# Patent

## Title
Spirocyclic CETP inhibitors

## Abstract

Compounds having the structure of Formula (I), including pharmaceutically acceptable salts of the compounds, are CETP inhibitors and may be useful for raising HDL-cholesterol and reducing LDL-cholesterol in human patients and for treating or preventing atherosclerosis. The chemical compounds that are disclosed cholesterol ester transfer protein (CETP) and are expected to have utility in raising HDL-C, lowering LDL-C, and in the treatment and prevention of atherosclerosis.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a U.S. National Phase application under 35 U.S.C. §371 of PCT Application No. PCT/US2013/075531, filed Dec. 17, 2013, which claims priority under 35 U.S.C. §119(e) from U.S. Provisional Application No. 61/739,436, filed Dec. 19, 2012.
FIELD OF THE INVENTION
This invention relates to chemical compounds that inhibit cholesterol ester transfer protein (CETP) and are expected to have utility in raising HDL-C, lowering LDL-C, and in the treatment and prevention of atherosclerosis.
BACKGROUND OF THE INVENTION
Atherosclerosis and its clinical consequences, including coronary heart disease (CHD), stroke and peripheral vascular disease, represent a truly enormous burden to the health care systems of the industrialized world. In the United States alone, approximately 13 million patients have been diagnosed with CHD, and greater than one half million deaths are attributed to CHD each year. Further, this toll is expected to grow over the next quarter century as an epidemic in obesity and diabetes continues to grow.
It has long been recognized that in mammals, variations in circulating lipoprotein profiles correlate with the risk of atherosclerosis and CHD. The clinical success of HMG-CoA reductase inhibitors, especially the statins, in reducing coronary events is based on the reduction of circulating low density lipoprotein cholesterol (LDL-C), levels of which correlate directly with an increased risk for atherosclerosis. More recently, epidemiologic studies have demonstrated an inverse relationship between high density lipoprotein cholesterol (HDL-C) levels and atherosclerosis, leading to the conclusion that low serum HDL-C levels are associated with an increased risk for CHD.
Metabolic control of lipoprotein levels is a complex and dynamic process involving many factors. One important metabolic control in man is the cholesteryl ester transfer protein (CETP), a plasma glycoprotein that catalyzes the movement of cholesteryl esters from HDL to the apoB containing lipoproteins, especially VLDL (see Hesler, C. B., et. al. (1987) Purification and characterization of human plasma cholesteryl ester transfer protein. J. Biol. Chem. 262(5), 2275-2282)). Under physiological conditions, the net reaction is a heteroexchange in which CETP carries triglyceride to HDL from the apoB lipoprotein and transports cholesterol ester from HDL to the apoB lipoprotein.
In humans, CETP plays a role in reverse cholesterol transport, the process whereby cholesterol is returned to the liver from peripheral tissues. Intriguingly, many animals do not possess CETP, including animals that have high HDL levels and are known to be resistant to coronary heart disease, such as rodents (see Guyard-Dangremont, V., et. al., (1998) Phospholipid and cholesteryl ester transfer activities in plasma from 14 vertebrate species. Relation to atherogenesis susceptibility, Comp. Biochem. Physiol. B Biochem. Mol. Biol. 120(3), 517-525). Numerous epidemiologic studies correlating the effects of natural variation in CETP activity with respect to coronary heart disease risk have been performed, including studies on a small number of known human null mutations (see Hirano, K.-I., Yamashita, S. and Matsuzawa, Y. (2000) Pros and cons of inhibiting cholesteryl ester transfer protein, Curr. Opin. Lipidol. 11(6), 589-596). These studies have clearly demonstrated an inverse correlation between plasma HDL-C concentration and CETP activity (see Inazu, A., et. al. (2000) Cholesteryl ester transfer protein and atherosclerosis, Curr. Opin. Lipidol. 11(4), 389-396), leading to the hypothesis that pharmacologic inhibition of CETP lipid transfer activity may be beneficial to humans by increasing levels of HDL-C while lowering LDL-C.
Despite the significant therapeutic advance that statins such as simvastatin and atorvastatin represent, statins only achieve a risk reduction of approximately one-third in the treatment and prevention of atherosclerosis and ensuing atherosclerotic disease events. Currently, few pharmacologic therapies are available that favorably raise circulating levels of HDL-C. Certain statins and some fibrates offer modest HDL-C gains. Niacin provides an effective therapy for raising HDL-C but suffers from patient compliance issues, due in part to side effects such as flushing. Drugs that inhibit CETP (CETP inhibitors) have been under development with the expectation that they will effectively raise HDL cholesterol levels and also reduce the incidence of atherosclerosis in patients. Torcetrapib was the first drug that was tested in a long-term outcomes clinical trial. The clinical trial of torcetrapib was terminated early due to a higher incidence of mortality in patients to whom torcetrapib and atorvastatin were administered concomitantly compared with patients who were treated with atorvastatin alone. The cause of the increased mortality is not completely understood, but it is not believed to be associated with the CETP inhibiting effects of the drug. Dalcetrapib was recently tested in a Phase III outcomes trial, which was terminated early because the interim data did not show a clinical benefit. There were no safety issues detected for dalcetrapib.
Anacetrapib is currently the only CETP inhibitor being tested in a large scale Phase III clinical outcomes trial. Data from the recently completed DEFINE Phase II/III trial of anacetrapib are promising. Patients who were treated with anacetrapib along with baseline statin therapy showed an increase of HDL-C of 138% and a decrease of LDL-C of 40% compared with patients who were treated with just a statin. See: N. Engl. J. Med. 2010: 363: 2406-15. The DEFINE study was not carried out on a large enough scale to serve as a pivotal outcomes trial, but the data in the DEFINE trial were sufficient to indicate that an increase in mortality for patients treated with anacetrapib is unlikely. Additional drug candidates are in development. Evacetrapib currently appears to be the next CETP inhibitor that will proceed to a Phase III outcomes trial. Additional compounds are being sought that may have properties that are advantageous compared with the CETP inhibitors that have so far been studied or are currently being studied. Such properties may include, for example, higher potency, reduced off-target activity, better pharmacodynamics, higher bioavailability, or a reduced food effect compared with many of the highly lipophilic compounds that have so far been studied. “Food effect” refers to the variability in exposure to the active drug that occurs depending on when the patient had last eaten, whether or not the drug is administered with food, and the fat content of the food.
SUMMARY OF THE INVENTION
The compound of Formula I, or a pharmaceutically acceptable salt thereof, is a potent CETP inhibitor, having the utilities described herein:




In Formula I, each R is independently H or —C1-C3alkyl;
R1 is a cyclic substituent group connected to the oxazolidinone ring by a spirocyclic connection, wherein R1 is —C3-C8 cycloalkyl in which 1-2 carbons are optionally replaced by O, —NH—, —N(C1-C3alkyl)-, S, or —S(O)2—, wherein R1 is optionally substituted with 1-4 substituent groups which are halogen, —OH, —CN, —C1-C3 alkyl optionally substituted with 1-5 halogens, or —OC1-C3 alkyl optionally substituted with 1-5 halogens;
A and B are each A1 or A2, wherein one of A and B is A1 and the other of A and B is A2,
A1 has the structure:




wherein D is selected from the group consisting of:

    
    
        (a) phenyl, and
        (b) HET(1);
    
    


wherein ring D comprises at least two carbon atoms that are bonded to each other, wherein one of the two carbon atoms that are bonded to each other in ring D is connected to the group A3 and the other of the two carbon atoms that are bonded to each other in ring D is connected to the remainder of the structure of Formula I, so that A3 and the remainder of the structure of formula I are ortho to each other on ring D;
A3 is

    
    
        (a) phenyl, or
        (b) HET(1), wherein
    
    


A3 is optionally substituted with 1-5 substituent groups independently selected from Ra and optionally 1-2 groups X;
HET(1) is a monocyclic 3-8-membered heterocyclic or heteroaromatic ring having 1-4 heteroatom groups independently selected from N, NH, S, O, —S(O)—, —S(O)2—, and —N(O)—, wherein HET(1) optionally comprises 1-4 double bonds and a carbonyl group;
A2 is

    
    
        (a) phenyl,
        (b) HET(1), or
        (c) —C3-C8 cycloalkyl optionally having 1-3 double bonds;
    
    


wherein A2 is optionally substituted with 1-5 substituent groups independently selected from Ra;
Each Ra is independently selected from the group consisting of —C1-C6 alkyl, —C2-C6 alkenyl, —C2-C6 alkynyl, —OC1-C6alkyl, —OC2-C6alkenyl, —OC2-C6alkynyl, —C(═O)C1-C6alkyl, —C(═O)H, —OH, —NR6R7, —C(═O)NR6R7, —NR6C(═O)OC1-C6 alkyl, —NR6C(═O)NR6R7, —S(O)xC1-C6 alkyl, —S(O)yNR6R7, —NR6S(O)yNR6R7, —NR6S(O)yC1-C6 alkyl, halogen, —CN, —NO2, or HET(1), wherein HET(1) is optionally substituted with 1-3 substituent groups which are independently halogen, CH3, CF3, —OCH3, or —OCF3;
wherein for compounds in which Ra is selected from the group consisting of —C1-C6 alkyl, —C2-C6 alkenyl, —C2-C6 alkynyl, —OC1-C6alkyl, —OC2-C6 alkenyl, —OC2-C6 alkynyl, —C(═O)C1-C6alkyl, —NR6C(═O)OC1-C6 alkyl, —S(O)xC1-C6 alkyl, and —NR6S(O)yC1-C6 alkyl, Ra is optionally substituted with 1-11 halogens;
X is (a) C3-C8cycloalkyl which optionally comprises 1-2 double bonds and which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens and 1-2 groups —OH; (b) C1-C5 alkyl which is optionally substituted with 1-2 groups D1, optionally with 1-2 groups —OH, and optionally with 1-11 halogens; (c) phenyl which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens and 1-2 groups —OH; (d) HET1 which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens; or (e) D1;
D1 is —CO2R8, —C(O)NR6R7, SO2NR6R7, or HET(1);
R6 and R7 are each independently H or —C1-5alkyl;
R8 is H or —C1-5alkyl optionally substituted with 1-7 halogens;
p is an integer from 0-4;
x is 0, 1, or 2; and
y is 1 or 2.
In the compound of formula I and formula II (defined later) and in subgroups and other embodiments of the invention, alkyl groups and substituents based on alkyl groups, such as alkoxy, may be linear or branched unless otherwise indicated.
In general, references to the compound(s) of formula I or II are meant to also include subsets of compounds of formula I or II as may be defined herein, and also are meant to include the specific numbered examples provided herein.


DETAILED DESCRIPTION OF THE INVENTION
In further embodiments of the invention, the substituent groups defined above may have alternative values independent of one another, as written below. Such embodiments include pharmaceutically acceptable salts when such salts are possible.
In some embodiments, B is A1 and A is A2.
In some embodiments, B is A2 and A is A1.
In some embodiments, each R is H or CH3. In some embodiments, R is H.
In some embodiments, p is an integer from 0-2.
In some embodiments, p is an integer from 0-2.
In some embodiments, R1 is a cyclopropyl or cyclobutyl group connected to the oxazolidinone group of formula I by a spirocyclic linkage, wherein the cyclopropyl or cyclobutyl group is optionally substituted with 1-2 substituents which are halogen, CH3, CF3, —OCH3, or —OCF3.
In some embodiments, D and A3 are each independently phenyl, pyridyl, isoxazolyl, thienyl, imidazolyl, furyl, pyrrolyl, pyrazolyl, N-oxido-pyridyl, 1,3-thiazolyl, 1,3-oxazolyl, 1,2,4-triazolyl, tetrazolyl, pyridazinyl, pyrimidinyl, or pyrazinyl.
In some embodiments, A2 is phenyl, thienyl, imidazolyl, 1,3-thiazolyl, 1,3-oxazolyl, pyrrolyl, pyrazolyl, isoxazolyl, furyl, 1,2,4-triazolyl, tetrazolyl, pyridyl, N-oxido-pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, tetrahydropyranyl, or tetrahydrofuryl; wherein D, A3, and A2 are substituted as in Claim 1.
In some embodiments, B is A1, wherein A1 has the structure of formula II:




wherein Y1 and Y2 are each N or —CH—, and —CH— is optionally substituted with Ra2 in place of H;
Ra1 is (a) a 3-7-membered heterocycle having 1-2 heteroatoms which are independently N, S or O, wherein the heterocycle optionally has 1-3 double bonds and is optionally substituted with 1-3 substituent groups which are independently halogen, CH3, CF3, —OCH3, or —OCF3; (b) —NR6R7; (c) —N(C1-C3 alkyl)(SO2C1-C3alkyl); (d) C1-C3 alkyl optionally substituted with 1-3 halogens; (e) —OC1-C3 alkyl optionally substituted with 1-3 halogens; and (f) —SC1-C3 alkyl optionally substituted with 1-3 halogens;
Each Ra2 is optionally halogen, CH3, CF3, —OCH3, or —OCF3;
A is A2, wherein A2 is phenyl or pyridyl optionally substituted with 1-3 groups which are independently halogen, CH3, CF3, —OCH3, —OCF3, or —CN;
A3 is phenyl or pyridyl, which is optionally substituted with one group X and with 1-4 substituent groups which are independently halogen, C1-C3 alkyl optionally substituted with 1-3 halogens, and —OC1-C3 alkyl optionally substituted with 1-3 halogens;
X is (a) C3-C8cycloalkyl which optionally comprises 1-2 double bonds and which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens and 1-2 groups —OH; (b) C1-C5 alkyl which is optionally substituted with 1-2 groups D1, optionally with 1-2 groups —OH, and optionally with 1-11 halogens; (c) phenyl which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens and 1-2 groups —OH; (d) HET1 which is optionally substituted with 1-2 groups D1 and optionally with 1-5 substituent groups which are halogen, C1-C5 alkyl, —OC1-C5 alkyl, —CN, or —OH, wherein C1-C5 alkyl and —OC1-C5 alkyl are optionally substituted with 1-11 halogens; or (e) D1;
D1 is —CO2R8;
R6 and R7 are each independently H or —C1-3alkyl;
R8 is H or —C1-3alkyl optionally substituted with 1-5 halogens; and
d is 0, 1 or 2.
In some embodiments, R1 is unsubstituted cyclopropyl or cyclobutyl.
In some embodiments, B is A1, wherein A1 has the structure of formula II:




wherein Y1 and Y2 are each —CH— or —N—;
A is A2 wherein A2 is 3,5-bis-trifluoromethylphenyl;
Ra1 is 3-fluoroazetidinyl, 4-morpholinyl, CF3, —N(CH3)2, —N(CH3)(SO2CH3), or CH3S—,
A3 is




wherein Y3 is —CH— or —N—;
X is 3-cyclobutyl-D1, 3-cyclobutyl-CH2OH, 2-methyl-4-phenyl-D1, 2-methyl-4-phenyl-CH2OH, —CH2CH2D1, —CH2OH, or isopropyl, wherein D1 is —CO2R8;
R8 is H or —CH3;
Rb is —OCH3 or halogen;
d is 0; and
e is an integer from 0-3.
DEFINITIONS AND ABBREVIATIONS
“Ac” is acetyl, which is CH3C(═O)—.
“Alkyl” means saturated carbon chains which may be linear or branched or combinations thereof, unless the carbon chain is defined otherwise. Other groups having the prefix “alk”, such as alkoxy and alkanoyl, also may be linear or branched or combinations thereof, unless the carbon chain is defined otherwise. Examples of alkyl groups include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like.
“Alkylene” groups are alkyl groups that are difunctional rather than monofunctional. For example, methyl is an alkyl group and methylene (—CH2—) is the corresponding alkylene group. Alkyl groups that are shown as difunctional are alkylene groups, even if they are referred to as alkyl groups.
“Alkenyl” means carbon chains which contain at least one carbon-carbon double bond, and which may be linear or branched or combinations thereof. Examples of alkenyl include vinyl, allyl, isopropenyl, pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2-methyl-2-butenyl, and the like.
“Alkynyl” means carbon chains which contain at least one carbon-carbon triple bond, and which may be linear or branched or combinations thereof. Examples of alkynyl include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl and the like.
“Cycloalkyl” means a saturated carbocyclic ring having from 3 to 8 carbon atoms, unless otherwise stated. The term also includes a cycloalkyl ring fused to an aryl group. Examples of cycloalkyl include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. “Cycloalkenyl” means a non-aromatic carbocyclic ring having one or more double binds, but less than the number of double bonds that are required for the cycloalkenyl to be aromatic.
“Aryl” when used to describe a substituent or group in a structure means a monocyclic or bicyclic compound in which the rings are aromatic and which contain only carbon ring atoms. The term “aryl” can also refer to an aryl group that is fused to a cycloalkyl or heterocycle. Preferred “aryls” are phenyl and naphthyl. Phenyl is generally the most preferred aryl group.
“Heterocycle” or “heterocyclic” means a fully or partially saturated or aromatic cyclic compound containing 1 or more heteroatom groups which may be one or more of N, S, O, S(O), S(O)2, or (N)R, and may have one or more double bonds, where R is H or a substituent group. In general, when heterocycles are defined herein, the definition will include the number of ring members, the number of double bonds (if any), and the specific heteroatoms. The heterocycles in some cases will be aromatic, depending on the number of double bonds (e.g. 6-membered ring with 3 double bonds). Aromatic heterocycles are also referred to as heteroaromatics. S(O), S(O)2, and N(R) are referred to as heteroatom groups, and each heteroatom group is counted as one ring member, as is also the case for N, S, and O.
“Benzoheterocycle” represents a phenyl ring fused to a heterocyclic ring. Examples include indole, benzofuran, 2,3-dihydrobenzofuran and quinoline.
“Boc” is tert-butoxycarbonyl.
“n-BuLi” is n-butyl lithium.
“Celite®” is a trade name for diatomaceous earth.
“DBU” is 1,8-diazabicyclo[5.4.0]undec-7-ene.
“D-Epoxone” is a commercial epoxidation catalyst.
“DIPEA” and “DIEA” are N,N-diisopropylethylamine.
“DCM” is dichloromethane.
“DIBAL” or “DIBAL-H” is diisobutylaluminum hydride.
“DMF” is N,N-dimethylformamide.
“DMAP” is 4-dimethylaminopyridine.
“DMSO” is dimethyl sulfoxide.
“DOPC” is 1,2-dioleoyl-sn-glycero-3-phosphocholine.
“EDTA” is ethylenediaminetetraacetic acid.
“EtOAc” is ethyl acetate.
“EtOH” is ethanol.
“Halogen” includes fluorine, chlorine, bromine and iodine.
“HATU” is O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate, a peptide coupling reagent.
“HPLC” is high pressure liquid chromatography.
“IPA” is isopropyl alcohol.
“LAH” is lithium aluminum hydride.
“LCMS” is liquid chromatograpy-mass specrtrometry.
“LiHMDS” is lithium hexamethyldisilazide.
“Me” represents methyl.
“MeCN” is acetonitrile.
“MeOH” is methanol.
“Ms Cl” is methanesulfonyl chloride
“MS-ESI” is electrospray ionization mass spectrometry.
“NMP” is N-methyl-2-pyrrolidone.
“OXONE®” is a commercial persulfate oxidizing agent from DuPont.
“PEG” is poly(ethylene glycol).
“RBF” is a round bottom flask.
“Rochelle's salt” is potassium sodium tartrate.
“RT” is an abbreviation for room temperature.
“SFC” is supercritical fluid chromatography.
“SM” is starting material.
“TEA” is triethylamine.
“TFA” is trifluoroacetic acid.
“THF” is tetrahydrofuran.
“TLC” is thin layer chromatography.
The term “composition,” as in pharmaceutical composition, is intended to encompass a product comprising the active ingredient(s) and the inert ingredient(s) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients. Accordingly, the pharmaceutical compositions of the present invention encompass any composition made by admixing a compound of Formula I or II and a pharmaceutically acceptable carrier.
The substituent “tetrazole” means a 2H-tetrazol-5-yl substituent group and tautomers thereof.
Optical Isomers-Diastereomers-Geometric Isomers-Tautomers
The compounds disclosed herein generally have at least one asymmetric center, and can thus occur as pure stereoisomers and as mixtures of stereoisomers, including racemates, racemic mixtures, single enantiomers, mixtures of enantiomers, diastereomeric mixtures and individual diastereomers in all ratios. The invention includes all possible enantiomers and diastereomers and mixtures of two or more stereoisomers, for example mixtures of enantiomers and/or diastereomers, in all ratios. Thus, enantiomers are a subject of the invention in enantiomerically pure form, both as levorotatory and as dextrorotatory antipodes, in the form of racemates and in the form of mixtures of the two enantiomers in all ratios. In the case of a cis/trans isomerism the invention includes both the cis form and the trans form as well as mixtures of these forms in all ratios. The preparation of individual stereoisomers can be carried out, if desired, by separation of a mixture by customary methods, for example by chromatography or crystallization, by the use of stereochemically uniform starting materials for the synthesis or by stereoselective synthesis. Optionally a derivatization can be carried out before a separation of stereoisomers. The separation of a mixture of stereoisomers can be carried out at an intermediate step during the synthesis of a compound of Formula I or II or it can be done on a final racemic product. Absolute stereochemistry may be determined by X-ray crystallography of crystalline products or crystalline intermediates which are derivatized, if necessary, with a reagent containing a stereogenic center of known configuration. Where compounds of this invention are capable of tautomerization, all individual tautomers as well as mixtures thereof are included in the scope of this invention. The present invention includes all such isomers, as well as salts, solvates (which includes hydrates) and solvated salts of such racemates, enantiomers, diastereomers and tautomers and mixtures thereof.
Different stereoisomers having the same 2-dimensional chemical structure may have different levels of activity with respect to CETP inhibition, so that some stereoisomers may have higher activity than others. The compounds that are potent inhibitors of CETP may have utility in patients for raising HDL-C, lowering LDL-C, treating dyslipidemia, and for preventing, treating or delaying the onset of conditions that are related to atherosclerosis. Stereoisomers that have little or no activity may have utility as research tools for better understanding CETP inhibition. All stereoisomers and mixtures of stereoisomers of the claimed compounds thus have utility. The compounds of Formula I or II may also occur as atropisomers (rotamers) due to hindered rotation, which may be observable by NMR spectroscopy, and in some cases may be stable enough with respect to conversion by bond rotation to other atropisomers that they can be isolated and assayed.
Salts
When the compounds of Formula I contain one or more acidic or basic groups the invention also includes the corresponding pharmaceutically acceptable salts. Thus, the compounds of Formula I which contain acidic groups can be used according to the invention, for example, as alkali metal salts, alkaline earth metal salts or as ammonium salts. Examples of such salts include but are not limited to sodium salts, potassium salts, calcium salts, magnesium salts or salts with ammonia or organic amines such as, for example, ethylamine, ethanolamine, triethanolamine or amino acids. Compounds of Formula I which contain one or more basic groups, i.e. groups which can be protonated, can be used according to the invention in the form of their acid addition salts with inorganic or organic acids as, for example but not limited to, salts with hydrogen chloride, hydrogen bromide, phosphoric acid, sulfuric acid, nitric acid, benzenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acids, oxalic acid, acetic acid, trifluoroacetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, diethylacetic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfaminic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid, etc. If the compounds of Formula I simultaneously contain acidic and basic groups in the molecule the invention also includes, in addition to the salt forms mentioned, inner salts or betaines (zwitterions). Salts can be obtained from the compounds of Formula I by customary methods which are known to the person skilled in the art, for example by combination with an organic or inorganic acid or base in a solvent or dispersant, or by anion exchange or cation exchange from other salts. The present invention also includes all salts of the compounds of Formula I which, owing to low physiological compatibility, are not directly suitable for use in pharmaceuticals but which can be used, for example, as intermediates for chemical reactions or for the preparation of pharmaceutically acceptable salts.
Furthermore, compounds of the present invention may exist in one or more amorphous forms and/or one or more crystalline forms, and as such all amorphous and crystalline forms and mixtures thereof of the compounds of Formula I are intended to be included within the scope of the present invention. In addition, some of the compounds of the instant invention may form solvates with water (i.e., a hydrate) or common organic solvents. Such solvates and hydrates, particularly the pharmaceutically acceptable solvates and hydrates, of the instant compounds are likewise encompassed within the scope of this invention, along with un-solvated and anhydrous forms.
It will be understood that, as used herein, references to the compounds of Formula I and II and to the examples are meant to also include the pharmaceutically acceptable salts and prodrugs, where such salts and prodrugs are possible.
Prodrugs
Prodrugs, which are compounds that are converted to the compound of Formula I or II as they are being administered to a patient or after they have been administered to a patient, are also compounds of formula I or II in the sense that they provide the claimed pharmaceutically active drug moiety to the patient.
Isotopes
In the compounds of Formula I and Formula II, the atoms may exhibit their natural isotopic abundances, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is meant to include all suitable isotopic variations of the compounds of generic Formula I and Formula II. For example, different isotopic forms of hydrogen (H) include protium (1H) and deuterium (2H). Protium is the predominant hydrogen isotope found in nature. Enriching for deuterium may afford certain therapeutic advantages, such as increasing in vivo half-life or reducing dosage requirements, or may provide a compound useful as a standard for characterization of biological samples. Isotopically-enriched compounds within generic Formula I and II can be prepared without undue experimentation by conventional techniques well known to those skilled in the art or by processes analogous to those described in the Schemes and Examples herein using appropriate isotopically-enriched reagents and/or intermediates.
Utilities
The compounds disclosed herein, including pharmaceutically acceptable salts thereof, are potent inhibitors of CETP. The compounds may therefore be useful in treating mammalian patients, preferably human patients, having diseases and conditions that are treated by inhibition of CETP.
One aspect of the present invention provides a method for treating or reducing the risk of developing a disease or condition that may be treated or prevented by inhibition of CETP by administering a therapeutically effective amount of the compound of Formula I or II to a patient in need of treatment. The patient is a human or mammal, but is most often a human. A “therapeutically effective amount” is the amount of compound that is effective in obtaining a desired clinical outcome in the treatment of a specific disease.
Diseases or conditions that may be treated with the compounds of Formula I or Formula II, or which the patient may have a reduced risk of developing as a result of being treated with the compounds of Formula I or Formula II, include: atherosclerosis, peripheral vascular disease, dyslipidemia, hyperbetalipoproteinemia, hypoalphalipoproteinemia, hypercholesterolemia, hypertriglyceridemia, familial-hypercholesterolemia, cardiovascular disorders, angina, ischemia, cardiac ischemia, stroke, myocardial infarction, reperfusion injury, angioplastic restenosis, hypertension, vascular complications of diabetes, obesity, endotoxemia, and metabolic syndrome. There are reports in the scientific literature that suggest that inhibition of CETP may have utility in preventing or slowing the development of Alzheimer's disease. The compounds of Formula I and II may therefore have utility in preventing or delaying the progression of Alzheimer's disease or other neurodegenerative diseases.
The compounds disclosed herein are particularly effective in raising HDL-C and/or increasing the ratio of HDL-C to LDL-C. The compounds may also be effective in reducing LDL-C, and may be effective in treating dyslipidemia. These changes in HDL-C and LDL-C may be beneficial in treating atherosclerosis, reducing or delaying the development of atherosclerosis, reducing the risk of developing atherosclerosis, or preventing atherosclerosis. The compounds disclosed herein may thus be beneficial in treating atherosclerosis, reducing or delaying the development of atherosclerosis, reducing the risk of developing atherosclerosis, or preventing atherosclerosis.
Likely indications for atherosclerosis and dyslipidemia using the compounds described herein are written below, where the drug product is titled “CETP inhibitor:”
Atherosclerosis. In patients at high risk of cardiovascular events because of existing coronary, cerebrovascular, or peripheral vascular disease, CETP inhibitor co-administered with an HMG-CoA reductase inhibitor is indicated to reduce the risk of coronary mortality, myocardial infarction, coronary revascularization procedures, ischemic stroke, and cardiovascular death.
Dyslipidemia. CETP inhibitor co-administered with a statin is indicated to reduce elevated LDL-C, apolipoprotein B (ApoB), lipoprotein a (Lp(a)), non-HDL-C, and total cholesterol; and increase HDL-C and apolipoprotein A-1 (Apo A-1) in patients with mixed or primary dyslipidemia.
Administration and Dose Ranges
Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dose of the compounds described herein. For example, oral, rectal, topical, parenteral, ocular, pulmonary, nasal, and the like may be employed. Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like. Preferably the compound of Formula I or II is administered orally.
When treating the diseases for which the compound of Formula I or II is indicated, generally satisfactory results are expected when the compound of Formula I or II is administered at a daily dosage of from about 0.1 milligram to about 1000 milligram in one dose daily or divided into more than one dose per day.
Oral administration will usually be carried out using tablets. Examples of doses in tablets include 0.1 mg, 0.5 mg, 1 mg, 2 mg, 5 mg, 10 mg, 25 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 275 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, and 1000 mg. Other oral forms can also have the same dosages (e.g. capsules). A preferred dose is likely in the range of 50-200 mg.
Pharmaceutical Compositions
Another aspect of the present invention provides pharmaceutical compositions which comprise the compound of Formula I or II and a pharmaceutically acceptable carrier. The pharmaceutical compositions of the present invention comprise the compound of Formula I or II or a pharmaceutically acceptable salt as an active ingredient, as well as a pharmaceutically acceptable carrier and optionally other therapeutic ingredients. The term “pharmaceutically acceptable salts” refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic bases or acids and organic bases or acids. A pharmaceutical composition may also comprise a prodrug, or a pharmaceutically acceptable salt thereof, if a prodrug is administered. A pharmaceutical composition may also consist essentially of the compound of Formula I or II, or a pharmaceutically acceptable salt of the compound, and a pharmaceutically acceptable carrier, without other therapeutic ingredients.
Pharmaceutical compositions may be formulated to be suitable for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous), ocular (ophthalmic), pulmonary (nasal or buccal inhalation), or nasal administration, although the most suitable route in any given case will depend on the nature and severity of the conditions being treated and on the nature of the active ingredient. They may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the art of pharmacy.
In practical use, the compound of Formula I or II can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g., oral or parenteral (including intravenous). In preparing the compositions for oral dosage form, any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations, such as, for example, suspensions, elixirs and solutions; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations such as, for example, powders, hard and soft capsules and tablets, with the solid oral preparations being preferred over the liquid preparations.
Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques. Such compositions and preparations should contain at least 0.1 percent of active compound. The percentage of active compound in these compositions may, of course, be varied and may conveniently be between about 2 percent to about 60 percent of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that an effective dosage will be obtained. The active compound can also be administered intranasally as, for example, liquid drops or spray.
The tablets, pills, capsules, and the like may also contain a binder such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose or saccharin. When a dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as a fatty oil.
Various other materials may be present as coatings or to modify the physical form of the dosage unit. For instance, tablets may be coated with shellac, sugar or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl and propylparabens as preservatives, a dye and a flavoring such as cherry or orange flavor.
The compound of formula I or II may also be administered parenterally. Solutions or suspensions of the compound can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols and mixtures thereof in oils. Under ordinary conditions of storage and use, these preparations may contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), suitable mixtures thereof, and vegetable oils.
Combination Therapy
The compound of Formula I or II, including pharmaceutically acceptable salts thereof, may be used in pharmaceutical combinations with other drugs that may also be useful in the treatment or amelioration of the diseases or conditions for which the compound of Formula I or II is useful. Such other drugs may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with the compound of Formula I or II. When the compound of Formula I or II is used contemporaneously with one or more other drugs, a pharmaceutical composition in unit dosage form containing such other drugs and the compound of Formula I or II is preferred. However, the combination therapy also includes therapies in which the compound of Formula I or II and one or more other drugs are administered concomitantly, on the same or different schedules. The drugs that are administered, whether alone or in combination with other drugs, include the free-acid, free-base, and pharmaceutically acceptable salt forms, pro-drug forms (including but not limited to esters), and salts of pro-drugs of the medicinal agents where chemically possible
When oral formulations are used, the drugs may be combined into a single combination tablet or other oral dosage form, or the drugs may be packaged together as separate tablets or other oral dosage forms. It is also contemplated that when used in combination with one or more other active ingredients, the compound of formula I or II and the other active ingredients may be used in lower doses than when each is used singly. Accordingly, the pharmaceutical compositions of the compound of formula I or II include those that contain one or more other active ingredients, in addition to the compound of Formula I or II.
The compound of Formula I or II will likely be approved initially for coadministration with a statin, which could be administered in the form of a fixed dose combination of the compound of formula I or II and a statin. Additional drugs may also be administered in combination with the compound of Formula I or II and the statin, either by coadministration or in a fixed dose combination. The compound of formula I or II and the drugs that are administered with it may be administered as pharmaceutically acceptable salts, as prodrugs, or otherwise formulated for immediate release, extended release, or controlled release, as necessary.
Examples of statins that may be administered in combination with the compound of Formula I or II include, but are not limited to, (i) simvastatin and lovastatin which are marketed as ZOCOR® and MEVACOR® in lactone prodrug form and function as inhibitors after administration, and (ii) dihydroxy open ring acid HMG-CoA reductase inhibitors such as atorvastatin (particularly the calcium salt sold in LIPITOR®), rosuvastatin (particularly the calcium salt sold in CRESTOR®), pravastatin (particularly the sodium salt sold in PRAVACHOL®), fluvastatin (particularly the sodium salt sold in LESCOL®), and pitavastatin (particularly the calcium salt sold in LIVALO®), and (iii) other statins that may yet be developed. Preferred statins for combination therapy include atorvastatin, rosuvastatin, and simvasatin, as described above.
Cholesterol absorption inhibitors, and particularly ezetimibe (ZETIA®), as well as other cholesterol asorption inhibitors, such as stanol esters, beta-sitosterol, sterol glycosides such as tiqueside, and other azetidinones, may be administered with the compound of Formula I or II, generally with a statin, as described above. The preferred cholesterol absorbtion inhibitor is ezetimibe. Combinations of the compound of formula I or II with a statin and a cholesterol inhibitor, such as ezetimibe, are also contemplated. Preferred 3-component combinations include combinations of the compound of formula I or II with simvastatin, atorvastatin, or rosuvastatin in combination with ezetimibe, where the statins may be salt forms or prodrugs as described above. The combination of simvastatin with ezetimibe is currently marketed as VYTORIN®.
Other cholesterol reducing drugs that may be coadministered with the compound of formula I or II in addition to HMG-CoA reductase inhibitors (statins) and cholesterol absorption inhibitors include (i) bile acid sequestrants, as for example cholestyramine, colestipol, dialkylaminoalkyl derivatives of a cross-linked dextran, Colestid®, and LoCholest®, (ii) niacin and related compounds, such as nicotinyl alcohol, nicotinamide, and nicotinic acid or a salt thereof, in an immediate release or extended release form, which may optionally be in the forme of a combination with a DP-1 antagonist, such as laropiprant (TREDAPTIVE®); (iii) PPARα agonists, such as gemfibrozil and fenofibric acid derivatives (fibrates), including clofibrate, fenofibrate, bezafibrate, ciprofibrate, and etofibrate, (iv) acyl CoA: cholesterol acyltransferase (ACAT) inhibitors, such as avasimibe and melinamide, and including selective ACAT-1 and ACAT-2 inhibitors and dual inhibitors, (v) phenolic anti-oxidants, such as probucol, (vi) microsomal triglyceride transfer protein (MTP)/ApoB secretion inhibitors, (vii) anti-oxidant vitamins, such as vitamins C and E and beta carotene, (viii) thyromimetics, (ix) LDL (low density lipoprotein) receptor inducers, (x) platelet aggregation inhibitors, for example glycoprotein IIb/IIIa fibrinogen receptor antagonists and aspirin, (xi) vitamin B 12 (also known as cyanocobalamin), (xii) folic acid or a pharmaceutically acceptable salt or ester thereof, such as the sodium salt and the methylglucamine salt, (xiii) FXR and LXR ligands, including both inhibitors and agonists, (xiv) agents that enhance ABCA1 gene expression, (xv) ileal bile acid transporters, and (xvi) niacin receptor agonists (e.g. acipimox and acifran) and partial agonists.
Finally the compound of formula I or II can be combined with compounds that are useful for treating other diseases, such as diabetes, hypertension and obesity, as well as other anti-atherosclerotic compounds. Such combinations may be used to treat one or more of such diseases as diabetes, obesity, atherosclerosis, and dyslipidemia, or more than one of the diseases associated with metabolic syndrome. The combinations may exhibit synergistic activity in treating these diseases, allowing for the possibility of administering reduced doses of active ingredients, such as doses that otherwise might be sub-therapeutic.
Examples of other active ingredients that may be administered in combination with a compound of formula I or II include, but are not limited to, compounds that are primarily anti-diabetic compounds, including:
(a) PPAR gamma agonists and partial agonists, including glitazones and non-glitazones (e.g. pioglitazone, englitazone, MCC-555, rosiglitazone, balaglitazone, netoglitazone, T-131, LY-300512, LY-818, and compounds described in WO 02/060388, WO 02/08188, WO 2004/019869, WO 2004/020409, WO 2004/020408, and WO2004/066963);
(b) biguanides such as metformin, phenformin, and pharmaceutically acceptable salts thereof, in particular metformin hydrochloride and extended release formulations thereof, such as GlumetzaTM, FortametTM, and GlucophageXRTM;
(c) protein tyrosine phosphatase-1B (PTP-1B) inhibitors, such as ISIS-113715 and TTP814;
(d) dipeptidyl peptidase IV (DP-IV) inhibitors, including sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, dutogliptin, teneligliptin, MK-3102, and gemigliptin;
(e) insulin or insulin mimetics, such as for example insulin lispro, insulin glargine, insulin detemir, insulin glulisine, insulin degludec, SBS 1000, insulin zinc suspension, and oral and inhalable formulations of insulin and insulin analogs;
(f) sulfonylureas, such as tolbutamide, glipizide, glimepiride, acetohexamide, chlorpropamide, glibenclamide, and related materials;
(g) α-glucosidase inhibitors (such as acarbose, adiposine; camiglibose; emiglitate; miglitol; voglibose; pradimicin-Q; and salbostatin);
(h) PPARα/γ dual agonists, such as muraglitazar, tesaglitazar, farglitazar, and naveglitazar;
(i) PPARδ agonists such as GW501516 and those disclosed in WO97/28149;
(j) glucagon receptor antagonists;
(k) GLP-1; GLP-1 derivatives; GLP-1 mimetics, GLP-1 analogs, and GLP-1 receptor agonists, such as exendins, e.g. exenatide (BYETTA), dulaglutide, semaglutide, albiglutide, liraglutide, lixisenatide, and taspoglutide, including intranasal, tranxsdermal, and once weekly fomulations thereof, and oxyntomodulin analogs and derivatives, and non-peptidyl GLP-1 receptor agonists;
(l) GIP-1;
(m) amylin and amylin analogs (e.g. pramlintide);
(n) Non-sulfonylurea insulin secretagogues, such as the meglitinides (e.g. glimepiride, mitiglinide, meglitinide, nateglinide, and rapeglinide); and
(o) leptin and leptin derivatives and agonists.
Preferred combinations with antidiabetic compounds include combinations of the compounds disclosed herein with DP-IV inhibitors (sitagliptin, vildagliptin, saxagliptin, alogliptin, linagliptin, dutogliptin, teneligliptin, omarigliptin, and gemigliptin), combinations with biguanides, and combinations with both a DP-IV inhibitor and a biguanide. The preferred DP-IV inhibitor is sitagliptin, and the preferred biguanide is metformin in the formulations and salt forms described above.
Other active ingredients that may be used in combination with the compound of formula I or II include antiobesity compounds, including 5-HT(serotonin) inhibitors, neuropeptide Y5 (NPY5) inhibitors, melanocortin 4 receptor (Mc4r) agonists, cannabinoid receptor 1 (CB-1) antagonists/inverse agonists, and β3 adrenergic receptor agonists. These are listed in more detail later in this section.
These other active ingredients also include active ingredients that are used to treat inflammatory conditions, such as aspirin, non-steroidal anti-inflammatory drugs, glucocorticoids, azulfidine, and selective cyclooxygenase-2 (COX-2) inhibitors, including etoricoxib, celecoxib, rofecoxib, and Bextra.
Antihypertensive compounds may also be used advantageously in combination therapy with the compound of formula I or II. Examples of antihypertensive compounds that may be used with the compound of formula I or II include thiazide-like diuretics, e.g., hydrochlorothiazide (HCTZ or HCT); angiotensin converting enzyme inhibitors (e.g., alacepril, benazepril, captopril, ceronapril, cilazapril, delapril, enalapril, enalaprilat, fosinopril, imidapril, lisinopril, moveltipril, perindopril, quinapril, ramipril, spirapril, temocapril, or trandolapril); dual inhibitors of angiotensin converting enzyme (ACE) and neutral endopeptidase (NEP) such as omapatrilat, sampatrilat and fasidotril; angiotensin II receptor antagonists, also known as angiotensin receptor blockers or ARBs, which may be in free-base, free-acid, salt or pro-drug form, such as azilsartan, e.g., azilsartan medoxomil potassium (EDARBI®), candesartan, e.g., candesartan cilexetil (ATACAND®), eprosartan, e.g., eprosartan mesylate (TEVETAN®), irbesartan (AVAPRO®), losartan, e.g., losartan potassium (COZAAR®), olmesartan, e.g., olmesartan medoximil (BENICAR®), telmisartan (MICARDIS®), valsartan (DIOVAN®), and any of these drugs used in combination with a thiazide-like diuretic such as hydrochlorothiazide (e.g., HYZAAR®, DIOVAN HCT®, ATACAND HCT®), etc.); potassium sparing diuretics such as amiloride HCl, spironolactone, epleranone, triamterene, each with or without HCTZ; carbonic anhydrase inhibitors, such as acetazolamide; neutral endopeptidase inhibitors (e.g., thiorphan and phosphoramidon); aldosterone antagonists; aldosterone synthase inhibitors; renin inhibitors (e.g. urea derivatives of di- and tri-peptides (See U.S. Pat. No. 5,116,835), amino acids and derivatives (U.S. Pat. Nos. 5,095,119 and 5,104,869), amino acid chains linked by non-peptidic bonds (U.S. Pat. No. 5,114,937), di- and tri-peptide derivatives (U.S. Pat. No. 5,106,835), peptidyl amino diols (U.S. Pat. Nos. 5,063,208 and 4,845,079) and peptidyl beta-aminoacyl aminodiol carbamates (U.S. Pat. No. 5,089,471); also, a variety of other peptide analogs as disclosed in the following U.S. Pat. Nos. 5,071,837; 5,064,965; 5,063,207; 5,036,054; 5,036,053; 5,034,512 and 4,894,437, and small molecule renin inhibitors (including diol sulfonamides and sulfinyls (U.S. Pat. No. 5,098,924), N-morpholino derivatives (U.S. Pat. No. 5,055,466), N-heterocyclic alcohols (U.S. Pat. No. 4,885,292) and pyrolimidazolones (U.S. Pat. No. 5,075,451); also, pepstatin derivatives (U.S. Pat. No. 4,980,283) and fluoro- and chloro-derivatives of statone-containing peptides (U.S. Pat. No. 5,066,643); enalkrein; RO 42-5892; A 65317; CP 80794; ES 1005; ES 8891; SQ 34017; aliskiren (2(S),4(S),5(S),7(S)—N-(2-carbamoyl-2-methylpropyl)-5-amino-4-hydroxy-2,7-diisopropyl-8-[4-methoxy-3-(3-methoxypropoxy)-phenyl]-octanamid hemifumarate) SPP600, SPP630 and SPP635); endothelin receptor antagonists; vasodilators (e.g. nitroprusside); calcium channel blockers (e.g., amlodipine, nifedipine, verapamil, diltiazem, felodipine, gallopamil, niludipine, nimodipine, nicardipine, bepridil, nisoldipine); potassium channel activators (e.g., nicorandil, pinacidil, cromakalim, minoxidil, aprilkalim, loprazolam); sympatholitics; beta-adrenergic blocking drugs (e.g., acebutolol, atenolol, betaxolol, bisoprolol, carvedilol, metoprolol, metoprolol tartate, nadolol, propranolol, sotalol, timolol); alpha adrenergic blocking drugs (e.g., doxazocin, prazocin or alpha methyldopa); central alpha adrenergic agonists; peripheral vasodilators (e.g. hydralazine); and nitrates or nitric oxide donating compounds, e.g. isosorbide mononitrate.
Preferred antihypertensives that may be used in combination with the CETP inhibitors disclosed herein include one or more of an angiotensin II antagonist (losartan), an ACE inhibitor (enalapril or captopril), and hydrochlorothiazide.
Anti-obesity compounds may be administered in combination with the compounds of Formula I or Formula II, including: (1) growth hormone secretagogues and growth hormone secretagogue receptor agonists/antagonists, such as NN703 and hexarelin; (2) protein tyrosine phosphatase-1B (PTP-1B) inhibitors; (3) cannabinoid receptor ligands, such as cannabinoid CB1 receptor antagonists or inverse agonists, such as rimonabant (Sanofi Synthelabo), AMT-251, and SR-14778 and SR 141716A (Sanofi Synthelabo), SLV-319 (Solvay), BAY 65-2520 (Bayer); (4) anti-obesity serotonergic agents, such as fenfluramine, dexfenfluramine, phentermine, and sibutramine; (5) β3-adrenoreceptor agonists, such as AD9677/TAK677 (Dainippon/Takeda), CL-316,243, SB 418790, BRL-37344, L-796568, BMS-196085, BRL-35135A, CGP12177A, BTA-243, Trecadrine, Zeneca D7114, and SR 59119A; (6) pancreatic lipase inhibitors, such as orlistat (Xenical®), Triton WR1339, RHC80267, lipstatin, tetrahydrolipstatin, teasaponin, and diethylumbelliferyl phosphate; (7) neuropeptide Y1 antagonists, such as BIBP3226, J-115814, BIBO 3304, LY-357897, CP-671906, and GI-264879A; (8) neuropeptide Y5 antagonists, such as GW-569180A, GW-594884A, GW-587081X, GW-548118X, FR226928, FR 240662, FR252384, 1229U91, GI-264879A, CGP71683A, LY-377897, PD-160170, SR-120562A, SR-120819A and JCF-104; (9) melanin-concentrating hormone (MCH) receptor antagonists; (10) melanin-concentrating hormone 1 receptor (MCH1R) antagonists, such as T-226296 (Takeda); (11) melanin-concentrating hormone 2 receptor (MCH2R) agonist/antagonists; (12) orexin-1 receptor antagonists, such as SB-334867-A; (13) melanocortin agonists, such as Melanotan II; (14) other Mc4r (melanocortin 4 receptor) agonists, such as CHIR86036 (Chiron), ME-10142, and ME-10145 (Melacure), CHIR86036 (Chiron); PT-141, and PT-14 (Palatin); (15) 5HT-2 agonists; (16) 5HT2C (serotonin receptor 2C) agonists, such as BVT933, DPCA37215, WAY161503, and R-1065; (17) galanin antagonists; (18) CCK agonists; (19) CCK-A (cholecystokinin-A) agonists, such as AR-R 15849, GI 181771, JMV-180, A-71378, A-71623 and SR146131; (20) GLP-1 agonists; (21) corticotropin-releasing hormone agonists; (22) histamine receptor-3 (H3) modulators; (23) histamine receptor-3 (H3) antagonists/inverse agonists, such as hioperamide, 3-(1H-imidazol-4-yl)propyl N-(4-pentenyl)carbamate, clobenpropit, iodophenpropit, imoproxifan, and GT2394 (Gliatech); (24) β-hydroxy steroid dehydrogenase-1 inhibitors (11β-HSD-1 inhibitors), such as BVT 3498 and, BVT 2733, (25) PDE (phosphodiesterase) inhibitors, such as theophylline, pentoxifylline, zaprinast, sildenafil, amrinone, milrinone, cilostamide, rolipram, and cilomilast; (26) phosphodiesterase-3B (PDE3B) inhibitors; (27) NE (norepinephrine) transport inhibitors, such as GW 320659, despiramine, talsupram, and nomifensine; (28) ghrelin receptor antagonists; (29) leptin, including recombinant human leptin (PEG-OB, Hoffman La Roche) and recombinant methionyl human leptin (Amgen); (30) leptin derivatives; (31) BRS3 (bombesin receptor subtype 3) agonists such as [D-Phe6,beta-Ala11,Phe13,Nle14]Bn(6-14) and [D-Phe6,Phe13]Bn(6-13)propylamide; (32) CNTF (Ciliary neurotrophic factors), such as GI-181771 (Glaxo-SmithKline), SR146131 (Sanofi Synthelabo), butabindide, PD170,292, and PD 149164 (Pfizer); (33) CNTF derivatives, such as axokine (Regeneron); (34) monoamine reuptake inhibitors, such as sibutramine; (35) UCP-1 (uncoupling protein-1, 2, or 3) activators, such as phytanic acid, 4-[(E)-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-napthalenyl)-1-propenyl]benzoic acid (TTNPB), and retinoic acid; (36) thyroid hormone β □agonists, such as KB-2611 (KaroBioBMS); (37) FAS (fatty acid synthase) inhibitors, such as Cerulenin and C75; (38) DGAT1 (diacylglycerol acyltransferase 1) inhibitors; (39) DGAT2 (diacylglycerol acyltransferase 2) inhibitors; (40) ACC2 (acetyl-CoA carboxylase-2) inhibitors; (41) glucocorticoid antagonists; (42) acyl-estrogens, such as oleoyl-estrone; (43) dicarboxylate transporter inhibitors; (44) peptide YY, PYY 3-36, peptide YY analogs, derivatives, and fragments such as BIM-43073D, BIM-43004C, (45) Neuropeptide Y2 (NPY2) receptor agonists such NPY3-36, N acetyl [Leu(28,31)] NPY 24-36, TASP-V, and cyclo-(28/32)-Ac-[Lys28-Glu32]-(25-36)-pNPY; (46) Neuropeptide Y4 (NPY4) agonists such as pancreatic peptide (PP); (47) Neuropeptide Y1 (NPY1) antagonists such as BIBP3226, J-115814, BIBO 3304, LY-357897, CP-671906, and GI-264879A; (48) Opioid antagonists, such as nalmefene (Revex®), 3-methoxynaltrexone, naloxone, and naltrexone; (49) glucose transporter inhibitors; (50) phosphate transporter inhibitors; (51) 5-HT (serotonin) inhibitors; (52) beta-blockers; (53) Neurokinin-1 receptor antagonists (NK-1 antagonists); (54) clobenzorex; (55) cloforex; (56) clominorex; (57) clortermine; (58) cyclexedrine; (59) dextroamphetamine; (60) diphemethoxidine, (61) N-ethylamphetamine; (62) fenbutrazate; (63) fenisorex; (64) fenproporex; (65) fludorex; (66) fluminorex; (67) furfurylmethylamphetamine; (68) levamfetamine; (69) levophacetoperane; (70) mefenorex; (71) metamfepramone; (72) methamphetamine; (73) norpseudoephedrine; (74) pentorex; (75) phendimetrazine; (76) phenmetrazine; (77) picilorex; (78) phytopharm 57; (79) zonisamide, (80) aminorex; (81) amphechloral; (82) amphetamine; (83) benzphetamine; and (84) chlorphentermine.
The combination therapies described above which use the compounds of Formula I or Formula II may also be useful in the treatment of the metabolic syndrome. According to one widely used definition, a patient having metabolic syndrome is characterized as having three or more symptoms selected from the following group of five symptoms: (1) abdominal obesity; (2) hypertriglyceridemia; (3) low high-density lipoprotein cholesterol (HDL); (4) high blood pressure; and (5) elevated fasting glucose, which may be in the range characteristic of Type 2 diabetes if the patient is also diabetic. Each of these symptoms is defined clinically in the Third Report of the National Cholesterol Education Program Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III, or ATP III), National Institutes of Health, 2001, NIH Publication No. 01-3670. Patients with metabolic syndrome have an increased risk of developing the macrovascular and microvascular complications that are listed above, including atherosclerosis and coronary heart disease. The combinations described above may ameliorate more than one symptom of metabolic syndrome concurrently (e.g. two symptoms, three symptoms, four symptoms, or all five of the symptoms).
ASSAYS
Protocol: Scintillation Proximity Assay (SPA) for CETP Activity
First, low density lipoprotein (LDL) (Meridian) was biotinylated by incubating LDL with biotin for 1 hour on ice, after which it was dialyzed to remove free biotin. Then compounds at varying concentrations were incubated with 15 nM CETP (reagent production group, In Vitro Pharmacology, MRL Rahway) and 50 ug/ml of the biotinylated LDL in 50 mM HEPES, 150 mM NaCl, pH 7.4, for 1 hour at 37° C. The reaction was started by adding 3H-cholesterol ester high density lipoprotein (HDL) (American Radiochemicals Corp) at a concentration of  ̃0.6 nM. The reaction proceeded for 2 hours at 37° C., after which time it was quenched by the addition of 12% acetic acid. PVT streptavadin-coated scintillation proximity beads, which had been brought to room temperature, were then added at a concentration of 4 mg/ml. The assay was then mixed and counted after one half hour in a Microbeta plate reader.
In Vitro Radioactive Assays of CETP-Catalyzed CE and TG Transfer (RTA Assay)
Reagents and sources are: [3H] cholesteryl oleate (GE #TRK.886), [3H] Triolein (Perkin-Elmer NET-431), Butylated hydroxyl toluene (Aldrich, #D4740-4), DOPC (Sigma, #P6354), Sodium Bromide (Fisher scientific #S255-500), PEG 8000 (Fisher, #BP233-1), and human HDL (Intracel Corp #RP-036).
An in vitro assay for determining IC50's to identify compounds that inhibit CETP transfer activity is performed based on a modification of a published method (Morton and Zilversmit, (1981) A plasma inhibitor of triglyceride and cholesteryl ester transfer activities, J. Biol. Chem. 256(23), 11992-11995). The ability of inhibitors to alter CETP activity is performed using two different assays: one using recombinant CETP and one using an endogenous plasma source of CETP. Both assays measure the transfer of [3H] cholesteryl oleate or [3H] triolein from exogenous LDL to HDL.
Radiolabeled donor particles are generated by first combining 100 μl of 200 μM butylated hydroxyl toluene in CHCl3, 216 μL of 21.57 mM DOPC in EtOH, and either 500 μCi [3H]-triolein (Perkin Elmer #NET-431) or 500 μCi [3H]-cholesteryl oleate (GE #TRK886) in a glass tube. Reagents are mixed, dried under nitrogen, and then resuspended in 2 mL of 50 mM Tris, 27 μM EDTA at pH 7.4. After a brief vortex, the solution is sonicated until clear and mixed with 20 mL of fresh human serum. The mixture is incubated overnight at 37° C. The [3H] labeled LDL substrate is separated at 1.063 g/ml density by sequential ultracentrifugal flotation in NaBr according to the method of Havel, Eder, et al., 1955, and Chapman, Goldstein, et al., 1981. Once isolated the particles are dialyzed 3× in CETP buffer (50 mM Tris, pH 7.4, 100 mM NaCl, 1 mM EDTA). Human HDL is purchased from Intracel and used as the acceptor particles.
Transfer assays are performed in a 96-well v-bottom polypropylene plate. For the RTA using recombinant CETP (2% RTA), an assay cocktail is prepared with the final concentrations 128 μg/mL HDL, 20 nM rCETP, 2% human serum, and 1×CETP buffer. 1 μL of each test compound diluted in DMSO is added to 47 μL of assay cocktail per well and incubated at 37° C. for 1 hour. To initiate the transfer reaction, 2 μL radiolabeled LDL is added. After an additional 60 min of incubation at 37° C., the transfer action is terminated by precipitation of LDL with an equal volume of 20% W/V PEG 8000. The plates are centrifuged at 2000 rpm for 30 minutes at 4° C. A 40 μL aliquot of the HDL-containing supernatant is transferred to a Packard OptiplateTM with 200 μL of MicroScintTM 20. After mixing, plates are counted by liquid scintillation. Counts present in the supernatant for blanks (wells containing only HDL acceptor, CETP buffer and DMSO) are subtracted from those containing test compounds and used to correct for non-specific transfer.
For the transfer assay using endogenous CETP from serum (95% RTA), the same procedure is used except that human serum is added such that a final concentration of serum of 95% of the total assay volume is achieved, yielding a concentration of approximately 15 nM endogenous CETP in the assay. This is then combined with HDL and CETP buffer and the reaction proceeds as above and is terminated as described.
Comparison of the counts of samples with inhibitors to an uninhibited (DMSO only) positive control yield a percent inhibition. A plot of percent inhibition vs. log of inhibitor concentration, fit to a Sigmoidal 4 parameter equation is used to calculate IC50.
EXAMPLES
The following schemes and examples are provided so that the invention will be more fully appreciated and understood. These schemes and examples are illustrative and are not to be construed as limiting the invention in any way. The claims appended hereto define the scope of the invention.
Starting materials are commercially available or are made using known procedures or as shown below. The examples may be synthesized according to the general schemes provided below and through the synthetic intermediates that are described. The data reported for the examples below were generally obtained using the RTA assay in 95% human serum. The IC50's for the examples using this assay are in the range of about 55-6299 nM. Preferred compounds have an IC50 less than about 500 nM. More preferred compounds have an IC50 less than about 200 nM, and very preferred compounds have an IC50 less than about 100 nM. When compounds of Formula I or Formula II are mentioned herein, such compounds include compounds defined generically by Formula I or II and also the specific examples disclosed herein.




Intermediate A is prepared from a commercially available N-Cbz protected amino acid (Scheme A). Formation of the corresponding Weinreb amide and reaction with a known Grignard reagent provided the ketone adduct. Subsequent reduction and and treatment with sodium hydride results in the synthesis of intermediate A.




Intermediate A1 and A2
(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one and (7S)-7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one (scheme A)
Step 1: To a stirred solution of 1-{[(benzyloxy)carbonyl]amino}cyclopropanecarboxylic acid (143.0 g, 610 mmol), triethylamine (405 mL, 2.90 mol) and HATU (346.6 g, 910 mmol) in DMF (1.5 L) under nitrogen was added N,O-dimethylhydroxylamine hydrochloride (119 g, 1.22 mol). The reaction suspension was stirred at room temperature for 3 h. The reaction was diluted with ethyl acetate (6000 mL) and washed with H2O (3000 mL), brine, dried over Na2SO4, filtered and concentrated. The residue was purified by column chromatography on silica gel to give benzyl {1-[methoxy(methyl)carbamoyl]cyclopropyl}carbamate as slightly yellow solid. 1H NMR (400 MHz, CDCl3)□δ 7.34-7.35 (m, 5H), 5.53 (br, 1H), 5.12 (s, 2H), 3.65 (s, 3H), 3.15 (s, 3H), 1.46-1.49 (m, 2H), 1.03-1.15 (m, 2H).
Step 2: A room temperature solution of isopropylmagnesium chloride (2 M, 497 mL, 0.98 mol) was added over a 30 min period to a −10° C. solution of 3,5-bis(trifluoromethyl)bromobenzene (260 g, 0.89 mmol) in 400 mL of anhydrous THF, such that the reaction temperature never exceeded −5° C. The mixture was stirred at −10° C. for 1 h. To a stirred solution of benzyl {1-[methoxy(methyl)carbamoyl]cyclopropyl}carbamate (103 g, 370 mmol) in THF (500 mL) under nitrogen at −15° C. was added a solution of isopropylmagnesium chloride lithium chloride complex (284 mL, 370 mmol). After stirring at −15° C. under nitrogen for 15 min, 3,5-bis(trifluoromethyl)phenylmagnesium bromide ( ̃1000 mL, 0.89 mmol) was added. The reaction was allowed to warm up to room temperature overnight. The mixture was quenched with aqueous saturated NH4Cl (1000 mL) and 2 N HCl to pH  ̃5 and extracted with ethyl acetate (3×1000 mL). The solid was filtered and washed with petroleum ether (200 mL) to give benzyl (1-{[3,5-bis(trifluoromethyl)phenyl]carbonyl}cyclopropyl)carbamate as a slightly yellow solid. MS ESI calc'd. for C20H16F6NO3 [M+H]+ 432.1, found 432.3.
Step 3: To a stirred solution of benzyl (1-{[3,5-bis(trifluoromethyl)phenyl]carbonyl}cyclopropyl)carbamate (131.5 g, 300 mmol) in THF (2573 mL) at −70° C. under nitrogen was added a solution of lithium aluminum hydride (485 mL, 485 mmol). The resulting mixture was stirred at −70° C. for 1 h. The mixture was quenched with aqueous saturated Na2SO4.10H2O (100 mL), diluted with EtOAc (3000 mL), and evaporated under reduced pressure to give crude benzyl (1-{[3,5-bis(trifluoromethyl)phenyl](hydroxy)methyl}cyclopropyl)carbamate as a slightly yellow solid which is used in the next step without further purification. 1H NMR (400 MHz, CDCl3) δ 7.81-7.84 (m, 3H), 7.26-7.38 (m, 5H), 4.97-5.16 (m, 4H), 4.38-4.40 (m, 1H), 1.21-1.27 (m, 1H), 1.06-1.15 (m, 2H), 0.86-0.92 (m, 1H).
Step 4: To a stirred solution of benzyl (1-{[3,5-bis(trifluoromethyl)phenyl](hydroxy)methyl}cyclopropyl)carbamate (133 g, 0.307 mol) in THF (1500 mL) at 0° C. under nitrogen was added sodium hydride (60%, 27 g, 0.675 mol). The resulting suspension was stirred at room temperature for 16 h. The reaction was quenched with ice-cold 1 N HCl and concentrated. The residue was purified by column chromatography on silica gel to give 7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one as a colorless solid. 1H NMR (400 MHz, CDCl3)□ δ 7.92 (s, 1H), 7.77 (s, 2H), 6.11 (s, 1H), 5.62 (s, 1H), 1.13-1.20 (m, 1H), 0.81-0.88 (m, 2H), 0.27-0.34 (m, 1H).
Step 5: A solution of racemic 7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one (502 g, 1.54 mol) in isopropanol ( ̃100 mg/mL) was subject to purification by chiral SFC chromatography (Column: ChiralPak AS-10μ 50×300 mm, 38° C. Mobile Phase: 10% IPA/CO2. Flow rate: 200 mL/min. Wavelength: 210 nm) to give:
(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one (Rt 5.6 min) and
(7S)-7-[3,5-bis(trifluoromethyl)phenyl]-6-oxa-4-azaspiro[2.4]heptan-5-one (Rt=6.9 min).
1H NMR and MS spectra for both enantiomers match that of the racemic material.




Intermediate A3
8-[3,5-bis(trifluoromethyl)phenyl]-7-oxa-5-azaspiro[3.4]octan-6-one (scheme B)
Step 1: To a stirred solution of 1-{[(benzyloxy)carbonyl]amino}cyclobutanecarboxylic acid (2.0 g, 8.02 mmol), HATU (4.58 g, 12.04 mmol) and triethylamine (5.59 mL, 40.1 mmol) in DMF (35 mL) under nitrogen was added N,O-dimethylhydroxylamine hydrochloride (1.565 g, 16.05 mmol). The reaction suspension was stirred at room temperature for 2.5 h after which the reaction was diluted with ethyl acetate (200 mL) and water. The organic layer was washed with water (2×100 mL) and brine, and was dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to yield benzyl {1-[methoxy(methyl)carbamoyl]cyclobutyl}carbamate as slightly yellow solid. MS ESI calc′d. for C15H21N2O4 [M+H]+ 293.1, found 293.3.
Step 2: To a stirred solution of benzyl {1-[methoxy(methyl)carbamoyl]cyclobutyl}carbamate in THF (70 mL) under nitrogen at −20° C. was added a solution of 3,5-bis(trifluoromethyl)phenylmagnesium bromide (0.5 M in THF, 33.1 mL, 16.56 mmol) dropwise. The resulting mixture was stirred at for 1 h and then at room temperature for 3 h under nitrogen. The reaction was cooled to 0° C. and additional 3,5-bis(trifluoromethyl)phenylmagnesium bromide (15 mL, 7.5 mmol) was added. After stirring at 0° C. for 1 h under nitrogen, the reaction mixture was quenched with 1N HCl. The mixture was diluted with saturated aqueous NH4Cl and ethyl acetate. The organic was dried over sodium sulfate, filtered and then evaporated under reduced pressure. The residue was purified by column chromatography on silica gel to yield benzyl (1-{[3,5-bis(trifluoromethyl)phenyl]carbonyl}cyclobutyl)carbamate. Note: unreacted starting material was recovered (1.5 g). MS ESI calc'd. for C14H11F6NO3 [M-benzyl+H]+ 355.1, found 355.2.
Step 3: To a stirred solution of benzyl (1-{[3,5-bis(trifluoromethyl)phenyl]carbonyl}cyclobutyl)carbamate (570 mg, 1.280 mmol) in THF (11 mL) under nitrogen at −70° C. was added a solution of LiAlH4 (0.5 M THF, 1.408 mL, 1.408 mmol) dropwise. The resulting mixture was stirred at −70° C. for 1 h. The reaction was quenched with saturated aqueous sodium sulfate and was diluted with EtOAc. The mixture was stirred for 30 minutes at room temperature and filtered before the organic was separated and concentrated. The resultant residue was purified by column chromatography on silica gel to give benzyl (1-{[3,5-bis(trifluoromethyl)phenyl](hydroxy)methyl}cyclobutyl)carbamate as a white crystalline solid. MS ESI calc'd. for C21H20F6NO3 [M+H]+ 448.4, found 448.3.
Step 4: To a stirred solution of benzyl (1-{[3,5-bis(trifluoromethyl)phenyl](hydroxy)methyl}cyclobutyl)carbamate (480 mg, 1.073 mmol) in THF (10 mL) at 0° C. under nitrogen was added sodium hydride (94 mg, 2.360 mmol). The resulting suspension was stirred at room temperature for 3 h. The reaction was cooled to 0° C. and was quenched with saturated aqueous NH4Cl. The reaction was partitioned with ethyl acetate and the combined organic extracts were dried over sodium sulfate before being filtered and concentrating to dryness. The residue was purified by column chromatography on silica gel to yield 8-[3,5-bis(trifluoromethyl)phenyl]-7-oxa-5-azaspiro[3.4]octan-6-one as a colorless solid. MS ESI calc'd. for C14H12F6NO2 [M+H]+ 340.1, found 340.3. 1H NMR (500 MHz, CDCl3) δ 1.30-2.62 (m, 6H), 5.56 (s, 1H), 6.32 (br, 1H), 7.88 (s, 2H), 7.98 (s, 1H).







Intermediate B is prepared in a three part procedure, according to scheme B. In part A, the condensation product of commercially available starting materials provides an appropriately functionalized pyrimidine. A two-step procedure reduces the pendant carboxylic acid to the benzyl alcohol. In part B, a commercially available carboxylic acid is transformed to the corresponding methyl ester before coupling with a boronic acid to yield a biaryl product. Iodination and metal-halogen exchange allows the installation of the boronic acid functionality. In part C, a palladium-catalyzed coupling of the products from part A and B yields intermediate B.




Intermediate B
methyl 3′-[4-(hydroxymethyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (scheme B)
Part A, Step 1: Into a 50-L 4-necked round-bottom flask was placed a solution of (2Z)-2,3-dibromo-4-oxobut-2-enoic acid (5000 g, 18.42 mol) in water (30 L) and bis((methylsulfanyl)methanimidamide) sulfate (5429 g, 19.50 mol). This was followed by the addition of triethylamine (5917 g, 58.47 mol) dropwise with stirring at <10° C. over 120 min. The resulting solution was stirred for 3 h at room temperature. After being left undisturbed for 72 hours, the pH value of the solution was adjusted to 2 with HCl (3 M). The solid was collected by filtration, washed with water and dried in air. This resulted in crude 5-bromo-2-(methylsulfanyl)pyrimidine-4-carboxylic acid as a yellow solid.
Part A, Step 2: Into a 10-L 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed a solution of 5-bromo-2-(methylsulfanyl)pyrimidine-4-carboxylic acid (500 g, 2.01 mol) in dichloromethane (4000 mL) and N,N-dimethylformamide (14.6 g, 199.75 mmol). This was followed by the addition of oxalic dichloride (508 g, 4.00 mol) dropwise with stirring at 0-5° C. over 120 min. The resulting solution was stirred overnight at room temperature. Removal of all volatiles under reduced pressure resulted in crude 5-bromo-2-(methylsulfanyl)pyrimidine-4-carbonyl chloride as a black solid.
Part A, Step 3: Into a 10000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed a solution of LiBH4 (53.6 g, 1.20 equiv) in tetrahydrofuran (1000 mL). This was followed by the addition of a solution of 5-bromo-2-(methylsulfanyl)pyrimidine-4-carbonyl chloride (544 g, 2.03 mol) in tetrahydrofuran/MeCN (2500/1500 mL) dropwise with stirring at <−70° C. over 3 hr. The resulting solution was stirred for 2 h at −70° C., then quenched by the addition of 2000 mL of HCl (1.0 N). The pH value of the solution was adjusted to 12 with sodium carbonate (aq. 2.0 N). The resulting solution was extracted with ethyl acetate (2×2000 mL). The organic layers were combined, washed with brine (2000 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue purified by silica gel chromatography to result in 5-bromo-2-(methylsulfanyl)pyrimidin-4-yl]methanol as a yellow solid.
Part B, Step 1: Into a 10000-mL 4-necked round-bottom flask was placed a solution of 4-bromo-3-methylbenzoic acid (500 g, 2.33 mol) in methanol (5000 mL). This was followed by the addition of thionyl chloride (556 g, 4.67 mol) dropwise with stirring at <10° C. over 120 min. The resulting solution was heated to reflux for 5 h in an oil bath. The resulting mixture was cooled and concentrated under vacuum.
Part B, Step 2: Into a 10000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed a solution of (4-methoxyphenyl)boronic acid (350 g, 2.30 mol) in 1,4-dioxane (3500 mL), methyl 4-bromo-3-methylbenzoate (527 g, 2.30 mol), Pd(PPh3)4 (79.8 g, 69.06 mmol), cesium carbonate (1501 g, 4.60 mol), and water (850 mL). The resulting solution was stirred overnight at 100° C. in an oil bath. The reaction was then cooled and quenched by the addition of water (3000 mL). The resulting solution was extracted with ethyl acetate (2×2000 mL). The organic layers were combined, washed with brine (2000 mL), dried, and concentrated under vacuum. The residue was purified by silica gel chromatography to result in methyl 4-(4-methoxyphenyl)-3-methylbenzoate as a red solid.
Part B, Step 3: Into a 10000-mL 4-necked round-bottom flask was placed methyl 4-(4-methoxyphenyl)-3-methylbenzoate (470 g, 1.83 mol), Ag2SO4 (572.8 g, 1.84 mol), iodine (466.3 g, 1.84 mol), methanol (4000 mL) and ethyl acetate (1000 mL). The resulting solution was stirred for 3 h at room temperature. The resulting mixture was concentrated under vacuum. The residue was diluted with ethyl acetate/brine (3000/1500 mL). The solid was filtered out and the filtrate was extracted with ethyl acetate (2×1000 mL). The organic layers were combined, dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in methyl 4-(3-iodo-4-methoxyphenyl)-3-methylbenzoate as a yellow solid.
Part B, Step 4: Into a 10000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed a solution of methyl 4-(3-iodo-4-methoxyphenyl)-3-methylbenzoate (600 g, 1.57 mol) in tetrahydrofuran (5000 mL). This was followed by the addition of isopropyl magnesium chloride (960 mL, 1.20 equiv) dropwise with stirring at <−25° C. over 60 min. The reaction was maintained for 1 h at −15° C., followed by addition of trimethyl borate (329.2 g, 3.17 mol) dropwise with stirring at <−20° C. over 30 min. The resulting solution was stirred for 60 min at <0° C., then quenched by the addition of 5000 mL of H3PO4 (aq. 1.0 M). The bulk of THF was removed under reduced pressure and the resultant solid was collected by filtration and washed with toluene (2×200 mL). This resulted in [2-methoxy-5-[4-(methoxycarbonyl)-2-methylphenyl]phenyl]boronic acid as a yellow solid. Part C: Into a 3000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed [5-bromo-2-(methylsulfanyl)pyrimidin-4-yl]methanol (130 g, 560 mmol), [2-methoxy-5-[4-(methoxycarbonyl)-2-methylphenyl]phenyl]boronic acid (87.5 g), Pd(PPh3)4 (32.3, 28 mmol), cesium carbonate (365.1 g), water (300 mL), 1,4-dioxane (1200 mL). The resulting solution was stirred for 5 hour at 100° C., then a second batch of [2-methoxy-5-[4-(methoxycarbonyl)-2-methylphenyl]phenyl]boronic acid (50 g) was added to the reaction. A third batch of [2-methoxy-5-[4-(methoxycarbonyl)-2-methylphenyl]phenyl]boronic acid (37 g) was added after 2 hours. The resulting solution was stirred for 3 h at 100° C. The reaction mixture was cooled to room temperature and diluted with water (1000 mL). The resulting solution was extracted with ethyl acetate (2×1000 mL). The organic layers were combined, washed with brine (1000 mL), dried over anhydrous sodium sulfate and concentrated under vacuum. The residue was purified by silica gel chromatography to yield methyl 4-[3-[4-(hydroxymethyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4-methoxyphenyl]-3-methylbenzoate as a yellow solid. MS ESI calc'd. for C22H23N2O4S [M+H]+ 411, found 411. 1H NMR (300 MHz, CDCl3) δ 8.398 (1H, s), 7.976 (1H, s), 7.916 (2H, d), 7.410 (1H, d), 7.284-7.325 (1H, m), 7.066-7.114 (2H, m), 4.583 (2H, s), 3.954 (3H, s), 3.853 (3H, s), 2.663 (3H, s), 2.367 (3H, s).




Intermediate C is prepared from a commercially available carboxylic acid
(Scheme C). Formation of the methyl ester and subsequent treatment with sodium iodide and TMSCl provides the corresponding 2-pyridyl iodide. A copper-mediated trifluoromethylation reaction completes the carbon framework. Reduction of the ester to the benzylic alcohol provides intermediate C.




Intermediate C
[3-chloro-6-(trifluoromethyl)pyridin-2-yl]methanol (scheme C)
Step 1: Into a 20-L 4-necked round-bottom flask was placed 3,6-dichloropyridine-2-carboxylic acid (1000 g, 5.21 mol) and methanol (10000 mL). This was followed by the dropwise addition of sulfuric acid (100 mL) with stirring. The resulting solution was stirred for 4 h at 80° C. The reaction mixture was cooled with a water/ice bath to room temperature and was then concentrated under reduced pressure. The residue was quenched by the addition of water (3000 mL). The resulting solution was extracted with ethyl acetate (2×4000 mL) and the organic layers were combined, dried over anhydrous sodium sulfate and concentrated under vacuum. This resulted in methyl 3,6-dichloropyridine-2-carboxylate as a white solid.
Step 2: Into a 10-L 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed acetonitrile (6000 mL), chlorotrimethylsilane (227.5 g, 2.09 mol), sodium iodide (790 g, 5.26 mol) and methyl 3,6-dichloropyridine-2-carboxylate (360 g, 1.75 mol). After stirring for 2 h at 80° C., the reaction mixture was poured into water. The resulting solution was extracted with ethyl acetate and the organic layers were combined, dried over anhydrous sodium sulfate and concentrated under reduced pressure. This resulted in methyl 3-chloro-6-iodopyridine-2-carboxylate as a white solid.
Step 3: Into a 3000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed NMP (1800 mL), potassium fluoride (46.9 g, 807.27 mmol), copper (I) iodide (164 g, 861.12 mmol) and trimethyl(trifluoromethyl)silane (102 mL, 1.20 equiv). To this was added methyl 3-chloro-6-iodopyridine-2-carboxylate (160 g, 537.85 mmol) at 50° C. The resulting solution was stirred for 16 h at 50° C. The reaction mixture was quenched by the addition of water/ice. The resulting solution was extracted with ethyl acetate (2×1000 mL) and the organic layers were combined, dried over anhydrous sodium sulfate and concentrated in vacuo. This resulted in methyl 3-chloro-6-(trifluoromethyl)pyridine-2-carboxylate as a yellow solid.
Step 4: Into a 10000-mL 4-necked round-bottom flask, purged and maintained with an inert atmosphere of nitrogen, was placed methyl 3-chloro-6-(trifluoromethyl)pyridine-2-carboxylate (360 g, 1.50 mol) and dichloromethane (4000 mL). This was followed by the dropwise addition of DIBAL (25% in toluene, 1740 g, 12.23 mol) with stirring at −30° C. The resulting solution was stirred for 1 h at −30° C. and was then quenched by the addition of water (120 mL) and NaOH (15% in water, 120 mL). The organic was separated, dried over anhydrous sodium sulfate, and concentrated under vacuum. The residue was purified by silica gel chromatography to yield [3-chloro-6-(trifluoromethyl)pyridin-2-yl]methanol as a white solid. MS ESI calc'd. for C7H6ClF3NO [M+H]+ 212, found 212. 1H NMR (400 MHz, CDCl3) δ 3.884 (1H, d), 4.845 (2H, s), 7.575-7.602 (1H, d, J=8.1 Hz), 7.836-7.863 (1H, d, J=8.1 Hz).




Intermediates of the type in scheme D are synthesized from mesylate formed from a prepared or known benzylic alcohol (scheme D). Displacement of the mesylate by the appropriate oxazolidinone results in intermediate D. In Scheme D, the letter “A” can represent either CH or N in the structures.




Intermediates D1 and D2
methyl 3′-[4-({(8S)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate and methyl 3′-[4-({(8R)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (scheme D)
Step 1: To a stirred, cooled mixture of methyl 3′-[4-(hydroxymethyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (3.0 g, 7.31 mmol) and triethylamine (2.04 mL, 14.62 mmol) in dichloromethane (30 mL) under nitrogen at 0° C. was added MsCl (0.854 mL, 10.96 mmol). The mixture was stirred at 0° C. for 3 h before the reaction was diluted with ethyl acetate and water. The organic layer was washed with water and brine, was dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to yield methyl 4′-methoxy-2-methyl-3′-[2-(methylsulfanyl)-4-{[(methylsulfonyl)oxy]methyl}pyrimidin-5-yl]biphenyl-4-carboxylate as slightly yellow solid. (M+H). MS ESI calc'd. for C23H25N2O6S2 [M+H]+ 489.1, found 489.2.
Step 2: To a stirred solution of 8-[3,5-bis(trifluoromethyl)phenyl]-7-oxa-5-azaspiro[3.4]octan-6-one (290 mg, 0.855 mmol) in THF (6 mL) at 0° C. was added sodium hydride (60%, 41.0 mg, 1.026 mmol). After stirring for 15 min, a solution of 4′-methoxy-2-methyl-3′-[2-(methylsulfanyl)-4-{[(methylsulfonyl)oxy]methyl}pyrimidin-5-yl]biphenyl-4-carboxylate (501 mg, 1.026 mmol) in THF (5 mL) was added. The reaction suspension was stirred at room temperature overnight and was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate and water (70 mL) and the organic layer was washed with water (70 mL), brine, dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography to yield methyl 3′-[4-{8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate as solid. MS ESI calc'd. for C36H32F6N3O5S [M+H]+ 732.2, found 732.5. 1H NMR (500 MHz, CDCl3) δ 1.12-1.56 (m, 4H), 2.08-2.24 (m, 2H), 2.38 (s, 3H), 2.60 (s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 4.38-4.70 (m, 2H), 5.42-5.65 (s, 1H), 7.12-7.46 (m, 4H), 7.88-7.98 (m, 5H), 8.38 (s, 1H).
Step 3: The racemic material was purified by chiral SFC (Column: Chiral Technology 1A-H 30×250 mm, 35° C. Mobile Phase: 20% MeOH/CO2. Flow rate: 70 mL/min. Wavelength: 220 nm) to provide:

    Methyl 3′-[4-({(8S)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (intermediate D1, Rt=5.9 min, 246 mg, 0.336 mmol) and
    Methyl 3′-[4-({(8R)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (intermediate D2, Rt=8.6 min, 239 mg, 0.327 mmol). RTA (95% HS): 913 nM


1H NMR and MS spectra for both enantiomers match that of the racemic material.
The following intermediates in table 1 were prepared according to scheme D using the procedure outlined in the synthesis of intermediate D1 and D2 utilizing commercially available or prepared benzylic alcohols. As an alternative reagent, Na or LiHMDS can be utilized in step 2. Step 3 can be omitted if the oxazolidinone is chiral or racemic material is desired.










TABLE 1








Exact Mass [M + H]+


Int
Structure
IUPAC Name
or 1H NMR







D3




7-[3,5- bis(trifluoromethyl)phenyl]- 4-({5-[4-fluoro-2-methoxy- 5-(propan-2-yl)phenyl]-2- (methylsulfanyl)pyrimidin-4- yl}methyl)-6-oxa-4- azaspiro[2.4]heptan-5-one
Calc'd 630.2, found 630.3


 


D4




8-[3,5- bis(trifluoromethyl)phenyl]- 5-({5-[4-fluoro-2-methoxy- 5-(propan-2-yl)phenyl]-2- (methylsulfanyl)pyrimidin-4- yl}methyl)-7-oxa-5- azaspiro[3.4]octan-6-one
Calc'd 644.2, found 644.4


 


D5




methyl 3′-[4-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-2- (methylsulfanyl)pyrimidin-5- yl]-4′-methoxy-2- methylbiphenyl-4- carboxylate
Calc'd 718.2, found 718.4


 


D6




(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 4-{[3-chloro-6- (trifluoromethyl)pyridin-2- yl]methyl}-6-oxa-4- azaspiro[2.4]heptan-5-one
Calc'd 519.0, found 519.3













Intermediate E was prepared via iodination and subsequent Miyaura borylation from commercially available starting materials.




Intermediate E
methyl 3-[4-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propanoate
Step 1: A 3-neck 5 L RBF equipped with mechanical stirrer, thermometer, and a nitrogen bubbler, was charged with 3-(4-methoxyphenyl)propionic acid methyl ester (100 g, 515 mmol), silver sulfate (161 g, 515 mmol) and iodine (131 g, 515 mmol) in methanol (2 L). The reaction mixture was stirred vigorously at room temperature for 1 hour. The reaction was filtered through Solka Floc (ethyl acetate wash). The filtrate was concentrated and the residue was taken up in ethyl acetate (4 L). The organic was washed with water, saturated aq. NaHSO3 (50 mL), and brine (50 mL) before drying over Na2SO4, filtering, and concentrating to dryness. The crude reaction was purified by column chromatography to yield methyl 3-(3-iodo-4-methoxyphenyl)propanoate as a clear oil. MS ESI calc'd. for C11H14IO3 [M+H]+ 321.0, found 321.0.
Step 2: A 3-neck 12 L RBF equipped with mechanical stirrer, thermometer, nitrogen bubbler, condenser and addition funnel, was charged with methyl 3-(3-iodo-4-methoxyphenyl)propanoate (155 g, 484 mmol), bis(pinacolato)diboron (154 g, 605 mmol), and potassium acetate (95 g, 48.4 mmol) in DMSO (3 L) and dioxane (0.9 L). The system was degassed three times with nitrogen gas before the addition of dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloromethane adduct (39.5 g, 48.4 mmol). The system was degassed three times and was then heated to 50° C. for 1 hour. The temperature was raised to 80° C. and the reaction was stirred overnight. The reaction was diluted with ethyl acetate (4 L) and was partitioned with water and then with brine. The organic was dried over MgSO4, filtered, and concentrated in vacuo. The crude reaction was purified by column chromatography to yield methyl 3-[4-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]propanoate as a tan solid. MS ESI calc'd. for C17H26BO5 [M+H]+ 321.2, found 321.2.




Preparation of intermediate F begins with formation of a tert-butyl ester which is then subjected to a Miyuara coupling to obtain the corresponding boronic ester. Suzuki coupling with a commercially available 5-bromo-3-chloro-2-methoxypyridine yields the coupled chloride. A second Miyaura coupling provides the desired boronic ester intermediate F.




Intermediate F
tert-butyl 4-[6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl]-3-methylbenzoate
Step 1: To a 250 mL RBF was added 4-bromo-3-methylbenzoic acid (10 g, 46.5 mmol), DMAP (8.52 g, 69.8 mmol) and tert-butyl alcohol (100 mL). Di-tert-butyl dicarbonate (12.96 mL, 55.8 mmol) was added via a syringe to the solution, which caused vigorous bubbling, foaming and the loss of some material. The remaining reaction mixture was heated at 70° C. overnight. The reaction was cooled to room temperature and the volatiles were removed under reduced pressure. Crude material was diluted with ethyl acetate:hexanes (1:4, 200 mL) and was washed sequentially with 5% aqueous KOH (200 mL) and saturated aqueous ammonium chloride (2×100 mL). The organics were dried over sodium sulfate, filtered and concentrated before purification by column chromatography. tert-Butyl 4-bromo-3-methylbenzoate was isolated as a colorless oil. 1H NMR (500 MHz, CDCl3) δ 7.87 (s, 1H), 7.67 (d, J=8.3 Hz, 1H), 7.60 (d, J=8.2 Hz, 1H), 2.47 (s, 3H), 1.62 (s, 9H).
Step 2: To a 250 mL RBF was loaded 1,1′-bis(di-tert-butylphosphino)ferrocene palladium dichloride (0.317 g, 0.487 mmol), tert-butyl 4-bromo-3-methylbenzoate (6.6 g, 24.34 mmol), bis(pinacolato)diboron (7.42 g, 29.2 mmol), potassium acetate (5.97 g, 60.9 mmol) and dioxane (25 mL). The system was flushed with nitrogen and was heated at 125° C. overnight. The reaction was cooled to room temperature, diluted with ethyl acetate:hexanes (1:9, 120 mL), washed sequentially with water (150 mL) and then brine (50 mL). The organics were dried over sodium sulfate, filtered and concentrated before purification by column chromatography. tert-Butyl 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate was isolated as a crystalline solid. Note 1H NMR indicated it is about 70% pure. 1H NMR (500 MHz, CDCl3) δ 7.8 (m, 3H), 2.60 (s, 3H), 1.58 (s, 9H), 1.39 (s, 12H).
Step 3: To a 250 mL RBF was added 5-bromo-3-chloro-2-methoxypyridine (1.5 g, 6.74 mmol), tribasic potassium phosphate (2.86 g, 13.5 mmol), [1,1′-bis(diphenylphosphino)ferrocene]palladium(II) dichloromethane adduct (0.275 g, 0.337 mmol), tert-butyl 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (2.27 g, 7.13 mmol), dioxane (50 mL) and water (3 mL). The flask was sealed and was stirred at 80° C. overnight. The reaction was cooled to room temperature, diluted with ethyl acetate, washed with water, filtered and concentrated. The resultant residue was purified by column chromatography to yield tert-butyl 4-(5-chloro-6-methoxypyridin-3-yl)-3-methylbenzoate. MS ESI calc'd. for C18H21ClNO3 [M+H]+ 334.1, found 334.0.
Step 4: To a 250 mL RBF was added yield tert-butyl 4-(5-chloro-6-methoxypyridin-3-yl)-3-methylbenzoate (4.5 g, 13.5 mmol), bis(pinacolato)diboron (6.85 g, 27.0 mmol), potassium acetate (3.97 g, 40.4 mmol), and chloro(2-dicyclohexylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl)[2-(2′-amino-1,1′-biphenyl)]palladium(II) (0.212 g, 0.27 mmol) followed by anhydrous dioxane (50 mL). The system was evacuated and backfilled with nitrogen (3×s) and was heated to 120° C. for 2 hours. The mixture was cooled, filtered over celite (ethyl acetate wash) and was concentrated. The residue was purified by column chromatography to afford tert-butyl 4-[6-methoxy-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-3-yl]-3-methylbenzoate as a solid. MS ESI calc'd. for C24H33BNO5 [M+H]+ 426.2, found 426.0.
General Synthetic Schemes
Representative compounds of the present invention can be synthesized according to the general schemes outlined below as well as the representative examples that follow. Since the schemes are illustrative, the invention should not be construed as being limited by the chemical reactions and conditions expressed. The preparation of the various starting materials used in the schemes is well within the skill of a person versed in the art.




In accordance with Scheme 1, a cross-coupling reaction between intermediate D and an appropriately functionalized boronic acid/ester provides compounds of the general formula (I). In cases where an ester is present in the final compound, a saponification or hydrolysis may be additionally carried out. In Scheme 1, the letter “A” can represent either CH or N in the structures.




In accordance with Scheme 2, a compound with a pendant methyl ester is transformed to a primary alcohol via reaction with a metal hydride such as LAH or L-selectride to provide a compound of the general formula (I).




In accordance with Scheme 3, intermediate D is transformed to a sulfone and a subsequent displacement by an amine provides a compound of the general formula (I). If an ester moiety is present in the molecule, a saponification with base may be further carried out.




In accordance with Scheme 4, a compound of the general formula (I) is synthesized from mesylate formation from a prepared or known benzylic alcohol and subsequent displacement of the mesylate by an oxazolidinone.
EXAMPLES
The following schemes and examples are provided so that the invention will be more fully appreciated and understood. Starting materials are made using known procedures or as shown below.
Example 1




trans-3-{3-[2-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-6-(trifluoromethyl)pyridin-3-yl]-4-methoxyphenyl}cyclobutanecarboxylic acid (Scheme 1)
Step 1: To a solution of (7R)-7-[3,5-bis(trifluoromethyl)phenyl]-4-{[3-chloro-6-(trifluoromethyl)pyridin-2-yl]methyl}-6-oxa-4-azaspiro[2.4]heptan-5-one (100 mg, 0.193 mmol) in THF (2.0 mL) was added methyl trans-3-[4-methoxy-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]cyclobutanecarboxylate (100 mg, 0.289 mmol), 1,1′-bis(di-tert-butylphosphino)ferrocene palladium dichloride (12.6 mg, 0.019 mmol), and aqueous potassium carbonate (0.289 mL, 2.0 M). The mixture was degassed, flushed with nitrogen and heated to 90° C. 2 h. The reaction was diluted with ethyl acetate, washed with water, brine, dried over sodium sulfate, filtered and concentrated. The crude product was purified by column chromatography to afford methyl trans-3-{3-[2-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-6-(trifluoromethyl)pyridin-3-yl]-4-methoxyphenyl}cyclobutanecarboxylate. MS ESI calc'd. for C33H28F9N2O5 [M+H]+ 703.2, found 703.2.
Step 2: To a stirred solution of methyl trans-3-{3-[2-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-6-(trifluoromethyl)pyridin-3-yl]-4-methoxyphenyl}cyclobutanecarboxylate (60 mg, 0.085 mmol) in dioxane (1.7 mL) was added an aqueous solution of lithium hydroxide (0.85 mL, 0.5 M). The mixture was heated at 60° C. until complete by LCMS. The reaction mixture was cooled and acidified by the addition of TFA (0.066 mL, 0.85 mmol). Volatiles were removed under reduced pressure and the residue was purified by preparative HPLC reverse phase (C-18) to give trans-3-{3-[2-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-6-(trifluoromethyl)pyridin-3-yl]-4-methoxyphenyl}cyclobutanecarboxylic acid as a white solid following lyophilization. MS ESI calc'd. for C32H26F9N2O5 [M+H]+ 689.2, found 689.2. 1H NMR (500 MHz, CD3OD) δ 8.03 (s, 1H), 8.00 (br s, 2H), 7.78 (m, 2H), 7.40 (m, 1H), 7.13 (br s, 1H), 7.11 (d, 1H, J=8.7 Hz), 5.81 (m, 1H), 4.38 (m, 1H), 4.16 (m, 1H), 3.80 (s, 3H), 3.74 (m, 1H), 3.12 (m, 1H), 2.65 (m, 2H), 2.45 (m, 2H), 1.03 (m, 1H), 0.69 (m, 2H), 0.06 (m, 1H). RTA (95% HS): 962 nM
The following compounds in table 2 were prepared according to general scheme 1 using the procedure outlined for example 1 utilizing intermediate D and known boronic acids or esters in step 1.











TABLE 2









Exact





IC50
Mass


Ex
Structure
IUPAC Name
(nM)
[M + H]+



















2




3′-[2-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-6- (trifluoromethyl)pyridine-3- yl]-4′-methoxy-2- methylbiphenyl-4-carboxylic acid
212
Calc'd 725.2, found 725.5


 


3




5′-[2-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-6- (trifluoromethyl)pyridin-3- yl]-2′-fluoro-4′-methoxy-2- methylbiphenyl-4-carboxylic acid
119
Calc'd 743.2, found 743.5


 


4




4-[2′-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-2-methoxy-6′- (trifluoromethyl)-3,3′- bipyridin-5-yl]-3- methylbenzoic acid
173
Calc'd 726.2, found 726.5


 


5




3-{3-[2-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-6- (trifluoromethyl)pyridin-3- yl]-4- methoxyphenyl}propanoic acid
3357
Calc'd 663.2, found 663.5









Example 6




(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-4-{[3-{5-[trans-3-(hydroxymethyl)cyclobutyl]-2-methoxyphenyl}-6-(trifluoromethyl)pyridin-2-yl]methyl}-6-oxa-4-azaspiro[2.4]heptan-5-one (Scheme 2)
To a cooled, stirred solution of methyl trans-3-{3-[2-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-6-(trifluoromethyl)pyridin-3-yl]-4-methoxyphenyl}cyclobutanecarboxylate (58 mg, 0.083 mmol) in THF (1.5 mL) was added a solution of L-Selectride (0.206 mL, 1.0 M in THF) dropwise via syringe at 0° C. The reaction mixture was stirred at 0° C. for 30 min then quenched with TFA (0.050 mL, 0.649 mmol). The mixture was poured into saturated sodium bicarbonate and extracted with ethyl acetate. Combined extracts were washed with brine, dried over sodium sulfate, filtered and concentrated. The resultant oil was purified by preparative HPLC reverse phase (C-18) to give (7R)-7-[3,5-bis(trifluoromethyl)phenyl]-4-{[3-{5-[trans-3-(hydroxymethyl)cyclobutyl]-2-methoxyphenyl}-6-(trifluoromethyl)pyridin-2-yl]methyl}-6-oxa-4-azaspiro[2.4]heptan-5-one as a white solid following lyophilization. MS ESI calc'd. for C32H28F9N2O4 [M+H]+ 675.2, found 675.3. 1H NMR (500 MHz, CD3OD) δ 8.03 (s, 1H), 7.99 (br s, 2H), 7.77 (m, 2H), 7.40 (m, 1H), 7.13 (br s, 1H), 7.09 (d, 1H, J=8.6 Hz), 5.81 (m, 1H), 4.34 (m, 1H), 4.15 (m, 1H), 3.80 (s, 3H), 3.71 (d, 2H, J=7.2 Hz), 3.60 (m, 1H), 2.46 (m, 1H), 2.25 (m, 4H), 1.03 (m, 1H), 0.69 (m, 2H), 0.06 (m, 1H). RTA (95% HS): 1850 nM.
The following compounds in table 3 were prepared according to general scheme 2 using the procedure outlined for example 6. An alternative reducing agent may be lithium aluminium hydride.











TABLE 3









Exact





IC50
Mass


ex
Structure
IUPAC Name
(nM)
[M + H]+







7




(8R)-8-[3,5- bis(trifluoromethyl)phenyl]-5- ({2-(3-fluoroazetidin-1-yl)-5- [4′-(hydroxymethyl)-4- methoxy-2′-methylbiphenyl-3- yl]pyrimidin-4-yl}methyl)-7- oxa-5-azaspiro[3.4]octan-6-one
68
Calc'd 731.2, found 731.5









Examples 8 and 9




3′-[4-({(7S)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid and 3′-[4-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid (Scheme 3)
Step 1: To a stirred solution of methyl 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (335 mg, 0.467 mmol) in THF (6 mL) under nitrogen was added a solution of OXONE (717 mg, 1.167 mmol) in water (0.8 mL). The resulting reaction was heated at 50° C. for 20 h. The mixture was diluted with ethyl acetate (50 mL), washed with water (3×50 mL), dried (sodium sulfate), filtered and evaporated under reduced pressure to yield crude methyl 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(methylsulfonyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate. MS ESI calc'd. for C35H30F6N3O7S [M+H]+ 750.2, found 750.3.
Step 2: To a stirred solution of methyl 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(methylsulfonyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (340 mg, 0.454 mmol) in THF (10 mL) under nitrogen was added a solution of dimethylamine (2.27 mL, 4.54 mmol). The resulting solution was stirred at room temperature overnight. Volatiles were removed under reduced pressure and the residue was purified by column chromatography on silica gel to give methyl 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate as slightly yellow solid. MS ESI calc'd. for C36H33F6N4O5 [M+H]+ 715.2, found 715.3.
Step 3: To a stirred solution of methyl 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (300 mg, 0.420 mmol) in dioxane (10 mL) under nitrogen was added a solution of lithium hydroxide (1M in water, 2.52 mL, 2.52 mmol). The resulting mixture was stirred at room temperature overnight. Following acidification with TFA, the reaction was diluted with ethyl acetate (100 mL) and water (50 mL). The organic layer was washed with brine, dried over sodium sulfate, filtered and concentrated. The residue was purified by preparative HPLC Reverse phase (C-18), eluting to give 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid as a colorless solid. 1H NMR (500 MHz, CDCl3) δ 0.03-1.01 (m, 4H), 2.36 (s, 3H), 3.26 (s, 6H), 3.85 (s, 3H), 4.03-4.20 (m, 2H), 5.60 (s, 1H), 7.02-7.03 (d, J=8.5 Hz, 1H), 7.13-7.14 (d, J=2.0 Hz, 1H), 7.27-7.37 (m, 1H), 7.38 (d, J=2.1 Hz, 1H), 7.77 (s, 2H), 7.89-7.97 (m, 3H), 8.15 (s, 1H) ppm. MS ESI calc'd. for C35H31F6N4O5 [M+H]+ 701.2, found 701.3.
Step 4: Racemic 3′-[4-({7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid was purified by chiral SFC (Column: Chiral Technology AD-H 30×250 mm, 35° C. Mobile Phase: 25% IPA/CO2 with (Et)2NH as a modifier. Flow rate: 70 mL/min. Wavelength: 220 nm) to provide:
3′-[4-({(7S)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid (example 8) SPA: 6299 nM; and
3′-[4-({(7R)-7-[3,5-bis(trifluoromethyl)phenyl]-5-oxo-6-oxa-4-azaspiro[2.4]hept-4-yl}methyl)-2-(dimethylamino)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylic acid (example 9). RTA (95% HS): 55.5 nM.
1H NMR and MS spectra for both enantiomers match that of the racemic material.
The following compounds in table 4 were prepared according to general scheme 3 using the procedure outlined for examples 8 or 9 utilizing intermediate D in step 1 and commercially available amines in step 2. Step 3 and/or 4 may be omitted if no ester moiety is present in the molecule or the material is not racemic.











TABLE 4









Exact





IC50
Mass


ex
Structure
IUPAC Name
(nM)
[M + H]+



















10




7-[3,5- bis(trifluoromethyl)phenyl]- 4-({2-(dimethylamino)-5-[4- fluoro-2-methoxy-5-(1- methylethyl)phenyl] pyrimidin-4-yl} methyl)-6-oxa-4- azaspiro[2.4]heptan-5-one
345
Calc'd 627.2, found 627.3


 


11




8-[3,5- bis(trifluoromethyl)phenyl]- 5-({2-(dimethylamino)-5[4- fluoro-2-methoxy-5-(1- methylethyl)phenyl] pyrimidin-4-yl} methyl)-7-oxa-5- azaspiro[3.4]octan-6-one
318
Calc'd 641.2, found 641.4


 


12




3′-[4-({(8R)-8-[3,5- bis(trifluoromethyl)phenyl]- 6-oxo-7-oxa-5- azaspiro[3.4]oct-5- yl}methyl)-2-(3- fluoroazetidin-1- yl)pyrimidin-5-yl]-4′- methoxy-2-methylbiphenyl- 4-carboxylic acid
73
Calc'd 745.2, found 745.5


 


13




3′-[4-({(8R)-8-[3,5- bis(trifluoromethyl)phenyl]- 6-oxo-7-oxa-5- azaspiro[3.4]oct-5- yl}methyl)-2-morpholin-4- ylpyrimidin-5-yl]-4′- methoxy-2-methylbiphenyl- 4-carboxylic acid
242
Calc'd 757.2, found 757.5


 


14




3′-[4-({(8R)-8-[3,5- bis(trifluoromethyl)phenyl]- 6-oxo-7-oxa-5- azaspiro[3.4]oct-5- yl}methyl)-2-[methyl (methylsulfonyl)amino] pyrimidin-5-yl}-4′- methoxy-2-methylbiphenyl- 4-carboxylic acid
561
Calc'd 779.2, found 779.5


 


15




3′-[4-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-2-(3- fluoroazetidin-1- yl)pyrimidin-5-yl]-4′- methoxy-2-methylbiphenyl- 4-carboxylic acid
104
Calc'd 731.2, found 731.5


 


16




3′-[4-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-2-morpholin-4- ylpyrimidin-5-yl]-4′- methoxy-2-methylbiphenyl- 4-carboxylic acid
272
Calc'd 743.2, found 743.5









Example 17




methyl 3′-[4-({(8R)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (scheme 4)
Step 1: To a stirred, cooled mixture of methyl 3′-[4-(hydroxymethyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (3.0 g, 7.31 mmol) and triethylamine (2.04 mL, 14.62 mmol) in dichloromethane (30 mL) under nitrogen at 0° C. was added MsCl (0.854 mL, 10.96 mmol). The mixture was stirred at 0° C. for 3 h before the reaction was diluted with ethyl acetate and water. The organic layer was washed with water and brine, was dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography on silica gel to yield methyl 4′-methoxy-2-methyl-3′-[2-(methylsulfanyl)-4-{[(methylsulfonyl)oxy]methyl}pyrimidin-5-yl]biphenyl-4-carboxylate as slightly yellow solid. (M+H). MS ESI calc'd. for C23H25N2O6S2 [M+H]+ 489.1, found 489.2.
Step 2: To a stirred solution of 8-[3,5-bis(trifluoromethyl)phenyl]-7-oxa-5-azaspiro[3.4]octan-6-one (290 mg, 0.855 mmol) in THF (6 mL) at 0° C. was added sodium hydride (60%, 41.0 mg, 1.026 mmol). After stirring for 15 min, a solution of 4′-methoxy-2-methyl-3′-[2-(methylsulfanyl)-4-{[(methylsulfonyl)oxy]methyl}pyrimidin-5-yl]biphenyl-4-carboxylate (501 mg, 1.026 mmol) in THF (5 mL) was added. The reaction suspension was stirred at room temperature overnight and was quenched with saturated aqueous ammonium chloride. The mixture was diluted with ethyl acetate and water (70 mL) and the organic layer was washed with water (70 mL), brine, dried over sodium sulfate, filtered and concentrated. The residue was purified by column chromatography to yield methyl 3′-[4-({8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate as solid. MS ESI calc'd. for C36H32F6N3O5S [M+H]+ 732.2, found 732.5. 1H NMR (500 MHz, CDCl3) δ 1.12-1.56 (m, 4H), 2.08-2.24 (m, 2H), 2.38 (s, 3H), 2.60 (s, 3H), 3.86 (s, 3H), 3.88 (s, 3H), 4.38-4.70 (m, 2H), 5.42-5.65 (s, 1H), 7.12-7.46 (m, 4H), 7.88-7.98 (m, 5H), 8.38 (s, 1H) ppm.
Step 3: The racemic material was purified by chiral SFC (Column: Chiral Technology 1A-H 30×250 mm, 35° C. Mobile Phase: 20% MeOH/CO2. Flow rate: 70 mL/min. Wavelength: (220 nm) to provide:

    Methyl 3′-[4-({(8S)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (intermediate D1, Rt=5.9 min) and
    Methyl 3′-[4-({(8R)-8-[3,5-bis(trifluoromethyl)phenyl]-6-oxo-7-oxa-5-azaspiro[3.4]oct-5-yl}methyl)-2-(methylsulfanyl)pyrimidin-5-yl]-4′-methoxy-2-methylbiphenyl-4-carboxylate (intermediate D2, Rt=8.6 min). RTA (95% HS): 913 nM


1H NMR and MS spectra for both enantiomers match that of the racemic material.
The following compounds in table 5 were prepared according to general scheme 4 using the procedure outlined for example 17 utilizing intermediate D in step 1 and commercially available amines in step 2. Step 3 and/or 4 may be omitted if no ester moiety is present in the molecule or the material is not racemic.











TABLE 5









Exact





IC50
Mass


ex
Structure
IUPAC Name
(nM)
[M + H]+







18




methyl 3′-[4-({(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 5-oxo-6-oxa-4- azaspiro[2.4]hept-4- yl}methyl)-2- (methylsulfanyl)pyrimidin-5- yl]-4′-methoxy-2- methylbiphenyl-4- carboxylate
596
Calc'd 718.2, found 718.4


 


19




(7R)-7-[3,5- bis(trifluoromethyl)phenyl]- 4-{[4′-fluoro-2′-methoxy-5′- (1-methylethyl)-4- (trifluoromethyl)biphenyl-2- yl]methyl}-6-oxa-4- azaspiro[2.4]heptan-5-one
361
Calc'd 650.2, found 650.0

## Claims

1. A compound having formula I, or a pharmaceutically acceptable salt thereof,



Wherein each R is H;
R1 is unsubstituted cyclopropyl or cyclobutyl;
B is A1, wherein A1 has the structure of formula II:




wherein Y1 and Y2 are each —CH— or —N—, with the proviso that Y1 and Y2 are not both —CH—;
R1a is 3-fluoroazetidinyl, 4-morpholinyl, CF3, —N(CH3)2, —N(CH3)(SO2CH3), or CH3S—;
A is A2, wherein A2 is 3,5-bis-trifluoromethylphenyl;
A3 is




wherein Y3 is —CH— or —N—;
X is 3-cyclobutyl-D1, 3-cyclobutyl-CH2OH, 2-methyl-4-phenyl-D1,
2-methyl-4-phenyl-CH2OH, —CH2CH2D1, —CH2OH, or isopropyl, wherein D1 is —CO2R8;
R8 is H or —CH3;
Rb is —OCH3 or halogen;
d is 0; and
e is an integer from 0-3.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, having the structure below:

3. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.