mmol) was added via syringe, and the resulting mixture was concentrated under reduced pressure. Methanol (20 mL) and acetic acid (540 μL, 9.43 mmol) were added sequentially. After 20 min, the resulting mixture was concentrated under reduced pressure, and diethyl ether (100 mL), ethyl acetate (25 mL), and saturated aqueous sodium bicarbonate solution (20 mL) were added sequentially. The organic layer was washed with water (60 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 15% ethyl acetate in hexanes) to give intermediate 62-2. LCMS: 179.0 [M−H]−.

Triphenylmethyl chloride (1.17 g, 4.20 mmol) was added to a vigorously stirred mixture of ((3S,7aS)-3-(((tert-butyldimethylsilyl)oxy)methyl)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (1.00 g, 3.50 mmol), triethylamine (770 μL, 5.25 mmol), 4-(dimethylamino)pyridine (85.6 mg, 701 μmol), and dichloromethane (5.0 mL) at room temperature, and the resulting mixture was heated to 40° C. After 95 min, the resulting mixture was heated to 65° C. After 120 min, the resulting mixture was cooled to room temperature, and diethyl ether (100 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (100 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 5% methanol in dichloromethane) to give intermediate 63-1. LCMS: 528.3.

Tetrabutylammonium fluoride solution (1.0 M in tetrahydrofuran, 10.5 mL, 11 mmol) was added via syringe to a stirred solution of intermediate 63-1 (1.16 g, 2.20 mmol) in tetrahydrofuran (2.0 mL) at room temperature. After 30 min, the resulting mixture was heated to 50° C. After 15 min, the resulting mixture was cooled to room temperature, and diethyl ether (100 mL), ethyl acetate (20 mL), saturated aqueous ammonium chloride solution (2.0 mL), and saturated aqueous sodium carbonate solution (10 mL) were added sequentially. The organic layer was washed with water (2×100 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 100% methanol in dichloromethane) to give intermediate 63-2. LCMS: 414.2.

Trimethylphosphine solution (1.0 M in tetrahydrofuran, 2.70 mL, 2.7 mmol) was added over 2 min via syringe to a stirred mixture of intermediate 63-2 (447 mg, 1.08 mmol), di-tert-butyl (E)-diazene-1,2-dicarboxylate (622 mg, 2.70 mmol), 1,1,1,3,3,3-hexafluoro-2-(trifluoromethyl)propan-2-ol (753 μL, 5.40 mmol), and tetrahydrofuran (4.0 mL) at 0° C., and the resulting mixture was warmed to room temperature. After 7 min, the resulting mixture was heated to 70° C. After 53 min, the resulting mixture was heated to 90° C. After 40 min, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 5% methanol in dichloromethane) to give intermediate 63-3. LCMS: 632.2.

Concentrated hydrochloric acid (346 μL, 4.2 mmol) was added via syringe to a stirred solution of intermediate 63-3 (524 mg, 830 μmol) in methanol (4.0 mL) at room temperature, and the resulting mixture was heated to 60° C. After 70 min, the resulting mixture was cooled to room temperature. Triethylamine (1.0 mL) was added via syringe, and the resulting mixture was concentrated under reduced pressure. Saturated aqueous sodium carbonate (10 mL) and water were added sequentially. The aqueous layer was extracted with dichloromethane (4×35 mL). The combined organic layers were dried over anhydrous magnesium sulfate, were filtered, and were concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 100% methanol in dichloromethane) to give intermediate 63-4. LCMS: 390.1.

Intermediate 63-5 was synthesized in a manner similar to intermediate 13-6 using intermediate 27-5 instead of intermediate 13-5. LCMS: 560.0.

Intermediate 63-6 was synthesized in a manner similar to intermediate 52-7 using intermediate 63-5 instead of intermediate 52-4. LCMS: 830.5.

Intermediate 63-7 was synthesized in a manner similar to intermediate 13-9 using intermediate 63-6 instead of intermediate 13-8. LCMS: 862.4.

Intermediate 64-1 was synthesized in a manner similar to 27-4 using intermediate 61-1 instead of intermediate 27-3. LCMS: 387.0.

Intermediate 64-2 was synthesized in a manner similar to 27-5 using intermediate 64-1 instead of intermediate 27-4. LCMS: 252.9.

Intermediate 64-3 was synthesized in a manner similar to 63-5 using intermediate 64-2 instead of intermediate 13-6. LCMS: 572.4, 574.0.

Intermediate 64-4 was synthesized in a manner similar to 63-7 using intermediate 64-3 instead of intermediate 63-5. LCMS: 876.4.

Intermediate 65-1 was synthesized in a manner similar to intermediate 7-5 using allyl bromide instead of 3-chloro-2-(methoxymethoxy)prop-1-ene. LCMS: 357.1.

Intermediate 67-1 was synthesized in a manner similar to intermediate 35-1 using intermediate 63-7 instead of intermediate 13-9. LCMS: 870.5.

Intermediate 67-2 was synthesized in a manner similar to intermediate 35-2 using intermediate 67-1 instead of intermediate 35-1. LCMS: 868.5.

Intermediate 67-3 was synthesized in a manner similar to intermediate 35-3 using intermediate 67-2 instead of intermediate 35-2 and using (R)-3-fluoropiperidine instead of 1,4-oxazepane. LCMS: 955.6.

Intermediate 67-4 was synthesized in a manner similar to intermediate 35-4 using intermediate 67-3 instead of intermediate 35-3. LCMS: 799.4.

Intermediate 71-1 was synthesized in a manner similar to intermediate 53-2 using cyclopropyl magnesium bromide instead of methyl magnesium bromide. LCMS: 439.2 [M+Na]+.

Example 73-1 was synthesized in a manner similar to intermediate 30-4 using methyl 2-bromo-5-chlorobenzoate instead of methyl 2-bromo-4,5-difluorobenzoate. LCMS: 529.3.

Tetramethyltin (65.5 μL, 473 μmol) was added via syringe to a vigorously stirred mixture of intermediate 73-1 (50.0 mg, 94.5 μmol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (6.9 mg, 9.5 μmol), cesium carbonate (7.7 mg, 24 mol), and N,N-dimethylformamide (0.7 mL) at room temperature, and the resulting mixture was heated to 90° C. After 90 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 ml), ethyl acetate (20 mL), and saturated aqueous ammonium chloride solution (1 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 7% ethyl acetate in hexanes) to give intermediate 73-2. LCMS: 509.1.

To a vigorously stirred solution of Intermediate 27-5 (1.00 g, 4.20 mmol) and (2,5-dioxopyrrolidin-1-yl) 2-trimethylsilylethyl carbonate (1.14 g, 4.41 mmol) in dichloromethane (13.9 mL) was added 4-methylmorpholine (0.46 mL, 4.20 mmol) slowly with stirring at room temperature. The resulted solution was stirred at room temperature for 72 hours. The mixture was washed with water (15 mL) and extracted with ethyl acetate (3×30 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure to give a liquid. The liquid was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 75-1. LCMS: 405.1 [M+Na]+.

To a vigorously stirred solution of Intermediate 75-1 (50.0 mg, 0.13 mmol) in tetrahydrofuran (1.5 mL) was added the solution of 9-borabicyclo[3.3.1]nonane (0.50 M in tetrahydrofuran, 0.4 mL, 0.20 mmol) dropwise at 0° C. The resulted solution was stirred at 50° C. for 45 minutes. Then hydrogen peroxide (50% wt in water, 0.2 mL, 0.13 mmol) and sodium hydroxide (20 mg, 0.33 mmol) were added at room temperature and the resulting mixture was stirred at room temperature for 30 minutes then quenched with saturated sodium thiosulfate solution (1.5 mL) at 0° C. The mixture was extracted with dichloromethane (3×10 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure to give a liquid. The liquid was purified by silica gel column chromatography (0 to 100% ethyl acetate in hexanes) to give Intermediate 75-2. LCMS: 400.7 [M+H]+, 423.1 [M+Na]+.

To a vigorously stirred solution of Intermediate 75-2 (0.55 g, 1.37 mmol) in dichloromethane (5 mL) under nitrogen was added Dess Martin periodinane (0.64 g, 1.51 mmol) at room temperature. The mixture was stirred at room temperature for 12 hours before saturated aqueous sodium bicarbonate (10 mL) was added. The mixture was extracted with ethyl acetate (3×10 mL) and the combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 50% ethyl acetate in hexanes) to give Intermediate 75-3. LCMS: 421.1 [M+Na]+.

To a vigorously stirred solution of methyltriphenylphosphonium bromide (1.29 g, 3.62 mmol) in tetrahydrofuran (5.8 mL) at room temperature was added KHMDS solution (1.0 M in tetrahydrofuran, 3.30 mL, 3.30 mmol) dropwise to afford a solution. The mixture was stirred for 1 hour at room temperature and was cooled to −78° C. whereupon a solution of Intermediate 75-3 (0.44 g, 1.10 mmol) in tetrahydrofuran (5.8 mL) was added dropwise over 20 minutes. The resulting solution was allowed to gradually warm to room temperature and stir for 3 hours. The mixture was quenched with methanol (10 mL) and stirred for 15 min. Saturated aqueous ammonium chloride solution (12 mL) was added and the mixture was extracted with ethyl acetate (3×12 mL). The combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 40% ethyl acetate in hexanes) to give Intermediate 75-4. LCMS: 419.2 [M+Na]+.

Cesium fluoride (95.8 mg, 0.63 mmol) was added to a vigorously stirred solution of Intermediate 75-4 (50.0 mg, 0.13 mmol) in N,N-dimethylformamide (0.8 mL) at room temperature. The resulting mixture was stirred at 90° C. for 30 minutes. After cooled to room temperature, the mixture was filtered, washed by dichloromethane (5 mL), and the filtrate was concentrated under reduced pressure to give crude product of Intermediate 75-5, which was used for the next step without purification. LCMS: 253.0.

A mixture of Intermediate 75-5 (230 mg, 0.911 mmol), Intermediate 53-1 (342 mg, 0.957 mmol) and DIPEA (384 mg, 2.97 mmol) in DCM (1.3 mL) was stirred at room temperature for 24 minutes. The mixture was purified by flash column chromatography on silica gel (0 to 80% EtOAc in hexanes) to give Intermediate 75-6. LCMS: 574.0.

To a vigorously stirred solution of Intermediate 75-6 (30.0 mg, 0.052 mmol) in anhydrous 1,4-dioxane (0.5 mL), the solution of 9-borabicyclo[3.3.1]nonane (0.50 M in tetrahydrofuran, 0.12 mL, 0.06 mmol) was added at room temperature. The resulting solution was stirred at 50° C. for 1 hour before it was cooled to room temperature. The solution was transferred to a reaction vial containing Pd(dppf)Cl2(3.70 mg, 0.0052 mmol), potassium phosphate (36.6 mg, 0.16 mmol) and degassed water (0.1 mL) at rt under nitrogen atmosphere. The reaction mixture was stirred at 90° C. for 10 minutes before it was cooled to room temperature. The mixture was washed with water (2 mL) and extracted with ethyl acetate (3×5 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 75-7. LCMS: 494.1.

Intermediate 75-8 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 75-7 instead of Intermediate 13-8. LCMS: 526.1.

Intermediate 75-9 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 75-8 instead of Intermediate 53-9. LCMS: 591.2.

Intermediate 75-10 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 75-9 instead of Intermediate 17-9. LCMS: 941.3.

Intermediate 75-11 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 75-10 instead of Intermediate 13-10. LCMS: 784.9.

3-Chloroperoxybenzoic acid (77% wt, 4590 mg, 20.5 mmol) was added in two equal portions over 5 min to a vigorously stirred solution of Intermediate 115-1 (7.30 g, 9.31 mmol) in dichloromethane (131 mL) at 0° C. After 125 min, the resulting mixture was warmed to room temperature. The residue was purified by flash column chromatography on silica gel (0% to 75% ethyl acetate in hexanes) to give Intermediate 76-0. LCMS: 816.4.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 1.84 mL, 1.84 mmol) was added over 1 min via syringe to a stirred mixture of Intermediate 76-0 (1000 mg, 1.23 mmol), [(2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-8-yl]methanol (725.4 mg, 4.56 mmol), and 2-methyltetrahydrofuran (25.8 mL) 0° C. After 2 hours, ethyl acetate (20 mL), and water (5 mL) were added sequentially at 0° C. The aqueous layer was washed with ethyl acetate (3×20 mL). The organic layers were combined and dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 76-1. LCMS: 881.1.

Intermediate 76-1 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 75-9 instead of Intermediate 17-9. LCMS: 881.1.

Cesium fluoride (37.8 mg, 0.249 mmol) was added to a vigorously stirred solution of Intermediate 76-1 (21.9 mg, 24.9 μmol) in N,N-dimethylformamide (0.6 mL) at room temperature. After 1 hour, diethyl ether (4 mL), ethyl acetate (2 mL), and saturated aqueous sodium bicarbonate solution (1 mL) were added sequentially. The organic layer was washed with water (2×4 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 to 60% ethyl acetate in hexanes) to give Intermediate 76-2. LCMS: 724.9.

Intermediate 77-1 was synthesized in a manner similar to intermediate 27-3 using 8-(tert-butyl) 2-ethyl (1S,2R,5R)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate instead of 8-(tert-butyl) 2-ethyl (1S,2S,5R)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate. LCMS: 375.2

Methoxymethyl(triphenyl)phosphonium bromide (1.03 g, 2.67 mmol) was dissolved in 10 ml of THF and cooled to −78° C. To it was added KHMDS solution (1 M in THF, 2.67 ml, 2.67 mmol). The reaction mixture was stirred vigorously 15 minutes then warmed to 0° C. and stirred vigorously for 30 minutes. To it was dropwise added a solution of 03-benzyl 08-tert-butyl (1S,2R,5R)-2-formyl-3,8-diazabicyclo [3.2.1] octane-3,8-dicarboxylate (500 mg, 1.34 mmol) in 10 ml of THF. After stirring vigorously for 8 minutes, the mixture was warmed to room temperature, and stirred for overnight. The mixture was diluted with ethyl ether and quenched the reaction with water. The organic layer was separated and the aqueous phase extracted with EtOAc. The combined organic layers were washed with water, brine, dried over MgSO4and concentrated under reduced pressure. Purification by column chromatography over silica gel gave intermediate 77-2. LCMS: 403.3.

Intermediate 77-3 was synthesized in a manner similar to intermediate 27-5 using intermediate 77-2 instead of intermediate 27-4. LCMS: 269.2

Intermediate 77-4 was synthesized in a manner similar to intermediate 17-8 using intermediate 77-3 instead of intermediate 17-7. LCMS: 687.2

Intermediate 77-4 (22 mg, 0.032 mmol) was dissolved in 1 ml of DCM and to it was added 0.1 ml of HCl solution (4 N, in 1,4-dioxane). It was concentrated to dryness after 5 minutes; then dissolved in 1.5 ml of THF and to it was added sodium borohydride (12.1 mg, 0.32 mmol), stirred at room temperature for 1 hour. The reaction mixture was diluted with ethyl acetate and washed with saturated sodium bicarbonate and water, dried (MgSO4), filtered and concentrated. The residue was purified by RP-HPLC. LCMS: 674.2

Intermediate 77-6 was synthesized in a manner similar to intermediate 18-4 using intermediate 77-5 instead of intermediate 18-3. LCMS: 593.3

Intermediate 77-7 was synthesized in a manner similar to intermediate 2-1 using intermediate 77-6 instead of intermediate 17-9. LCMS: 943.6

Intermediate 78-1 was synthesized in a manner similar to intermediate 67-3 using 4,4-dimethyl-1,4-azasilepane hydrochloride instead of (R)-3-fluoropiperidine. LCMS: 995.6.

Intermediate 78-2 was synthesized in a manner similar to intermediate 35-4. LCMS: 839.5.

To a solution of [Ir(cod)Cl]2(25.6 mg, 34.9 μmol) and 1,2-bis(diphenylphosphino)ethane(27.8 mg, 69.8 μmol) in THF (2 mL) was added pinacolborane (223 mg, 1.75 mmol) and intermediate 64-3 (200 mg, 349 μmol). The reaction mixture was stirred under N2atmosphere at 70° C. overnight before it was quenched with MeOH (0.2 mL). After evaporation of the volatile components, the residue was purified with flash chromatography on silica gel (0 to 30% ethyl acetate in hexanes) to give the boronate intermediate. Pd(dppf)Cl2(12 mg, 0.17 mmol) and potassium phosphate tribasic monohydrate (120 mg, 0.514 mmol) were added to a vial containing the boronate intermediate. The reaction vial was flushed with N2. Dioxane (2 mL) and water (0.5) were added, and the resulting mixture was stirred at 90° C. under N2for 20 min before it was cooled to rt. Water (10 mL) was added and the mixture was extracted with EtOAc (3×10 mL). The combined organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified with flash chromatography on silica gel (0 to 50% ethyl acetate in hexanes) to give the intermediate 79-1. LCMS: 494.0 [M+H]+.

Intermediate 79-2 was synthesized in a manner similar to Intermediate 75-10 using Intermediate 79-1 instead of Intermediate 75-9. LCMS: 844.4 [M+H]+.

Intermediate 79-3 was synthesized in a manner similar to Intermediate 75-8 using Intermediate 79-2 instead of Intermediate 75-7. LCMS: 876.3 [M+H]+.

Intermediate 79-4 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 79-3 instead of Intermediate 75-8. LCMS: 941.0 [M+H]+.

Intermediate 79-5 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 79-4 instead of Intermediate 75-11. LCMS: 784.9 [M+H]+

Intermediate 80-1 was synthesized in a manner similar to Intermediate 53-2 using ethyl magnesium bromide instead of methyl magnesium bromide. LCMS: 427.2 [M+Na]+.

Intermediate 80-2 was synthesized in a manner similar to Intermediate 53-3 using Intermediate 80-1 instead of Intermediate 53-2. LCMS: 520.6 [M+H]+, 541.9 [M+Na]+.

Intermediate 80-3 was synthesized in a manner similar to Intermediate 53-4 using Intermediate 80-2 instead of Intermediate 53-3. LCMS: 385.3.

Intermediate 80-4 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 80-3 instead of Intermediate 53-4. LCMS: 706.4.

Intermediate 80-5 was synthesized in a manner similar to Intermediate 53-6 using Intermediate 80-4 instead of Intermediate 53-5. LCMS: 592.0.

Intermediate 80-6 was synthesized in a manner similar to Intermediate 53-7 using Intermediate 80-5 instead of Intermediate 53-6. LCMS: 510.6.

Intermediate 80-7 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 80-6 instead of Intermediate 13-7. LCMS: 861.0.

Intermediate 80-8 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 80-7 instead of Intermediate 13-8. LCMS: 892.8.

Intermediate 80-9 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 80-8 instead of Intermediate 13-9. LCMS: 957.7.

Intermediate 80-10 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 80-9 instead of Intermediate 13-10. LCMS: 801.1.

Intermediate 81-1 was synthesized in a manner similar to intermediate 27-6 using intermediate 75-5 instead of intermediate 27-5. LCMS: 671.2

The reaction mixture of 81-1 (20 mg, 0.0299 mmol), Pd(dppf)Cl2(4.37 mg, 0.006 mmol), and cesium carbonate (29.2 mg, 0.089 mmol) in THF (2 mL) and water (0.5 mL) was stirred under N2atmosphere at 90° C. for 30 minutes before it was cooled to room temperature. Water was added, and the mixture was extracted with EtOAc. The combined organic phase was washed with brine, dried over with Na2SO4, filtered and concentrated. The residue was purified by reverse phase preparative HPLC (0.1% trifluoroacetic acid in acetonitrile/water) to give intermediate 81-2. LCMS: 589.3

Intermediate 81-3 was synthesized in a manner similar to Intermediate 13-8 using intermediate 81-2 instead of intermediate 13-7 and using 2-(7,8-difluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane instead of ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane. LCMS: 777.4

Lithium bis(trimethylsilyl)amide (1.0 M in tetrahydrofuran, 0.2 mL, 0.2 mmol) was added dropwise to a vigorously stirred solution of Intermediate 27-3 (37.4 mg, 0.1 mmol) and difluoromethylsulfonylbenzene (19.2 mg, 0.1 mmol) in tetrahydrofuran (0.6 mL) at −78° C. The reaction mixture was stirred at −78° C. for 1 hour before it was quenched with saturated aqueous ammonium chloride solution (2 mL) at −78° C. The mixture was extracted with ethyl acetate (3×50 mL). The combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 70% ethyl acetate in hexanes) to give Intermediate 82-1. LCMS: 589.1 [M+Na]+.

Intermediate 82-2 was synthesized in a manner similar to Intermediate 53-4 using Intermediate 82-1 instead of Intermediate 53-3. LCMS: 432.9.

Sodium hydrogen phosphate (37.3 mg, 0.26 mmol) and sodium mercury amalgam (57.7 mg, 0.26 mmol) were added to a vigorously stirred solution of Intermediate 82-2 (18.6 mg, 0.043 mmol) in methanol (1.0 mL) at −41° C. The reaction mixture was warmed up to room temperature and stirred at room temperature overnight before it was filtered through celite. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 10% methanol in dichloromethane) to give Intermediate 82-3. LCMS: 292.9.

Intermediate 82-4 was synthesized in a manner similar to Intermediate 53-3 using Intermediate 82-3 instead of Intermediate 53-2. LCMS: 407.2.

Intermediate 82-5 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 82-4 instead of Intermediate 53-4. LCMS: 728.0.

Intermediate 82-6 was synthesized in a manner similar to Intermediate 53-6 using Intermediate 82-5 instead of Intermediate 53-5. LCMS: 613.8.

Intermediate 82-7 was synthesized in a manner similar to Intermediate 53-7 using Intermediate 82-6 instead of Intermediate 53-6. LCMS: 532.4.

Intermediate 82-8 was synthesized in a manner similar to Intermediate 53-8 using Intermediate 82-7 instead of Intermediate 53-7. LCMS: 886.2.

Intermediate 82-9 was synthesized in a manner similar to Intermediate 53-9 using Intermediate 82-8 instead of Intermediate 53-8. LCMS: 914.3.

Intermediate 82-10 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 82-9 instead of Intermediate 53-9. LCMS: 979.6.

Intermediate 82-11 was synthesized in a manner similar to Intermediate 53-11 using Intermediate 82-10 instead of Intermediate 53-10. LCMS: 823.0.

Intermediate 84-1 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 64-3 instead of Intermediate 75-6. LCMS: 494.6 [M+H]+

Intermediate 84-2 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 84-1 instead of Intermediate 75-9. LCMS: 784.9 [M+H]+

Intermediate 84-3 was synthesized in a manner similar to Intermediate 75-8 using Intermediate 84-2 instead of Intermediate 75-7. LCMS: 816.6 [M+H]+

Intermediate 84-4 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 84-3 instead of Intermediate 75-8. LCMS: 881.6 [M+H]+

Intermediate 84-5 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 84-4 instead of Intermediate 75-10. LCMS: 725.0 [M+H]+

To an oven dried flask was added (3S,6S)-5-tert-butoxycarbonyl-2,2-difluoro-5-azaspiro[2.4]heptane-6-carboxylic acid (2.5 g, 9 mmol) and anhydrous methanol (20 mL) under Ar. (Trimethylsilyl)diazomethane (2M in diethyl ether, 6.7 mL, 14 mmol) was added dropwise. Additional (trimethylsilyl)diazomethane was added until a persistent yellow color was achieved. The reaction was quenched with 2N hydrochloric acid. The mixture was concentrated in vacuo to remove methanol then ethyl acetate was added. The mixture was washed with brine, dried over magnesium sulfate, filtered and concentrated in vacuo. The resulting material was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0 to 60%) to afford Intermediate 85-1. LCMS[M+Na]+: 314.2.

Intermediate 85-1 (2.3 g, 7.8 mmol) was left on high vac overnight. The flask was purged/filled with argon several times and then anhydrous tetrahydrofuran (50 mL) was added. The solution was cooled to −78C and LiHMDS (1M in tetrahydrofuran, 9.4 mL, 9.4 mmol) was added dropwise. The resulting mixture was stirred at −78C for 1 h then anhydrous hexamethylphosphoramide (8.2 mL, 47 mmol) was added dropwise followed by 1-chloro-3-iodo-propane (1.3 mL, 13 mmol). The resulting solution was allowed to warm to room temperature in the bath and stirred overnight. The solution was diluted with diethyl ether and washed with saturated aqueous ammonium chloride, 0.1M hydrochloric acid, water and brine. The solution was then dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0 to 50%) to afford Intermediate 85-2.

Intermediate 85-2 (2.9 g, 7.8 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL) was added. The solution was stirred at room temperature for 30 min. Toluene (2 mL) was added and the solution was concentrated in vacuo and left on high vacuum for 30 min. The residue was taken up into N,N-dimethylformamide (10 mL). Potassium iodide (130 mg, 780 μmol) and potassium carbonate (5.4 g, 39 mmol) were added. The mixture was stirred vigorously overnight at 50 C. The mixture was diluted with water and extracted 3 times with ethyl acetate. The aqueous layer was saturated with sodium chloride and extracted 3 times with dichloromethane. The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by basic alumina column chromatography using a gradient of ethyl acetate in hexanes (0 to 50%) to afford Intermediate 85-3. LCMS: 232.2.

Intermediate 85-3 (971 mg, 4.2 mmol) was azeotroped from toluene and dissolved in tetrahydrofuran (3 mL). An oven dried vial was charged with 1M lithium aluminum hydride in tetrahydrofuran (6.3 mL, 6.3 mmol) under Ar. The solution was diluted to 0.5M with tetrahydrofuran and cooled to OC. The ester solution was added dropwise and the resulting solution was stirred at OC for 10 min. The solution was diluted with diethyl ether and 240 uL of water was added slowly at OC. 240 uL of 3N sodium hydroxide was added followed by 720 uL of water. The resulting mixture was stirred vigorously at room temperature for 15 minutes. Magnesium sulfate was added and vigorous stirring was continued for an additional 15 minutes. The mixture was filtered, rinsing thoroughly with ethyl acetate. The filtrate was concentrated in vacuo to give Intermediate 85-4. LCMS: 204.2.

Intermediate 85-4 (767 mg, 3.8 mmol) was azeotroped from toluene. Imidazole (771 mg, 11 mmol) was added and the mixture was dissolved in N,N-dimethylformamide (7 mL) and cooled to OC. tert-butylchlorodiphenylsilane (1.5 mL, 5.7 mmoL) was added and the resulting solution was stirred over night at room temperature. Water was added and the mixture was extracted twice with diethyl ether. The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a gradient of methanol in dichloromethane (0 to 10%) to give Intermediate 85-5. LCMS: 442.3.

Intermediate 85-6 was purified by chiral SFC to give Intermediate 85-6 as the minor diastereomer (first eluting peak).

Intermediate 85-6 (30 mg, 68 μmol) was dissolved in tetrahydrofuran (0.5 mL) and tetra-n-butylammonium fluoride (1M in tetrahydrofuran, 0.1 mL, 100 μmol) was added. The resulting solution was stirred for 24 h at room temperature. The reaction mixture was concentrated in vacuo and dissolved in 1 mL of toluene. ⅓ of the solution was used in the next step without purification.

A solution of Intermediate 85-7 (5 mg, 25 μmol) in toluene was added to a vial charged with Intermediate 84-3 (20 mg, 25 μmol). The solution was concentrated in vacuo and dissolved in tetrahydrofuran (0.5 mL). LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) was added and the resulting solution was stirred at room temperature for 1 h. The solution was diluted with ethyl acetate and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on basic alumina using a gradient of ethyl acetate in hexanes (0 to 100%) to give Intermediate 85-8. LCMS: 769.4.

Intermediate 86-1 was synthesized in a manner similar to Intermediate 75-1 using Intermediate 27-1 instead of Intermediate 27-5. LCMS: 409.1 [M+Na]+

Intermediate 86-3 was synthesized in a manner similar to Intermediate 53-2 using Intermediate 86-2 instead of Intermediate 27-3. LCMS: 401.2 [M+H]+

Intermediate 86-5 was synthesized in a manner similar to Intermediate 64-1 using Intermediate 86-4 and ethyltriphenylphosphonium bromide instead of Intermediate 61-1 and methyltriphenylphosphonium bromide LCMS: 433.1 [M+Na]+

Intermediate 86-6 was synthesized in a manner similar to Intermediate 75-5 using Intermediate 86-5 instead of Intermediate 75-4 and LCMS: 267.0 [M+H]+

Intermediate 86-7 was synthesized in a manner similar to Intermediate 64-3 using Intermediate 86-6 instead of Intermediate 64-2. LCMS: 586.7, 588.1 [M+H]+

Intermediate 86-8 was synthesized in a manner similar to Intermediate 84-1 using Intermediate 86-7 instead of Intermediate 64-3. LCMS: 508.5 [M+H]+

Intermediate 86-9 was synthesized in a manner similar to Intermediate 75-8 using Intermediate 86-8 instead of Intermediate 75-7. LCMS: 540.3 [M+H]+

Intermediate 86-10 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 86-9 instead of Intermediate 75-8. LCMS: 605.3 [M+H]+

Intermediate 86-11 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 86-10 instead of Intermediate 75-9. LCMS: 895.1 [M+H]+

Intermediate 86-12 was synthesized in a manner similar to Intermediate 76-2 using Intermediate 86-11 instead of Intermediate 76-1. LCMS: 739.0 [M+H]+

Intermediate 87-1 was synthesized in a manner similar to Intermediate 75-10 using Intermediate 86-10 instead of Intermediate 75-9. LCMS: 955.4 [M+H]+

Intermediate 87-2 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 87-1 instead of Intermediate 75-10. LCMS: 799.1 [M+H]+

Intermediate 90-1 was synthesized in a manner similar to Intermediate 53-2 using ethyl magnesium bromide instead of methyl magnesium bromide and using Intermediate 86-2 instead of Intermediate 27-3. LCMS: 437.1 [M+Na]+.

Intermediate 90-2 was synthesized in a manner similar to Intermediate 27-3 using Intermediate 90-1 instead of Intermediate 27-2. LCMS: 412.7.

Intermediate 90-3 was synthesized in a manner similar to Intermediate 27-4 using Intermediate 90-2 instead of Intermediate 27-3. LCMS: 410.8.

Intermediate 90-4 was synthesized in a manner similar to Intermediate 75-5 using Intermediate 90-3 instead of Intermediate 75-4. LCMS: 267.0.

Intermediate 90-5 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 90-4 instead of Intermediate 53-4. LCMS: 588.2.

Intermediate 90-6 and Intermediate 91-1 were synthesized in a manner similar to

Intermediate 90-7 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 90-6 instead of Intermediate 13-8. LCMS: 540.2.

Intermediate 90-8 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 90-7 instead of Intermediate 53-9. LCMS: 605.1.

Intermediate 90-9 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 90-8 instead of Intermediate 17-9. LCMS: 955.4.

Intermediate 90-10 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 90-9 instead of Intermediate 13-10. LCMS: 798.8.

Intermediate 91-2 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 91-1 instead of Intermediate 17-9. LCMS: 895.6.

Intermediate 91-3 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 91-2 instead of Intermediate 13-10. LCMS: 739.0.

Intermediate 92-1 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 91-1 instead of Intermediate 13-8. LCMS: 540.2.

Intermediate 92-2 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 92-1 instead of Intermediate 53-9. LCMS: 605.2.

Intermediate 92-3 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 92-2 instead of Intermediate 17-9. LCMS: 955.6.

Intermediate 92-4 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 92-3 instead of Intermediate 13-10. LCMS: 798.9.

Intermediate 93-1 was synthesized in a manner similar to Intermediate 27-4 using ethyltriphenylphosphonium bromide instead of methyltriphenylphosphonium bromide. LCMS: 419.2 [M+Na]+

Intermediate 93-2 was synthesized in a manner similar to Intermediate 75-5 using Intermediate 93-1 instead of Intermediate 75-4. LCMS: 253.0 [M+H]+

Intermediate 93-3 was synthesized in a manner similar to Intermediate 63-5 using Intermediate 93-2 instead of Intermediate 27-5. LCMS: 572.6, 574.1 [M+H]+

Intermediate 93-4 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 93-3 instead of Intermediate 75-6. S-methyl isomer is the slower eluted fraction in the silica gel chromatography purification. LCMS: 494.5 [M+H]+

Intermediate 93-5 was synthesized in a manner similar to Intermediate 86-9 using Intermediate 93-4 instead of Intermediate 86-8. LCMS: 526.2 [M+H]+

Intermediate 93-6 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 93-5 instead of Intermediate 75-8. LCMS: 591.2 [M+H]+

Intermediate 93-7 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 93-6 instead of Intermediate 75-9. LCMS: 881.3 [M+H]+

Intermediate 93-8 was synthesized in a manner similar to Intermediate 76-2 using Intermediate 93-7 instead of Intermediate 76-1. LCMS: 725.1 [M+H]+

Intermediate 94-1 was synthesized in a manner similar to Intermediate 75-10 using Intermediate 93-6 instead of Intermediate 75-9. LCMS: 941.2 [M+H]+

Intermediate 94-2 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 94-1 instead of Intermediate 75-10. LCMS: 785.0 [M+H]+

Intermediate 95-1 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 93-3 instead of Intermediate 75-6. R-methyl isomer was the faster eluted fraction in the silica gel chromatography purification. LCMS: 494.6 [M+H]+

Intermediate 95-2 was synthesized in a manner similar to Intermediate 86-9 using Intermediate 95-1 instead of Intermediate 86-8. LCMS: 526.2 [M+H]+

Intermediate 95-3 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 95-2 instead of Intermediate 75-8. LCMS: 591.2 [M+H]+

Intermediate 95-4 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 95-3 instead of Intermediate 75-9. LCMS: 881.3 [M+H]+

Intermediate 95-5 was synthesized in a manner similar to Intermediate 76-2 using Intermediate 95-4 instead of Intermediate 76-1. LCMS: 725.0 [M+H]+

Intermediate 96-1 was synthesized in a manner similar to Intermediate 75-10 using Intermediate 95-3 instead of Intermediate 75-9. LCMS: 941.6 [M+H]+

Intermediate 96-2 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 96-1 instead of Intermediate 75-10. LCMS: 785.0 [M+H]+

The solution of Intermediate 75-6 (30.0 mg, 0.052 mmol), Pd(dppf)Cl2 (7.66 mg, 0.0105 mmol), cesium carbonate (51.2 mg, 0.16 mmol) in anhydrous 1,4-dioxane (1.0 mL) and degassed water (0.2 mL) was vigorously stirred at 90° C. for 15 minutes before it was cooled to room temperature. The mixture was washed with water (2 mL) and extracted with ethyl acetate (3×5 mL). The organic layer was collected and combined, dried over magnesium sulfate, and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 97-1. LCMS: 492.1.

A vigorously stirred solution of Intermediate 97-1 (10.1 mg, 0.021 mmol) in anhydrous dichloromethane (0.5 mL), was bubbled with ozone at −78° C. for until the color of solution was changed to light yellow. The solution was then bubbled with nitrogen for 1 minute and dimethyl sulfide (1.8 μL, 0.025 mmol) was added at −78° C. The mixture was stirred at −78° C. before being warmed to room temperature. The mixture was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 97-2. LCMS: 494.0.

To a vigorously stirred solution of Intermediate 97-2 (5.8 mg, 0.012 mmol) in anhydrous dichloromethane (1.0 mL), was added diethylaminosulfur trifluoride at 0° C. The mixture was warmed to 30° C. and stirred at 30° C. for 24 hours before it was transferred into saturated sodium bicarbonate solution (2 mL). The resulting mixture was extracted with dichloromethane (3×5 mL). The organic layer was collected and combined, dried over magnesium sulfate, and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 20% ethyl acetate in hexanes) to give Intermediate 97-3. LCMS: 516.1.

Intermediate 97-4 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 97-3 instead of Intermediate 13-8. LCMS: 548.0.

Intermediate 97-5 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 97-4 instead of Intermediate 53-9. LCMS: 613.1.

Intermediate 97-6 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 97-5 instead of Intermediate 17-9. LCMS: 963.3.

Intermediate 97-7 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 97-6 instead of Intermediate 13-10. LCMS: 807.0.

Intermediate 98-1 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 97-5 instead of Intermediate 17-9. LCMS: 903.1.

Intermediate 98-2 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 98-1 instead of Intermediate 13-10. LCMS: 747.0.

Intermediate 99-1 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 63-5 instead of Intermediate 75-6. LCMS: 480.5 [M+H]+

Intermediate 99-2 was synthesized in a manner similar to Intermediate 79-2 using Intermediate 99-1 and Intermediate 7-5 instead of Intermediate 79-1 and ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane. LCMS: 734.4 [M+H]+

Intermediate 99-3 was synthesized in a manner similar to Intermediate 79-3 using Intermediate 99-2 instead of Intermediate 79-2. LCMS: 766.1 [M+H]+

Intermediate 99-4 was synthesized in a manner similar to Intermediate 79-4 using Intermediate 99-3 and ((1S,7a'S)-2,2-difluorodihydro-1′H,3′H-spiro[cyclopropane-1,2′-pyrrolizin]-7a′(5′H)-yl)methanol instead of Intermediate 79-3 and ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol. LCMS: 875.0 [M+H]+

Intermediate 100-1 was synthesized in a manner similar to intermediate 35-1 using intermediate 84-3 instead of intermediate 13-9. LCMS: 824.5.

Intermediate 100-2 was synthesized in a manner similar to intermediate 35-2 using intermediate 100-1 instead of intermediate 35-1. LCMS: 822.4.

Intermediate 100-3 was synthesized in a manner similar to intermediate 35-3 using intermediate 100-2 instead of intermediate 35-2. LCMS: 949.6.

Intermediate 100-4 was synthesized in a manner similar to intermediate 35-4 using intermediate 100-3 instead of intermediate 35-3. LCMS: 793.8.

Intermediate 102-1 was synthesized in a manner similar to intermediate 63-7 using intermediate 7-5 instead of ((2-fluoro-6-(methoxymethoxy)-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane. LCMS: 766.2.

Intermediate 103-1 was synthesized in a manner similar to intermediate 106-2 using (S)-3-(trifluoromethoxy)pyrrolidine instead of 4-(trifluoromethoxy)piperidine. LCMS: 240.300.

N,N-Diisopropylethylamine (430 μL, 2.50 mmol) was added via syringe to a stirred mixture of 1-(methoxycarbonyl)cyclopropane-1-carboxylic acid (120 mg, 830 μmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (467 mg, 1.20 mmol), and N,N-dimethylformamide (1.6 mL) at room temperature. After 30 min, the resulting mixture was added via syringe to 4-(trifluoromethoxy)piperidine hydrochloride (216 mg, 1.10 mmol). After 238 min, saturated aqueous sodium bicarbonate solution was added, and the aqueous layer was extracted with diethyl ether (4×5 mL). The combined organic layers were washed with brine (2×10 mL), were dried over anhydrous magnesium sulfate, were filtered, and were concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 100% ethyl acetate in hexanes) to give intermediate 106-1. LCMS: 296.0.

Lithium aluminum hydride solution (2.0 M in tetrahydrofuran, 700 μL, 1.4 mmol) was added dropwise via syringe to a stirred solution of intermediate 106-1 (204 mg, 690 μmol) in tetrahydrofuran (3.0 mL) at 0° C. After 34 min, water (50 μL), aqueous sodium hydroxide solution (6.0 M, 50 μL), and water (150 μL) were added sequentially, and the resulting mixture was filtered. The filtrate was concentrated under reduced pressure, and the residue was purified by flash column chromatography on basic alumina (0 to 100% ethyl acetate in hexanes) to give intermediate 106-2. LCMS: 254.1.

To a solution of intermediate 109-1 (80 mg, 138.50 umol, 1 eq) in MeOH (2 mL) was added HCl/MeOH (4 M) (0.5 mL). The mixture was stirred at 25° C. for 1 hr. The reaction solution was blow-dried with N2. The residue was diluted with H2O (5 mL) and washed with EtOAc (5 mL*3). The aqueous phase was filtered and was freeze-dried to give intermediate 109-2 as its HCl salt. LCMS: 336.1.

Intermediate 113-1 was prepared in a manner analogous to Intermediate 122-2 using (8-ethynyl-7-fluoronaphthalen-1-yl) pinacol boronate.

O1-tert-butyl O2-methyl (2S,4R)-4-fluoropyrrolidine-1,2-dicarboxylate (4.1 g, 17 mmol) was dissolved in tetrahydrofuran in an oven dried flask under argon. The solution was cooled to −78C and LiHMDS (1M in tetrahydrofuran, 20 mL, 20 mmol) was added dropwise. The resulting solution was stirred at −78C for 1 h. Hexamethylphosphoramide (29 mL, 165 mmol) was added dropwise followed by [1-(bromomethyl)cyclopropyl]methoxymethylbenzene (5.4 g, 21 mmol). The resulting solution was allowed to warm to room temperature and stirred for 3 days. The solution was diluted with diethyl ether and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0 to 40%) to provide Intermediate 113-2. LCMS [M+Na]+: 444.2.

Intermediate 113-2 (4.5 g, 11 mmol) was dissolved in ethanol (40 mL) and 10% palladium on carbon (1.1 g, 1.1 mmol) was added. The flask was purged and filled with hydrogen 3 times and the mixture was stirred at room temperature under hydrogen overnight. The reaction mixture was filtered through celite, rinsing with ethyl acetate, and concentrated in vacuo. The resulting residue was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0 to 80%) to provide Intermediate 113-3. LCMS [M+Na]+: 354.2.

Intermediate 113-3 (2.9 g, 8.9 mmol), triphenylphosphine (3.4 g, 13 mmol) and carbon tetrabromide (4.4 g, 13 mmol) were dissolved in dichloromethane (30 mL) and the resulting solution was stirred overnight at room temperature. The solution was diluted with hexanes and purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0 to 80%) to provide Intermediate 113-4. LCMS [M+Na]+: 416.1.

Intermediate 113-4 (3.5 g, 8.9 mmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (5 mL) was added. The solution was stirred at room temperature for 30 min. Toluene (2 mL) was added and the solution was concentrated in vacuo and left on high vacuum for 30 min. The residue was taken up into N,N-dimethylformamide (10 mL). Potassium iodide (150 mg, 890 mmol) and potassium carbonate (3.6 g, 27 mmol) were added. The mixture was stirred vigorously overnight at 50 C. The mixture was poured into a mixture of saturated aqueous sodium carbonate and brine and extracted with ethyl acetate. Before each extraction, the aqueous layer was saturated with sodium chloride. The aqueous layer was extracted twice more with ethyl acetate and 3 times with dichloromethane. The combined organic layers were dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by basic alumina column chromatography using a gradient of ethyl acetate in hexanes (0 to 100%) to afford Intermediate 113-5. LCMS: 214.1.

Intermediate 113-5 (98 mg, 460 μmol) was dissolved in tetrahydrofuran (1 mL). An oven dried vial was charged with 1M lithium aluminum hydride in tetrahydrofuran (6.3 mL, 6.3 mmol) under Ar. The solution was diluted to 0.5M with tetrahydrofuran and cooled to OC. The ester solution was added dropwise and the resulting solution was stirred at OC for 30 min. The solution was diluted with diethyl ether and 26 uL of water was added slowly at OC. 26 uL of 3N sodium hydroxide was added followed by 75 uL of water. The resulting mixture was stirred vigorously at room temperature for 15 minutes. Magnesium sulfate was added and vigorous stirring was continued for an additional 15 minutes. The mixture was filtered, rinsing thoroughly with ethyl acetate. The filtrate was concentrated in vacuo to give Intermediate 113-6. LCMS: 186.2.

Intermediate 113-1 (5 mg, 7.7 μmol) and Intermediate 113-6 (2.9 mg, 16 μmol) were azeotroped together from toluene and dissolved in 2-methyltetrahydrofuran (0.5 mL). LiHMDS (1M in tetrahydrofuran, 16 uL, 16 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with ethyl acetate and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give crude Intermediate 113-7 which was used directly in the next step without purification. LCMS: 737.4.

To a solution of intermediate 114-0 (60.00 mg, 139.13 umol, 1 eq) in THF (2 mL) was added NaBH4(42.11 mg, 1.11 mmol, 8 eq) and BF3Et2O (315.94 mg, 2.23 mmol, 274.73 uL, 16 eq) at 0° C. under N2. The mixture was stirred at 60° C. for 1 hr under N2. The reaction mixture was quenched by addition saturated aqueous NH4Cl solution (2 mL) at 0° C., and then extracted with EtOAc (2 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100*30 mm*5 um; mobile phase: [water(TFA)-ACN]; B %: 10%-40%, 10 min) to give intermediate 114-1, the absolute stereochemistry of which was assigned arbitrarily. LCMS: 376.0.

Intermediate 113-1 (5 mg, 7.7 μmol) and Intermediate 114-1 (7 mg, 14 μmol) were azeotroped together from toluene and dissolved in 2-methyltetrahydrofuran (0.5 mL). LiHMDS (1M in tetrahydrofuran, 31 uL, 31 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with ethyl acetate and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over sodium sulfate, filtered and concentrated in vacuo to give crude Intermediate 114-2 as a single unknown trans pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 927.4.

A vigorously stirred mixture of Intermediate 75-7 (181 mg, 0.367 mmol), ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (prepared according to WO 2021/041671) (249 mg, 0.550 mmol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (13.2 mg, 36.7 mol), potassium phosphate (253 mg, 1.10 mmol), tetrahydrofuran (5.4 mL), and degassed water (1.1 mL) was heated to 70° C. After 80 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 115-1. LCMS: 784.4.

Cesium fluoride (415 mg, 2.73 mmol) was added to a vigorously stirred solution of Intermediate 115-1 (214 mg, 0.273 mmol) in N,N-dimethylformamide (6.2 mL) at room temperature. After 40 min, diethyl ether (40 mL), ethyl acetate (20 mL), and saturated aqueous sodium bicarbonate solution (10 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 to 60% ethyl acetate in hexanes) to give Intermediate 115-2. LCMS: 628.2.

3-Chloroperoxybenzoic acid (77% wt, 123 mg, 0.55 mmol) was added in two equal portions over 5 min to a vigorously stirred solution of Intermediate 115-2 (157 mg, 0.25 mmol) in dichloromethane (3.5 mL) at 0° C. After 90 min, the resulting mixture was warmed to room temperature. The residue was purified by flash column chromatography on silica gel (0% to 75% ethyl acetate in hexanes) to give Intermediate 115-3. LCMS: 660.3.

Intermediate 116-1 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 53-7 instead of Intermediate 17-9. LCMS: 786.3.

Intermediate 116-2 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 116-1 instead of Intermediate 13-8. LCMS: 818.2.

Intermediate 116-3 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 116-2 instead of Intermediate 13-9. LCMS: 883.0.

Intermediate 116-4 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 116-3 instead of Intermediate 13-10. LCMS: 727.0.

To a solution of ((3S,7aS)-3-(((tert-butyldimethylsilyl)oxy)methyl)hexahydro-1H-pyrrolizin-7a-yl)methanol (830 mg, 2.91 mmol, 1 eq) in DCM (5 mL) was added triethylamine (588.36 mg, 5.81 mmol, 809.29 μL, 2 eq) and benzoyl chloride (612.99 mg, 4.36 mmol, 506.60 μL, 1.5 eq) at 0° C. The mixture was stirred at 25° C. for 0.5 hr. The unreacted benzoyl chloride was quenched by addition H2O (30 mL) at 0° C., and the resulting mixture was extracted with DCM (30 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography on silica gel (PE:EtOAc=1:0 to 0:1) to give intermediate 119-1. LCMS: 390.2.

Intermediate 120-1 was synthesized in a manner similar to intermediate 102-1 using intermediate 65-1 instead of intermediate 7-5. LCMS: 706.2.

Intermediate 121-1 was prepared in a manner analogous to Intermediate 85-8 using Intermediate 113-1 as the starting material. LCMS: 755.3.

A vial was charged with Intermediate 63-6 (850 mg, 1 mmol) and cesium fluoride (3.7 g, 25 mmol). N,N-dimethylformamide (17 mL) was added and the resulting mixture was stirred vigorously at room temperature for 30 min. The mixture was diluted with diethyl ether and ethyl acetate, and washed with saturated aqueous sodium bicarbonate, water, and brine. The solution was dried over magnesium sulfate, filtered and concentrated in vacuo. The residue was purified by flash column chromatography on silica gel using a gradient of ethyl acetate in hexanes (0-100%) to give Intermediate 122-1. LCMS 674.3.

Intermediate 122-2 was prepared in a manner analogous to Intermediate 13-9 using Intermediate 122-1 as the starting material. LCMS: 706.2.

Intermediate 122-3 was prepared in a manner analogous to Intermediate 85-8 using Intermediate 122-2 as the starting material. LCMS: 815.4.

Intermediate 75-2 (950 mg, 2.37 mmol) was dissolved in pyridine (20 ml) and cooled to 0° C. To it was added acetic anhydride (1.12 ml, 11.9 mmol) and 4-Dimethylaminopyridine (57.9 mg, 0.474 mmol). The reaction mixture was stirred at 0° C. for 10 minutes. Upon completion, it was concentrated to dryness under reduced pressure and purified by silica gel chromatography eluting with ethyl acetate in hexane to afford the title compound. LCMS: 465.3

Intermediate 123-2 was synthesized in a manner similar to intermediate 75-5 using intermediate 123-1 instead of intermediate 75-4. LCMS: 299.2

Intermediate 123-3 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 123-2 instead of Intermediate 53-4. LCMS: 619.3.

Intermediate 123-3 (270 mg, 0.436 mmol) was dissolved in THF (6 ml)/MeOH (4 ml)/water (2 ml) and to it was added lithium hydroxide monohydrate (91.5 mg, 2.18 mmol). The reaction mixture was stirred at room temperature for 10 minutes. Upon completion, it was partitioned between ethyl acetate and brine. The organic layer was separated, dried over magnesium sulfate, filtered and concentrated to dryness to afford the title compound. LCMS: 576.2.

Intermediate 123-4 (180 mg, 0.312 mmol), CuI (11.9 mg, 0.0124 mmol) and t-BuONa (60 mg, 0.624 mmol) were charged into a microwave tube and DMF (2 mL) was added. The system was evacuated and backfilled with argon three times. The reaction mixture was kept stirring at 100° C. for overnight. The cooled solution was diluted with ethyl acetate and washed with brine. The organic phase was dried over Na2SO4and concentrated in vacuo. The residue was purified by silica gel chromatography to afford the title compound. LCMS: 496.2.

Intermediate 123-6 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 123-5 instead of Intermediate 13-8. LCMS: 528.2.

Intermediate 123-7 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 123-6 instead of Intermediate 53-9. LCMS: 593.3.

Intermediate 123-8 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 123-7 instead of Intermediate 17-9. LCMS: 943.5.

Intermediate 124-1 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 123-7 instead of Intermediate 17-9. LCMS: 883.5.

Intermediate 122-2 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 129-1 (5.7 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 129-2 as a single unknown trans pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 987.4.

Intermediate 113-1 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 129-1 (5.7 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 130-1 as a single unknown trans pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 927.4.

Intermediate 132-1 was synthesized in a manner similar to Intermediate 75-5 using intermediate 170-1 instead of intermediate 27-5. LCMS: 253.2.

Intermediate 132-2 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 132-1 instead of Intermediate 53-4. LCMS: 572.1.

Intermediate 132-2 (57 mg, 0.1 mmol) was dissolved in dichloromethane (1 mL) and the stirred solution was evacuated and refilled with argon (3×). To this solution 1,2-bis(diphenylphosphino)ethane (12.2 mg, 0.03 mmol) was added, followed by bis(1,5-cyclooctadiene)diiridium(I) dichloride (10.3 mg, 0.015 mmol), after which the resulting mixture was again evacuated and refilled with nitrogen (3×). After stirring for 30 minutes at room temperature, the reaction mixture was cooled to 0° C. and a solution of pinacolborane (0.0229 ml, 0.015 mmol) in dichloromethane (0.5 mL) was added dropwise over 15 min. After the addition, the ice bath was removed, and the reaction was stirred for additional 90 minutes at room temperature. Upon completion, the reaction mixture was quenched with saturated aqueous NH4Cl solution, and the aqueous phase was extracted with dichloromethane. The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated. The crude mixture was purified by column chromatography (silica gel, 0→20% EtOAc in hexanes) to afford the title compound. LCMS: 702.2

Intermediate 132-3 (30 mg, 0.043 mmol) was dissolved in 1 mL dioxane and 0.5 mL water. Sodium carbonate (18.5 mg, 0.175 mmol) was added, and the mixture was degassed and backfilled with argon (3×). Pd(dppf)Cl2(4.6 mg, 0.006 mmol) was added, and the mixture was heated at 90° C. for 1 hour. The mixture was cooled to room temperature, diluted with EtOAc and washed with sat. aq. ammonium chloride solution. Combined organic fractions were washed with brine, dried and concentrated. The crude product was purified by column chromatography (silica gel, 0→20% EtOAc in hexanes) to afford the title compound. LCMS: 494.2

Intermediate 132-5 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 132-4 instead of Intermediate 13-8. LCMS: 526.2.

Intermediate 132-6 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 132-5 instead of Intermediate 53-9. LCMS: 591.3.

Intermediate 132-7 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 132-6 instead of Intermediate 17-9. LCMS: 942.6.

Intermediate 133-1 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 170-4 instead of Intermediate 13-8. LCMS: 512.2.

Intermediate 133-2 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 133-1 instead of Intermediate 53-9. LCMS: 578.3.

Intermediate 133-3 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 133-2 instead of Intermediate 17-9. LCMS: 927.6.

Intermediate 122-2 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 134-1 (4.8 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 134-2 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 987.4.

Intermediate 113-1 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 134-1 (4.8 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until full conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 135-1 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 927.4.

Intermediate 136-1 was synthesized in a manner similar to Intermediate 84-4 using (1-((1,1-difluoro-6-azaspiro[2.5]octan-6-yl)methyl)cyclopropyl)methanol instead of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol. LCMS: 953.3 [M+H]+

Intermediate 136-2 was synthesized in a manner similar to Intermediate 84-5 using Intermediate 136-1 instead of Intermediate 84-4. LCMS: 797.1 [M+H]+

Intermediate 137-0: LCMS: 282.1. The peak with longer Rt (Rt=1.454) was designated as intermediate 139-0, the absolute stereochemistry of which was assigned arbitrarily. Intermediate 139-0: LCMS: 282.1

To a mixture of intermediate 137-0(60 mg, 213.35 umol, 1 eq) in THF (3 mL) was added BF3·Et2O (484.49 mg, 3.41 mmol, 421.30 uL, 16 eq) at 0° C. under N2. The reaction mixture was stirred at 0° C. for 5 mins, then NaBH4(64.57 mg, 1.71 mmol, 8 eq) was added, the reaction mixture was stirred at 60° C. for 1 hr under N2. To the reaction mixture was added sat.aq. NH4Cl (10 mL) slowly and the reaction mixture was stirred at 0° C. for 0.5 hr under N2, and then extracted with EtOAc (10 mL*3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100*30 mm*5 um; mobile phase: [water(TFA)-ACN]; B %: 1%-20%, 10 min) to give intermediate 137-1, the absolute stereochemistry of which was assigned arbitrarily. LCMS: 226.1.

Intermediate 113-1 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 137-1 (4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until full conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 137-2 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 777.4.

Intermediate 122-2 (5 mg, 7.7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 137-1 (4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until full conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 138-1 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 837.4.

To a mixture of intermediate 139-0 (25 mg, 88.9 umol, 1 eq) and THF (2 mL) was added BF3·Et2O (201.9 mg, 1.4 mmol, 175.5 uL, 16 eq) at 0° C. under N2, the reaction mixture was stirred at 0° C. for 5 mins, then NaBH4(26.9 mg, 711.2 umol, 8 eq) was added, the reaction mixture was stirred at 60° C. for 1 hr under N2. To the reaction mixture was added sat. aq. NH4Cl (10 mL) slowly and stirred at 0° C. for 0.5 hr under N2, and then extracted with EtOAc (10 mL*3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100*30 mm*5 um; mobile phase: [water(TFA)-ACN]; B %: 1%-20%, 10 min) to give intermediate 139-1, the absolute stereochemistry of which was assigned arbitrarily. LCMS: 226.1.

Intermediate 122-2 (6 mg, 8 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 139-1 (4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 139-2 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 837.4.

Intermediate 113-1 (6 mg, 9 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 139-1 (4 mg, 13 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional 1M LiHMDS was added drop by drop until complete conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 140-1 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 777.4.

Intermediate 141-0, the absolute stereochemistry of which was assigned arbitrarily, and intermediate 141-1, the absolute stereochemistry of which was assigned arbitrarily, were synthesized in a manner similar to intermediate 137-1 and 139-1, respectively, using rac-(2S,7aR)-ethyl 2-hydroxy-5-oxohexahydro-1H-pyrrolizine-7a-carboxylate instead of rac-ethyl (2R,7aR)-2-hydroxy-5-oxotetrahydro-1H-pyrrolizine-7a(5H)-carboxylate. Intermediate 141-0: LCMS: 226.1. Intermediate 141-1: LCMS: 226.1.

Intermediate 122-2 (5 mg, 7.4 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 141-1 (4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional 1M LiHMDS was added until complete conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 141-2 as a single unknown trans pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 837.4.

Intermediate 113-1 (5 mg, 8 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 141-1 (4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional 1M LiHMDS was added until conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 142-1 as a single unknown trans pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 777.4.

A solution of benzyl 4-(hydroxymethyl)piperidine-1-carboxylate (500.1 mg, 2.0 mmol) and di-tert-butyl azodicarboxylate (2311.9 mg, 10.0 mmol) in tetrahydrofuran (9. mL) was stirred at 0° C. as 1 M trimethylphosphine in tetrahydrofuran (10 mL, 10 mmol) followed by nonafluoro-tert-BuOH (2.8 mL, 20.1 mmol) were added. After addition, the reaction mixture was stirred at 0° C. for 30 min and then at 70° C. overnight. The reaction mixture was diluted with ethyl acetate and the resulting solution was washed with sat'd NH4Cl (×2), sat'd NaHCO3(×2), and brine (×1). After the resulting organic solution was dried (MgSO4), and concentrated, the residue was purified by silica gel column chromatography eluting 0-45% ethyl acetate in hexane to give intermediate 143-1. LCMS: 468.2.

A mixture of benzyl intermediate 143-1 (264.1 mg, 0.565 mmol) and 10% palladium on carbon (28.1 mg) in ethanol (10 mL) was stirred under H2atmosphere. After 1 h, the reaction mixture was filtered and the filtrate was concentrated to give intermediate 143-2. LCMS: 334.1.

A mixture of intermediate 143-2 (0.57 mmol), 1-methoxycarbonylcyclopropanecarboxylic acid (88.3 mg, 0.613 mmol), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium-3-oxide hexafluorophosphate (331.1 mg, 0.87 mmol) in dimethylformamide (1.5 mL) was stirred at rt as diisopropylethylamine (0.34 mL, 2.0 mmol) was added. The resulting mixture was stirred at rt. After 3 d, the reaction mixture was diluted with ethyl acetate (25 mL) and washed with saturated NaHCO3(˜25 mL×1) and then water (˜25 mL×1). After the aqueous fractions were extracted with ethyl acetate (˜20 mL×1), the resulting organic fractions were combined, dried (MgSO4), filtered and concentrated. The residue was purified by silica gel column chromatography eluting 0-100% ethyl acetate in hexane to give intermediate 143-3. LCMS: 460.2.

A solution of methyl intermediate 143-3 (198.9 mg, 0.44 mmol) was stirred at 0° C. as 2 M lithium aluminum hydride (0.44 mL, 0.88 mmol) was added. The resulting mixture was stirred at rt for 3 h. The reaction mixture was stirred at 0° C. as sodium sulfate decahydrate (˜2.3 g). After the mixture was stirred for 10 min, it was diluted with ethyl acetate (˜20 mL), dried (Na2SO4), filtered, and concentrated. The residue was purified by silica gel column chromatography eluting 0-100% ethyl acetate in hexane to give intermediate 143-4. LCMS: 418.2.

A mixture of intermediate 122-2 (41.4 mg, 58.7 umol) and intermediate 143-4 (34.8 mg, 83.5 umol) was co-evaporated with toluene (×2) and dissolved in 2-methyltetrahydrofuran (0.7 mL). The solution was stirred at 0° C. as 1 M lithium bis(trimethylsilyl)amide was added dropwise. After 15 min, the reaction mixture was diluted with Sat'd NaHCO3and the product was extracted with ethyl acetate (×2). After the extracts were washed with water (×1), the organic fractions were combined, dried (MgSO4), and concentrated to give intermediate 143-5. LCMS: 1029.4.

Intermediate 122-2 (5 mg, 7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 144-1 (5.5 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 26 uL, 26 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until full conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 144-2 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 987.4.

Intermediate 113-1 (5 mg, 8 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 144-1 (6 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 26 uL, 26 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 145-1 as a single unknown cis pyrrolizidine isomer, which was used directly in the next step without purification. LCMS: 927.4.

Intermediate 146-1 was synthesized in a manner similar to Intermediate 84-4 using (1-((4-(trifluoromethyl)piperidin-1-yl)methyl)cyclopropyl)methanol instead of ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol. LCMS: 959.2 [M+H]+

Intermediate 146-2 was synthesized in a manner similar to Intermediate 84-5 using Intermediate 146-1 instead of Intermediate 84-4. LCMS: 803.1 [M+H]+

To a solution of intermediate 148-1 (130 mg, 232.30 umol, 1 eq) in EtOAc (1 mL) was added HCl/EtOAc (0.25 mL). The mixture was stirred at 25° C. for 1 hr. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (5 mL) and was washed with EtOAc (4 mL*3). The combined aqueous phases were lyophilized to give intermediate 148-2 as its HCl salt. LCMS: 318.1.

Intermediate 150-2 was synthesized in a manner similar to intermediate 119-4 using intermediate 150-1 instead of intermediate 119-3. LCMS: 322.0.

Intermediate 155-1 was synthesized in a manner similar to intermediate 148-2 using 2-chloro-5-(trifluoromethyl)pyrazine instead of 4-chloro-2-(trifluoromethyl)pyrimidine. LCMS: 318.0.

Intermediate 156-1 was synthesized in a manner similar to 64-1 using intermediate 86-4 instead of intermediate 61-1. LCMS: 419.1 [M+Na]+.

Intermediate 156-2 was synthesized in a manner similar to 75-2 using intermediate 156-1 instead of intermediate 75-1. This isomer is the faster eluted fraction. LCMS: 415.3 [M+H]+.

Intermediate 156-3 was synthesized in a manner similar to 75-3 using intermediate 156-2 instead of intermediate 75-2. LCMS: 413.3 [M+H]+.

Intermediate 156-4 was synthesized in a manner similar to 75-4 using intermediate 156-3 instead of intermediate 75-3. LCMS: 411.0 [M+H]+.

Intermediate 156-5 was synthesized in a manner similar to 75-5 using intermediate 156-4 instead of intermediate 75-4. LCMS: 267.1 [M+H]+.

Intermediate 156-6 was synthesized in a manner similar to 75-6 using intermediate 156-5 instead of intermediate 75-5. LCMS: 586.8, 588.2 [M+H]+.

Intermediate 156-7 was synthesized in a manner similar to 75-7 using intermediate 156-6 instead of intermediate 75-6. LCMS: 508.7 [M+H]+.

Intermediate 156-8 was synthesized in a manner similar to 76-1 using intermediate 156-7 instead of intermediate 75-9. LCMS: 798.5 [M+H]+.

Intermediate 156-9 was synthesized in a manner similar to 75-8 using intermediate 156-8 instead of intermediate 75-7. LCMS: 830.5 [M+H]+.

Intermediate 156-10 was synthesized in a manner similar to 75-9 using intermediate 156-9 instead of intermediate 75-8. LCMS: 895.1 [M+H]+.

Intermediate 156-11 was synthesized in a manner similar to 76-2 using intermediate 156-10 instead of intermediate 76-1. LCMS: 739.0 [M+H]+.

Intermediate 84-3 (5 mg, 6 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). 1,2,3,5,6,7-Hexahydropyrrolizin-8-ylmethanol (1.7 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 11 uL, 11 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added until complete conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 157-1, which was used directly in the next step without purification. LCMS: 863.5.

A vial was charged with Intermediate 157-1 (5.3 mg, 6 μmol) and cesium fluoride (22 mg, 150 μmol). N,N-dimethylformamide (0.5 mL) was added and the resulting mixture was stirred vigorously for 30 minutes. Acetic acid (0.37 uL, 6 μmol) and diethyl ether were added and the mixture was filtered and concentrated in vacuo to give crude Intermediate 157-2, which was used without purification in the next step. LCMS 707.4

Intermediate 158-1 was prepared in a manner analogous to Intermediate 113-6 using 05-tert-butyl 06-methyl (6S)-5-azaspiro[2.4]heptane-5,6-dicarboxylate as the starting material. LCMS: 194.2.

Intermediate 159-1 was synthesized in a manner similar to 75-2 using intermediate 156-1 instead of intermediate 75-1. This isomer is the slower eluted fraction. LCMS: 415.3 [M+H]+.

Intermediate 159-2 was synthesized in a manner similar to 75-3 using intermediate 159-1 instead of intermediate 75-2. LCMS: 413.3 [M+H]+.

Intermediate 159-3 was synthesized in a manner similar to 75-4 using intermediate 159-2 instead of intermediate 75-3. LCMS: 411.0 [M+H]+.

Intermediate 159-4 was synthesized in a manner similar to 75-5 using intermediate 159-3 instead of intermediate 75-4. LCMS: 267.1 [M+H]+.

Intermediate 159-5 was synthesized in a manner similar to 75-6 using intermediate 159-4 instead of intermediate 75-5. LCMS: 586.7, 588.2 [M+H]+.

Intermediate 159-6 was synthesized in a manner similar to 75-7 using intermediate 159-5 instead of intermediate 75-6. LCMS: 508.7 [M+H]+.

Intermediate 159-7 was synthesized in a manner similar to 76-1 using intermediate 159-6 instead of intermediate 75-9. LCMS: 798.5 [M+H]+.

Intermediate 159-8 was synthesized in a manner similar to 75-8 using intermediate 159-7 instead of intermediate 75-7. LCMS: 830.5 [M+H]+.

Intermediate 159-9 was synthesized in a manner similar to 75-9 using intermediate 159-8 instead of intermediate 75-8. LCMS: 895.1 [M+H]+.

Intermediate 159-10 was synthesized in a manner similar to 76-2 using intermediate 159-9 instead of intermediate 76-1. LCMS: 739.0 [M+H]+.

Intermediate 160-1 was synthesized in a manner similar to intermediate 148-2 using 2-chloro-3-(trifluoromethoxy)pyridine instead of 4-chloro-2-(trifluoromethyl)pyrimidine. LCMS: 333.1.

Intermediate 161-1 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 75-7 instead of Intermediate 13-7. LCMS: 844.6.

Intermediate 161-2 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 161-1 instead of Intermediate 13-8. LCMS: 876.3.

To a solution of intermediate 163-0 (170 mg, 343.04 umol, 1 eq) in EtOAc (2 mL) was added HCl/EtOAc (0.5 mL). The mixture was stirred at 25° C. for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100*30 mm*5 um; mobile phase: [water(HCl)-ACN]; B %: 1%-20%, 10 min) to give intermediate 163-1. LCMS: 254.1.

Intermediate 122-2 (5 mg, 7 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 163-1 (3.4 mg, 12 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 26 uL, 26 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 163-2, which was used directly in the next step without purification. LCMS: 865.4.

Intermediate 113-1 (6 mg, 9 μmol) was azeotroped from toluene then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). Intermediate 163-1 (3.7 mg, 13 μmol) was left on high vacuum overnight then dissolved in 2-methyl-tetrahydrofuran (0.5 mL). To the alcohol was added LiHMDS (1M in tetrahydrofuran, 26 uL, 26 μmol) and the resulting solution was added to the sulfone solution under argon atmosphere. The resulting solution was stirred at room temperature for 5 minutes. Additional LiHMDS was added dropwise until full conversion was observed by LCMS. The solution was diluted with ethyl acetate and diethyl ether and washed with a mixture of saturated aqueous sodium carbonate and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 164-1, which was used directly in the next step without purification. LCMS: 805.4.

Intermediate 122-2 (5.5 mg, 8 μmol) and Intermediate 165-1 (3.4 mg, 12 μmol) were azeotroped together from toluene with N,N-diisopropylethylamine (20 uL) and dissolved in 2-methyl-tetrahydrofuran (0.5 mL) under argon. LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with diethyl ether and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 165-2, which was used directly in the next step without purification. LCMS: 785.4.

Intermediate 113-1 (5.5 mg, 8 μmol) and Intermediate 165-1 (3.7 mg, 13 μmol) were azeotroped together from toluene with N,N-diisopropylethylamine (20 uL) and dissolved in 2-methyl-tetrahydrofuran (0.5 mL) under argon. LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with diethyl ether and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 166-1, which was used directly in the next step without purification. LCMS: 725.4.

To a mixture of methyl intermediate 167-0 (60 mg, 204.6 umol, 1 eq) in THF (2 mL) was added BF3·Et2O (464.6 mg, 3.3 mmol, 404.0 uL, 16 eq) at 0° C. under N2, the reaction mixture was stirred at 0° C. for 5 mins. Then NaBH4(61.9 mg, 1.6 mmol, 8 eq) was added, and the reaction mixture was stirred at 60° C. for 1 hr under N2. To the reaction mixture was added sat.aq. NH4Cl (10 mL) slowly and the reaction mixture was stirred at 0° C. for 0.5 hr under N2, and then extracted with EtOAc (10 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex Luna C18 100*30 mm*5 um; mobile phase: [water(TFA)-ACN]; B %: 1%-30%, 10 min) to give intermediate 167-1. LCMS: 252.1.

Intermediate 122-2 (5.5 mg, 8 mol) and Intermediate 167-1 (4.3 mg, 12 mol) were azeotroped together from toluene with N,N-diisopropylethylamine (20 uL) and dissolved in 2-methyl-tetrahydrofuran (0.5 mL) under argon. LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with diethyl ether and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 167-2, which was used directly in the next step without purification. LCMS: 863.4.

Intermediate 122-2 (5.5 mg, 8 μmol) and Intermediate 168-1 (4.5 mg, 12 μmol) were azeotroped together from toluene with N,N-diisopropylethylamine (20 uL) and dissolved in 2-methyl-tetrahydrofuran (0.5 mL) under argon. LiHMDS (1M in tetrahydrofuran, 27 uL, 27 μmol) was added and the resulting solution was stirred at room temperature for 5 minutes. The solution was diluted with diethyl ether and washed with water and brine. The organic layer was dried over magnesium sulfate, filtered and concentrated in vacuo to give crude Intermediate 168-2, which was used directly in the next step without purification. LCMS: 879.4.

Intermediate 169-1 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 80-6 instead of Intermediate 17-9. LCMS: 800.7.

Intermediate 169-2 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 169-1 instead of Intermediate 13-8. LCMS: 832.6.

Intermediate 169-3 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 169-2 instead of Intermediate 13-9. LCMS: 897.3.

Intermediate 169-4 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 169-3 instead of Intermediate 13-10. LCMS: 741.1.

Intermediate 170-1 was synthesized in a manner similar to Intermediate 27-5 using 8-(tert-butyl) 2-ethyl (1R,2R,5S)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate instead of 8-(tert-butyl) 2-ethyl (1R,2S,5S)-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate. LCMS: 239.2

Intermediate 170-2 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 170-1 instead of Intermediate 53-4. LCMS: 558.1

Intermediate 170-3 was synthesized in a manner similar to Intermediate 132-3 using Intermediate 170-2 instead of Intermediate 132-2. LCMS: 686.3

Intermediate 170-4 was synthesized in a manner similar to Intermediate 132-4 using Intermediate 170-3 instead of Intermediate 132-3. LCMS: 480.2

Intermediate 170-5 was synthesized in a manner similar to Intermediate 4-1 using Intermediate 170-4 instead of Intermediate 17-9. LCMS: 771.4.

Intermediate 170-6 was synthesized in a manner similar to Intermediate 28-2 using Intermediate 170-5 instead of Intermediate 28-1. LCMS: 615.3

Intermediate 170-7 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 170-6 instead of Intermediate 13-8. LCMS: 646.3.

Intermediate 170-8 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 170-7 instead of Intermediate 53-9. LCMS: 711.4

To a vigorously stirred solution of (ethyl)triphenylphosphonium bromide (0.782 g, 2.11 mmol) in tetrahydrofuran (10 mL) at room temperature was added KHMDS solution (1.0 M in tetrahydrofuran, 1.8 mL, 2.6 mmol) dropwise to afford a solution. The mixture was stirred for 1 hour at room temperature and was cooled to −78° C. whereupon a solution of Intermediate 75-3 (280 mg, 0.703 mmol) in tetrahydrofuran (5 mL) was added dropwise over 20 minutes. The resulting solution was allowed to gradually warm to room temperature and stir for 3 hours. The mixture was quenched with methanol (10 mL) and stirred for 15 min. Saturated aqueous ammonium chloride solution (50 mL) was added and the mixture was extracted with ethyl acetate (3×50 mL). The combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 40% ethyl acetate in hexanes) to give Intermediate 171-1 LCMS: 433.6 [M+Na]+.

Cesium fluoride (481 mg, 3.17 mmol) was added to a vigorously stirred solution of Intermediate 171-1 (260 mg, 0.63 mmol) in N,N-dimethylformamide (2.0 mL) at room temperature. The resulting mixture was stirred at 90° C. for 30 minutes. After cooled to room temperature, the mixture was diluted with EtOAc (30 mL), filtered, and the filtrate was concentrated under reduced pressure to give crude product of Intermediate 171-2, which was used for the next step without purification. LCMS: 267.1 [M+H]+.

To a vigorously stirred solution of Intermediate 171-3 (2.30 g, 3.92 mmol) in anhydrous 2-methyl THF (8 mL), the solution of 9-borabicyclo[3.3.1]nonane (0.50 M in tetrahydrofuran, 11.8 mL, 5.88 mmol) was added at room temperature. The resulting solution was stirred at 70° C. for 0.5 hour before it was cooled to room temperature. The solution was transferred to a reaction vial containing Pd(dppf)Cl2(247 mg, 0.35 mmol), potassium phosphate (2.23 g, 10.5 mmol) and degassed water (1.5 mL) at rt under nitrogen atmosphere. The reaction mixture was stirred at 90° C. for 10 minutes before it was cooled to room temperature. The mixture was washed with water (20 mL) and extracted with ethyl acetate (3×50 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give an inseparable mixture of 171-4, 174-1, and 182-1 as the faster eluted fraction, and 175-2 as the slower eluted fraction. LCMS: 508.4 [M+H]+.

A vigorously stirred mixture of 171-4, 174-1, and 182-1 (1.35 g, 2.66 mmol), ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (prepared according to WO 2021/041671) (2.40 g, 5.31 mol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (387 mg, 0.53 mmol), potassium phosphate (1.69 g, 7.97 mmol) in tetrahydrofuran (10 mL) and water (5 mL) was heated to 70° C. After 80 min, the resulting mixture was cooled to room temperature, and ethyl acetate (200 mL) were added sequentially. The organic layer was washed with brine (100 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give an inseparable mixture of 171-5 and 174-2 as the slower eluted fraction, and 182-2 as the faster eluted fraction. LCMS: 798.6 [M+H]+.

3-Chloroperoxybenzoic acid (77% wt, 535 mg, 2.39 mmol) was added in small portions over 5 min to a vigorously stirred solution of mixture of Intermediate 171-5 and 174-2 (760 mg, 0.95 mmol) in dichloromethane (15.0 mL) at 0° C. After 25 min, the resulting mixture was warmed to room temperature. After 60 min, the solution was filtered and the filtrated concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0% to 75% ethyl acetate in hexanes) to give Intermediate 171-6 as faster eluted fraction, and 174-3 as the slower eluted fraction. LCMS: 830.4 [M+H]+.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 452 μL, 452 μmol) was added over 1 min via syringe to a stirred mixture of Intermediate 171-6 (250 mg, 301 μmol), (2R,7aS)-2-Fluorotetrahydro-1H-pyrrolizine-7a(5H)-methano (125 mg, 785 μmol), and tetrahydrofuran (2.5 mL) at 0° C. After 30 min, ethyl acetate (20 mL), and saturated aqueous sodium chloride solution (20 mL) were added sequentially. The organic layer was washed with water (30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give the crude product of Intermediate 171-7. LCMS: 895.2 [M+H]+.

Cesium fluoride (687 mg, 4.52 mmol) was added to a vigorously stirred solution of Intermediate 171-7 (270 mg, 0.302 mmol) in N,N-dimethylformamide (5 mL) at room temperature. After 30 min, ethyl acetate (100 mL) was added. The mixture was filtered, and the filtrated was concentrated under reduced pressure to give the crude product of Intermediate 171-8. LCMS: 739.1 [M+H]+.

Sodium borohydride (118.5 mg, 4.9 mmol) was added to a vigorously stirred solution of Intermediate 97-2 (35.2 mg, 0.071 mmol) in methanol (0.45 mL) at 0° C. The reaction mixture was stirred at 0° C. for 25 minutes before it was quenched with water (1 mL) at 0° C. The mixture was extracted with ethyl acetate (3×5 mL). The combined organic phase was dried over magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0% to 80% ethyl acetate in hexanes) to give Intermediate 172-1 and Intermediate 173-1. LCMS: 496.6.

To a vigorously stirred solution of Intermediate 172-1 (61.0 mg, 0.12 mmol) in anhydrous dichloromethane (0.57 mL), was added diethylaminosulfur trifluoride at −78° C. The mixture was stirred at −78° C. for 1 hour before it was quenched with water (1 mL) and saturated aqueous sodium bicarbonate solution (1 mL) at −78° C. The resulting mixture was extracted with dichloromethane (3×5 mL). The organic layer was collected and combined, dried over magnesium sulfate, and concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 172-2. LCMS: 498.6.

Intermediate 172-3 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 172-2 instead of Intermediate 13-7. LCMS: 788.6.

Intermediate 172-4 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 172-3 instead of Intermediate 13-8. LCMS: 820.4.

Intermediate 172-5 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 172-4 instead of Intermediate 13-9. LCMS: 885.2.

Intermediate 172-6 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 172-5 instead of Intermediate 13-10. LCMS: 729.0.

Intermediate 173-2 was synthesized in a manner similar to Intermediate 172-2 using Intermediate 173-1 instead of Intermediate 172-1. LCMS: 498.2.

Intermediate 173-3 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 173-2 instead of Intermediate 13-7. LCMS: 788.5.

Intermediate 173-4 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 173-3 instead of Intermediate 13-8. LCMS: 820.2.

Intermediate 173-5 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 173-4 instead of Intermediate 13-9. LCMS: 885.2.

Intermediate 173-6 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 173-5 instead of Intermediate 13-10. LCMS: 728.9.

Intermediate 174-1 was synthesized in a manner similar to 75-7 using intermediate 171-3 instead of intermediate 75-6. LCMS: 508.3 [M+H]+.

Intermediate 174-2 was synthesized in a manner similar to 76-1 using intermediate 174-1 instead of intermediate 75-9. LCMS: 798.6 [M+H]+.

Intermediate 174-3 was synthesized in a manner similar to 75-8 using intermediate 174-2 instead of intermediate 75-7. LCMS: 830.4 [M+H]+.

Intermediate 174-4 was synthesized in a manner similar to 75-9 using intermediate 174-3 instead of intermediate 75-8. LCMS: 895.2 [M+H]+.

Intermediate 174-5 was synthesized in a manner similar to 76-2 using intermediate 174-4 instead of intermediate 76-1. LCMS: 739.1 [M+H]+.

Intermediate 175-1 was synthesized in a manner similar to 75-7 using intermediate 171-3 instead of intermediate 75-6. LCMS: 508.3 [M+H]+.

Intermediate 175-2 was synthesized in a manner similar to 76-1 using intermediate 175-1 instead of intermediate 75-9. LCMS: 798.6 [M+H]+.

Intermediate 175-3 was synthesized in a manner similar to 75-8 using intermediate 175-2 instead of intermediate 75-7. LCMS: 830.4 [M+H]+.

Intermediate 175-4 was synthesized in a manner similar to 75-9 using intermediate 175-3 instead of intermediate 75-8. LCMS: 895.2 [M+H]+.

Intermediate 175-5 was synthesized in a manner similar to 76-2 using intermediate 175-4 instead of intermediate 76-1. LCMS: 739.1 [M+H]+.

At 0° C., sodium hydride (60% in mineral oil, 73 mg, 1.79 mmol) was add to benzyl 4-hydroxypiperidine-1-carboxylate (211 mg, 0.897 mmol) in 2 mL DMF. 4-Bromo-2-(trifluoromethyl)pyrimidine 1 (204 mg, 0.897 mmol) was added after 10 minutes. The reaction was stirred at 0° C. for 30 minutes. The reaction was quenched by adding saturated bicarbonate solution, and the product was extracted with EtOAc. The organic layer was concentrated down and purified by silica column (eluting with EtOAc/hexane, 0-100%) to give intermediate 180-1. LCMS: 382.2.

A stirred mixture of intermediate 180-1 (81 mg, 0.21 mmol), EtOAc (10 mL), EtOH (10 mL), and palladium (10% wt on activated carbon, 30 mg) was placed under an atmosphere of hydrogen gas (balloon) at room temperature. After 1 h, the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to give intermediate 180-2. LCMS: 248.2.

In a round bottom flask, 1-((benzyloxy)methyl)cyclopropane-1-carbaldehyde (40 mg, 0.21 mmol) and intermediate 180-2 (52 mg, 0.21 mmol) were dissolved in 1.5 mL THF. The reaction was stirred at room temperature for 30 minutes, and sodium triacetoxyborohydride (89 mg, 0.42 mmol) was added followed by a drop of acetic acid. The reaction was stirred at room temperature overnight. A saturated bicarbonate solution was added to the reaction, and the product was extracted with ethyl acetate. The organic layer was concentrated down and purified by silica column (eluting with EtOAc/hexane, 0-100%) to give intermediate 180-3. LCMS: 422.2.

A stirred mixture of intermediate 180-3 (65 mg, 0.154 mmol), EtOAc (5 mL), EtOH (5 mL), palladium (10% wt on activated carbon, 20 mg) at room temperature was placed under an atmosphere of hydrogen gas (balloon). After 1 h, the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to give intermediate 180-4. LCMS: 332.2.

Intermediate 182-1 was synthesized in a manner similar to 75-7 using intermediate 171-3 instead of intermediate 75-6. LCMS: 508.3 [M+H]+.

Intermediate 182-2 was synthesized in a manner similar to 76-1 using intermediate 182-1 instead of intermediate 75-9. LCMS: 798.6 [M+H]+.

Intermediate 182-3 was synthesized in a manner similar to 75-8 using intermediate 182-2 instead of intermediate 75-7. LCMS: 830.4 [M+H]+.

Intermediate 182-4 was synthesized in a manner similar to 75-9 using intermediate 182-3 instead of intermediate 75-8. LCMS: 895.2 [M+H]+.

Intermediate 182-5 was synthesized in a manner similar to 76-2 using intermediate 182-4 instead of intermediate 76-1. LCMS: 739.1 [M+H]+.

8-(tert-butyl) 2-ethyl (1S,2S,5R)-4-oxo-3,8-diazabicyclo[3.2.1]octane-2,8-dicarboxylate (5 g, 16.8 mmol) was dissolved in dry THF (50 mL) under an atmosphere of nitrogen. After Cooling to 0° C., a solution of methyl magnesium bromide in THF (15.7 mL, 3.2 M solution in 2-MeTHF, 50.3 mmol) was added dropwise via syringe. The solution was allowed to warm to room temperature and stirred for 3 hours. The reaction mixture was slowly quenched with adding saturated ammonium chloride solution and water. The reaction mixture was extracted with EtOAc (3×), and the organic layers were combined and washed with saturated sodium chloride solution, and dried over magnesium sulfate. The solvents were removed by rotary evaporation. The residue was purified by SGC eluting with MeOH/DCM to afford intermediate 183-1. MS (m/z) 284.9 [M+H]+

A mixture of tert-butyl (1R,2S,5S)-2-(2-hydroxypropan-2-yl)-4-oxo-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (3500 mg, 12.3 mmol) and Martin Sulfurane (16.5 g, 24.6 mmol) in toluene (50 mL) was stirred at rt for 2 hours. The reaction mixture was evaporated and purified by SGC eluting with EtOAc/hex to afford intermediate 183-2. MS (m/z) 288.9 [M+Na]+

Intermediate 183-4 was synthesized in a manner similar to 64-3 using intermediate 183-3 instead of intermediate 64-2. LCMS: 573.0, 574.3 [M+H]+.

Intermediate 183-5 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 183-4 instead of Intermediate 75-6. Slower eluted fraction on silica gel column chromatography. LCMS: 494.5 [M+H]+

Intermediate 183-6 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 183-5 instead of Intermediate 75-9. LCMS: 784.8 [M+H]+

Intermediate 183-7 was synthesized in a manner similar to Intermediate 75-8 using Intermediate 183-6 instead of Intermediate 75-7. LCMS: 816.8 [M+H]+

Intermediate 183-8 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 183-7 instead of Intermediate 75-8. LCMS: 881.1 [M+H]+

Intermediate 183-9 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 183-8 instead of Intermediate 75-10. LCMS: 725.1 [M+H]+

Intermediate 184-1 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 183-4 instead of Intermediate 75-6. Faster eluted fraction on silica gel column chromatography. LCMS: 494.3 [M+H]+

Intermediate 184-2 was synthesized in a manner similar to Intermediate 76-1 using Intermediate 184-1 instead of Intermediate 75-9. LCMS: 784.5 [M+H]+

Intermediate 184-3 was synthesized in a manner similar to Intermediate 75-8 using Intermediate 184-2 instead of Intermediate 75-7. LCMS: 816.4 [M+H]+

Intermediate 184-4 was synthesized in a manner similar to Intermediate 75-9 using Intermediate 184-3 instead of Intermediate 75-8. LCMS: 881.1 [M+H]+

Intermediate 184-5 was synthesized in a manner similar to Intermediate 75-11 using Intermediate 184-4 instead of Intermediate 75-10. LCMS: 725.1 [M+H]+

Intermediate 186-1 was synthesized in a manner similar to Intermediate 13-8 using Intermediate 172-1 instead of Intermediate 13-7. LCMS: 786.7.

Intermediate 186-2 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 186-1 instead of Intermediate 13-8. LCMS: 818.3.

Intermediate 186-3 was synthesized in a manner similar to Intermediate 13-10 using Intermediate 186-2 instead of Intermediate 13-9. LCMS: 883.1.

Intermediate 186-4 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 186-3 instead of Intermediate 13-10. LCMS: 727.1.

To a vigorously stirred solution of Intermediate 75-1 (70.0 mg, 0.18 mmol) in anhydrous 2-methyltetrahydrofuran (2.1 mL), a solution of 9-borabicyclo[3.3.1]nonane (0.50 M in tetrahydrofuran, 0.55 mL, 0.28 mmol) was added at room temperature. The resulting solution was stirred at 50° C. for 100 minutes before it was cooled to room temperature. The solution was transferred to a reaction vial containing Pd(dppf)Cl2(13.0 mg, 0.018 mmol), potassium phosphate (213.5 mg, 0.92 mmol), bromoethylene (1.0 M in tetrahydrofuran, 0.80 mL, 0.80 mmol) and degassed water (0.29 mL) at room temperature under nitrogen atmosphere. The reaction mixture was stirred at 70° C. for 12 minutes before it was cooled to room temperature. The mixture was washed with water (2 mL) and extracted with ethyl acetate (3×5 mL). The organic layer was collected and combined, dried over magnesium sulfate, concentrated under reduced pressure. The crude was purified by silica gel column chromatography (0 to 50% ethyl acetate in hexanes) to give Intermediate 188-1. LCMS: 433.2 [M+Na]+.

Intermediate 188-2 was synthesized in a manner similar to Intermediate 75-5 using Intermediate 188-1 instead of Intermediate 75-4. LCMS: 267.0.

Intermediate 188-3 was synthesized in a manner similar to Intermediate 53-5 using Intermediate 188-2 instead of Intermediate 53-4. LCMS: 588.1.

Intermediate 188-4 was synthesized in a manner similar to Intermediate 75-7 using Intermediate 188-3 instead of Intermediate 75-6. LCMS: 508.3.

Intermediate 188-5 was synthesized in a manner similar to Intermediate 13-9 using Intermediate 188-4 instead of Intermediate 13-8. LCMS: 539.9.

Intermediate 188-6 was synthesized in a manner similar to Intermediate 53-10 using Intermediate 188-5 instead of Intermediate 53-9. LCMS: 605.0.

Intermediate 188-7 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 188-6 instead of Intermediate 17-9. LCMS: 895.1.

Intermediate 188-8 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 188-7 instead of Intermediate 13-10. LCMS: 739.1.

Intermediate 189-1 was synthesized in a manner similar to Intermediate 2-1 using Intermediate 188-6 instead of Intermediate 17-9. LCMS: 955.3.

Intermediate 189-2 was synthesized in a manner similar to Intermediate 13-11 using Intermediate 189-1 instead of Intermediate 13-10. LCMS: 799.0.

Benzyl bromide (633 μL, 5.32 mmol) was added to a stirred mixture of intermediate 27-5 (1.15 g, 4.84 mmol), N,N-diisopropylethylamine (1.22 mL, 7.01 mmol), and acetonitrile at room temperature, and the resulting mixture was heated to 80° C. After 40 min, the resulting mixture was heated to 90° C. After 17 h, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 12% ethyl acetate in hexanes) to give intermediate 190-1. LCMS: 329.2.

Secondary butyllithium solution (1.31 M in cyclohexane, 2.81 mL, 3.67 mmol) was added over 2 min via syringe to a vigorously stirred mixture of intermediate 190-1 (402 mg, 1.22 mmol), N,N,N′,N′-tetramethylethane-1,2-diamine (551 μL, 3.67 mmol), and diethyl ether (4.5 mL) at −40° C. After 1 min, the resulting mixture was warmed to 0° C. After 35 min, the resulting mixture was cooled to −78° C. over 5 min, and iodomethane (267 μL, 4.29 mmol) was added via syringe. After 5 min, the resulting mixture was warmed to 0° C. After 90 min, triethylamine (3.0 mL) and saturated aqueous sodium bicarbonate solution (4.0 mL) were added sequentially, and the resulting mixture was warmed to room temperature. Diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially, and the organic layer was washed with a mixture of water and brine (3:1 v:v, 10 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 5.5% ethyl acetate in hexanes) to give intermediate 190-2. LCMS: 343.2.

Bis(1,5-cyclooctadiene)diiridium(I) dichloride (58.8 mg, 87.6 μmol) was added to a vigorously stirred mixture of intermediate 190-2 (200 mg, 584 μmol), ethylenebis(diphenylphosphine) (69.8 mg, 175 μmol), and dichloromethane (0.2 mL) at room temperature. After 8 min, the resulting mixture was cooled to 0° C. over 3 min. 4,4,5,5-Tetramethyl-1,3,2-dioxaborolane (169 μmol, 1.17 mmol) was added via syringe over 10 min, and the resulting mixture was warmed to room temperature. After 5 h, the resulting mixture was purified by flash column chromatography on silica gel (0 to 40% ethyl acetate in hexanes) to give intermediate 190-3. LCMS: 471.3.

A vigorously agitated mixture of intermediate 190-3 (223 mg, 474 μmol), palladium(II) hydroxide (20% wt on activated carbon, 49.9 mg, 71 μmol), and tetrahydrofuran (3.0 mL) at room temperature in a glass shaker was placed under an atmosphere of hydrogen gas (50 psi). After 16.5 h, the resulting mixture was filtered through celite. The filtrate was concentrated under reduced pressure. Dichloromethane (1.0 mL) and N,N-diisopropylethylamine (248 μL, 1.42 mmol) were added sequentially, and the resulting mixture was stirred at room temperature. Intermediate 53-1 (180 mg, 474 μmol) was added. After 40 min, saturated aqueous ammonium chloride solution (10 mL), aqueous hydrogen chloride solution (2.0 M, 1.0 mL), diethyl ether (40 mL), and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (15 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. [1,1′-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (34.7 mg, 47.4 μmol) and 1,4-dioxane (4.5 mL) were added sequentially, and the resulting mixture was vigorously stirred and was sparged with nitrogen gas. After 15 min, sparging was ceased, and saturated aqueous sodium carbonate solution (2.0 M, 949 μL, 1.9 mmol) was added via syringe. The resulting mixture was heated to 90° C. After 50 min, the resulting mixture was heated to 110° C. After 70 min, the resulting mixture was cooled to room temperature, and diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (20 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. The residue was purified by flash column chromatography on silica gel (0 to 24% ethyl acetate in hexanes) to give intermediate 190-4. LCMS: 494.2.

Intermediate 190-5 was synthesized in a manner similar to intermediate 52-7 using intermediate 190-4 instead of intermediate 52-6. LCMS: 844.1.

Intermediate 192-1 was synthesized in a manner similar to intermediate 52-4 using intermediate 75-5 instead of intermediate 27-5. LCMS: 668.2.

A solution of intermediate 192-1 (109.9 mg, 164 umol; azeotroped with toluene 3×) in tetrahydrofuran (0.4 mL) was stirred at rt as 0.5 M 9-borabicyclo[3.3.1]nonane in tetrahydrofuran (0.99 mL) was added. The resulting solution was heated at 60° C. heating block for 90 min and then cooled to rt. To the reaction mixture was added water (0.4 mL) and stirred at rt for 1 h. To the resulting solution was added 1.5 M aqueous tripotassium phosphate (1.10 mL, 1.64 umol) and the mixture was purged with Ar gas for 15 min. To this degassed vial were added ferrouscyclopenta-1,3-dien-1-yl(diphenyl)phosphane dichloropalladium (12.74 mg, 17.4 umol), and dioxane (2.1 mL). The resulting mixture was purged with Ar gas for 15 min again. The resulting solution was heated at 90° C. heating block for 1 h and then cooled to rt. The reaction mixture was diluted with saturated sodium bicarbonate (˜5 mL), and the product was extracted with ethyl acetate (˜10 mL×3). The extracts were combined, dried (MgSO4), and concentrated. The residue was purified by silica gel column chromatography eluting 40-100% ethyl acetate in hexane followed by 0-20% MeOH in ethyl acetate to give intermediate 192-2. LCMS: 590.3.

A mixture of intermediate 192-2 (9.01 mg, 15.3 umol), 2-[2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthyl]ethynyl-triisopropyl-silane (9.52 mg, 21.0 umol), and cataCXium A Pd G3(2.23 mg, 3.06 umol) in 1,4-dioxane (0.4 mL) and 1.5 M aqueous tripotassium phosphate (0.11 mL) was purged with Ar gas for 15 min and then reacted at 120° C. for 30 min in uW reactor. The reaction mixture was diluted with saturated sodium bicarbonate (˜25 mL) and the product was extracted with ethyl acetate (˜20 mL×2). The combined extracts were dried (MgSO4), and concentrated to give intermediate 192-3. LCMS: 880.2.

Intermediate 193-1 was synthesized in a manner similar to intermediate 192-3 using intermediate 64-2 instead of intermediate 75-5 and using N-(6-fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)ethynyl)naphthalen-2-yl)-1,1-diphenylmethanimine (WO2022170999) instead of 2-[2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthyl]ethynyl-triisopropyl-silane. LCMS: 1059.3.

To a mixture of intermediate 193-1 (16.17 mg, 15.3 umol) and CsF (35.24 mg, 232 umol) was added DMF (0.6 mL) and the resulting solution was stirred for 1 h at rt. After the reaction mixture was diluted with saturated sodium bicarbonate solution (˜10 mL), water (˜15 mL), the product was extracted with ethyl acetate (˜20 mL×2). The organic extracts were washed with water (˜25 mL×1), combined, dried (MgSO4), and concentrated. The resulting residue was purified by silica gel column chromatography eluting 0-20% MeOH in dichloromethane to give intermediate 193-2. LCMS: 903.5.

To a vial charged with intermediate 194-1 (224 mg, 0.523 mmol) and 2-Me-THF (3 ml), 9-BBN (1.96 ml, 0.98 mmol, 0.5 M in THF) was added under N2atmosphere. The reaction mixture was stirred at 50° C. Upon full consumption of the starting material (about 100 minutes), the mixture was cooled to room temperature. To another vial, Pd(dppf)Cl2(37 mg, 0.052 mmol) and K3PO4(610 mg, 2.62 mmol) were added, evacuate and refill with N2three times, to it was added degassed water (1.5 ml), the crude hydroboration solution and vinyl bromide solution (2.3 ml, 2.3 mmol, 1 M in THF). The resulting mixture was stirred at 70° C. for 20 minutes and cooled to room temperature. It was partitioned between ethyl acetate and water. The organic layer was dried over sodium sulfate, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with ethyl acetate/hexane to afford intermediate 194-2. LCMS: 433.2 (M+Na).

To the vial charged with compound intermediate 194-2 (130 mg, 0.32 mmol) and cesium fluoride (240 mg, 1.58 mmol). To it was added DMF (1.3 ml). The mixture was stirred at 70° C. for 4 hours. Upon the completion of this reaction, the mixture was filtered thru a celite pad the filtrate was concentrated to give intermediate 194-3. LCMS: 267.2

To a vial charged with intermediate 194-3 (100 mg, 0.375 mmol) and Intermediate 53-1 (134 mg, 0.375 mmol), dichloromethane (1 ml) was added under N2protection and cooled to 0° C. To it was added N, N-diisopropylethylamine (0.196 ml, 1.13 mmol). The resulting mixture was stirred under 0° C. for 12 min before loaded onto ISCO flash column directly and eluting with ethyl acetate/hexane to afford intermediate 194-4. LCMS: 588.2.

Intermediate 194-4 (100 mg, 0.17 mmol) was dissolved in acetonitrile (3 mL) and purged with argon. To it was added triethylamine (0.071 ml, 0.511 mmol) and tetrakis(triphenylphosphine)palladium(0) (19.7 mg, 0.017 mmol). The reaction mixture was stirred at 100° C. for 1-hour. Upon completion, it cooled to room temperature and concentrated to dryness. The residue was Purify by silica gel chromatography eluting with ethyl acetate and hexane to afford intermediate 194-5. LCMS: 506.2.

Intermediate 194-5 (91 mg, 0.18 mmol) was dissolved in 5 ml of ethyl acetate and was sparged under an argon atmosphere. platinum(iv) oxide (20.4 mg, 0.0899 mmol) was added, and the mixture was sparged under a hydrogen atmosphere (1 atm, balloon). The mixture was stirred vigorously for 4 hours. after sparged with argon, it was filtered through a pad of Celite®. The Celite® was washed with ethyl acetate. The filtrate was concentrated to dryness and purified by RP-HPLC to afford intermediate 194-6 (methylated carbon stereocenter stereochemistry assigned arbitrarily). LCMS: 508.2.

Intermediate 194-7 was synthesized in a manner similar to intermediate 13-9 using intermediate 194-6 instead of intermediate 13-8. LCMS: 540.2.

Azeotrope compound 194-7 (41 mg, 0.0759 mmol) and [(2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-8-yl]methanol (24.2 mg, 0.152 mmol) with 1 mL toluene 2 times, then dissolve in THF (1 ml). Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 114 μL, 0.114 mmol) was added under 0° C. The reactions were stirred at 0° C. for 15 min. Upon completion, the mixture was diluted with ethyl acetate and quenched with H2O under 0° C. and extract with ethyl acetate. Organic layer was dried over sodium sulfate and conc in vacuo. The residue was purified with reverse phase prep HPLC to afford intermediate 194-8. LCMS: 605.2

N-(6-Fluoro-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-5-((triisopropylsilyl)ethynyl)naphthalen-2-yl)-1,1-diphenylmethanimine (150 mg, 0.24 mmol) was dissolved in 1 mL ethyl acetate. A solution of HCl in 1,4-dixoane (4.0M, 0.59 ml, 24 mmol) was added followed by water (0.0085 ml, 0.47 mmol) to facilitate hydrolysis. After 1-hour, desired product precipitated out from the solution. Hexane was added and the resulting mixture was filtered to give intermediate 194-9. LCMS: 468.3.

Intermediate 194-10 was synthesized in a manner similar to intermediate 4-1 using intermediate 194-8 and intermediate 194-9 instead of intermediate 17-9 and ((2-fluoro-8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)naphthalen-1-yl)ethynyl)triisopropylsilane (WO2021041671). LCMS: 910.6.

Cesium fluoride (92 mg, 606 μmol) was added to a vigorously stirred solution of intermediate 194-10 (23 mg, 25.3 μmol) in N,N-dimethylformamide (0.6 mL) at room temperature. After 30 min, diethyl ether (40 mL) and ethyl acetate (20 mL) were added sequentially. The organic layer was washed with water (2×40 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduce pressure to afford intermediate 194-11. LCMS: 754.4.

Intermediate 195-1 was synthesized in a manner similar to 171-7 using intermediate 165-1 instead of intermediate ((2R,7aS)-2-fluorotetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol. LCMS: 909.3.

Intermediate 195-2 was synthesized in a manner similar to 171-8 using intermediate 195-1 instead of intermediate 171-7. LCMS: 753.1.

To a mixture of intermediate 63-2 (300 mg, 725 μmol) in THF (1.5 mL) was added t-BuOK solution (1.0 M in tetrahydrofuran, 1.09 mL, 1.1 mmol), and the resulting mixture was degassed and purged with N23 times at 0° C. for 0.5 hr. 3-Chloro-5-(trifluoromethyl)pyridazine (158.9 mg, 870.5 μmol) was added, the reaction mixture was stirred at 25° C. for 1 hr under N2atmosphere. The unreacted alkoxides were quenched by addition saturated aqueous H2O (4 mL) at 0° C., and the aqueous layer was extracted with EtOAc (4 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by purified by flash silica gel chromatography (12 g Silica Flash Column, Eluent of 0˜20% Dichloromethane/Methanol gradient@80 mL/min) to give intermediate 198-0. LCMS: 560.2.

To a solution of intermediate 198-0 (300 mg, 536 μmol) in EtOAc (0.5 mL) was added HCV/EtOAc (0.5 mL). The mixture was stirred at 25° C. for 0.5 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [water(NH4HCO3)-ACN]; gradient: 5%-30% B over 8 min) to give intermediate 198-1. LCMS: 318.1.

To a solution of intermediate 63-2 (50.00 mg, 120.90 μmol) in THF (2.00 mL) was added t-BuOK (1 M, 181.36 μL) at 0° C. under N2. The mixture was stirred at 0° C. for 0.5 hr, then 4-bromo-3-chloro-6-(trifluoromethyl)pyridazine (47.41 mg, 181.36 μmol) was added to the mixture, and the mixture was stirred at 25° C. for 0.5 hr. The reaction mixture was quenched by addition saturated aqueous NH4Cl solution (20.00 mL) at 0° C., and then extracted with EtOAc (20.00 mL*3). The combined organic layers were dried over Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; gradient: 60%-90% B over 8.0 min) to give intermediate 199-1. LCMS: 594.3.

To a solution of intermediate 199-1 (150.00 mg, 252.50 μmol) in MeOH (1.00 mL) was added Pd/C (150.00 mg, 10% purity) under Ar. Then triethylamine (127.75 mg, 1.26 mmol, 175.72 μL) was added to the mixture at 0° C. The suspension was degassed under vacuum and purged with H2several times. The mixture was stirred under H2(15 psi) at 25° C. for 1 hr. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give intermediate 199-2. LCMS 560.3.

To a solution of intermediate 199-2 (150.00 mg, 268.04 μmol) in EtOAc (1.00 mL) was added HCl/EtOAc (1.00 mL, 4M). The mixture was stirred at 25° C. for 1 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 5%-45% B over 8.0 min) to give intermediate 199-3. LCMS: 318.1.

To a solution of 4-chloro-6-(trifluoromethyl)pyrimidine (1 g, 5.48 mmol) in MeOH (10 mL) was added NaSH (921.42 mg, 16.44 mmol). The mixture was stirred at 25° C. for 12 hrs. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (15 mL) and extracted with EtOAc (20 mL*3). The combined organic layers were washed with brine (10 mL*2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ethyl acetate in petroleum ether) to afford intermediate 201-1.1H NMR (400 MHz, DMSO-d6) δ ppm 14.66 (br s, 1H), 8.63-8.25 (m, 1H), 7.54 (s, 1H).

To a solution of intermediate 63-2 (300.00 mg, 725.43 μmol) in DCM (3.00 mL) was added TEA (293.62 mg, 2.90 mmol, 403.88 μL) and methanesulfonyl chloride (249.30 mg, 2.18 mmol, 168.44 μL) at 0° C. The mixture was stirred at 25° C. for 1 hr. The reaction mixture was quenched with NaHCO3(10%, 10 ml), and then extracted with DCM (10.00 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give intermediate 201-2. LCMS: 492.3.

To a solution of intermediate 201-2 (300 mg, 610.20 μmol) in DMF (5 mL) was added TEA (185.24 mg, 1.83 mmol, 254.80 μL) and 6-(trifluoromethyl)pyrimidine-4-thiol (219.86 mg, 1.22 mmol). The mixture was stirred at 70° C. for 12 hrs. The reaction mixture was concentrated under reduced pressure. The residue was diluted with H2O (15 mL) and extracted with EtOAc (20 mL*3). The combined organic layers were washed with brine (10 mL*2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (PE:EtOAc=3:1) to afford intermediate 201-3. LCMS: 576.4.

To a solution of intermediate 201-3 (240 mg, 416.89 μmol) in EtOAc (1 mL) was added HCV/EtOAc (1 mL). The mixture was stirred at 25° C. for 0.2 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [water(NH4HCO3)-ACN]; B %: 20%-50%, 8 min) to afford intermediate 201-4. LCMS: 334.1.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 27.7 μL, 27.7 μmol) was added over 1 min via syringe to a stirred mixture of Intermediate 171-6 (11.5 mg, 13.9 mol), ((3S,7aS)-3-(((6-(trifluoromethyl)pyrimidin-4-yl)oxy)methyl)tetrahydro-1H-pyrrolizin-7a(5H)-yl)methanol (11.5 mg, 36.1 μmol), and 2-methyltetrahydrofuran (0.24 mL) at 0° C. After 1 hour, ethyl acetate (2 mL), and water (0.5 mL) were added sequentially at 0° C. The aqueous layer was washed with ethyl acetate (3×2 mL). The organic layers were combined and dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 203-1. LCMS: 1053.3.

Cesium fluoride (31.6 mg, 0.208 mmol) was added to a vigorously stirred solution of Intermediate 203-1 (14.6 mg, 13.9 μmol) in N,N-dimethylformamide (0.2 mL) at room temperature. After 1 hour, diethyl ether (4 mL), ethyl acetate (2 mL), and saturated aqueous sodium bicarbonate solution (1 mL) were added sequentially. The organic layer was washed with water (2×4 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure to give Intermediate 203-2. LCMS: 897.2.

To a solution of 1-bromo-2-fluoro-3-nitro-benzene (70 g, 318.19 mmol) in H2SO4(20 mL) was added NBS (56.63 g, 318.19 mmol). The mixture was stirred at 50° C. for 3 hrs. The reaction mixture was quenched with ice water (1000 mL) and extracted with EtOAc (700 ml*3). The combined organic phase was dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by purified by flash silica gel chromatography (100 g Silica Flash Column, Eluent of 0˜10% Ethyl acetate/Petroleum ether gradient@80 mL/min) to give intermediate 204-1.1H NMR (400 MHz, CDCl3) δ 8.06 (dd, J=2.4, 5.9 Hz, 1H), 7.93 (dd, J=3.0, 5.5 Hz, 1H).

To a solution of intermediate 204-1 (70 g, 234.20 mmol) and Fe (39.24 g, 702.60 mmol) in H2O (30 mL) was added HCl (120 mL) at 25° C. The mixture was stirred at 100° C. for 1 hr. The reaction mixture was filtered, the filter cake was extracted with EtOAc (200 mL), and the combined filtrates were concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (100 g Silica Flash Column, Eluent of 0˜20% Ethyl acetate/Petroleum ether gradient@80 mL/min) to give intermediate 204-2. LCMS: 269.9.

To a solution of H2SO4(50 mL) in 50° C. was added intermediate 204-3 (60 g, 176.50 mmol) in portions, then the mixture was stirred at 70° C. for 1 hr. The reaction mixture was cooled to room temperature and slowly added to ice water (200 ml), and the resulting mixture was filtered. The filter cake was concentrated under reduced pressure to give intermediate 204-4. LCMS: 323.9.

To a solution of intermediate 204-4 (20 g, 61.94 mmol) in NaOH (2 M, 309.68 mL) was added H2O2(35.11 g, 309.68 mmol, 29.76 mL, 30% purity) at 0° C. The mixture was stirred at 25° C. for 16 hrs. The reaction mixture was adjusted to pH=2 with 1M HCl (200 ml), then the reaction mixture was filtered, and the filter cake concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Phenomenex luna c18 250 mm*100 mm*10 um; mobile phase: [water(FA)-ACN]; B %: 25%-55%, min) to give intermediate 204-5. LCMS: 313.8.

To a solution of intermediate 204-5 (11 g, 35.15 mmol) in H2SO4(50 mL) was added NCS (9.39 g, 70.31 mmol) at 25° C. The mixture was stirred at 80° C. for 12 hrs. The reaction mixture was added to ice H2O (50 ml), then the reaction mixture was filtered, and the filter cake concentrated under reduced pressure to give intermediate 204-6. LCMS: 347.8.

To the solution of intermediate 204-6 (8.00 g, 23.03 mmol) in DCM (40.00 mL) was added SOCl2(16.44 g, 138.18 mmol, 10.04 mL) and DMF (168.34 mg, 2.30 mmol, 177.20 μL), the reaction mixture stirred at 60° C. for 5 hrs. The reaction mixture was concentrated under reduced pressure to give a residue, then in acetone (40.00 mL) was added NH4SCN (2.63 g, 34.55 mmol, 2.63 mL), the reaction mixture was stirred at 25° C. for 2 hrs. The reaction mixture was filtered, the filter cake was washed with H2O (100.00 ml) and 10% NaOH (30 ml) then the solid was triturated with MeOH (50.00 ml) and ACN (50.00 ml) to give intermediate 204-7.

To a mixture of intermediate 204-7 (1.00 g, 2.57 mmol) in MeOH (20.00 mL) and H2O (10.00 mL) was added NaOH (205.94 mg, 5.15 mmol) and Mel (730.81 mg, 5.15 mmol, 320.53 μL) at 25° C., the reaction mixture was stirred at 25° C. for 30 mins. H2O (20.00 ml) was added to the reaction mixture, and the reaction mixture was adjusted to pH=6 with 1 N HCl (10.00 ml) then the reaction mixture was filtered, and the filter cake was triturated with MeOH (50.00 ml) and ACN (50.00 ml) at 25° C. The resulting filter cake was concentrated under reduced pressure to give intermediate 204-8. LCMS: 402.8.

Intermediate 204-9 was synthesized in a manner similar to 53-1 using intermediate intermediate 204-8 instead of intermediate 13-3. LCMS: 421.0.

Intermediate 204-10 was synthesized in a manner similar to 63-5 using intermediate 204-9-1 instead of intermediate 53-1. LCMS: 623.4.

The solution of 9-BBN (4.04 mL, 0.5 M) was added to the solution of intermediate 204-10 (950 mg, 1.53 mmol) in 2-MeTHF (5 mL) at rt under N2atmosphere. The reaction mixture was stirred at 50′C for 1 h before it was cooled to rt. The mixture was transferred via a syringe to a vial containing K3PO4(871 mg, 4.11 mmol), Pd(dtbpf)Cl2(136 mg, 0.2 mmol) and degassed water (1 mL) under N2atmosphere. The reaction mixture was stirred at 90′C for 20 min before it was cooled to rt. EtOAc (150 mL) was added and the mixture was washed with brine (150 mL). The organic phase was separated and dried over Na2SO4and filtered. After concentrated in vacuo, the residue was purified with ISCO (silica gel, EtOAc-hexanes 0-80%) to give intermediate 204-11. LCMS: 543.7.

Intermediate 204-13 was synthesized in a manner similar to 63-7 using intermediate 204-12 instead of intermediate 63-6. LCMS: 881.9.

Intermediate 204-14 was synthesized in a manner similar to 79-4 using intermediate 204-13 instead of intermediate 79-3. LCMS: 960.8.

Intermediate 204-15 was synthesized in a manner similar to 79-5 using intermediate 204-14 instead of intermediate 79-4. LCMS: 960.8.

To a solution of intermediate 63-2 (200.0 mg, 483.6 μmol) in THF (5.00 mL) was added t-BuOK solution (1.0 M in tetrahydrofuran, 483.6 μL, 480 μmol) at 0° C. under N2, and the resulting mixture was stirred at 0° C. for 0.5 hr, then 4-chloro-6-(difluoromethyl)pyrimidine (238.7 mg, 1.45 mmol) was added to the mixture. The mixture was stirred at 25° C. for 2 hrs under N2. The alkoxides were quenched by addition of saturated aqueous NH4Cl solution (15.00 mL) at 0° C., and the aqueous layer was extracted with EtOAc (30 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (Plate PE:EtOAc=3:1) to give intermediate 205-1. LCMS: 542.4.

To a solution of intermediate 205-1 (150.0 mg, 276.9 μmol) in EtOAc (2.00 mL) was added HCl/EtOAc (1.00 mL). The mixture was stirred at 25° C. for 0.5 hr. The reaction mixture was concentrated under N2to give a residue and then adjusted with 1 M NH3·H2O to pH 6-7. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 5%-50% B over 8.0 min) to give intermediate 205-2. LCMS: 300.2.

To a solution of 4-chloro-2-methylsulfanyl-pyrimidine-5-carbaldehyde (1.00 g, 5.30 mmol) in DCE (15 mL) was added DAST (2.56 g, 15.90 mmol, 2.10 mL). The mixture was stirred at 25° C. for 1 hr. The acids were quenched with sat. aq. NaHCO3(20 ml), and the aqueous layer was extracted with DCM (20 ml*3). The combined organic layers were washed with brine (30 mL*2), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (PE:EtOAc=10:1) to give intermediate 206-1.1H NMR (400 MHz, CDCl3) δ ppm 8.67 (s, 1H), 6.86 (t, J=54.2 Hz, 1H), 2.61 (s, 3H).

To a solution of intermediate 63-2 (500 mg, 1.21 mmol) in THF (5 mL) was added t-BuOK solution (1.0 M in tetrahydrofuran, 1.81 mL, 1.8 mmol) at 0° C. After 30 min, intermediate 206-1 (305.6 mg, 1.45 mmol) was added. The mixture was stirred at 25° C. for 1 hr under N2. The alkoxides were quenched by addition of saturated aqueous NH4Cl solution (10 mL) at 0° C., and the aqueous layer was extracted with EtOAc (20 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by prep-TLC (PE:EtOAc=3:1) to give intermediate 206-2. LCMS: 588.3.

To a solution of intermediate 206-3 (50.0 mg, 92.3 μmol) in EtOAc (1 mL) was added HCl/EtOAc (1 mL). The mixture was stirred at 25° C. for 0.2 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 5%-35% B over 8.0 min) to give intermediate 206-4. LCMS: 300.1.

To a solution of intermediate 63-2 (950 mg, 2.30 mmol) in THF (4 mL) was added NaH (183.8 mg, 4.6 mmol, 60% purity) at 0° C. for 0.5 hr. To the mixture was added 2-chloro-3-(difluoromethyl)pyrazine (491.4 mg, 2.99 mmol), and the resulting mixture was stirred at 40° C. for 12 hrs. The unreacted bases was quenched by addition saturated aqueous NH4Cl solution (5 mL) at 0° C., and the aqueous layer was extracted with EtOAc (5 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (4 g Silica Flash Column, Eluent of 0˜20% Ethyl acetate/Petroleum ether gradient@40 mL/min) to give intermediate 207-1. LCMS: 542.3.

To a solution of intermediate 207-1 (250 mg, 462 μmol) in EtOAc (1 mL) was added HCl/EtOAc (1 mL). The mixture was stirred at 20° C. for 0.5 hr. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 1%-25% B over 8.0 min) to give intermediate 207-2. LCMS: 300.1.

Lithium bis(trimethylsilyl)amide solution (1.0 M in tetrahydrofuran, 54.6 μL, 54.6 μmol) was added to a stirred reaction mixture of intermediate 115-3 (20.0 mg, 30.3 μmol), intermediate 207-2 (13.6 mg, 45.5 μmol), and tetrahydrofuran (1 mL) at 0° C. Upon completion (about 5 min), added brine and extracted with ethyl acetate. The organic layer was washed with aqueous sodium carbonate and water, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by RP-HPLC (15% to 90% 0.1% TFA in MeCN/0.1% TFA in H2O). Fractions containing the product were pooled and lyophilized to yield intermediate 207-3. LCMS: 865.5.

To a solution intermediate 63-2 (140.0 mg, 338.5 μmol) in THF (2 mL) was added t-BuOK (1.0 M in tetrahydrofuran, 507.80 μL, 510 μmol) at 0° C. under N2, and the mixture was stirred at 0° C. for 0.5 hr, then 4-chloro-2-(difluoromethyl)pyrimidine (83.6 mg, 508 μmol) was added to the mixture. The mixture was stirred at 25° C. for 1 hr under N2. The alkoxides were quenched by addition of saturated aqueous NH4Cl (10 mL) at 0° C., and then extracted with EtOAc (15 mL*3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by preparative TLC (PE:EtOAc=0:1) to give intermediate 208-1. LCMS: 542.4.

To a solution of intermediate 208-1 (300.0 mg, 553.9 μmol) in EtOAc (1.50 mL) was added HCl/EtOAc (1.50 mL). The mixture was stirred at 25° C. for 0.5 hr. The reaction mixture was concentrated under N2to give a residue and then adjusted with 1 M NH3·H2O to pH=6-7. The residue was purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 mm*10 um; mobile phase: [H2O (10 mM NH4HCO3)-ACN]; gradient: 10%-50% B over 8.0 min) to give intermediate 208-2. LCMS: 300.1.

Compound 5 and Compound 6 were synthesized in a manner similar to Compound 1 using Intermediate 5-6 instead of 2-(7,8-difluoro-3-(methoxymethoxy)naphthalen-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane.

Aqeuous potassium phosphate solution (1.5 M, 288 μL, 430 μmol) was added via syringe to a vigorously stirred mixture of Intermediate 17-9 (50.0 mg, 86.4 μmol), Intermediate 8-1 (31.7 mg, 86.4 μmol), [(di(1-adamantyl)-butylphosphine)-2-(2′-amino-1,1′-biphenyl)]palladium(II) methanesulfonate (3.1 mg, 43 μmol), and tetrahydrofuran (1.0 mL) at room temperature, and the resulting mixture was heated to 70° C. After 35 min, the resulting mixture was heated to 90° C. after 4 h, the resulting mixture was cooled to room temperature, and ethyl acetate (50 mL) was added. The organic layer was washed with a mixture of water and brine (3:1 v:v, 30 mL), was dried over anhydrous magnesium sulfate, was filtered, and was concentrated under reduced pressure. Dichloromethane (3.0 mL) and trifluoroacetic acid (1.5 mL) were added sequentially, and the resulting mixture was heated to 50° C. After 15 min, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. The residue was purified by reverse phase preparative HPLC (0.1% acetic acid in acetonitrile/water) to give Compound 8 and Compound 9.

Trimethylphosphine solution (1.0 M in tetrahydrofuran, 63.7 μL, 64 μmol) was added via syringe to a stirred mixture of Intermediate 36-3 (10.0 mg, 12.7 μmol), di-tert-butyl (E)-diazene-1,2-dicarboxylate (14.8 mg, 63.7 μmol), 1,1,1,3,3,3-hexafluoro-2-(trifluoromethyl) propan-2-ol (14.2 μL, 102 μmol), and 2-methyltetrahydrofuran (0.8 mL) at 0° C., and the resulting mixture was warmed to room temperature. After 60 min, the resulting mixture was heated to 90° C. After 75 min, the resulting mixture was cooled to room temperature and was concentrated under reduced pressure. Acetonitrile (0.3 mL) was added via syringe, and the resulting mixture was vigorously stirred and was cooled to 0° C. Hydrogen chloride solution (4.0 M in 1,4-dioxane, 500 μL, 2.0 mmol) was added via syringe. After 90 min, the resulting mixture was purified by reverse phase preparative HPLC (0.1% acetic acid in acetonitrile/water) to give Compound 55 and Compound 56.

IV. Biological Examples

Compounds were tested for binding to GDP-loaded KRAS G12D in a 384-well assay format using a TR-FRET probe displacement assay in buffer consisting of 50 mM Hepes (pH 7.4), 150 mM NaCl, 5 mM MgCl2and 0.005% Tween-20. 0.5 nM enzyme was used in this assay with 0.25 nM Eu-streptavidin and 200 nM (2×KD) Cy-5 labelled probe. Compounds were serially diluted (1:3) in DMSO. The LabCyte ECHO Acoustic dispenser system was used to pre-spot the assay plates (384-well Non-Binding Surface plates, Corning, Catalog #3824) with 50 nL of compound. The compounds were pre-incubated with 5 μL of 2× final enzyme concentration for 30 minutes before adding 5 μL of 2× final concentration of Eu-streptavidin and TR-FRET probe (10 μL final reaction volume). The plates were incubated at room temperature for 2 hours before measuring TR-FRET ratio on the Envision plate reader. IC50values were defined as the compound concentration that causes a 50% decrease in TR-FRET ratio and were calculated using a sigmoidal dose-response model to generate curve fits.

Example B. 2D Cell Viability Assay

Compounds were tested in a 384-well format for their ability to inhibit the viability of GP2D (KRAS G12D) cells in 2D assays. Compounds were serially diluted (1:3) in DMSO. The LabCyte ECHO Acoustic dispenser system was used to pre-spot assay plates with 200 nL of test molecule per well. 1000 cells/well (40 μL volume per well) in RPMI medium with 10% FBS, and Penicillin-Streptomycin-Glutamine were plated in pre-spotted 384-well plates (Greiner, Catalog #781076) on the BioTEK EL406 liquid dispenser with a 5 μL dispensing cassette (BioTek 7170011). The plates were incubated at 37° C., 5% CO2for 4 days before addition of CellTiter-Glo (CTG) reagent and measurement of luminescence signal. EC50values were defined as the compound concentration that causes a 50% decrease in luminescence signal and were calculated using a sigmoidal dose-response model to generate curve fits.

Example C. 3D Cell Viability Assay

Compounds were tested in a 384-well format for their ability to inhibit the viability of AsPC-1 (KRAS G12D) cells in 3D assays. On day 0, 1000 cells/well (80 μL volume per well) in DMEM with 10% FBS, and Penicillin-Streptomycin-Glutamine were plated in 384-well Ultra-Low Attachment Spheroid Microplates (Corning, Catalog #3830) on the BioTEK EL406 liquid dispenser with a 5 μL dispensing cassette (BioTek 7170011). The plates were incubated at 37° C., 5% CO2for 3 days to allow spheroid formation. On day 3, compounds were serially diluted (1:3) in DMSO. The Biomek FX was used to add 400 nL of test molecule per well. The plates were incubated at 37° C., 5% CO2for 4 days before addition of CellTiter-Glo 3D (Promega, Catalog #9683) reagent and measurement of luminescence signal. EC50values were defined as the compound concentration that causes a 50% decrease in luminescence signal and were calculated using a sigmoidal dose-response model to generate curve fits.

Biological Data

Provided below in Table 2 is data related to compounds disclosed herein.

TABLE 2Biological Data for Compounds Disclosed HereinG12DG12DG12D2D3DExampleIC50EC50EC50No.Structure(nM)(nM)(nM)10.51917020.2642322416044459253250N/A6270N/A70.616368254N/A90.520771018250N/A11156N/A126130N/A130.42171434530N/A170.44918018>1000>10000N/A195903300N/A207210>10002121003100N/A220.10.6223193124117190270.7429280.67N/A29560>1000N/A300.211260314900>1000N/A320.22N/A330.33N/A340.22N/A350.3617360.24N/A3719N/A380.32847390.448964021974410.6384222895430.65N/A4427N/A450.34N/A46910>1000N/A473400>1000N/A482200>1000N/A490.23714050324N/A51298N/A520.117530.12—54342—550.50.8—561350—570.628—6126520—62779—630.20.2—640.10.3—6589450—66860>1000—670.081—680.050.6—690.10.6—700.11—71481—72510>1000—73———7416310—750.060.170.5760.142777683—780.23.91579715.3800.12.22810.22.8188211.4104.4833.267.4843.927.7856.858860.514.464870.40.20.38836.5254.68950.9379.1901001000911001000921.917.352932.526.682940.10.62.2950.631.9200960.30.51.9970.27.7559812.6124.2990.67.411010031.8372.910112.7175.21023.339.56801035.362.61040.88.7531055.3434.91060.21.98.21077.24601081.5449.81090.11.20.81100.336.1511110.769.93601120.3412701130.27.35711412510001150.111.51001160.66251170.18.51180.472.71192.4124.7390120336.910001210.37122<0.10.12.9123<0.10.531240.52681125416.61000126294.41271.688.91280.24.1591292.713.313010010013155.31001320.22.2501330.11.11340.52.23113588.110013617.110013724.51001380.317.21390.10.64.21403.775.91410.20.43.71423.694.81431.93.111014483.110014510010014615.6100147<0.11.3191480.10.77.7149<0.10.31500.247.21510.151520.15.41530.34.31540.111550.10.35.21561001001576.429.415812.51001591001001600.21.4161610.30.3231620.40.51630.15.11640.338165<0.10.73.91660.33.5110167<0.10.7321680.20.5241690.826.519017018.1100<1,000171<0.11.91201723.565.3480173<0.14.21001745.71001750.511.91501760.10.49.81770.11.5201780.737.41790.10.47.71800.85.11810.50.36.91820.4871831.9632701840.7191850.42.723.11860.2131870.9621880.21001890.10.62.91900.11.57.9191239.81101920.14.51930.054.919431701950.071.51960.11.81970.042.41980.022.41990.073.22000.052.92010.02102020.37642030.121.22040.11.32050.062.22060.041.62070.0612080.093.3

The present disclosure provides reference to various embodiments and techniques. However, it should be understood that many variations and modifications can be made while remaining within the spirit and scope of the present disclosure. The description is made with the understanding that it is to be considered an exemplification of the claimed subject matter, and is not intended to limit the appended claims to the specific embodiments illustrated.