HSD17B13 inhibitors and / or degradation agents

Abstract

Described herein are compounds of Formula I, wherein the variables are defined herein; their use as HSD17B13 inhibitors and / or degradation agents, pharmaceutical compositions containing said compounds and their use for the treatment of e.g. NAFLD and NASH. # imgabs0 #

Description

Technical Field

[0001] The present application provides compounds that are inhibitors and / or degraders of hydroxysteroid 17β-dehydrogenase 13 (HSD17B13), pharmaceutical compositions containing such compounds, and their use for treating conditions, diseases, or disorders associated with HSD17B13 activity. Background Art

[0002] Hydroxysteroid 17β-dehydrogenase 13 (HSD17B13) is a steroid dehydrogenase family enzyme associated with hepatic lipid droplets. From 2018 to the present, multiple human genetic variants of HSD17B13 have been identified that can prevent NASH progression, wherein these human variants lead to reduced liver inflammation, ballooning, and fibrosis. Abul-Husn et al., 2018 reported that truncated variants are over-enriched in individuals with simple steatosis and under-enriched in individuals with NASH and NASH+fibrosis, which means that they prevent disease progression (Abul-Husn et al., "Protein-Truncating HSD17B13 Variant and Protection from Chronic Liver Disease", N Engl J Med 2018; 378: 1096-1106). Later that year, Kozlitina et al. reported a second truncated variant with reduced allele frequency in Africans and Hispanics with chronic liver disease (Kozlitina et al., "HSD17B13 and Chronic Liver Disease in Blacks and Hispanics", N Engl J Med 2018; 379: 1876-1877). In 2019, Ma et al. found that the coding variant P260S was associated with reduced inflammation and ballooning. HSD17B13 expression has been shown to be significantly upregulated in humans with nonalcoholic fatty liver disease (NAFLD) (Ma et al., "17-Beta Hydroxysteroid Dehydrogenase 13IsaHepatic Retinol Dehydrogenase Associated With Histological Features of Nonalcoholic Fatty Liver Disease", Hepatology 2019; 69 (4): 1504-1519). A mouse model placed on a pro-NASH diet also demonstrated upregulation of the protein. Therefore, it is speculated that inhibition or degradation of HSD17B13 enzyme activity may slow or prevent the progression of liver diseases such as non-alcoholic fatty liver disease (NAFLD) (including NASH (non-alcoholic steatohepatitis)), liver inflammation, fibrosis, cirrhosis, and the development of hepatocellular carcinoma.

[0003] Despite some early studies related to HSD17B13, there remains a need for agents that have HSD17B13 inhibitory / degrading activity and that can be used to treat, prevent, or alleviate the manifestations of the diseases described herein. Summary of the Invention

[0004] The present application relates to a compound of formula I or a pharmaceutically acceptable salt of said compound,

[0005]

[0006] in

[0007] A is -NH-C(O)- or heteroaryl having 1, 2, 3 or 4 heteroatoms selected from O, N and S, wherein A is optionally substituted by one or two R 4 replace;

[0008] B is absent or is H, aryl, heteroaryl, heterocyclyl, fluorine, chlorine, bromine, oxo, cyano, hydroxy, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) fluoroalkyl, (C1-C6) alkoxy, or (C1-C6) fluoroalkoxy, wherein the heteroaryl or heterocyclyl has 1, 2 or 3 heteroatoms selected from O, N and S, and wherein B is optionally replaced by one or two R 5 replace;

[0009] C does not exist or is H, -NH-C(O)-R 7 、-S(O)2-R 7 、-OS(O)2-R 7 , fluorine, chlorine, bromine, oxo, cyano, hydroxy, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy, (C3-C6) cyclic ether, (C1-C6) fluoroalkyl, (C1-C6) fluoroalkoxy, aryl, heteroaryl or heterocyclic group, wherein the heteroaryl or heterocyclic group has 1, 2 or 3 heteroatoms selected from O, N and S, and wherein C is optionally replaced by one, two or three R 6 replace;

[0010] R 1 、R 2 and R 3 are each independently selected from H and fluorine;

[0011] Each R 4 、R 5 and R 6 independently selected from oxo, hydroxy, chlorine, fluorine, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)fluoroalkyl, (C3-C6)cycloalkyl, and heterocyclyl having 1, 2 or 3 heteroatoms selected from O and N;

[0012] R 7 is hydroxy, chloro, fluoro, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)fluoroalkyl, or (C3-C6)cycloalkyl; and

[0013] n is 0, 1 or 2.

[0014] The present application also relates to methods for treating the following diseases by administering to a human in need of such treatment a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound: fatty liver, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, nonalcoholic steatohepatitis with liver fibrosis, nonalcoholic steatohepatitis with cirrhosis, nonalcoholic steatohepatitis with cirrhosis, hepatocellular carcinoma, alcoholic fatty liver disease, alcoholic steatohepatitis, hepatitis B, hepatitis C, biliary cirrhosis, renal clear cell carcinoma, head and neck squamous cell carcinoma, colorectal adenocarcinoma, mesothelioma, gastric adenocarcinoma, adrenocortical carcinoma, renal papillary cell carcinoma, cervical and endocervical cancer, bladder urothelial carcinoma, lung adenocarcinoma, type I diabetes, idiopathic type I diabetes (Type Ib), latent autoimmune diabetes in adults (LADA), early-onset type 2 diabetes (EOD), atypical diabetes of the young (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, restenosis after angioplasty, peripheral vascular disease, intermittent claudication, postprandial lipemia, metabolic acidosis, ketosis, arthritis, diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic neuropathy, skin and connective tissue disorders, foot ulcers and ulcerative colitis, endothelial dysfunction and impaired vascular compliance, kidney disease, end-stage renal disease, chronic kidney disease at risk of progression, and maple syrup urine disease.

[0015] The present application also relates to a method of reducing cirrhosis, decompensated cirrhosis, progression to a Model for End-Stage Liver Disease (MELD) score >= 15, liver transplantation, death (liver-related), and hepatocellular carcinoma by administering a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound to a human in need of such treatment.

[0016] The present application also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.

[0017] The present application also relates to a pharmaceutical combination composition, which comprises: a therapeutically effective amount of a composition having:

[0018] a first compound, wherein the first compound is a compound of Formula I or a pharmaceutically acceptable salt of the compound;

[0019] A second compound, wherein the second compound is an anti-diabetic agent, a therapeutic agent for non-alcoholic steatohepatitis, a therapeutic agent for non-alcoholic fatty liver disease, or an anti-heart failure therapeutic agent, and a pharmaceutically acceptable carrier, vehicle, or diluent.

[0020] It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed. DETAILED DESCRIPTION

[0021] The present application may be understood more readily by reference to the following detailed description of exemplary embodiments of the invention and the Examples included therein.

[0022] It should be understood that the present invention is not limited to specific synthetic preparation methods, which can of course vary. It should also be understood that the terms used herein are used only for the purpose of describing specific embodiments and are not intended to be limiting. In this specification and the claims that follow, reference will be made to a number of terms that should be defined as having the following meanings:

[0023] As used herein in the specification, "a" or "an" may mean one or more. As used herein in one or more claims, when used in conjunction with the word "comprising," the word "a" or "an" may mean one or more than one. As used herein, "another" may mean at least a second or more.

[0024] The term "about" is intended to be a relative term, indicating an approximation of ±10%, in one embodiment ±5%, in another embodiment ±2% of the nominal value to which it refers. For the purposes of this disclosure, this approximation level is appropriate unless a more stringent range is explicitly stated for the value.

[0025] The term "and / or" means one or more. For example, "X and / or Y" should be understood to mean "X and Y" or "X or Y", and should be considered to provide clear support for both meanings or either meaning. Similarly, when more than two expressions are listed, as in "X, Y and / or Z", it should be understood to mean i) "X and Y", "X, Y and Z", "X and Z", or "Y and Z", or ii) "X or Y or Z", and should be considered to provide clear support for all meanings.

[0026] The term "alkyl" (alone or in combination) means a group of formula C n H 2n+1 An acyclic saturated hydrocarbon group which may be straight or branched. Examples of such groups include methyl, ethyl, n-propyl, isopropyl, butyl, sec-butyl, isobutyl, and tert-butyl. The carbon atom content of the alkyl group and various other hydrocarbon-containing moieties is indicated by prefixes designating the upper and lower limits on the number of carbon atoms in the moiety, i.e., the prefix C i -C j A moiety designating an integer "i" to an integer "j" number of carbon atoms, inclusive. Thus, for example, C1-C3 alkyl refers to an alkyl group of one to three carbon atoms, inclusive.

[0027] "Fluoroalkyl" means an alkyl group as defined herein substituted with one, two, or three fluorine atoms. Exemplary (C1) fluoroalkyl compounds include fluoromethyl, difluoromethyl, and trifluoromethyl; exemplary (C2) fluoroalkyl compounds include 1-fluoroethyl, 2-fluoroethyl, 1,1-difluoroethyl, 1,2-difluoroethyl, 1,1,1-trifluoroethyl, 1,1,2-trifluoroethyl, and the like.

[0028] "Cycloalkyl" refers to a non-aromatic ring of formula C n H 2n-1 Examples of such carbocycles include cyclopropyl and cyclobutyl.

[0029] "Fluorocycloalkyl" means a non-aromatic cycloalkyl ring as defined herein that is substituted with one, two, or three fluorine atoms. Exemplary (C3) fluorocycloalkyl compounds include fluorocyclopropyl, difluorocyclopropyl, and trifluorocyclopropyl; exemplary (C4) fluorocycloalkyl compounds include 1-fluorocyclobutyl, 2-fluorocyclobutyl, 1,1-difluorocyclobutyl, 1,2-difluorocyclobutyl, 1,1,1-trifluorocyclobutyl, 1,1,2-trifluorocyclobutyl, and the like.

[0030] "Alkoxy" means a straight or branched chain saturated alkyl group bonded through an oxygen group. Exemplary such alkoxy groups (assuming the specified length encompasses a specific example) are methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentoxy, isopentoxy, neopentoxy, tert-pentoxy, hexoxy, isohexoxy, heptoxy, and octoxy.

[0031] "Fluoroalkoxy" means an alkoxy group, as defined herein, substituted with one, two, or three fluorine atoms. Exemplary (C1) fluoroalkoxy compounds include fluoromethoxy, difluoromethoxy, and trifluoromethoxy; exemplary (C2) fluoroalkyl compounds include 1-fluoroethoxy, 2-fluoroethoxy, 1,1-difluoroethoxy, 1,2-difluoroethoxy, 1,1,1-trifluoroethoxy, 1,1,2-trifluoroethoxy, and the like.

[0032] "Halogen" refers to bromine, chlorine, fluorine or iodine.

[0033] The term "heteroaryl" refers to a monovalent or divalent group containing at least one aromatic ring and at least one heteroatom (e.g., 1 to 5 heteroatoms, each independently N, O, or S) ring member. The total number of ring members may be indicated (e.g., 5- to 10-membered heteroaryl). A heteroaryl group may include two fused rings, at least one of which is aromatic and the other of which is aromatic, saturated, or partially unsaturated, and at least one of the fused rings contains a heteroatom.

[0034] When "ene" is added to the end of a term after "base" to form a new term, the new term refers to a diradical formed by removing one hydrogen atom from the original term from which the new term was derived. For example, alkylene refers to a diradical formed by removing one hydrogen atom from an alkyl group, and "methylene" refers to a divalent radical -CH2- obtained by removing one hydrogen atom from a methyl group. Further examples of such diradicals include, but are not limited to, alkenylene, alkynylene, cycloalkylene, phenylene, heterocyclylene, and heteroarylene, which are derived from alkenyl, alkynyl, cycloalkyl, phenyl, heterocyclyl, and heteroarylene. "C 1-3 Non-limiting examples of "alkylene" include: -CH2-, -CH(CH3)-, -CH2-CH2-, -CH2-CH2-CH2-, -CH(CH3)-CH2-, and -CH(CH2CH3)-. For cyclic moieties, removal of a hydrogen may occur at any atom of sufficient valence.

[0035] As used herein, "compound" includes any pharmaceutically acceptable derivative or variant, including conformational isomers (e.g., cis and trans isomers), atropisomers (i.e., stereoisomers due to hindered rotation), and all optical isomers (e.g., enantiomers and diastereomers), racemic mixtures, diastereomeric mixtures, and other mixtures of such isomers, as well as solvates, hydrates, isomorphs, polymorphs, tautomers, esters, salt forms, and prodrugs. The expression "prodrug" refers to a compound that is a drug precursor that releases the drug in vivo after administration via some chemical or physiological process (e.g., the prodrug is converted to the desired drug form upon reaching physiological pH or by enzymatic action). Exemplary prodrugs release the corresponding free acid upon cleavage, and such hydrolyzable ester residues of compounds of Formula I include, but are not limited to, those having a carboxyl moiety in which the free hydrogen is replaced by (C1-C4)alkyl, (C2-C7)alkanoyloxymethyl, 1-(alkanoyloxy)ethyl having 4 to 9 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having 5 to 10 carbon atoms, alkoxycarbonyloxymethyl having 3 to 6 carbon atoms, 1-(alkoxycarbonyloxy)ethyl having 4 to 7 carbon atoms, 1-methyl-1-(alkanoyloxy)ethyl having 5 to 8 carbon atoms, -(alkoxycarbonyloxy)ethyl, N-(alkoxycarbonyl)aminomethyl having 3 to 9 carbon atoms, 1-(N-(alkoxycarbonyl)amino)ethyl having 4 to 10 carbon atoms, 3-phthalidyl, 4-crotonolactone, γ-butyrolactone-4-yl, di-N,N-(C1-C2)alkylamino(C2-C3)alkyl (such as β-dimethylaminoethyl), carbamoyl-(C1-C2)alkyl, N,N-di(C1-C2)alkylcarbamoyl-(C1-C2)alkyl, and piperidinyl-, pyrrolidinyl- or morpholino(C2-C3)alkyl.

[0036] As used herein, arrows or wavy lines It indicates the point of attachment of a substituent to another group.

[0037] As used herein, "deuterium enrichment factor" means the ratio between the abundance of deuterium and the natural abundance of deuterium (each relative to the abundance of hydrogen). In certain embodiments, the deuterium enrichment factor for atomic positions designated as having deuterium is typically at least 1000 (15% deuterium incorporation), at least 2000 (30% deuterium incorporation), at least 3000 (45% deuterium incorporation), at least 3500 (52.5% deuterium incorporation), at least 3500 (52.5% deuterium incorporation at each designated deuterium atom), at least 4000 (60% deuterium incorporation), or at least 5000 (70% deuterium incorporation). , at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0038] The term "mammal" refers to humans, livestock, or companion animals.

[0039] The term "companion animal" or "companion animals" refers to animals kept as pets or domestic animals. Examples of companion animals include dogs, cats, and rodents (including hamsters, guinea pigs, gerbils, etc.), rabbits, and ferrets.

[0040] The term "livestock" refers to animals raised or raised in an agricultural environment for the purpose of producing products such as food or fiber, or for their labor. In some embodiments, livestock are suitable for consumption by mammals (e.g., humans). Examples of livestock animals include cattle, goats, horses, pigs, sheep (including lambs), and rabbits.

[0041] "Patient" refers to warm-blooded animals such as guinea pigs, minipigs, mice, rats, gerbils, cats, rabbits, dogs, cows, goats, sheep, horses, monkeys, chimpanzees, and humans.

[0042] The term "treating" or "treatment" means to relieve the symptoms associated with a disease, disorder, or condition, or to stop the further progression or worsening of those symptoms. Depending on the patient's disease and condition, as used herein, the term "treatment" may include one or more of curative, palliative, and preventative treatments. Treatment may also include administering a pharmaceutical formulation in combination with other therapies.

[0043] "Therapeutically effective amount" means an amount of a compound of the invention that (i) treats or prevents a particular disease, condition, or disorder, (ii) alleviates, ameliorates, or eliminates one or more symptoms of a particular disease, condition, or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition, or disorder as described herein.

[0044] The term "pharmaceutically acceptable" means a substance (e.g., a compound of the invention) and any salt thereof, or a composition containing a substance or salt of the invention that is suitable for administration to a patient. Unless otherwise indicated, when referring to a compound of Formula I, it should be understood that pharmaceutically acceptable salts of the compound are also contemplated.

[0045] In embodiments of the compound or a pharmaceutically acceptable salt of said compound, the compound has Formula IA

[0046]

[0047] In an embodiment of the compound or a pharmaceutically acceptable salt of said compound, the compound has the formula IB

[0048]

[0049] In an embodiment of the compound or a pharmaceutically acceptable salt of said compound, R 2 It's F.

[0050] In embodiments of the compound or a pharmaceutically acceptable salt of the compound, A is thiazolyl, pyrazolyl, oxazolyl, imidazolyl, isoxazolyl, isothiazolyl, imidazotriazinyl, imidazopyridazinyl, imidazopyridinyl, benzimidazolyl, benzothiazolyl, purinyl, pyridopyridazinyl, quinazolinyl, indazolyl, imidazopyridinyl, benzoxazolyl, pyrazolopyridinyl, isoindolinonyl, triazolyl, or oxadiazolyl.

[0051] In another embodiment of the compound, A is

[0052]

[0053] In another embodiment of the compound or a pharmaceutically acceptable salt of the compound, B is absent or is H, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, piperazinyl, quinoxalinyl, phenyl, triazolyl, thiazolyl, thiadiazolyl, oxazolyl, imidazolyl, indazolyl, (C1-C6)alkyl, (C1-C6)fluoroalkyl, (C1-C6)alkoxy, bromo, chloro, fluoro, or oxo, and wherein B is optionally substituted with one or two fluoro, oxo, hydroxy, (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)fluoroalkyl, (C1-C6)alkoxy, or (C3-C6)cyclic ether.

[0054] In another embodiment of the compound or a pharmaceutically acceptable salt of the compound, B is pyrimidinyl, (C1-C3)fluoroalkyl-substituted pyrimidinyl, (C1-C3)alkyl-substituted pyrazolyl, methoxy-substituted pyridazinyl, difluoromethyl-substituted pyrazinyl, trifluoromethyl-substituted pyrimidinyl, or methoxy-substituted pyrimidinyl.

[0055] In another embodiment of the compound or a pharmaceutically acceptable salt of the compound, C is absent or is H, pyridyl, piperazinyl, oxolanyl, (C3-C6)cycloalkyl, (C1-C6)alkyl, (C1-C6)fluoroalkyl, (C1-C6)alkoxy, cyano, bromo, chloro, fluoro, or oxo, and wherein C is optionally substituted with one, two, or three fluoro, oxo, hydroxy, (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)fluoroalkyl, or (C1-C6)alkoxy.

[0056] In another embodiment of the compound or a pharmaceutically acceptable salt of the compound, C is absent or is pyridinyl, piperazinyl, (C3-C6)cycloalkyl, (C1-C6)alkyl, (C1-C6)fluoroalkyl; and wherein C is optionally substituted with one, two or three fluoro, oxo, hydroxy, or (C1-C6)alkyl.

[0057] In compound embodiments, the compound is: 2,3,5-trifluoro-4-hydroxy-N-[(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide; 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide; 2,3,5-trifluoro-4-hydroxy-N-({4-[6-(pyrimidin-5-yl)bicyclo[2.2.2]octan-1-yl)methyl]benzamide. 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide; 2,3,5-trifluoro-4-hydroxy-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]oct-1-yl}methyl)benzamide; N-[(4-{5-[5-(difluoromethyl)-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide; )pyrazin-2-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]-3,5-difluoro-4-hydroxybenzamide; 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide; 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[6-(2-methoxypyrimidin-5-yl)-2H-pyrazolo[4,3-c]pyridin-2-yl)- ]cyclohexyl}methyl)benzamide; 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(piperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]oct-1-yl)methyl]benzamide; or 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]oct-1-yl)methyl]benzamide; or a pharmaceutically acceptable salt of the compound.

[0058] In a compound embodiment, the compound is 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide or a pharmaceutically acceptable salt of said compound.

[0059] In an embodiment of the present invention, a method for treating fatty liver, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, nonalcoholic steatohepatitis with liver fibrosis, nonalcoholic steatohepatitis with cirrhosis, or nonalcoholic steatohepatitis with cirrhosis and hepatocellular carcinoma comprises administering to a human in need of such treatment a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound.

[0060] In an embodiment of the invention, the method comprises treating nonalcoholic steatohepatitis.

[0061] In an embodiment of the present invention, the pharmaceutical composition comprises a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.

[0062] In an embodiment of the present invention, the pharmaceutical combination composition comprises a therapeutically effective amount of a composition comprising: a first compound, wherein the first compound is a compound of Formula I or a pharmaceutically acceptable salt of the compound; a second compound, wherein the second compound is an antidiabetic agent, a non-alcoholic steatohepatitis therapeutic agent, a non-alcoholic fatty liver disease therapeutic agent, or an anti-heart failure therapeutic agent; and a pharmaceutically acceptable carrier, vehicle, or diluent.

[0063] In an embodiment of the present invention, the nonalcoholic steatohepatitis therapeutic agent or nonalcoholic fatty liver disease therapeutic agent in the pharmaceutical combination composition is an ACC inhibitor, a KHK inhibitor, a DGAT-2 inhibitor, a FXR agonist, metformin, an incretin analog, or an incretin receptor modulator.

[0064] In embodiments of the present disclosure, the antidiabetic agent is a SGLT-2 inhibitor, metformin, an incretin mimetic, an incretin receptor modulator, a DPP-4 inhibitor, or a PPAR agonist.

[0065] The present invention includes compounds of the present invention that are targeting protein ligands covalently linked to E3 ligase ligands or ligands known to interact with the ubiquitin proteasome system (Degrons) through linkers of varying lengths and functions. When so linked to Degrons, the compounds of the present invention are referred to herein as bifunctional compounds of the present invention. The compounds of the present invention are generally referred to as targeting ligands within these bifunctional compounds of the present invention. These bifunctional compounds of the present invention can be used as therapeutic agents for treating various diseases, including various liver diseases.

[0066] The bifunctional compounds of the present invention have the following general structure: Degron-Linker-Targeting Ligand, wherein the Linker is covalently bound to at least one Degron and at least one Targeting Ligand, wherein the Degron is a compound capable of binding to a ubiquitin ligase such as an E3 ubiquitin ligase (e.g., cereblon (CRBN), von Hippel-Lindau (VHL), etc.), and the Targeting Ligand is capable of binding to one or more proteins HSD17B13 and is a compound of the present invention as presented in any embodiment described herein. Such bifunctional compounds of the present invention are generally presented as compounds of Formula II, wherein L is a Linker and D is a Degron:

[0067]

[0068] Degron is small in size and highly effective in recruiting targeted proteins for degradation. Degron is a compound that is used to connect targeted proteins to ubiquitin ligases for proteasomal degradation through linkers and targeting ligands. In certain embodiments, Degron is a compound capable of binding to or binding to ubiquitin ligases. In other embodiments, Degron is a compound capable of binding to or binding to E3 ubiquitin ligases (including cereblon), wherein Degron is thalidomide, lenalidomide, pomalidomide, or ibedomide, or the newer IMiD CRBN ligands disclosed in WO2019 / 060693, WO2019 / 140387, WO2019 / 236483, or their analogs. In other embodiments, Degron is a compound capable of binding to or binding to E3 ubiquitin ligases (including von Hippel-Lindau ligands). See, for example, WO2020 / 092907; WO2013106643; Buckley et al., J. Am. Chem. Soc. 2012, 134, 4465-4468, "Targeting the VonHippel-Lindau E3 Ubiquitin Ligase Using Small Molecules to Disrupt the VHL / Hif-1alpha Interaction", Soares et al., J. Med. Chem. 2019, 61, 599-618, "Group-BasedOptimization of Potent and Cell-Active Inhibitors of the von Hippel–Lindau(VHL)E3 Ubiquitin Ligase: Structure–Activity Relationships Leading to the Chemical Probe (2S, 4R) -1- ((S) -2- (1-Cyanocyclopropanecarboxamido) -3, 3-dimethylbutanoyl) -4-hydroxy-N- (4- (4-methylthiazol-5-yl) benzyl) pyrrolidine-2-carboxamide (VH298)". In other embodiments, Degron is a compound that can bind to or bind to E3 ubiquitin ligases, including inhibitor of apoptosis protein ligases (IAP1, IAP2, XIAP).See, for example, Itoh et al., J. Am. Chem. Soc. 2010, 132, 5820-5826 "Protein Knockdown Using Methyl Bestatin-Ligand Hybrid Molecules: Design and Synthesis of Inducers of Ubiquitination-Mediated Degradation of Cellular Retinoic Acid-Binding Proteins', Mares et al., Commun. Biol. 2020, 3, 140, "Extended pharmacodynamic responses observed upon PROTAC-mediated degradation of RIPK2', and Tinworth et al., ACS Chem. Biol. 2019, 14, 342-347, "PROTAC-Mediated Degradation of Bruton's Tyrosine Kinase Is Inhibited by Covalent In other embodiments, Degron is a compound that can bind to or bind to other ubiquitin proteasome proteins that can be induced to degrade (including but not limited to Hsp70 / 90 molecular chaperone complex (WO2020 / 207395), Usp14 (WO2019 / 238886), UchL5 (WO2019238816), BILO (WO201719705) and Rpn11 (WO2019 / 238817)).

[0069] In certain embodiments, linkers are designed and optimized based on SAR (structure activity relationship) and X-ray crystallography of the attachment position of the targeting ligand relative to the linker.

[0070] In certain embodiments, the optimal linker length and composition vary depending on the target and can be estimated based on the X-ray structure of the original targeting ligand bound to its target. The linker length and composition can also be changed to adjust metabolic stability and pharmacokinetic (PK) and pharmacodynamic (PD) parameters.

[0071] In certain embodiments, where a target ligand binds multiple targets, selectivity can be achieved by varying the linker length, where the ligand binds some of its targets in different binding pockets (e.g., deeper or shallower binding pockets than others).

[0072] The linker ("L") provides a covalent link between the targeting ligand and the Degron. The linker has two terminal groups, one of which is connected to the Degron and the other is connected to the targeting ligand. The structure of the linker may not be critical, as long as it does not substantially interfere with the activity of the targeting ligand or the Degron. In some embodiments, the linker is an alkyl chain (e.g., having 2-20 alkyl units), or a polyethylene glycol (PEG) chain (CH2CH2-O or (O-CH2CH2)). In other embodiments, the linker may be an alkylene chain, a PEG chain, or a divalent alkylene chain, any of which may be interrupted and / or terminated (at one or both ends) in at least one of the following: -O-, -S-, -N(R L )–, –C=C–, –C(O)–, –C(O)O–, –OC(O)–, –OC(O)O–, –C(NOR L )–、–C(O)N(R L )–、–C(O)N(R L )C(O)–、–C(O)N(R L )C(O)N(R L )–、–N(R L )C(O)–、–N(R L )C(O)N(R L )–、–N(R L )C(O)O–、–OC(O)N(R L )–、–C(NR L )–,–N(R L )C(NR L )–、–C(NR L )N(R L )–、–N(R L )C(NR L )N(R L )–, –OB(CH3)O–, –S(O)2–, –OS(O)–, –S(O)O–, –S(O)–, –OS(O)2–, –S(O)2O–, –N(R L )S(O)2–、–S(O)2N(R L )–、–N(R L )S(O)–、–S(O)N(R L )–、–N(R L )S(O)2N(R L )–, –N(R’)S(O)N(R’)–, C 3-12 carbocyclene, 3- to 12-membered heterocyclene, 5- to 12-membered heteroarylene, or arylene, or any combination thereof, wherein RL Is H or C 1-6 Alkyl groups, wherein the interrupting groups and one or both terminal groups may be the same or different.

[0073] In some embodiments, the linker can be a C terminated NH group. 1-10 Alkylene chain, where nitrogen is also bound to the Degron, or the linker can be C 1-10 Alkylene chain or PEG chain having 1-8 PEG units and interrupted or terminated with -(CH2) n’ -C(O)-NH-, wherein n' is 0 to about 5. With respect to linkers, "carbocyclylene" refers to a divalent carbocyclic group that is optionally substituted. "Heterocyclylene" refers to a divalent heterocyclyl group that may be optionally substituted. "Heteroarylene" refers to a divalent heteroaryl group that may be optionally substituted.

[0074] Non-limiting examples of linkers include -(CH2) n’ -、-(CH2CH2-O) n” -(CH2) n’ -C(O)-, (CH2) n’ -C(O)-N(R L )-(CH2CH2-O) n” -(CH2) n’ -C(O)-, -(CH2CH2-O) n” -(CH2) n’ -N(R L )-C(O)-、-(CH2CH2-O) n” -(CH2) n’ -C(O)-N(R L )-、-(CH2) n’ -phenylene-N(R L )-C(O)-(CH2) n’ -、-N(R L )-(CH2) n’ -O-phenylene-(CH2) n” -N(R L )-(CH2) n’ -、-(CH2) n’ -C(O)-N(R L )-phenylene-C(O)-、-N(R L )-(CH2) n’ -phenylene-(CH2) n” -Heterocyclylene-, -(CH2) n’ -phenylene-N(R L )-C(O)-(CH2CH2-O) n” -(CH2)n’ -、-(CH2) n’ -phenylene-(CH2) n” -Heterocyclylene-(CH2) n” C(O)-N(R L )-(CH2) n’ -、-(CH2) n’ -phenylene-O-(CH2) n’ -Heterocyclylene-(CH2) n’ -、-(CH2) n’ -phenylene-(CH2) n’ -Heterocyclylene-(CH2) n’ -O-, -(CH2) n’ -Heterocyclylene-(CH2) n’ , where R L Is H or C 1-6 alkyl; n' is 0 to about 10; and n" is 1 to about 10.

[0075] Another embodiment includes a compound selected from any one of the Examples described herein, or a pharmaceutically acceptable salt thereof.

[0076] Another embodiment includes a prodrug of any one of the Examples described herein, or a pharmaceutically acceptable salt thereof.

[0077] Another embodiment includes a phosphate prodrug of any one of the Examples described herein, or a pharmaceutically acceptable salt thereof.

[0078] Another embodiment includes any novel class of intermediates described in the general schemes or examples.

[0079] Another embodiment includes any of the novel specific intermediates described in the formulations described herein and in the Examples.

[0080] Another embodiment comprises any of the novel methods described herein.

[0081] All pharmaceutically acceptable isotopically labeled compounds of Formula I, wherein one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number usually found in nature, are within the scope of the present application.

[0082] Examples of suitable isotopes for inclusion in the compounds of the present invention include hydrogen (e.g. 2 H and 3 H), carbon (such as 11 C. 13 C and 14 C), chlorine (such as 36 Cl), fluorine (such as 18F), nitrogen (such as 13 N and 15 N), oxygen (such as 15 O. 17 O and 18 O), and sulfur (such as 35 S) isotopes.

[0083] Certain isotopically-labeled compounds of Formula I (e.g., those into which a radioactive isotope is incorporated) are useful in drug and / or substrate tissue distribution studies. 3 H) and carbon-14 (i.e. 14 C) is particularly useful for this purpose due to its ease of introduction and readily available detection means.

[0084] Using heavier isotopes such as deuterium (i.e. 2 H) substitutions may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances.

[0085] Using positron-emitting isotopes (such as 11 C. 18 F. 15 O and 13 N) substitutions can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy.

[0086] Isotopically labeled compounds of Formula I can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples and Preparations, using an appropriate isotopically labeled reagent in place of the non-labeled reagent previously employed.

[0087] Certain compounds of Formula I and intermediates described herein may exist in more than one crystalline form (often referred to as a "polymorph"). Polymorphs can be prepared by crystallization under various conditions, for example, using different solvents or different solvent mixtures for recrystallization; crystallization at different temperatures; and / or various cooling modes, ranging from very fast to very slow cooling during crystallization. Polymorphs can also be obtained by heating or melting a compound followed by gradual or rapid cooling. The presence of polymorphs can be determined by solid probe NMR spectroscopy, IR spectroscopy, differential scanning calorimetry, powder X-ray diffraction, or such other techniques.

[0088] Salts encompassed within the term "pharmaceutically acceptable salts" refer to compounds of the invention generally prepared by reacting a free base or free acid with a suitable organic or inorganic acid, or a suitable organic or inorganic base, respectively, to provide salts of the compounds of the invention suitable for administration to a patient. Basic salts are preferred, however, some compounds can also form acid salts. Suitable acid addition salts are formed from acids that form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate / carbonate, bisulfate / sulfate, borate, camphorsulfonate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronate, hexafluorophosphate, hyphenate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, methanesulfonate, methylsulfate, naphthylate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, pyroglutamate, sucrose, stearate, succinate, tannate, tartrate, tosylate, trifluoroacetate, and xinofoate.

[0089] Suitable basic salts are formed from bases that form non-toxic salts. Examples include aluminum salts, arginine salts, calcium salts, choline salts, diethylamine salts, glycine salts, lysine salts, magnesium salts, meglumine salts, ethanolamine salts, potassium salts, sodium salts, tromethamine salts, and zinc salts. Hemi-salts of acids and bases, such as hemisulfates and hemicalcium salts, can also be formed. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002).

[0090] Hemi-salts of acids and bases can also be formed, such as hemisulphate and hemicalcium salts. For a review of suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley-VCH, 2002).

[0091] Pharmaceutically acceptable salts of compounds of Formula I can be prepared by one or more of the following three methods:

[0092] (i) by reacting a compound of formula I with a desired acid or base;

[0093] (ii) by removing an acid- or base-labile protecting group from a suitable precursor of the compound of the invention, or by ring-opening a suitable cyclic precursor (e.g., a lactone or lactam) using the desired acid or base; or

[0094] (iii) by converting one salt of the compound of the present invention into another salt via reaction with an appropriate acid or base or with the aid of a suitable ion exchange column.

[0095] All three reactions are typically carried out in solution. The resulting salt can be precipitated and collected by filtration, or can be recovered by evaporating the solvent. The degree of ionization of the resulting salt can vary from fully ionized to almost non-ionized.

[0096] The compounds of Formula I and their pharmaceutically acceptable salts can exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of Formula I or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable solvent molecules (e.g., ethanol). When the solvent is water, the term "hydrate" is used.

[0097] The currently accepted classification system for organic hydrates is one that defines isolated site, channel, or metal ion coordination hydrates—see KR Morris, Polymorphism in Pharmaceutical Solids (HG Brittain, Marcel Dekker, ed., 1995). Isolated site hydrates are hydrates in which water molecules are isolated from direct contact with each other by intervening organic molecules. In channel hydrates, water molecules are located in lattice channels where they are in close proximity to other water molecules. In metal ion coordination hydrates, water molecules are bonded to the metal ion.

[0098] When the solvent or water is tightly bound, the complex can have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water / solvent content can depend on humidity and drying conditions. In this case, non-stoichiometry becomes the norm.

[0099] Also included within the scope of the present invention are multi-component complexes (other than salts and solvates) in which the drug and at least one other component are present in stoichiometric or non-stoichiometric amounts. Complexes of this type include clathrates (drug-host inclusion complexes) and co-crystals. The latter are typically defined as crystalline complexes of neutral molecular components bound together by non-covalent interactions, but can also be complexes of neutral molecules with salts. Co-crystals can be prepared by melt crystallization, recrystallization from a solvent, or physical grinding of the components together - see Chem Commun, 17, 1889-1896, O. Almarsson and MJ Zaworotko (2004). For a general review of multi-component complexes, see J Pharm Sci, 64(8), 1269-1288, Haleblian (August 1975).

[0100] Also included within the scope of the present invention are active metabolites of the compounds of Formula I (prodrugs), that is, compounds formed in vivo following administration of the drug, typically by oxidation or dealkylation. Some examples of metabolites according to the present invention include:

[0101] (i) in the case where the compound of formula I contains a methyl group, its hydroxymethyl derivative (-CH3->-CH2OH), and

[0102] (ii) In case the compound of formula I contains an alkoxy group, its hydroxy derivative (-OR->-OH).

[0103] The compounds of the present invention can exist in a solid state continuum from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which a material lacks long-range order at the molecular level and can exhibit the physical properties of a solid or liquid depending on the temperature. Typically, such materials do not give a characteristic X-ray diffraction pattern and, although exhibiting the properties of a solid, are more formally described as liquids. Upon heating, a change from solid to liquid properties occurs, characterized by a change in state, typically a second-order change ("glass transition"). The term "crystalline" refers to a solid phase in which a material has a regular, ordered internal structure at the molecular level and gives a characteristic X-ray diffraction pattern with well-defined peaks. When fully heated, such materials will also exhibit the properties of a liquid, but the change from solid to liquid is characterized by a phase transition, typically a first-order change ("melting point").

[0104] When subjected to suitable conditions, the compounds of Formula I can also exist in a mesomorphic state (mesogenic phase or liquid crystal). The mesomorphic state is an intermediate state between a true crystalline state and a true liquid state (melt or solution). The mesomorphism caused by temperature changes is described as "thermotropic", while the mesomorphism caused by the addition of a second component (such as water or another solvent) is described as "lyotropic". Compounds with the potential to form lyotropic mesomorphic phases are described as "amphiphilic" and are formed by having ions (such as -COO - Na + 、-COO - K + , or -SO3 - Na + ) or non-ionic (such as -N - N + (CH3)3) Molecular composition of the polar head group. For more information, see Crystals and the Polarizing Microscope, NH Hartshorne and A. Stuart, 4th edition (Edward Arnold, 1970).

[0105] The compounds of Formula I may exhibit polymorphism and / or one or more isomerisms (e.g., optical, geometric, or tautomerism). The compounds of Formula I may also be isotopically labeled. Such variations are implicit in the compounds of Formula I as defined with reference to their structural features and are therefore within the scope of the present invention.

[0106] The terms "concentrate", "evaporate" and "concentrate in vacuo" refer to the removal of solvent under reduced pressure on a rotary evaporator at a bath temperature below 60° C. The abbreviations "min" and "h" stand for "minutes" and "hours", respectively. The term "room temperature or ambient temperature" means a temperature of 18 to 25°C, "GCMS" means gas chromatography-mass spectrometry, "LCMS" means liquid chromatography-mass spectrometry, "UPLC" means ultra-high performance liquid chromatography, "SFC" means supercritical fluid chromatography, "HPLC" means high pressure liquid chromatography, "MPLC" means medium pressure liquid chromatography, "TLC" means thin layer chromatography, "MS" means mass spectrometry or mass spectroscopy or mass spectrometry, "NMR" means nuclear magnetic resonance spectroscopy, "DCM" means dichloromethane, "DMSO" means dimethyl sulfoxide, "DME" means 1,2-dimethoxyethane, "EtOAc" means ethyl acetate, "MeOH" means methanol, "Ph" means phenyl, "Pr" means propyl, "trityl" means triphenylmethyl, "ACN" means acetonitrile, "DEAD" means diethyl azodicarboxylate, and "DIAD" means diisopropyl azodicarboxylate.

[0107] In general, the compounds of the present invention can be prepared by methods including those similar to those known in the chemical arts, particularly according to the descriptions contained herein. Specific methods for making the compounds of the present invention are provided as further features of the present invention and are illustrated by the following reaction schemes. Other methods can be described in the experimental section. The following summarizes the specific synthetic routes for preparing the compounds of Formula I.

[0108] As used herein, the expressions "reaction inert solvent" and "inert solvent" refer to a solvent or mixtures thereof that do not interact with the starting materials, reagents, intermediates, or products in a manner that adversely affects the yield of the desired product.

[0109] First, it should be noted that when preparing compounds of Formula I, it should be noted that some preparation methods that can be used to prepare the compounds described herein may require protection of distal functional groups (e.g., primary amines, secondary amines, carboxyl groups in Formula I precursors). The need for such protection will vary depending on the nature of the distal functional group and the conditions of the preparation method. The need for such protection is easily determined by those skilled in the art. The use of such protection / deprotection methods is also within the skill of the art. For a general description of protecting groups and their uses, see TW Greene, Protective Groups in Organic Synthesis, John Wiley & Sons, New York, 1991.

[0110] For example, some compounds contain primary amine or carboxylic acid functional groups, which, if not protected, may interfere with reactions at other sites of the molecule. Therefore, such functional groups may be protected by suitable blocking groups that can be removed in subsequent steps. Suitable blocking groups for amine and carboxylic acid protection include those commonly used in peptide synthesis (such as N-tert-butyloxycarbonyl, benzyloxycarbonyl, and 9-fluorenylmethyleneoxycarbonyl for amines, and lower alkyl groups or benzyl esters for carboxylic acids), which are generally not chemically reactive under the described reaction conditions and can typically be removed without chemically altering other functional groups in the compound of Formula I.

[0111] The compounds and intermediates of formula I can contain asymmetric or chiral centers and therefore exist in different stereoisomeric forms. Unless otherwise indicated, all stereoisomeric forms of the compound and their mixtures (including racemic mixtures) are intended to be included herein. In addition, all geometric and positional isomers are included within the scope of the compound. For example, if the compound introduces a double bond or a fused ring, both cis and trans forms and mixtures are included within the scope of the present invention.

[0112] In addition, the compounds and intermediates of Formula I encompass all atropisomers and stereoisomeric mixtures thereof, including racemic mixtures. Atropisomers include those that can be separated into individual stereoisomers and maintain their stereoisomeric purity for varying lengths of time (including medium and long periods). Atropisomers also include those that cannot be readily separated into individual stereoisomers due to interconversion within a period of time (including short to medium periods).

[0113] The chiral compounds of the present invention (and chiral precursors thereof) can be obtained in enantiomerically enriched form using chromatography (typically high pressure liquid chromatography (HPLC) or supercritical fluid chromatography (SFC)) on a resin having an asymmetric stationary phase and a mobile phase consisting of a hydrocarbon (typically heptane or hexane) containing 0 to 50% (typically 2 to 20%) isopropanol and 0 to 5% alkylamine (typically 0.1% diethylamine (DEA) or isopropylamine). Concentration of the eluate yields an enriched mixture.

[0114] Diastereomeric mixtures can be separated into their individual diastereomers based on their physicochemical differences by methods well known to those skilled in the art (such as by chromatography and / or fractional crystallization). Enantiomers can be separated in the following manner: by reacting with an appropriate optically active compound (e.g., a chiral auxiliary such as a chiral alcohol or Moshe's acyl chloride), the enantiomeric mixture is converted into a diastereomeric mixture, the diastereomers are separated, and each diastereomer is converted (e.g., hydrolyzed) into the corresponding pure enantiomer. Enantiomers can also be separated by using a chiral HPLC column. Alternatively, specific stereoisomers can be synthesized by using optically active raw materials, by using an asymmetric synthesis of optically active reagents, substrates, catalysts or solvents, or by asymmetric transformation, one stereoisomer is converted into another stereoisomer.

[0115] Where a compound has two or more stereocenters and the absolute or relative stereochemistry is given in the name, the nomenclature R and S refers to each stereocenter numbered in ascending order (1, 2, 3, etc.) according to the conventional IUPAC numbering scheme for individual molecules. Where a compound has one or more stereocenters and the stereochemistry is not given in the name or structure, it is understood that the name or structure is intended to encompass all forms of the compound, including racemic forms.

[0116] The compounds of the present invention may contain olefin-like double bonds. When such bonds are present, the compounds of the present invention exist in cis and trans configurations, as well as mixtures thereof. The term "cis" refers to the orientation of two substituents relative to each other and the plane of the ring (both "up" or both "down"). Similarly, the term "trans" refers to the orientation of two substituents relative to each other and the plane of the ring (substituents are on opposite sides of the ring).

[0117] It is also possible that the intermediates and compounds of formula I can exist in different tautomeric forms, and all such forms are encompassed within the scope of the present invention. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can be mutually converted via a low energy barrier. For example, proton tautomers (also referred to as prototropic tautomers) include mutual conversions via proton migration, such as keto-enol and imine-enamine isomerizations. The specific example of proton tautomers is a tetrazole moiety, in which a proton can migrate between four ring nitrogens as follows.

[0118]

[0119] Valence tautomers include interconversions by reorganization of some of the bonding electrons.

[0120] Included within the scope of the compounds claimed herein are all stereoisomers, geometric isomers, and tautomeric forms of the compounds of Formula I, including compounds exhibiting more than one type of isomerism, and mixtures of one or more thereof. Also included are acid addition salts or base salts in which the counterion is optically active, such as D-lactate or L-lysine, or racemic, such as DL-tartrate or DL-arginine.

[0121] Compounds of Formula I can be prepared according to the general routes and examples provided herein.

[0122] General route

[0123] In general, the compounds of the present invention can be prepared by the methods described herein and similar methods known to those skilled in the art. Specific methods for making the compounds of the present invention are described in the following reaction schemes. Other methods are described in the experimental section. The routes and examples provided herein (including corresponding descriptions) are for illustration only. The substituents marked in Schemes 1-7 are as described in the application, wherein PMB is p-methoxybenzyl ether and Boc is tert-butyloxycarbonyl.

[0124] Route 1 refers to the preparation of a compound of formula IA. The compound of formula IA can be easily prepared by intermediates IV, VI and VIII. Intermediate IV can be prepared by the amide bond formation reaction between carboxylic acid intermediate II and amine intermediate III. Similarly, intermediates VI and VIII can be prepared by the amide bond formation reaction between intermediate II and intermediates V and VII, respectively. By in a suitable solvent (such as dichloromethane), in the presence of an activating reagent (such as O-(7-azabenzotriazole-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate; HATU) and a base (such as N, N-diisopropylethylamine), a carboxylic acid (such as II) and an amine (such as III, V or VII) are combined to achieve this type of amide bond formation reaction.

[0125]

[0126] Route 2 refers to the preparation of compounds of formula IA-1, IA-2, IA-3 and IA-4 from intermediate IV. The ester in intermediate IV can be hydrolyzed to obtain intermediate IX. The carboxylic acid functional group in intermediate IX can be converted into various heteroaryl ring systems by methods known to those skilled in the art. For example, intermediate IX can react with aminophenol (such as X) under suitable conditions to obtain a compound of formula IA-1 after removing the PMB protecting group. Alternatively, intermediate IX can be coupled with an intermediate of structure XI, and the resulting compound can be further dehydrated and deprotected to obtain a compound of formula IA-2. Those skilled in the art can also recognize that the carboxylic acid in intermediate IX can be converted into an alternative functional group, which can have additional functions for constructing other heteroaryl ring systems. For example, the carboxylic acid in compound IX can be converted into bromoketone by methods known in the art to obtain intermediate XII. Intermediate XII can react with aminopyridine (XIII) and then deprotect to prepare a compound of formula IA-3. Alternatively, the carboxylic acid in IX can be converted to a primary amide and then dehydrated to obtain a nitrile-containing intermediate of structure XIV. Intermediate XIV can be reacted with hydroxylamine to obtain compound XV. The compound of structure XV can be reacted with a carboxylic acid of structure XVI. The resulting compound can be dehydrated and deprotected to form an oxadiazole-containing compound of formula IA-4.

[0127]

[0128] Route 3 refers to the preparation of compounds of formula IA-5 and IA-6 by intermediate VI. The Boc protecting group in intermediate VI can be selectively removed to obtain intermediate XVII. Intermediate XVII can react with a nitroaldehyde-containing compound (XVIII) in the presence of a trialkylphosphine to obtain a compound of formula IA-5 after removing the PMB protecting group. Alternatively, compound XVII can react with a bromoester-containing compound (XIX) and then deprotect to obtain a compound of formula IA-6.

[0129]

[0130] Route 4 refers to the preparation of compounds of formula IA and IA-7 from intermediate VIII. The intermediate of structure VIII can react with aryl and heteroaryl halides (XX) in the presence of a suitable metal-containing catalyst and ligand to obtain a compound of formula IA after removing the PMB protecting group. Alternatively, sodium azide can be used to replace the bromide in intermediate VIII. The resulting intermediate can react with an alkyne-containing compound (XXI) in the presence of a copper catalyst to obtain a compound of formula IA-7.

[0131]

[0132] Route 5 refers to an alternative preparation of a compound of formula IA-5. In some cases, the compound can be prepared by the method described herein, which contains substituents that can be synthetically used to prepare an alternative compound of formula IA. For example, the intermediate of structure XXII can be prepared by the method described for preparing the compound of formula IA-5. The bromine substituent in intermediate XXII can be reacted with boronic acid (XXIII) or boric ester (XXIII) by Suzuki reaction to obtain a compound of formula IA-5. In addition, the compound of structure XXII can be reacted with an intermediate of structure XXIV, wherein BH represents a primary amine or a secondary amine. In this case, XXII and XXIV can react with each other under Buchwald reaction conditions to obtain a compound or another variant of formula IA-5. Alternatively, the bromine substituent in XXII can be converted to boronic acid (XXV; R=H) or boric ester (XXV; R=alkyl). The compound of structure XXV can react with aryl and heteroaryl halides of structure XXVI to obtain a compound of formula IA-5. Alternatively, compounds of structure XXV can be reacted with aromatic heterocycles (XXIV') bearing NH under Cham-Lam coupling conditions to yield compounds of formula IA-5. The transformation examples provided in Scheme 5 are not intended to be comprehensive. The examples provided are merely independent examples of synthetic sequences that can be used to modify the B- and C-substituents of compounds of formula IA. One skilled in the art will also recognize that similar transformations can be achieved using compounds containing alternative A-substituents as shown in Scheme 5.

[0133]

[0134] Route 6 refers to the preparation of a compound of formula IB. The compound of formula IB can be easily prepared by intermediates XXIX and XXX. Intermediate XXIX can be prepared by the amide bond formation reaction between carboxylic acid intermediate II and amine intermediate XXVII. Similarly, intermediate XXX can be prepared by the amide bond formation reaction between intermediate II and intermediate XXVIII. By in a suitable solvent (such as dichloromethane), in the presence of an activating reagent (such as O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; HATU) and a base (such as N,N-diisopropylethylamine), a carboxylic acid (such as II) and an amine (such as XXVII or XXVIII) are combined to achieve this type of amide bond formation reaction. By methods similar to those described for preparing the compound of formula IA by intermediate IV in routes 2 and 5, the preparation of the compound of formula IB can be achieved by intermediate XXIX. Likewise, the preparation of compounds of formula IB can be accomplished from intermediate XXX by methods analogous to those described for the preparation of compounds of formula IA from intermediate VI in Schemes 3 and 5.

[0135]

[0136] Route 7 refers to an alternative sequence of synthetic steps that can be used to prepare compounds of Formula IA or compounds of Formula IB. For example, intermediates such as XXXI, XXXII, or XXXIII can be converted to intermediates of structure XXXIV via the methods described herein. The amine intermediate of structure XXXIV can react with a carboxylic acid of structure II in an amide bond-forming reaction. The resulting product can be deprotected to obtain a compound of Formula IA. Similarly, intermediates such as XXXV and XXXVI can be converted to intermediates of structure XXXVII. The amine intermediate of structure XXXVII can react with a carboxylic acid of structure II and then deprotected to obtain a compound of Formula IB.

[0137]

[0138] The raw materials and reagents used for the above-mentioned compounds of formula I are also readily available or can be readily synthesized by those skilled in the art using conventional organic synthesis methods. For example, many of the compounds used herein are related to or derived from compounds of great scientific interest and commercial demand, and therefore many such compounds are commercially available, or have been reported in the literature, or can be readily prepared from other commonly available substances by methods reported in the literature.

[0139] The present application also relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I or a pharmaceutically acceptable salt of said compound and a pharmaceutically acceptable carrier, vehicle or diluent.

[0140] The compounds of the present invention may also be used in combination with other agents (e.g., anti-atherosclerotic drugs and anti-thrombotic drugs) to treat the diseases / conditions described herein. The present application also relates to a pharmaceutical combination composition comprising: a therapeutically effective amount of a composition having:

[0141] a first compound, which is a compound of any of Formula I or a pharmaceutically acceptable salt thereof;

[0142] a second compound which is a therapeutic agent for kidney disease, an antidiabetic agent, a therapeutic agent for nonalcoholic steatohepatitis, a therapeutic agent for nonalcoholic fatty liver disease, or an anti-heart failure therapeutic agent, and

[0143] Pharmaceutically acceptable carrier, vehicle or diluent.

[0144] In one embodiment, the agent treating kidney disease can be used to treat acute and / or chronic kidney disease.

[0145] In one embodiment, the agent treating nonalcoholic steatohepatitis or nonalcoholic fatty liver disease is an ACC inhibitor, a KHK inhibitor, a DGAT-2 inhibitor, a FXR agonist, a GLP-1R agonist, metformin, an incretin analog, or an incretin receptor modulator.

[0146] In another embodiment, the antidiabetic agent is a SGLT-2 inhibitor, metformin, an incretin mimetic, an incretin receptor modulator, a DPP-4 inhibitor, or a PPAR agonist.

[0147] In another embodiment, the antidiabetic agent is metformin, sitagliptin, or ergliflozin.

[0148] In another embodiment, the anti-heart failure agent is an ACE inhibitor, angiotensin receptor blocker, angiotensin receptor neprilysin inhibitor, beta adrenergic receptor blocker, calcium channel blocker, or vasodilator.

[0149] Combination Potion

[0150] The compounds can be administered alone or in combination with one or more additional therapeutic agents. "Administered in combination" or "combination therapy" means that the compound and one or more additional therapeutic agents are administered simultaneously to the mammal being treated. When administered in combination, each component can be administered simultaneously or sequentially in any order at different time points. Thus, the individual components can be administered separately but sufficiently close in time to provide the desired therapeutic effect. The phrases "administered in parallel," "administered together," "administered simultaneously," and "administered simultaneously" mean that the compounds are administered in combination. Thus, the methods of prevention and treatment described herein include the use of combination agents.

[0151] The combination agent is administered to a mammal in a therapeutically effective amount. "Therapeutically effective amount" means an amount of a compound of Formula I that is effective to treat the desired disease / condition (e.g., NASH, heart failure, kidney disease, or diabetes) when administered to a mammal alone or in combination with an additional therapeutic agent.

[0152] In view of the NASH / NAFLD activity of the compounds of the present invention, they can be co-administered with other agents for the treatment of non-alcoholic steatohepatitis (NASH) and / or non-alcoholic fatty liver disease (NAFLD) and associated diseases / conditions, such as orlistat, TZD and other insulin sensitizers, FGF21 analogs, metformin, ω-3-acid ethyl esters (e.g., lovastatin), fibrates, HMG-CoA reductase inhibitors (e.g., pravastatin, lovastatin, atorvastatin, simvastatin, fluvastatin, NK-104 (also known as pitavastatin or nivastatin or nisbastatin) and ZD-4522 (also known as rosuvastatin or atorvastatin or vivastatin), ezetimibe, proprotein convertase subtilisin Kexin PCSK9 inhibitors (e.g., evolocumab, alirocumab), probucol, ursodeoxycholic acid, TGR5 agonists, FXR agonists, vitamin E, betaine, pentoxifylline, CB1 antagonists, carnitine, N-acetylcysteine, reduced glutathione, lorcaserin, naltrexone in combination with buproprion, SGLT2 inhibitors (including dapagliflozin, canagliflozin, empagliflozin, togliflozin), , ergliflozin, ASP-1941, THR1474, TS-071, ISIS388626 and LX4211 and those in WO2010023594), phentermine, topiramate, GLP-1 receptor agonists, GIP receptor agonists, dual GLP-1 receptor / glucagon receptor agonists (i.e., OPK88003, MEDI0382, JNJ-64565111, NN9277, BI 456906), dual GLP-1 receptor / GIP receptor agonists (i.e., tesiparatide (LY3298176), NN9423), angiotensin receptor blockers, acetyl-CoA carboxylase (ACC) inhibitors, diacetylglycerol O-acyltransferase 1 (DGAT-1) inhibitors (such as those described in WO09016462 or WO2010086820, AZD7687 or LCQ908), diacetylglycerol O-acyltransferase 2 (DGAT-2) inhibitors, PNPLA3 inhibitors, FGF21 analogs, FGF19 analogs, PPAR agonists, FXR agonists, AMPK activators, SCD1 inhibitors, or MPO inhibitors.

[0153] Exemplary GLP-1 receptor agonists include liraglutide, albiglutide, exenatide, albiglutide, lixisenatide, dulaglutide, semaglutide, HM15211, LY3298176, Medi-0382, NN-9924, TTP-054, TTP-273, apegenatide, those described in WO2018109607, those described in PCT / IB2019 / 054867 filed June 11, 2019, including the following:

[0154] 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0155] 2-({4-[2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0156] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0157] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0158] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0159] 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0160] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0161] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-3-(1,3-oxazol-2-ylmethyl)-3H-imidazo[4,5-b]pyridine-5-carboxylic acid;

[0162] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0163] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-4-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0164] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(pyridin-3-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0165] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-5-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0166] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-1,2,3-triazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0167] 2-({4-[2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0168] 2-({4-[2-(4-chloro-2-fluorophenyl)-7-fluoro-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0169] 2-({4-[2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-(1,3-oxazol-2-ylmethyl)-1H-benzimidazole-6-carboxylic acid;

[0170] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0171] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0172] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-7-fluoro-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0173] 2-({4-[(2S)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0174] 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0175] 2-({4-[(2S)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0176] 2-({4-[(2R)-2-(4-cyano-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0177] 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0178] 2-({4-[(2R)-2-(4-chloro-2-fluorophenyl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(1-ethyl-1H-imidazol-5-yl)methyl]-1H-benzimidazole-6-carboxylic acid;

[0179] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0180] 2-({4-[(2S)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0181] 2-({4-[(2R)-2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0182] 2-({4-[2-(5-chloropyridin-2-yl)-2-methyl-1,3-benzodioxol-4-yl]piperidin-1-yl}methyl)-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid, DIAST-X2; and

[0183] 2-[(4-{6-[(4-cyano-2-fluorobenzyl)oxy]pyridin-2-yl}piperidin-1-yl)methyl]-1-[(2S)-oxetan-2-ylmethyl]-1H-benzimidazole-6-carboxylic acid;

[0184] or pharmaceutically acceptable salts thereof.

[0185] Exemplary ACC inhibitors include 4-(4-[(1-isopropyl-7-oxo-1,4,6,7-tetrahydro-1'H-spiro[indazole-5,4'-piperidinyl]-1'-yl)carbonyl]-6-methoxypyridin-2-yl)benzoic acid; and firsocostat (GS-0976) and pharmaceutically acceptable salts thereof.

[0186] Exemplary FXR agonists include tropifexor (2-[(1R,3R,5S)-3-({5-cyclopropyl-3-[2-(trifluoromethoxy)phenyl]-1,2-oxazol-4-yl}methoxy)-8-azabicyclo[3.2.1]octan-8-yl]-4-fluoro-1,3-benzothiazole-6-carboxylic acid); cilofexor (GS-9674); obeticholic acid; LY2562175; Met409; TERN-101; and EDP-305, and pharmaceutically acceptable salts thereof.

[0187] Exemplary DGAT2 inhibitors include (S)-2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-(tetrahydrofuran-3-yl)pyrimidine-5-carboxamide;

[0188] 2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-((3R,4S)-4-fluoropiperidin-3-yl)pyrimidine-5-carboxamide;

[0189] 2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-((3S,5S)-5-fluoropiperidin-3-yl)pyrimidine-5-carboxamide;

[0190] 2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-((3R,4S)-4-fluoropiperidin-3-yl)pyrimidine-5-carboxamide;

[0191] 2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-((3R,4R)-4-fluoropiperidin-3-yl)pyrimidine-5-carboxamide;

[0192] 2-(5-((3-ethoxy-5-fluoropyridin-2-yl)oxy)pyridin-3-yl)-N-((3R,4R)-4-fluoropiperidin-3-yl)pyrimidine-5-carboxamide; and

[0193] 2-(5-((3-ethoxypyridin-2-yl)oxy)pyridin-3-yl)-N-((3S,5S)-5-fluoropiperidin-3-yl)pyrimidine-5-carboxamide;

[0194] or pharmaceutically acceptable salts thereof.

[0195] Exemplary KHK inhibitors include [(1R,5S,6R)-3-{2-[(2S)-2-methylazetidin-1-yl]-6-(trifluoromethyl)pyrimidin-4-yl}-3-azabicyclo[3.1.0]hexan-6-yl]acetic acid and pharmaceutically acceptable salts thereof.

[0196] In view of the anti-diabetic activity of the compounds of the present invention, they can be co-administered with other anti-diabetic agents. Suitable anti-diabetic agents include insulin, metformin, GLP-1 receptor agonists (described above), acetyl-CoA carboxylase (ACC) inhibitors (described above), SGLT2 inhibitors (described above), monoacylglycerol-O-acyltransferase inhibitors, phosphodiesterase (PDE)-10 inhibitors, AMPK activators, sulfonylureas (e.g., acetophenonesulfonylcyclohexamide, chlorpropamide, termipramide, glibenclamide, glipizide, glyburide, glimepiride, gliclazide, glivalamide, gliquidone, glixolamide, tolazamide and tolbutamide), meglitinide, α-amylase inhibitors (e.g., amylase inhibitors, tretinoin and AL-3688), α-glucosidase hydrolase inhibitors (e.g., acarbose), α-glucosidase inhibitors (e.g., lipolysin, canaglitazone, emiglitazone, miglitol, voglibose, pramixin-Q, and saponin), PPARγ agonists (e.g., balaglitazone, ciglitazone, darglitazone, englitazone, isaglitazone, pioglitazone, and rosiglitazone), PPARα / γ agonists (e.g., CLX-0940, GW-1536, GW-1929, GW-2433, KRP-297, L-796449, LR-90, MK-0767, and SB-219994), protein tyrosine phosphatase-1B (PTP-1B) inhibitors (e.g., trodusquemine, hyrtiosal extract, and Zhang, S. et al., Drug Discovery Today, 12 (9 / 10), 373-381 (2007)), SIRT-1 activators (e.g., resveratrol, GSK2245840 or GSK184072), dipeptidyl peptidase IV (DPP-IV) inhibitors (e.g., those in WO2005116014, sitagliptin, vildagliptin, alogliptin, dulagliptin, linagliptin and saxagliptin), insulin secretagogues, fatty acid oxidation inhibitors, A2 antagonists, c-jun amino-terminal kinase (JNK) inhibitors, glucokinase activators ( GKa) (such as those described in WO2010103437, WO201010343f8, WO2010013161, WO2007122482, TTP-399, TTP-355, TTP-547, AZD1656, ARRY403, MK-0599, TAK-329, AZD5658 or GKM-001), insulin, insulin mimetics, glycogen phosphorylase inhibitors (e.g., GSK1362885), VPAC2 receptor agonists, glucagon receptor modulators (such as Demong, DEet al., Annual Reports in Medicinal Chemistry 2008, 43, 119-137), GPR119 modulators (particularly agonists such as those described in WO2010140092, WO2010128425, WO2010128414, WO2010106457, Jones, RM et al., Medicinal Chemistry 2009, 44, 149-170 (e.g., MBX-2982, GSK1292263, APD597 and PSN821)), FGF21 derivatives or analogs (such as those described in Kharitonenkov, A. et al., Current Opinion in Investigational Drugs 2009, 10(4)359-364), TGR5 (also known as GPBAR1) receptor modulators (particularly agonists such as those described in Zhong, M., Current Topics in Medicinal Chemistry, 2010, 10 (4), 386-396 and INT777), GPR40 agonists (such as those described in Medina, JC, Annual Reports in Medicinal Chemistry, 2008, 43, 75-85, including but not limited to TAK-875), GPR120 modulators (particularly agonists), high affinity nicotinic acid receptor (HM74A) activators, and SGLT1 inhibitors (such as GSK1614235). A further representative list of antidiabetic agents that can be combined with the compounds of the present application can be found, for example, on page 28, line 35 to page 30, line 19 of WO2011005611.

[0197] Other antidiabetic agents may include inhibitors or modulators of carnitine palmitoyltransferase, inhibitors of fructose 1,6-bisphosphatase, inhibitors of aldose reductase, mineralocorticoid receptor inhibitors, inhibitors of TORC2, inhibitors of CCR2 and / or CCR5, inhibitors of PKC isoforms (e.g., PKCα, PKCβ, PKCγ), inhibitors of fatty acid synthase, inhibitors of serine palmitoyltransferase, GPR81, GPR39, GPR43, GPR41, GPR105, Kv1.3, retinol binding protein 4, glucocorticoid receptor, modulators of somatostatin receptors (e.g., SSTR1, SSTR2, SSTR3 and SSTR5), inhibitors or modulators of PDHK2 or PDHK4, inhibitors of MAP4K4, modulators of the IL1 family (including IL1β), modulators of RXRα. Additionally, suitable antidiabetic agents include the mechanisms listed by Carpino, PA, Goodwin, B. Expert Opin. Ther. Pat, 2010, 20(12), 1627-51.

[0198] In view of the anti-heart failure activity of the compounds of the present application, they can be co-administered with the following other anti-heart failure agents: ACE inhibitors (e.g., captopril, enalapril, fosinopril, lisinopril, perindopril, quinapril, ramipril, trandolapril), angiotensin II receptor blockers (e.g., candesartan, losartan, valsartan), angiotensin receptor neprilysin inhibitors (sacubitril / valsartan), I f Channel blockers ivabradine, beta-adrenergic blockers (e.g., bisoprolol, metoprolol succinate, carvedilol), SGLT2 inhibitors, aldosterone antagonists (e.g., spironolactone, eplerenone), cardiac myosin activators (e.g., omecamtiv mecarbil), guanylate cyclase stimulators (e.g., vericigide), cardiac myosin inhibitors (e.g., mavacetal), SERCA2a activators (e.g., istaroxime), hydralazine and isosorbide dinitrate, diuretics (e.g., furosemide, bumetanide, torsemide, chlorothiazide, amiloride, hydrochlorothiazide, indapamide, metolazone, triamterene), or digoxin.

[0199] The compounds of Formula I can also be used in combination with antihypertensive agents, and such antihypertensive activity is readily determined by one skilled in the art according to standard assays (e.g., blood pressure measurements). Examples of suitable antihypertensive agents include: alpha-adrenergic blockers; beta-adrenergic blockers; calcium channel blockers (e.g., diltiazem, verapamil, nifedipine, and amlodipine); vasodilators (e.g., hydralazine), diuretics (e.g., chlorothiazide, hydrochlorothiazide, flumethiazide, hydroflumethiazide, bendroflumethiazide, methylchlorothiazide, trichlorothiazide, polythiazide, benzthiazide, ethacrynic acid triclosan, chlorpromazine ... tricrynafen, chlorthalidone, torsemide, furosemide, musolimine, bumetanide, triamtrenene, amiloride, spironolactone); renin inhibitors; ACE inhibitors (e.g., captopril, zofenopril, fosinopril, enalapril, ceranopril, cilazopril, delapril, penttopril, quinapril, ramipril, lisinopril); AT-1 receptor antagonists (e.g., losartan, irbesartan, valsartan); ET receptor antagonists (e.g., sitaxsentan, atrasentan, and compounds disclosed in U.S. Pat. Nos. 5,612,359 and 6,043,265); dual ET / AII antagonists (e.g., WO 00 / 01389); neutral endopeptidase (NEP) inhibitors; vasopeptidase inhibitors (dual NEP-ACE inhibitors) (e.g., gemotrilat and nitrates). An exemplary anti-anginal agent is ivabradine.

[0200] Examples of suitable calcium channel blockers (L-type or T-type) include diltiazem, verapamil, nifedipine, amlodipine and mibefradil.

[0201] Examples of suitable cardiac glycosides include digitalis and ouabain.

[0202] In one embodiment, the compound of formula I may be co-administered with one or more diuretics. Examples of suitable diuretics include (a) Henle loop diuretics, such as furosemide (eg, LASIX TM ), torasemide (such as DEMADEX TM ), bemetanide (such as BUMEX TM ) and ethacrynic acid (such as EDECRIN TM ); (b) thiazide diuretics, such as chlorothiazide (such as DIURIL TM , ESIDRIX TM or HYDRODIURIL TM ), hydrochlorothiazide (such as MICROZIDETM ORETIC TM ), benzylthiazide, hydroflumethiazide (such as SALURON TM ), bendroflumethiazide, methychlorthiazide, polythiazide, trichloromethiazide, and indapamide (such as LOZOL TM ); (c) benzyl lactam-type diuretics, such as chlorthalidone (such as HYGROTON TM ) and metolazone (such as ZAROXOLYN TM ); (d) quinazoline-type diuretics, such as quinethazone; and (e) potassium-sparing diuretics, such as triamterene (eg, DYRENIUM TM ) and amiloride (such as MIDAMOR TM or MODURETIC TM ).

[0203] In another embodiment, the compound of Formula I can be co-administered with a Henle loop diuretic. In yet another embodiment, the Henle loop diuretic is selected from furosemide and torsemide. In yet another embodiment, one or more compounds of Formula I can be co-administered with furosemide. In yet another embodiment, one or more compounds of Formula I can be co-administered with torsemide (which may optionally be a controlled release or modified release form of torsemide).

[0204] In another embodiment, the compound of Formula I can be co-administered with a thiazide diuretic. In yet another embodiment, the thiazide diuretic is selected from chlorothiazide and hydrochlorothiazide. In yet another embodiment, one or more compounds of Formula I can be co-administered with chlorothiazide. In yet another embodiment, one or more compounds of Formula I can be co-administered with hydrochlorothiazide.

[0205] In another embodiment, one or more compounds of Formula I may be co-administered with a benzyl lactam-type diuretic. In yet another embodiment, the benzyl lactam-type diuretic is chlorthalidone.

[0206] Examples of suitable mineralocorticoid receptor antagonists include spironolactone and eplerenone.

[0207] Examples of suitable phosphodiesterase inhibitors include: PDE III inhibitors (such as cilostazol); and PDE V inhibitors (such as sildenafil).

[0208] Those skilled in the art will recognize that the compounds of the present invention may also be used in combination with other cardiovascular or cerebrovascular treatments (including PCI, stent implantation, drug-eluting stents, stem cell therapy) and medical devices (such as implanted pacemakers, defibrillators or cardiac resynchronization therapy).

[0209] The compounds of Formula I can also be used in combination with drugs used to manage chronic kidney disease, including phosphate binders (e.g., sucrose ferric oxyhydroxide, sevelamer, calcium acetate), sodium bicarbonate, erythropoietin stimulators, oral or intravenous iron supplements (e.g., iron sucrose, ferric carboxymaltose, nanoiron oxide (ferumoxytol)), potassium binders, calcitriol, or SGLT2 inhibitors (e.g., dapagliflozin, empagliflozin, or other SGLT2 inhibitors cited herein).

[0210] In particular, when provided as a single dose unit, there may be chemical interactions between the active ingredients of the combination. For this reason, when the compound of formula I and the second therapeutic agent are combined in a single dose unit, they can be formulated so that although the active ingredients are combined in a single dose unit, the physical contact between the active ingredients is minimized (i.e., reduced). For example, an active ingredient can be enteric coated. By enteric coating an active ingredient, not only the contact between the active ingredients of the combination may be minimized, but also one of these components may be controlled to release in the gastrointestinal tract so that one of these components is not released in the stomach but released in the intestine. One of the active ingredients can also be used in the entire gastrointestinal tract to achieve sustained release and also for minimizing the physical contact between the active ingredients of the combination. In addition, the component of sustained release can be additionally enteric coated so that the release of this component occurs only in the intestine. Another method relates to the preparation of a combination product, wherein one component is coated with a continuous and / or enteric release polymer, and other components are also coated with a polymer (such as low viscosity grade hydroxypropyl methylcellulose (HPMC) or other appropriate materials as known in the art) to further separate the active ingredients. Polymer coatings are used to form additional barriers to interaction with other components.

[0211] Sustained release formulations can be used. Suitable examples of sustained release formulations include semipermeable matrices of solid hydrophobic polymers containing the compound of the present invention, which are in the form of molded objects, such as films or microcapsules. Examples of sustained release matrices include polyesters, hydrogels (e.g., poly(2-hydroxyethyl methacrylate) or poly(vinyl alcohol)), polylactides (U.S. Patent No. 3,773,919), copolymers of L-glutamic acid and 7-ethyl L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT®, and the like. TM those used in (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D-(-)-3-hydroxybutyric acid.

[0212] These and other ways of minimizing contact between the components of the combination, whether administered in a single dosage form or in separate forms but simultaneously in the same manner, will be readily apparent to those skilled in the art once armed with this disclosure.

[0213] In combination therapy treatment, the compound of the invention and the other drug therapy are administered to a mammal (eg, a human, male or female) by conventional methods.

[0214] The compounds of formula I of the present invention, their prodrugs, and salts of such compounds and prodrugs are suitable for therapeutic use as agents that inhibit and / or degrade HSD17B13 in mammals, particularly humans, and are therefore useful in treating various disorders involving such effects (e.g., those described herein).

[0215] Diseases / conditions that may be treated with compounds of Formula I include, but are not limited to, NASH / NAFLD, diabetes, kidney disease, and heart failure and related diseases / conditions.

[0216] Therefore, given the positive correlation between activation of HSD17B13 and the development of NASH / NAFLD and associated diseases / disorders, the compounds of Formula I of the present invention, their prodrugs, and salts of such compounds and prodrugs can be used to prevent, arrest and / or regress fatty liver, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, non-alcoholic steatohepatitis with liver fibrosis, non-alcoholic steatohepatitis with cirrhosis, or non-alcoholic steatohepatitis with cirrhosis, and hepatocellular carcinoma due to their pharmacological effects.

[0217] The administration of the compounds of the present invention can be carried out via any method of systemic and / or local delivery of the compounds of the present invention. These methods include oral routes, parenteral, intraduodenal routes, buccal, intranasal, etc. Generally speaking, the compounds of the present invention are administered orally, but, for example, in cases where oral administration is not suitable for the purpose or in cases where the patient cannot take the drug, parenteral administration (e.g., intravenous, intramuscular, subcutaneous or intramedullary) can be used.

[0218] For the administration of human patients, the oral daily dose of the compound herein can be in the range of 1mg to 5000mg, which certainly depends on the mode and frequency of administration, disease state, and the age and condition of the patient, etc. An oral daily dose in the range of 3mg to 3000mg can be used. Other oral daily doses are in the range of 5mg to 1000mg. For convenience, the compound of Formula I can be administered in unit dosage form. If necessary, the unit dosage form of daily multiple doses can be used to increase the daily total dose. The unit dosage form can, for example, be a tablet or capsule containing about 0.1, 0.5, 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 125, 150, 175, 200, 250, 500 or 1000mg of compound. The total daily dose may be administered in single or divided doses and may, at the physician's discretion, fall outside of the typical range given herein.

[0219] For administration to human patients, the daily infusion dose of the compounds herein may be in the range of 1 mg to 2000 mg, depending of course on the mode and frequency of administration, the disease state, and the age and condition of the patient. Additional daily infusion doses are in the range of 5 mg to 1000 mg. The total daily dose may be administered in a single dose or in divided doses and may fall outside the typical ranges given herein at the physician's discretion.

[0220] The compounds may also be administered to animals other than humans, for example, for the indications detailed above. The exact dosage of each active ingredient administered will vary depending on many factors, including but not limited to the type of animal and the type of disease state being treated, the age of the animal, and the route of administration.

[0221] The dosage of the combination agent used in combination with the compound of formula I is effective for the indication being treated. Such dosage can be determined by standard assays (such as those cited above and provided herein). The combination agent can be administered simultaneously or sequentially in any order.

[0222] These dosages are based on an average human subject weighing about 60 kg to 70 kg. A physician will readily be able to determine dosages for subjects whose weight falls outside this range, such as infants and the elderly.

[0223] The dosage regimen may be adjusted to provide the optimal desired response. For example, a single bolus may be administered, several divided doses may be administered over time, or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation. It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and consistency of dosage. As used herein, a unit dosage form refers to a physically discrete unit suitable as a unit dosage for a mammalian subject to be treated; each unit contains a predetermined amount of active compound calculated to produce the desired therapeutic effect in combination with the required pharmaceutical carrier. The specifications of the unit dosage forms of the present invention are subject to and directly depend on (a) the unique characteristics of the chemotherapeutic agent and the specific therapeutic or prophylactic effect to be achieved, and (b) the inherent limitations in the art of formulating such active compounds for treating individual sensitivities.

[0224] Therefore, it will be understood by those skilled in the art that, based on the disclosure provided herein, dosages and administration regimens are adjusted according to methods known in the therapeutic field. That is, a maximum tolerated dose can be easily established, and an effective amount that provides a detectable therapeutic benefit to a patient can also be determined, which can be a temporary requirement for administering each agent to provide a detectable therapeutic benefit to a patient. Therefore, although certain dosages and administration regimens are exemplified herein, these examples in no way limit the dosages and administration regimens that can be provided to a patient.

[0225] It should be noted that dosage values ​​can vary according to the type and severity of the condition to be alleviated, and can include single or multiple doses. It should also be understood that for any particular subject, the specific dosing regimen should be adjusted over time according to individual needs and the professional judgment of the person administering or supervising the administration of the composition, and the dosage ranges set forth herein are merely exemplary and are not intended to limit the scope or practice of the claimed composition. For example, the dosage can be adjusted based on pharmacokinetic or pharmacodynamic parameters (which may include clinical effects such as toxic effects and / or laboratory values). Therefore, as determined by the skilled person, intra-patient dose escalation can be used. Determining the appropriate dosage and regimen for the administration of chemotherapeutic agents is well known in the relevant art, and once the teachings disclosed herein are provided, it will be understood by the skilled person to be encompassed.

[0226] The present application further comprises the use of a compound of Formula I as a medicament (such as a unit dose tablet or a unit dose capsule). In another embodiment, the present application comprises the use of a compound of Formula I for the manufacture of a medicament (such as a unit dose tablet or a unit dose capsule) to treat one or more conditions previously identified in the section discussing methods of treatment above.

[0227] The pharmaceutical compositions of the present invention can be prepared, packaged, or sold in bulk as a single unit dose or as multiple single unit doses. As used herein, a "unit dose" is a discrete amount of a pharmaceutical composition containing a predetermined amount of an active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient to be administered to a subject, or a convenient fraction of such a dose, for example, one-half or one-third of such a dose.

[0228] The compounds of the present invention or combinations can be administered alone, but are typically administered in admixture with one or more suitable pharmaceutical excipients, adjuvants, diluents, or carriers known in the art and selected with respect to the intended route of administration and standard pharmaceutical practice. The compounds of the present invention or combinations can be formulated into dosage forms that provide immediate release, delayed release, regulated release, sustained release, pulsed release, or controlled release, depending on the desired route of administration and the specificity of the release profile (commensurate with therapeutic needs).

[0229] Pharmaceutical compositions contain a compound or combination of the invention in an amount typically in the range of about 1% to about 75%, 80%, 85%, 90% or even 95% (by weight) of the composition, typically in the range of about 1%, 2% or 3% to about 50%, 60% or 70%, more usually in the range of about 1%, 2% or 3% to less than 50%, such as about 25%, 30% or 35%.

[0230] Methods for preparing various pharmaceutical compositions having specific amounts of active compounds are known to those skilled in the art. For example, see Remington: The Practice of Pharmacy, Lippincott Williams and Wilkins, Baltimore Md. 20th edition supplement, 2000.

[0231] Compositions suitable for parenteral injection generally include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Examples of suitable aqueous and non-aqueous carriers or diluents (including solvents and vehicles) include water, ethanol, polyols (propylene glycol, polyethylene glycol, glycerol, etc.), suitable mixtures thereof, triglycerides (including vegetable oils such as olive oil), and injectable organic esters such as ethyl oleate. Preferred carriers are available from Condea Vista Co. (Cranford, NJ). Branded caprylic / capric esters with glycerin or propylene glycol (e.g. 812, 829, 840). Proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.

[0232] These compositions for parenteral injection may also contain excipients such as preservatives, wetting agents, emulsifiers, and dispersants. The prevention of microbial contamination of the composition may be achieved with various antibacterial and antifungal agents (e.g., parabens, chlorobutanol, phenol, sorbic acid, etc.). It may also be desirable to include isotonic agents such as sugars, sodium chloride, etc. The prolonged absorption of the injectable pharmaceutical composition may be achieved by using agents that delay absorption (e.g., aluminum monostearate and gelatin).

[0233] Solid dosage forms for oral administration include capsules, tablets, chewable tablets, lozenges, pills, powders, and multiparticulate formulations (granules). In such solid dosage forms, the compound of Formula I or combination is mixed with at least one inert excipient, diluent, or carrier. Suitable excipients, diluents, or carriers include materials such as sodium citrate or dicalcium phosphate and / or (a) one or more fillers or extenders (e.g., microcrystalline cellulose (which may be (e.g., starch, lactose, sucrose, mannitol, silicic acid, xylitol, sorbitol, dextrose, calcium hydrogen phosphate, dextrin, α-cyclodextrin, β-cyclodextrin, polyethylene glycol, medium chain fatty acids, titanium oxide, magnesium oxide, aluminum oxide, etc.); (b) one or more binders (e.g., carboxymethyl cellulose, methyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, gelatin, gum arabic, ethyl cellulose, polyvinyl alcohol, pullulan, pregelatinized starch, agar, tragacanth gum, alginates, gelatin, polyvinyl pyrrolidone, sucrose, gum arabic, etc.); (c) one or more humectants (e.g., glycerin, etc.); (d) one or more disintegrants (e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, sodium carbonate, sodium lauryl sulfate, sodium starch glycolate (can be available from Edward Mendell Co.), cross-linked polyvinyl pyrrolidone, cross-linked sodium carboxymethyl cellulose type A (can be (obtained), polyacrilin potassium (ion exchange resin), etc.); (e) one or more solution retarders (e.g., paraffin, etc.); (f) one or more absorption accelerators (e.g., quaternary ammonium compounds, etc.); (g) one or more wetting agents (e.g., cetyl alcohol, glyceryl monostearate, etc.); (h) one or more adsorbents (e.g., kaolin, bentonite, etc.); and / or (i) one or more lubricants (e.g., talc, calcium stearate, magnesium stearate, stearic acid, polyethylene glycol stearate, cetyl alcohol, talc, hydrogenated castor oil, sucrose fatty acid esters, dimethyl polysiloxane, microcrystalline wax, yellow beeswax, white beeswax, solid polyethylene glycol, sodium lauryl sulfate, etc.). In the case of capsules and tablets, the dosage form may also contain a buffer.

[0234] Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatin capsules using such excipients as lactose or sugar paste as well as high molecular weight polyethylene glycols.

[0235] Solid dosage forms such as tablets, dragees, capsules and granules can be prepared to have coatings and shells, such as enteric coatings and other coatings well known in the art. They can contain sunscreens and can also have such compositions that release the compound of Formula I and / or additional medicaments in a delayed manner. Examples of spendable embedding compositions are polymeric substances and waxes. If appropriate, the medicine can also be in a microencapsulated form with one or more of the above-mentioned excipients.

[0236] For tablets, the active agent typically accounts for less than 50% (by weight) of the formulation, for example less than about 10%, such as 5% or 2.5% by weight. The major portion of the formulation comprises fillers, diluents, disintegrants, lubricants and optional flavorings. The composition of these excipients is well known in the art. Often, the filler / diluent comprises a mixture of two or more of the following components: microcrystalline cellulose, mannitol, lactose (all types), starch and dicalcium phosphate. The filler / diluent mixture typically accounts for less than 98% of the formulation, and preferably less than 95%, for example 93.5%. Preferred disintegrants include Starch and sodium lauryl sulfate. When present, disintegrants typically comprise less than 10% or less than 5%, for example, about 3%, by weight of the formulation. A preferred lubricant is magnesium stearate. When present, lubricants typically comprise less than 5% or less than 3%, for example, about 1%, by weight of the formulation.

[0237] Tablets can be manufactured by standard tableting methods (such as direct compression or wet, dry or melt granulation, melt congealing and extrusion). The core can be single-layer or multi-layer and can be coated with appropriate outer coatings known in the art.

[0238] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compound or combination of Formula I, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, for example, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame seed oil, etc.), (commercially available from CONDEA Vista Co. (Cranford, NJ)), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, or mixtures thereof, and the like.

[0239] Besides such inert diluents, the composition may also include excipients such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0240] Oral liquid forms of the compounds or combinations of the present invention include solutions in which the active compound is completely dissolved. Examples of solvents include all pharmaceutically established solvents suitable for oral administration, particularly solvents in which the compounds of the present invention exhibit good solubility, such as polyethylene glycol, polypropylene glycol, edible oils, and glyceryl and glyceride-based systems. For example, glyceryl and glyceride-based systems may include, for example, the following branded products (and corresponding generic products): 355EP (tricaprylin / caprin, available from Abitec (Columbus Ohio)), Crodamol TM GTC / C (medium chain triglycerides, obtained from Croda (Cowick Hall, UK)) or Labrafac TM CC (medium chain triglycerides, obtained from Gattefosse), 500P (triacetin, also known as triacetin, obtained from Abitec), MCM (medium chain mono- and diglycerides from Abitec), 812 (caprylic / capric triglyceride from Condea, Cranford NJ), 829 (caprylic / capric / succinic triglyceride, available from Condea), 840 (propylene glycol dicaprylate / dicaprate, available from Condea), M1944CS (oleoyl macrogol-6 glyceride, obtained from Gattefosse), Peceol TM (glyceryl monooleate from Gattefosse) and 35-1 (glyceryl monooleate, obtained from Gattefosse). Of particular interest are the medium chain (about C8 to C 10 ) triglyceride oil. These solvents often constitute the major portion of the composition, i.e., greater than about 50% (by weight), typically greater than about 80%, such as about 95% or 99%. Adjuvants and additives may also be included with the solvents, primarily as taste masking agents, palatability and flavoring agents, antioxidants, stabilizers, texture and viscosity modifiers, and solubilizing agents.

[0241] In addition to the compound or combination of formula I, the suspension may further contain carriers such as suspending agents, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, or mixtures of these substances, and the like.

[0242] Compositions for rectal or vaginal administration preferably comprise suppositories, which can be prepared by mixing a compound or combination of Formula I with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol or a suppository wax, which is solid at ordinary room temperature but liquid at body temperature and therefore melts in the rectum or vaginal cavity to release the active ingredient(s).

[0243] Dosage forms for topical administration of a compound or combination of Formula I include ointments, creams, lotions, powders and sprays. The drug is mixed with a pharmaceutically acceptable excipient, diluent or carrier and any preservatives, buffers, or propellants that may be required.

[0244] Many compounds of the invention are poorly soluble in water, for example, less than about 1 μg / mL. Therefore, liquid compositions in solubilizing non-aqueous solvents (such as the medium chain triglyceride oils discussed above) are preferred dosage forms for these compounds.

[0245] Solid amorphous dispersions (including dispersions formed by spray drying) are also preferred dosage forms for the insoluble compounds of the present invention. "Solid amorphous dispersion" means a solid material in which at least a portion of the insoluble compound is in amorphous form and dispersed in a water-soluble polymer. "Amorphous" means that the insoluble compound is not crystalline. "Crystallized" means that the compound exhibits a three-dimensional long-range order of at least 100 repeating units in each dimension. Therefore, the term amorphous is not only intended to include substantially disordered materials, but also includes materials that may have a certain smaller degree of order but the order is less than three dimensions and / or the order is only within a short distance. Amorphous materials can be characterized by techniques known in the art such as powder x-ray diffraction (PXRD) crystallography, solid-state NMR, or thermal techniques such as differential scanning calorimetry (DSC).

[0246] Preferably, at least a majority (i.e., at least about 60% by weight) of the poorly soluble compound in the solid amorphous dispersion is amorphous. The compound may be present in relatively pure amorphous domains or regions within the solid amorphous dispersion as a solid solution of the compound uniformly distributed throughout the polymer or any combination of these states or those intermediate thereto. Preferably, the solid amorphous dispersion is substantially uniform so that the amorphous compound is as uniformly dispersed throughout the polymer as possible. As used herein, "substantially uniform" means that the fraction of the compound present in relatively pure amorphous domains or regions within the solid amorphous dispersion is relatively small, approximately less than 20% by weight of the total amount of drug, and preferably less than 10% by weight.

[0247] Water-soluble polymers suitable for use in solid amorphous dispersions should be inert, pharmaceutically acceptable in the sense that they do not chemically react with the poorly soluble compound in an adverse manner, and have at least some solubility in aqueous solution at physiologically relevant pH (e.g., 1-8). The polymer can be neutral or ionizable and should have a water solubility of at least 0.1 mg / mL over at least a portion of the pH range of 1-8.

[0248] Water-soluble polymers suitable for use with compounds of Formula I can be cellulosic or non-cellulosic. The polymer can be neutral or ionizable in aqueous solution. Of these, ionizable and cellulosic polymers are preferred, with ionizable cellulosic polymers being more preferred.

[0249] Exemplary water-soluble polymers include hydroxypropyl methylcellulose acetate succinate (HPMCAS), hydroxypropyl methylcellulose (HPMC), hydroxypropyl methylcellulose phthalate (HPMCP), carboxymethyl ethylcellulose (CMEC), cellulose acetate phthalate (CAP), cellulose acetate trimellitate (CAT), polyvinyl pyrrolidone (PVP), hydroxypropyl cellulose (HPC), methylcellulose (MC), block copolymers of ethylene oxide and propylene oxide (PEO / PPO, also known as poloxamers), and mixtures thereof. Particularly preferred polymers include HPMCAS, HPMC, HPMCP, CMEC, CAP, CAT, PVP, poloxamers, and mixtures thereof. Most preferred is HPMCAS. See European Patent Application Publication No. 0 901 786 A2, the disclosure of which is incorporated herein by reference.

[0250] Solid amorphous dispersions can be prepared according to any method for forming solid amorphous dispersions that results in at least a majority (at least 60% by weight) of the poorly soluble compound being in an amorphous state. Such methods include mechanical, thermal, and solvent methods. Example mechanical methods include grinding and extrusion; melt methods, including high temperature melt methods, solvent-modified melt methods, and melt-condensation methods; and solvent methods, including non-solvent precipitation, spraying, and spray drying. See, for example, the following U.S. Patents, the relevant disclosures of which are incorporated herein by reference: Nos. 5,456,923 and 5,939,099, which describe dispersions formed by extrusion; Nos. 5,340,591 and 4,673,564, which describe dispersions formed by grinding methods; and Nos. 5,707,646 and 4,894,235, which describe dispersions formed by melt-condensation methods. In a preferred method, a solid amorphous dispersion is formed by spray drying, as disclosed in European Patent Application Publication No. 0 901 786 A2. In this method, the compound and polymer are dissolved in a solvent (such as acetone or methanol), and the solvent is then rapidly removed from the solution by spray drying to form a solid amorphous dispersion. The solid amorphous dispersion can be prepared to contain up to about 99% by weight of the compound, for example, 1%, 5%, 10%, 25%, 50%, 75%, 95%, or 98% by weight, as desired.

[0251] The solid dispersion itself can be used as a dosage form, or it can serve as a manufactured-use-product (MUP) in the preparation of other dosage forms such as capsules, tablets, solutions, or suspensions. An example of an aqueous suspension is an aqueous suspension of a 1:1 (w / w) compound / HPMCAS-HF spray-dried dispersion containing 2.5 mg / mL of the compound in 2% polysorbate-80. Solid dispersions for tablets or capsules are typically mixed with other excipients or adjuvants typically present in such dosage forms. For example, an exemplary filler for a capsule contains a 2:1 (w / w) compound / HPMCAS-MF spray-dried dispersion (60%), lactose (fast flow) (15%), microcrystalline cellulose (e.g., Avicel 1000). (R0-102) )(15.8%), sodium starch glycolate (7%), sodium lauryl sulfate (2%) and magnesium stearate (1%).

[0252] HPMCAS polymers can be used as Aqoat (R)-LF 、Aqoat (R)-MF and Aqoat (R)-HF Available from Shin-Etsu Chemical Co., LTD (Tokyo, Japan) in low, medium and high grades. The higher MF and HF grades are generally preferred.

[0253] The compound of formula I or a pharmaceutically acceptable salt of said compound can be used to treat non-human animals. Administration of the compound of formula I and in combination with another effective agent for treating the relevant condition can be achieved orally or parenterally.

[0254] The amount of the compound of Formula I or the combination of the compound of Formula I and another effective agent is administered so that an effective dose is obtained. Generally, the daily dose for oral administration to animals is from about 0.01 to about 1,000 mg / kg body weight, for example, from about 0.01 to about 300 mg / kg or from about 0.01 to about 100 mg / kg or from about 0.01 to about 50 mg / kg body weight, or from about 0.01 to about 25 mg / kg, or from about 0.01 to about 10 mg / kg or from about 0.01 to about 5 mg / kg.

[0255] Conveniently, the compound of formula I (or combination) can be carried in drinking water so that a therapeutic dose of the compound is taken with the daily water supply. The compound can be metered directly into the drinking water, preferably in the form of a liquid, water-soluble concentrate (such as an aqueous solution of a water-soluble salt).

[0256] Conveniently, the compound (or combination) of Formula I can also be added directly to the feed as is or in the form of an animal feed supplement (also referred to as a premix or concentrate). Premixes or concentrates of the compound in an excipient, diluent or carrier are more commonly used to include the agent in the feed. Suitable excipients, diluents or carriers are liquid or solid as needed, such as water, various coarse meals (such as alfalfa meal, soybean meal, cottonseed oil meal, linseed oil meal, corn cob meal and corn flour, molasses, urea, bone meal) and mineral mixtures commonly used in poultry feed. Particularly effective excipients, diluents or carriers are the corresponding animal feed itself; that is, a small portion of such feed. The carrier facilitates the uniform distribution of the compound in the finished feed blended with the premix. Preferably, the compound is fully blended into the premix and then blended into the feed. In this regard, the compound can be dispersed or dissolved in a suitable oily vehicle (such as soybean oil, corn oil, cottonseed oil, etc.) or a volatile organic solvent and then blended with the carrier. It will be appreciated that the proportions of the compound in the concentrate can vary widely, as the amount of the compound in the finished feed can be adjusted by blending the appropriate proportions of the premix with the feed to obtain the desired level of the compound.

[0257] As described above, feed manufacturers can blend high-potency concentrates with protein carriers (such as soybean oil meal and other coarse meals described above) to produce concentrated supplements suitable for direct feeding to animals. In such cases, the animals are allowed to eat their usual diet. Alternatively, such concentrated supplements can be added directly to feed to produce a nutritionally balanced finished feed containing therapeutically effective levels of the compound. The mixture is thoroughly blended using standard procedures (such as in a twin-drum mixer) to ensure uniformity.

[0258] If the supplement is used as a top dressing on the feed, it also helps to ensure uniformity of distribution of the compound throughout the top of the fed feed.

[0259] Drinking water and feed effective to increase lean mass deposition and improve lean to fat ratio are typically prepared by mixing a compound of Formula I with a sufficient amount of the animal feed to provide from about 0.001 to about 500 ppm of the compound in the feed or water.

[0260] Preferred medicated pig, cattle, sheep and goat feeds typically contain from about 1 to about 400 grams of a compound of formula I (or combination) per ton of feed, with the optimum amount for these animals typically being from about 50 to about 300 grams per ton of feed.

[0261] Preferred poultry and domestic pet feeds typically contain from about 1 to about 400 grams, and preferably from about 10 to about 400 grams, of the compound (or combination) per ton of feed.

[0262] For parenteral administration to animals, the compound of formula I (or combination) may be prepared in the form of a paste or pellet and typically administered as an implant under the skin of the animal's head or ear, where an increase in lean muscle mass and an improvement in the lean to fat ratio are sought.

[0263] Paste formulations can be prepared by dispersing the drug in pharmaceutically acceptable oils (such as peanut oil, sesame oil, corn oil, etc.).

[0264] Pellets containing an effective amount of a compound of Formula I, pharmaceutical composition, or combination can be prepared by mixing the compound of Formula I or combination with a diluent (such as carbowax, carnauba wax, etc.), and a lubricant (such as magnesium stearate or calcium stearate) can be added to improve the granulation process.

[0265] Of course, it is recognized that more than one bolus may be administered to an animal to achieve the desired dosage level that provides increased lean mass deposition and improved desired lean to fat ratio. Additionally, implants may be administered periodically during the animal's treatment period to maintain appropriate drug levels in the animal.

[0266] Liposomes containing these agents and / or compounds of the present invention are prepared by methods known in the art, as described in U.S. Patent Nos. 4,485,045 and 4,544,545. Liposomes with improved circulation time are disclosed in U.S. Patent No. 5,013,556. Particularly useful liposomes can be generated by a lipid composition comprising phosphatidylcholine, cholesterol, and PEG-derived phosphatidylethanolamine (PEG-PE) using a reverse phase evaporation method. Liposomes are extruded through a filter with a defined pore size to produce liposomes with a desired diameter.

[0267] These agents and / or compounds of the present invention may also be embedded in microcapsules (e.g., hydroxymethylcellulose or gelatin microcapsules and polymethyl methacrylate microcapsules) prepared, for example, by coacervation techniques or by interfacial polymerization, colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles and nanocapsules), or in coarse emulsions. Such techniques are disclosed in Remington, The Science and Practice of Pharmacy, 20th edition, Mack Publishing (2000).

[0268] Preparations for intravenous administration must be sterile. This is easily achieved, for example, by filtration through a sterile filtration membrane. The compounds of the invention are typically placed in a container with a sterile access port, such as an intravenous fluid bag or a vial with a stopper pierceable by a hypodermic needle.

[0269] Suitable emulsions may include commercially available fat emulsions (e.g. Infonutrol TM 、 and Lipiphysan TM ) to prepare. The active ingredient can be dissolved in a premixed emulsion composition, or alternatively, it can be dissolved in an oil (e.g., soybean oil, safflower oil, cottonseed oil, sesame oil, corn oil or almond oil) and in an emulsion formed when mixed with a phospholipid (e.g., lecithin, soybean lecithin or soybean lecithin) and water. It will be appreciated that other ingredients, such as glycerol or glucose, can be added to adjust the tension of the emulsion. Suitable emulsions typically contain up to 20% oil, for example 5% to 20%. Fat emulsions can contain fat droplets of 0.1 to 1.0 μm, particularly 0.1 to 0.5 μm, and have a pH in the range of 5.5 to 8.0.

[0270] The emulsion composition may be prepared by combining the compound of the present invention with Intralipid TM or those prepared by mixing their components (soybean oil, lecithin, glycerin and water).

[0271] Compositions for inhalation or insufflation include solutions and suspensions in pharmaceutically acceptable aqueous or organic solvents or mixtures thereof, and powders. Liquid or solid compositions may contain suitable pharmaceutically acceptable excipients as described above. In some embodiments, compositions are administered by oral or nasal respiratory route to produce local or systemic effects. Compositions in preferably sterile pharmaceutically acceptable solvents can be atomized using gas. Atomized solutions can be breathed directly from an atomizing device, or the atomizing device can be attached to a mask, tent, or intermittent positive pressure breathing machine. Solutions, suspensions, or powder compositions can be administered preferably by oral or nasal administration from a device that delivers the formulation in an appropriate manner.

[0272] The compounds herein can be formulated for oral, buccal, intranasal, parenteral (eg, intravenous, intramuscular or subcutaneous) or rectal administration, or in a form suitable for administration by inhalation. The compounds of the present invention can also be formulated for sustained delivery.

[0273] Methods for preparing various pharmaceutical compositions having specific amounts of active ingredients are known to those skilled in the art or will be apparent in light of this disclosure. For examples of methods for preparing pharmaceutical compositions, see Remington's Pharmaceutical Sciences, 20th edition (Lippincott Williams & Wilkins, 2000).

[0274] The pharmaceutical compositions according to the present invention may contain 0.1% to 95% by weight of one or more compounds of the present invention, preferably 1% to 70%. In any case, the composition to be administered will contain an amount of one or more compounds according to the present invention that is effective in treating the disease / disorder of the subject being treated.

[0275] Since one aspect of the application relates to treating diseases / disorders described herein with a combination of active ingredients that can be applied separately, the present invention further relates to combining separate pharmaceutical compositions in the form of a kit. The kit may include a composition comprising a compound of Formula I, or it may contain at least two separate pharmaceutical compositions: a compound of Formula I, a prodrug thereof, or a salt of such a compound or prodrug, and a second compound as described above. The kit includes tools for accommodating separate compositions, such as containers, sub-bottles, or sub-foil packs. Typically, the kit includes instructions for use of separate components. When separate components are preferably applied in different dosage forms (e.g., oral and parenteral), applied at different dosage intervals, or when the prescribing physician desires to titrate the individual components of the combination, the kit form is particularly advantageous.

[0276] The example of this test kit is so-called blister pack. Blister pack is well known in the packaging industry and is widely used for packaging pharmaceutical unit dosage forms (tablets, capsules, etc.). Blister pack is usually composed of a sheet of relatively hard material, which is covered with a foil of preferably transparent plastic material. During the packaging process, a recess is formed in the plastic foil. The recess has the size and shape of the tablet or capsule to be packaged. Next, the tablet or capsule is placed in the recess and a relatively hard sheet of material is sealed on the plastic foil on the face of the foil opposite to the direction in which the recess is formed. Therefore, the tablet or capsule is sealed in the recess between the plastic foil and the sheet. Preferably, the strength of the sheet allows the tablet or capsule to be taken out from the blister pack by manually applying pressure to the recess, thereby forming an opening in the sheet at the position of the recess. The tablet or capsule can then be taken out via the opening.

[0277] It may be desirable to provide a memory aid on the kit, for example, in the form of numbers next to the tablets or capsules, whereby the numbers correspond to the days of the regimen on which the tablets or capsules so specified should be taken. Another example of such a memory aid is a calendar printed on a card, for example, as follows: "Week 1, Monday, Tuesday, etc. ... Week 2, Monday, Tuesday ... " etc. Other variations of the memory aid will be apparent. A "daily dose" can be a single tablet or capsule or several pills or capsules to be taken on a given day. Moreover, a daily dose of a compound of Formula I can consist of one tablet or capsule, while an optional daily dose of a second compound can consist of several tablets or capsules, or vice versa. The memory aid should reflect this.

[0278] In another specific embodiment of the present invention, a dispenser is provided that is designed to dispense daily doses one at a time in the order of their intended use. Preferably, the dispenser is equipped with a memory aid to further promote compliance with the regimen. An example of such a memory aid is a mechanical counter that indicates the number of daily doses that have been dispensed. Another example of such a memory aid is a battery-powered microchip memory coupled to a liquid crystal reader or an audible reminder signal, for example, which reads out the date the last daily dose has been taken and / or reminds the date when the next dose is to be taken.

[0279] Moreover, since one aspect of the present application relates to treating the diseases / conditions described herein with a combination of active ingredients that can be administered in conjunction, the present invention also relates to combining separate pharmaceutical compositions into a single dosage form, such as (but not limited to) a single tablet or capsule, a double or multi-layer tablet or capsule, or by using segregated components or compartments within a tablet or capsule.

[0280] The active ingredient can be delivered as a solution in an aqueous or non-aqueous vehicle, with or without additional solvents, cosolvents, excipients, or complexing agents, selected from pharmaceutically acceptable diluents, excipients, vehicles, or carriers.

[0281] The active ingredient can be formulated together with pharmaceutically acceptable excipients as a solid dispersion or a self-emulsifying drug delivery system (SEDDS).

[0282] The active ingredient can be formulated as an immediate release or modified release tablet or capsule. Alternatively, the active ingredient can be delivered as the sole active ingredient within a capsule shell, without additional excipients.

[0283] Experimental procedures

[0284] The synthesis of various compounds of the present invention is shown below. The methods shown in these examples can be used alone or in combination with techniques generally known in the art to prepare additional compounds within the scope of the present invention. All starting materials in these preparations and examples are commercially available or can be prepared by methods known in the art or as described herein.

[0285] Reactions were carried out in air or, when oxygen or moisture sensitive reagents or intermediates were employed, under an inert atmosphere (nitrogen or argon). When appropriate, reaction equipment was dried using a heat gun under dynamic vacuum and anhydrous solvents (Sure-Seal® from Aldrich Chemical Company, Milwaukee, Wisconsin) were used. TM product or DriSolv® from EMD Chemicals (Gibbstown, NJ). TM product). In some cases, commercial solvents were passed through a The column was heated to a temperature of 100 °C for 1 % to 100 ppm by weight for dichloromethane, toluene, N,N-dimethylformamide, and tetrahydrofuran; and the following water QC standards were achieved: a) <100 ppm by weight for dichloromethane, toluene, N,N-dimethylformamide, and tetrahydrofuran; b) <180 ppm by weight for methanol, ethanol, 1,4-dioxane, and diisopropylamine. For very sensitive reactions, the solvent was further treated with sodium metal, calcium hydride, or molecular sieves and distilled immediately before use. Other commercial solvents and reagents were used without further purification. For syntheses citing procedures from other examples or methods, reaction conditions (reaction time and temperature) may vary. The product was typically dried under vacuum before further reaction or submission for biological testing.

[0286] When indicated, the reaction was heated by microwave irradiation using a Biotage Initiator or Personal Chemistry EmrysOptimizer microwave instrument. The reaction process was monitored by thin layer chromatography (TLC), liquid chromatography-mass spectrometry (LCMS), high performance liquid chromatography (HPLC) and / or gas chromatography-mass spectrometry (GCMS). TLC was performed on pre-coated silica gel plates using a fluorescent indicator (254 nm excitation wavelength) and visualized under UV light and / or with I , KMnO , CoCl , phosphomolybdic acid, or ammonium molybdate cerium stain. LCMS data were obtained using Leap Technologies automatic samplers, Gemini C18 columns, acetonitrile / water gradients, and trifluoroacetic acid, formic acid, or ammonium hydroxide modifiers on an Agilent 1100 series instrument. Column eluent was analyzed using a Waters ZQ mass spectrometer scanned in both positive and negative ion modes from 100 to 1200 Da. Other similar instruments were also used. HPLC data are typically acquired on an Agilent 1100 series instrument using a Gemini or XBridge C18 column, acetonitrile / water gradient, and trifluoroacetic acid or ammonium hydroxide modifier. GCMS data are acquired using a Hewlett Packard 6890 column oven using an HP 6890 injector, an HP-1 column (12 m × 0.2 mm × 0.33 μm), and helium carrier gas. Samples are analyzed on an HP 5973 mass selective detector scanned from 50 to 550 Da using electron ionization. Purification is performed by medium performance liquid chromatography (MPLC) using an Isco CombiFlash Companion, AnaLogix IntelliFlash 280, Biotage SP1, or Biotage Isolera One instrument and pre-packed Isco RediSep or Biotage Snap silica gel columns. Chiral purification is typically performed using Berger or Thar instruments; ChiralPAK-AD, -AS, -IC, Chiralcel-OD, or -OJ columns; and mixtures of CO2 with methanol, ethanol, propan-2-ol, or acetonitrile (alone or modified with trifluoroacetic acid or propan-2-amine) by chiral supercritical fluid chromatography (SFC). UV detection is used to initiate fraction collection. For syntheses citing procedures in other Examples or Methods, purification may vary: Generally, the solvents and solvent ratios used for the eluent / gradient are selected to provide the appropriate R f s or retention time.

[0287] Mass spectrometry data were reported by LCMS analysis. Mass spectrometry (MS) was performed using atmospheric pressure chemical ionization (APCI), electrospray ionization (ESI), electron impact ionization (EI), or electron scattering (ES) ionization sources. Proton nuclear magnetic spectroscopy ( 1 HNMR chemical shifts are given in parts per million downfield from tetramethylsilane and were recorded on 300, 400, 500, or 600 MHz Varian, Bruker, or Jeol spectrometers. Chemical shifts are expressed in parts per million (ppm, d) referenced to the residual peak of the deuterated solvent (chloroform, 7.26 ppm; CD2HOD, 3.31 ppm; acetonitrile-d2, 1.94 ppm; dimethyl sulfoxide-d5, 2.50 ppm; DHO, 4.79 ppm). Peak shapes are described as follows: s, singlet; d, doublet; t, triplet; q, quartet; quin, quintet; m, multiplet; br s, broad singlet; app, apparent. Analytical SFC data were acquired on a Berger analytical instrument as described above. Optical rotation data were acquired on a PerkinElmer 343 polarimeter using a 1 dm cell. Silica gel chromatography was primarily performed using medium pressure Biotage or ISCO systems using prepacked columns from various commercial suppliers including Biotage and ISCO. Microanalyses were performed by Quantitative Technologies Inc. and were within 0.4% of the calculated values.

[0288] Unless otherwise indicated, chemical reactions were performed at room temperature (approximately 23 degrees Celsius).

[0289] Unless otherwise noted, all reactants were obtained commercially without further purification or were prepared using methods known in the literature.

[0290] Hydrogenations can be performed in a Parr Shaker under pressurized hydrogen or in a Thales-nano H-Cube flow hydrogenation apparatus under full hydrogen and a flow rate of 1 to 2 mL / min at the indicated temperature.

[0291] HPLC, UPLC, LCMS, GCMS, and SFC retention times were measured using the methods mentioned in the procedures.

[0292] In some embodiments, chiral separations are performed to separate the enantiomers or diastereomers of a particular compound of the invention (in some embodiments, the separated enantiomers are designated as ENANT-1 and ENANT-2 based on their order of elution; similarly, the separated diastereomers are designated as DIAST-1 and DIAST-2 based on their order of elution). In some instances, the optical rotation of the enantiomers is measured using a polarimeter. Based on their observed optical rotation data (or their specific optical rotation data), the enantiomer that rotates clockwise is designated as the (+)-enantiomer, and the enantiomer that rotates counterclockwise is designated as the (-)-enantiomer. Racemic compounds are indicated by the absence of a drawn or described stereochemistry, or by the presence of (+ / -) adjacent to the structure; in the latter case, the indicated stereochemistry represents only one of the two enantiomers that constitute the racemic mixture.

[0293] The following compounds and intermediates are named using the naming conventions provided by ACD / ChemSketch 2017.2.1 (File Version C40H41, Build 99535, Advanced Chemistry Development, Inc., Toronto, Ontario, Canada). The naming conventions provided by ACD / ChemSketch 2017.2.1 are well known to those skilled in the art and are believed to be generally consistent with the IUPAC (International Union for Pure and Applied Chemistry) recommendations for organic chemical nomenclature and the CAS index rules.

[0294] Example

[0295] Preparation Example P1: 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzoic acid (P1)

[0296]

[0297] Step 1: Synthesis of methyl 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzoate (C1).

[0298] To a solution of sodium hydride (60% dispersion in mineral oil; 1.60 g, 40.0 mmol) in tetrahydrofuran (200 mL) at 0°C was added (4-methoxyphenyl)methanol (5.25 g, 38.0 mmol). After the reaction mixture was stirred at 0°C for 30 minutes, a solution of methyl 3,4,5-trifluorobenzoate (7.00 g, 36.8 mmol) in tetrahydrofuran (50 mL) was added, and the reaction mixture was then allowed to warm to 25°C and stirred for 1 hour. It was then quenched by the addition of saturated aqueous ammonium chloride solution, and the aqueous layer was extracted with ethyl acetate; the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo to provide C1 (11.2 g) as a solid. This material was directly carried into the next step.

[0299] Step 2: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzoic acid (P1)

[0300] To a solution of C1 (obtained from the previous step; 11.2 g, ≤36.3 mmol) in methanol (200 mL) was added a solution of sodium hydroxide (4.36 g, 109 mmol) in water (20 mL), and the reaction mixture was subsequently stirred at 26 ° C for 4 hours. It was then concentrated in vacuo, and the aqueous residue was washed with dichloromethane (2×150 mL). After the aqueous layer was acidified to pH 5, it was extracted with dichloromethane (3×300 mL), and the three dichloromethane layers were combined and washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered and concentrated under reduced pressure to give P1 as a white solid. Yield: 10 g, 34 mmol, 92% over 2 steps. 1 H NMR (400MHz, DMSO-d6)d 7.61–7.53(m,2H),7.34(d,J=8.6Hz,2H),6.92(d,J=8.7Hz,2H),5.20(s,2H),3.74(s,3H).

[0301] Preparation Example P2: 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzoic acid (P2)

[0302]

[0303] Step 1: Synthesis of ethyl 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzoate (C2)

[0304] 1-(Chloromethyl)-4-methoxybenzene (40.1 g, 256 mmol) was added to a mixture of ethyl 2,3,5-trifluoro-4-hydroxybenzoate (51.3 g, 233 mmol) and potassium carbonate (64.3 g, 465 mmol) in acetonitrile (100 mL). After the reaction mixture was stirred at 80° C. for 16 hours, LCMS analysis indicated conversion to C2: LCMS m / z 363.1 [M+Na + The solids were removed by filtration, and the filtrate was concentrated in vacuo to provide C2 as a yellow oil. Yield: 71.0 g, 209 mmol, 90%.

[0305] Step 2: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzoic acid (P2)

[0306] To a solution of C2 (71.0 g, 209 mmol) in methanol (500 mL) was added aqueous sodium hydroxide solution (3 M; 300 mL). After the reaction mixture was stirred at 50 ° C for 4 hours, it was concentrated in vacuo. The aqueous residue was acidified by adding 1 M hydrochloric acid, and the resulting solid was collected by filtration to give P2 as a white solid. Yield: 51.7 g, 166 mmol, 79%. LCMS m / z 335.1 [M + Na + ]. 1 H NMR (400MHz, DMSO-d6)d 7.38–7.29(m,1H),7.34(d,J=8.6Hz,2H),6.93(d,J=8.7Hz,2H),5.18(s,2H),3.75(s,3H).

[0307] Preparation Example P3: N-{[(1r,4r)-4-aminocyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P3)

[0308]

[0309]

[0310] Step 1: Synthesis of tert-butyl [(1r,4r)-4-({3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)cyclohexyl]carbamate (C3)

[0311] To a solution of P1 (19.3 g, 65.6 mmol), N,N-diisopropylethylamine (25.4 g, 197 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 27.5 g, 72.3 mmol) in dichloromethane (700 mL) was added tert-butyl [(1r,4r)-4-(aminomethyl)cyclohexyl]carbamate (15.0 g, 65.7 mmol). After the reaction mixture was stirred at 25 °C for 16 hours, LCMS analysis indicated the presence of C3: LCMS m / z 527.3 [M+Na + After filtration, the filter cake was washed with water and a mixture of dichloromethane and ethyl acetate to afford C3 as a white solid. Yield: 26.5 g, 52.5 mmol, 80%. 1 H NMR (400MHz, DMSO-d6)d 8.48(br t,J=6Hz,1H),7.63–7.53(m,2H),7.33(d,J=8.6Hz,2H),6.92(d,J=8.7Hz,2H),6.67(br d,J=8.0Hz,1H),5.16(s,2H),3.74(s,3H),3.22–3.10(m,1H),3.06(dd,J=6.1,6.1Hz,2H) ,1.83–1.65(m,4H),1.48–1.36(m,1H),1.36(s,9H),1.16–1.02(m,2H),1.00–0.85(m,2H).

[0312] Step 2: Synthesis of N-{[(1r,4r)-4-aminocyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P3)

[0313] To a solution of C3 (21.5 g, 42.6 mmol) and pyridine (27.0 g, 341 mmol) in dichloromethane (500 mL) at 0 ° C., trimethylsilyl trifluoromethanesulfonate (37.9 g, 170 mmol) was added dropwise. After the reaction mixture was stirred at 25 ° C. for 16 hours, an aqueous sodium bicarbonate solution (100 mL) was added and the mixture was filtered. The filter cake was washed with water and a mixture of dichloromethane and ethyl acetate to provide P3 as a white solid. Yield: 10.0 g, 24.7 mmol, 58%. LCMS m / z 405.3 [M + H] + . 1H NMR(400MHz,DMSO-d6)d 8.50(br t,J=6Hz,1H),7.71–7.41(m,4H),7.33(d,J=8.2Hz,2H),6.92(d,J=8.2Hz,2H),5.17(s,2H),3.74(s,3H),3.09(dd,J=6Hz ,2H),3.00–2.86(m,1H),1.96–1.85(m,2H),1.82–1.70(m,2H),1.54–1.38(m,1H),1.31–1.15(m,2H),1.07–0.92(m,2H).

[0314] Preparation Example P4: (1r,4r)-4-({3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)cyclohexane-1-carboxylic acid (P4)

[0315]

[0316] Step 1: Synthesis of methyl (1r,4r)-4-({3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)cyclohexane-1-carboxylate (C4).

[0317] To a solution of P1 (18.0 g, 61.2 mmol), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (14.1 g, 73.5 mmol) and 1H-benzotriazole-1-ol (9.92 g, 73.4 mmol) in dichloromethane (500 mL) was added triethylamine (7.41 g, 73.2 mmol) and (1r, 4r)-4-(aminomethyl)cyclohexane-1-carboxylic acid methyl ester (10.5 g, 61.3 mmol). After the reaction mixture was stirred at 28 ° C for 4 hours, it was extracted with dichloromethane. The combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, concentrated in vacuo, and purified via silica gel chromatography (eluent: 6% methanol / dichloromethane) to provide C4 as a white solid. Yield: 22.0 g, 49.2 mmol, 80%. LCMS m / z 448.2 [M+H] + .

[0318] Step 2: Synthesis of (1r,4r)-4-({3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)cyclohexane-1-carboxylic acid (P4).

[0319] A solution of sodium hydroxide (8.05 g, 201 mmol) in water (20 mL) was added to a solution of C4 (18.0 g, 40.2 mmol) in methanol (200 mL). The reaction mixture was stirred at 26 ° C for 6 hours, then the methanol was removed under reduced pressure, and the aqueous residue was washed with dichloromethane (2 × 20 mL). The aqueous layer was then adjusted to pH 5 and extracted with dichloromethane (3 × 50 mL); the three extracts were combined, washed with saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and concentrated in vacuo to give P4 as a white solid. Yield: 14.0 g, 32.3 mmol, 80%. LCMSm / z 434.2 [M + H] + . 1 H NMR(400MHz,DMSO-d6)d 8.50(br t,J=5.7Hz,1H),7.64–7.54(m,2H),7.33(d,J=8.5Hz,2H),6.92(d,J=8.4Hz,2H),5.16(s,2H),3.74(s,3H),3.08(dd,J=6,6 Hz,2H),2.18–2.05(m,1H),1.95–1.82(m,2H),1.80–1.68(m,2H),1.54–1.39(m,1H),1.33–1.16(m,2H),1.02–0.85(m,2H).

[0320] Preparation Example P5: 3,5-difluoro-N-{[(1r,4r)-4-(N-hydroxycarbamimidoyl)cyclohexyl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P5)

[0321]

[0322] Step 1: Synthesis of N-{[(1r,4r)-4-cyanocyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C5).

[0323] A solution of hydrogen chloride in 1,4-dioxane (4 M; 50 mL, 200 mmol) was added to a solution of tert-butyl {[(1r,4r)-4-cyanocyclohexyl]methyl}carbamate (4.86 g, 20.4 mmol) in tetrahydrofuran (50 mL), and the mixture was stirred at room temperature overnight. After removing the solvent via concentration under reduced pressure, the residue was triturated with diethyl ether to provide (1r,4r)-4-(aminomethyl)cyclohexane-1-carbonitrile hydrochloride.

[0324] O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 95%, 8.16g, 20.4mmol) is added to a dichloromethane (113mL) solution of P1 (5.0g, 17mmol). After the mixture is stirred for 1 hour, it is treated with N,N-diisopropylethylamine (8.88mL, 51.0mmol) and (1r,4r)-4-(aminomethyl)cyclohexane-1-carbonitrile hydrochloride obtained above. The reaction mixture is stirred at room temperature for 3 days, then washed with water, 1M hydrochloric acid, water, saturated sodium bicarbonate aqueous solution and saturated sodium chloride aqueous solution, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue is dissolved in a mixture of very small amounts of hot 10:1 ethyl acetate and heptane; after cooling to room temperature, it is filtered, and the filtrate is concentrated under reduced pressure. Silica gel chromatography provides C5 as a white solid. Yield: 5.80 g, 14.0 mmol, 82%. LCMS m / z 415.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6)d 8.50(br t,J=5.8Hz,1H),7.62–7.53(m,2H),7.33(d,J=8.6Hz,2H),6.92(d,J=8.6Hz,2H),5.16(s,2H),3.74(s,3H),3.08(dd,J =6,6Hz,2H),2.62(tt,J=11.9,3.6Hz,1H),2.04–1.95(m,2H),1.77–1.67(m,2H),1.60–1.37(m,3H),1.04–0.89(m,2H).

[0325] Step 2: Synthesis of 3,5-difluoro-N-{[(1r,4r)-4-(N-hydroxycarbamimidoyl)cyclohexyl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P5).

[0326] Hydroxylamine hydrochloride (8.38g, 121mmol) and triethylamine (16.8mL, 121mmol) are added to a solution of C5 (5.00g, 12.1mmol) in methanol (50mL). The reaction mixture is heated at 50°C for 24 hours, then cooled to room temperature and concentrated in vacuo. The residue is distributed between water (100mL) and ethyl acetate (100mL), and the mixture is vigorously stirred for 15 minutes. Filter, then rinse the collected solid with water (50mL) and ethyl acetate (50mL) to provide P5 as a white solid. Yield: 4.50g, 10.1mmol, 83%. LCMSm / z 448.4[M+H] + . 1H NMR (400 MHz, DMSO-d6), characteristic peaks: d 9.42 (s, 1H), 8.47 (br t, J = 5.8 Hz, 1H), 8.19 (br s, 1H), 7.81 (br s, 1H), 7.63–7.54 (m, 2H), 7.33 (d, J = 8.6 Hz, 2H), 6.92 (d, J = 8.7 Hz, 2H), 5.16 (s, 2H), 3.74 (s, 3H), 3.09 (dd, J = 6.6 Hz, 2H), 2.5–2.40 (m, 1H, hypothetical; partially obscured by solvent peak), 1.96–1.84 (m, 2H), 1.56–1.41 (m, 1H), 1.02–0.87 (m, 2H)

[0327] Preparation Example P6: N-{[(1s,4s)-4-bromocyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P6)

[0328]

[0329] Step 1: Synthesis of tert-butyl {[(1s,4s)-4-bromocyclohexyl]methyl}carbamate (C6).

[0330] To a solution of tert-butyl {[(1r,4r)-4-hydroxycyclohexyl]methyl}carbamate (5.00 g, 21.8 mmol) in dichloromethane (150 mL) at 0° C. was added carbon tetrabromide (10.8 g, 32.6 mmol). Triphenylphosphine (8.58 g, 32.7 mmol) was added in portions and the reaction mixture was stirred at 25° C. for 48 hours. After the solvent was removed in vacuo, C6 was purified by silica gel chromatography (gradient: 0% to 20% ethyl acetate / petroleum ether). Yield: 1.30 g, 4.45 mmol, 20%. LCMS m / z 314.1 (bromine isotope pattern observed) [M+Na + ]. 1 H NMR(400MHz,DMSO-d6)d6.86(br t,J=6.0Hz,1H),4.78–4.71(m,1H),2.82(dd,J=6,6Hz,2H),1.99–1.89(m,2H),1.87–1.75(m,2H),1.57–1.26(m,5H),1.37(s,9H).

[0331] Step 2: Synthesis of 1-[(1s,4s)-4-bromocyclohexyl]methanamine hydrochloride (C7).

[0332] To a solution of C6 (1.30 g, 4.45 mmol) in dichloromethane (20 mL) was added a solution of hydrogen chloride in 1,4-dioxane (4 M; 15 mL). After the reaction mixture was stirred at 25 °C for 2.5 hours, LCMS analysis indicated conversion to C7: LCMS m / z 192.1 (bromine isotope pattern observed) [M+H] + The solvent was removed in vacuo to afford C7 (900 mg), which was used directly in the next step.

[0333] Step 3: Synthesis of N-{[(1s,4s)-4-bromocyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P6).

[0334] To a solution of P1 (1.65 g, 5.61 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 2.67 g, 7.02 mmol) and N,N-diisopropylethylamine (1.82 g, 14.1 mmol) in dichloromethane (80 mL) was added C7 (from the previous step; 900 mg, ≤4.45 mmol), and the reaction mixture was stirred at room temperature for 3 hours. After the reaction mixture was concentrated in vacuo, silica gel chromatography (gradient: 0% to 30% ethyl acetate / petroleum ether) provided P6. Yield: 1.40 g, 2.99 mmol, 67% over 2 steps. LCMS m / z 490.0 (bromine isotope pattern [M+Na] observed) was obtained. + ]. 1 H NMR(400MHz,DMSO-d6)d 8.55(br t,J=5.8Hz,1H),7.63–7.54(m,2H),7.33(d,J=8.7Hz,2H),6.92(d,J=8.6Hz,2H),5.17(s,2H),4.80–4.72(m,1H), 3.74(s,3H),3.15(dd,J=6,6Hz,2H),2.02–1.91(m,2H),1.89–1.77(m,2H),1.70–1.53(m,3H),1.47–1.33(m,2H).

[0335] Preparation Example P7: 3,5-difluoro-N-{[4-(N-hydroxycarbamimidoyl)bicyclo[2.2.2]oct-1-yl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P7)

[0336]

[0337] Step 1: Synthesis of tert-butyl [(4-carbamoylbicyclo[2.2.2]oct-1-yl)methyl]carbamate (C8).

[0338] To a solution of 4-{[(tert-butoxycarbonyl)amino]methyl}bicyclo[2.2.2]octane-1-carboxylic acid (1.50 g, 5.29 mmol) in dichloromethane (20 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 3.02 g, 7.94 mmol), N,N-diisopropylethylamine (2.05 g, 15.9 mmol) and aqueous ammonium hydroxide (0.3 M; 22.9 mL, 6.87 mmol). After the reaction mixture was stirred at 25 ° C for 2 hours, it was diluted with dichloromethane (25 mL), washed sequentially with water (2×20 mL) and saturated aqueous sodium chloride solution (20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Trituration with water (20 mL) gave C8 as a white solid. Yield: 1.20 g, 4.25 mmol, 80%. LCMS m / z 283.2 [M+H] + .

[0339] Step 2: Synthesis of tert-butyl [(4-cyanobicyclo[2.2.2]octan-1-yl)methyl]carbamate (C9).

[0340] (Methoxycarbonylsulfamoyl)triethylammonium hydroxide inner salt (Burgess reagent; 1.86 g, 7.81 mmol) was added to a solution of C8 (1.10 g, 3.90 mmol) in a mixture of pyridine (15 mL) and dichloromethane (10 mL). After the reaction mixture was stirred at 25 ° C for 2 hours, it was concentrated in vacuo; the residue was diluted with water (30 mL) and extracted with dichloromethane (2×20 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (2×20 mL), dried over sodium sulfate, filtered, and concentrated under reduced pressure to provide C9 as a white solid. Yield: 1.00 g, 3.78 mmol, 97%. LCMS m / z 209.2 [(M-2-methylprop-1-ene)+H] + . 1 H NMR (400MHz, DMSO-d6) 6.80 (br t, J=6.4Hz, 1H), 2.66 (d, J=6.4Hz, 2H), 1.86–1.76 (m, 6H), 1.36 (s, 9H), 1.36–1.27 (m, 6H).

[0341] Step 3: Synthesis of 4-(aminomethyl)bicyclo[2.2.2]octane-1-carbonitrile hydrochloride (C10)

[0342] To a 0°C solution of C9 (1.00 g, 3.78 mmol) in dichloromethane (15 mL) was added a solution of hydrogen chloride in 1,4-dioxane (4 M; 3.8 mL, 15 mmol), and the reaction mixture was stirred at 25°C for 16 hours. The solvent was removed in vacuo to afford C10 as a white solid. Yield: 750 mg, 3.74 mmol, 99%. LCMS m / z 165.2 [M+H] + .

[0343] Step 4: Synthesis of N-[(4-cyanobicyclo[2.2.2]octan-1-yl)methyl]-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C11)

[0344] O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 1.60 g, 4.21 mmol) and N,N-diisopropylethylamine (1.81 g, 14.0 mmol) were added to a solution of P1 (1.13 g, 3.84 mmol) in N,N-dimethylformamide (10 mL). After the reaction mixture was stirred at 25 ° C for 10 minutes, C10 (700 mg, 3.49 mmol) was added and stirring was continued at 25 ° C for 4 hours. Water (25 mL) was then added, and the resulting mixture was extracted with ethyl acetate (2×25 mL); the combined organic layers were washed sequentially with water (2×10 mL) and saturated aqueous sodium chloride solution (2×10 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 50% ethyl acetate / petroleum ether) provided C11 as a light yellow solid. Yield: 1.29 g, 2.93 mmol, 84%. LCMS m / z 441.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)d 8.36(br t,J=6.3Hz,1H),7.64–7.54(m,2H),7.34(d,J=8.7Hz,2H),6.92(d,J=8.6Hz,2H),5. 17(s,2H),3.75(s,3H),3.01(d,J=6.2Hz,2H),1.87–1.78(m,6H),1.46–1.36(m,6H).

[0345] Step 5: Synthesis of 3,5-difluoro-N-{[4-(N-hydroxycarbamimidoyl)bicyclo[2.2.2]oct-1-yl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P7)

[0346] To a solution of C11 (1.20 g, 2.72 mmol) in methanol (25 mL) was added hydroxylamine hydrochloride (1.14 g, 16.4 mmol) and N,N-diisopropylethylamine (2.82 g, 21.8 mmol), and the reaction mixture was stirred at 70 ° C for 16 hours. The solvent was removed in vacuo to provide a residue that was purified via silica gel chromatography (gradient: 0% to 5% methanol / dichloromethane) to give P7 as a white solid. Yield: 748 mg, 1.58 mmol, 58%. LCMS m / z 474.2 [M + H] + . 1 H NMR(400MHz, DMSO-d6)d 8.85(s,1H),8.30(br t,J=6.2Hz,1H),7.64–7.55(m,2H),7.34(d,J=8.7Hz,2H),6.92(d,J=8.7Hz,2H),5.16(s,2H),5.11(br s, 2H), 3.74 (s, 3H), 3.01 (d, J = 6.2Hz, 2H), 1.66–1.56 (m, 6H), 1.41–1.31 (m, 6H).

[0347] Preparation Example P8: N-[(4-aminobicyclo[2.2.2]oct-1-yl)methyl]-2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P8)

[0348]

[0349] Step 1: Synthesis of tert-butyl [4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]oct-1-yl]carbamate (C12).

[0350] N, N-diisopropylethylamine (826mg, 6.39mmol) is added to a solution of P2 (1.00g, 3.20mmol) and O-(7-azabenzotriazole-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (HATU; 1.46g, 3.84mmol) in N, N-dimethylformamide (20mL). After the mixture was stirred at 25°C for 2 minutes, tert-butyl [4-(aminomethyl)bicyclo[2.2.2]octan-1-yl]carbamate (855mg, 3.36mmol) was added and stirring was continued at 20°C for 1 hour. The reaction mixture was then extracted with ethyl acetate (2×50mL), and the combined organic layers were dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (eluent: 1:1 petroleum ether / ethyl acetate) provided C12 as a white solid. Yield: 1.35g, 2.46mmol, 77%. LCMS m / z 549.3 [M+H] + . 1 H NMR(400MHz, DMSO-d6)d 8.25(t,J=6.3Hz,1H),7.35(d,J=8.6Hz,2H),7.30(ddd,J=10.9,6.0,2.3Hz,1H),6.94(d,J=8.7Hz,2H),6.32(br s,1H),5.21(s,2H),3.75(s,3H),2.96(d,J=6.2Hz,2H),1.76–1.64(m,6H),1.46–1.37(m,6H),1.35(s,9H).

[0351] Step 2: Synthesis of N-[(4-aminobicyclo[2.2.2]oct-1-yl)methyl]-2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P8)

[0352] To a solution of C12 (1.30 g, 2.37 mmol) and pyridine (1.50 g, 19.0 mmol) in dichloromethane (20 mL) was added trimethylsilyl trifluoromethanesulfonate (3.69 g, 16.6 mmol), and the reaction mixture was stirred at 20 ° C for 30 minutes. Aqueous sodium bicarbonate (2 M; 50 mL) was then added, and the resulting mixture was extracted with dichloromethane (2×50 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (30 mL), dried over sodium sulfate, filtered, and concentrated in vacuo; purified using silica gel chromatography (gradient: 13% to 17% methanol / dichloromethane) to provide P8 as a white solid. Yield: 765 mg, 1.71 mmol, 72%. LCMS m / z 449.2 [M+H] + . 1HNMR(400MHz,chloroform-d)d 7.57(ddd,J=11.8,6.8,2.3Hz,1H),7.33(d,J=8.6Hz,2H),6.87(d,J=8.5Hz,2H),6.55–6.44( m,1H),5.24(s,2H),3.80(s,3H),3.23(d,J=6.1Hz,2H),1.71–1.60(m,6H),1.59–1.49(m,6H).

[0353] Preparation Example P9: 4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxylic acid (P9)

[0354]

[0355] Step 1: Synthesis of methyl 4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxylate (C13).

[0356] To a solution of P2 (8.00 g, 25.6 mmol) and methyl 4-(aminomethyl)bicyclo[2.2.2]octane-1-carboxylate (5.05 g, 25.6 mmol) in N,N-dimethylformamide (60 mL) was added N,N-diisopropylethylamine (4.97 g, 38.4 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 11.7 g, 30.8 mmol). After the reaction mixture was stirred at room temperature for 4 hours, LCMS analysis indicated conversion to C13: LCMS m / z 492.2 [M+H] + The reaction mixture was poured into ice water, and the solid was collected by filtration and washed with water to provide C13 as a gray solid. Yield: 11.6 g, 23.6 mmol, 92%.

[0357] Step 2: Synthesis of 4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxylic acid (P9).

[0358] A solution of C13 (11.6 g, 23.6 mmol) in methanol (120 mL) was treated with an aqueous sodium hydroxide solution (3 M; 120 mL). The reaction mixture was stirred at 50 ° C for 6 hours and then acidified by adding hydrochloric acid. The resulting solid was collected by filtration and washed with water, then suspended in a mixture of ethyl acetate and methanol (10: 1 ratio, 80 mL). It was stirred at 80 ° C and slowly treated with methanol until a solution was obtained, which was then allowed to cool to room temperature. The resulting precipitate was collected by filtration and washed with ethyl acetate to obtain P9 as a white solid. Yield: 9.0 g, 18.8 mmol, 80%. LCMSm / z 478.1[M+H] + . 1 H NMR(400MHz,DMSO-d6)d 8.26(br t,J=6.2Hz,1H),7.42–7.23(m,3H),6.94(d,J=8.3Hz,2H),5.21(s,2H), 3.75(s,3H),2.98(d,J=6.2Hz,2H),1.71–1.55(m,6H),1.44–1.29(m,6H.

[0359] Preparation Example P10: 2,3,5-trifluoro-N-{[4-(N-hydroxycarbamimidoyl)bicyclo[2.2.2]oct-1-yl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P10)

[0360]

[0361] Step 1: Synthesis of 4-nitrophenyl 4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxylate (C14).

[0362] To a suspension of P9 (962 mg, 2.01 mmol) in dichloromethane (8 mL) at 0°C was added 4-nitrophenyl chloroformate (425 mg, 2.11 mmol) followed by triethylamine (0.842 mL, 6.04 mmol). The reaction mixture was allowed to warm to room temperature and then stirred at room temperature overnight before being concentrated in vacuo to provide C14 (1.20 g) as a solid. This material was carried directly to the next step. LCMS m / z 599.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d6), characteristic peaks: d 5.21 (s, 2H), 3.75 (s, 3H), 3.05 (d, J = 6.3 Hz, 2H), 1.93–1.83 (m, 6H), 1.53–1.43 (m, 6H).

[0363] Step 2: Synthesis of 4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxamide (C15).

[0364] A solution of C14 (from the previous step; 1.20 g, ≤2.01 mmol) in N,N-dimethylformamide (10 mL) was treated with concentrated ammonium hydroxide (14.5 M; 0.415 mL, 6.02 mmol), and the reaction mixture was stirred at room temperature for 5 hours. It was then added to water (100 mL), and the resulting mixture was extracted with ethyl acetate (3×80 mL); the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo to provide C15 as an off-white solid. Yield: 919 mg, 1.93 mmol, 96% over 2 steps. LCMS m / z 477.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6)d 8.27(br t,J=6Hz,1H),7.35(d,J=8.6Hz,2H),7.30(ddd,J=11.0,6.1,2.4Hz,1H),6.94(d,J=8.6Hz,2H),6.88(br s,1H),6.66(br s,1H),5.21(s,2H),3.75(s,3H),2.99(d,J=6.2Hz,2H),1.66–1.57(m,6H),1.41–1.33(m,6H).

[0365] Step 3: Synthesis of N-[(4-cyanobicyclo[2.2.2]octan-1-yl)methyl]-2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C16).

[0366] To a solution of C15 (797 mg, 1.67 mmol) in ethyl acetate (10 mL) was added (methoxycarbonylsulfamoyl)triethylammonium hydroxide inner salt (Burgess reagent; 997 mg, 4.18 mmol). The reaction mixture was stirred at room temperature overnight, then diluted with ethyl acetate (40 mL) and washed sequentially with water (2×30 mL) and saturated aqueous sodium chloride solution (30 mL). The organic layer was then dried over sodium sulfate, filtered, and concentrated in vacuo to afford C16 as a solid. Yield: 658 mg, 1.44 mmol, 86%. LCMS m / z 459.3 [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.32(br t,J=6.3Hz,1H),7.35(d,J=8.7Hz,2H),7.34–7.28(m,1H),6.94(d,J=8.6Hz,2H),5. 21(s,2H),3.75(s,3H),2.99(d,J=6.3Hz,2H),1.88–1.79(m,6H),1.46–1.37(m,6H).

[0367] Step 4: Synthesis of 2,3,5-trifluoro-N-{[4-(N-hydroxycarbamimidoyl)bicyclo[2.2.2]octan-1-yl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (P10).

[0368] To a suspension of C16 (658 mg, 1.44 mmol) in methanol (8.0 mL) was added triethylamine (0.440 mL, 3.16 mmol) followed by hydroxylamine hydrochloride (219 mg, 3.15 mmol). No reaction was observed within a few hours at room temperature. Hydroxylamine hydrochloride (219 mg, 3.15 mmol) was added again and the reaction mixture was heated at 50 ° C for 24 hours. After cooling, it was diluted with ethyl acetate (30 mL) and washed with water (2×40 mL) and saturated sodium chloride aqueous solution (30 mL) in sequence. The organic layer was then dried over sodium sulfate, filtered, and concentrated in vacuo to provide P10 as a solid. Yield: 330 mg, 0.671 mmol, 47%. LCMSm / z 492.4[M+H] + . 1 H NMR(400MHz,DMSO-d6)d 11.82(br s,1H),10.64(br s,1H),8.59(v br s,1H),8.33(br t,J=6.3Hz,1H),7.35(d,J=8.6Hz,2H),7.34–7.28(m,1H),6.94(d,J=8.6Hz,2H),5. 21(s,2H),3.75(s,3H),3.03(d,J=6.3Hz,2H),1.78–1.67(m,6H),1.49–1.38(m,6H).

[0369] Preparation Example P11: tert-Butyl {[(1r,4r)-4-(7-bromoimidazo[1,2-a]pyridin-2-yl)cyclohexyl]methyl}carbamate (P11)

[0370]

[0371]

[0372] Step 1: Synthesis of tert-butyl ({(1r,4r)-4-[methoxy(methyl)carbamoyl]cyclohexyl}methyl)carbamate (C17).

[0373] To a solution of (1r,4r)-4-{[(tert-butoxycarbonyl)amino]methyl}cyclohexane-1-carboxylic acid (10.2 g, 39.6 mmol) in N,N-dimethylformamide (100 mL) was added N,O-dimethylhydroxylamine hydrochloride (4.66 g, 47.8 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 19.7 g, 51.8 mmol) and triethylamine (16.7 mL, 120 mmol). After the reaction mixture was stirred at room temperature overnight, LCMS analysis indicated the formation of C17: LCMS m / z 301.5 [M+H] + In a pilot reaction run on a smaller scale, the reaction mixture was then concentrated under reduced pressure, diluted with a 1:1 mixture of ethyl acetate and dichloromethane, and filtered; the filtrate was concentrated in vacuo to provide C17. The product from the 39.6 mmol scale reaction was combined with the product of a similar reaction performed using (1r,4r)-4-{[(tert-butoxycarbonyl)amino]methyl}cyclohexane-1-carboxylic acid (9.50 g, 36.9 mmol) to provide C17 as an oil. Combined yield: 22.8 g, 75.9 mmol, 99%. 1 H NMR(500MHz,chloroform-d)d 4.56(br s,1H),3.68(s,3H),3.16(s,3H),2.98(br d,J=6.4Hz,2H),2.69–2.57(m,1H),1.87–1.77(m,4H),1.57–1.38(m,3H),1.44(s,9H),1.05–0.94(m,2H).

[0374] Step 2: Synthesis of tert-butyl {[(1r,4r)-4-acetylcyclohexyl]methyl}carbamate (C18).

[0375] Methylmagnesium bromide (3.0 M; 81.7 mL, 245 mmol) was added dropwise to a 0° C. solution of C17 (23.0 g, 76.6 mmol) in tetrahydrofuran (219 mL), and the reaction mixture was then allowed to warm to room temperature. After 2 hours, it was cooled to 0° C., treated with water (50 mL), and then diluted with ethyl acetate. The aqueous layer was extracted twice with ethyl acetate, and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (eluent: 0%, then 10%, then 25%, then 50% ethyl acetate / heptane) gave C18 as a solid. Yield: 13.3 g, 52.1 mmol, 68%. 1 H NMR(500MHz,chloroform-d)d 4.56(br s,1H),2.98(br dd,J=6,6Hz,2H),2.28(tt,J=12.2,3.5Hz,1H),2.13(s,3H),1.98–1.91(m,2H),1.88 –1.81(m,2H),1.44(s,9H),1.44–1.37(m,1H),1.37–1.26(m,2H),1.02–0.92(m,2H).

[0376] Step 3: Synthesis of tert-butyl {[(1r,4r)-4-(bromoacetyl)cyclohexyl]methyl}carbamate (C19).

[0377] Bromine (2.57mL, 50.2mmol) is added in batches to a methanol (158mL) solution of C18 (12.1g, 47.4mmol) at 0°C. After the mixture is stirred at 0°C for 1 hour and at room temperature for 1 hour, N, N-diisopropylethylamine (29.6mL, 170mmol) is added in batches. Stirring is continued at room temperature for 20 minutes, then the mixture is concentrated in vacuo and merged with the product of a similar reaction carried out using C18 (1.03g, 4.03mmol). Silica gel chromatography (eluent: 0%, then 10%, then 25% ethyl acetate / heptane) provides C19 as a solid. Combined yield: 9.07g, 27.1mmol, 53%. 1 H NMR (500 MHz, CHLOROFORM-d) d 4.56 (br s, 1H), 3.95 (s, 2H), 2.99 (dd, J = 6, 6 Hz, 2H), 2.68 (tt, J = 12.1, 3.4 Hz, 1H), 2.00–1.92 (m, 2H), 1.90–1.82 (m, 2H), 1.48–1.34 (m, 3H), 1.44 (s, 9H), 1.06–0.95 (m, 2H).

[0378] Step 4: Synthesis of tert-butyl {[(1r,4r)-4-(7-bromoimidazo[1,2-a]pyridin-2-yl)cyclohexyl]methyl}carbamate (P11).

[0379] A suspension of C19 (1.00 g, 2.99 mmol) and 4- bromopyridine -2- amine (1.04 g, 6.01 mmol) in ethanol (20 mL) was heated at 70 ° C overnight. After the reaction mixture was cooled to room temperature, it was poured into water (150 mL) under stirring and stirred for 20 minutes. The solid was collected via filtration and washed with water to obtain P11 as a white solid. Yield: 976 mg, 2.39 mmol, 80%. LCMSm / z 408.2 (bromine isotope pattern observed) [M + H] + . 1 H NMR (400MHz, DMSO-d6)d 8.41(br d,J=7.1Hz,1H),7.75(br d,J=2.0Hz,1H),7.69(s,1H),6.97(dd,J=7.2,2.0Hz,1H),6.83(br t,J=5.9Hz,1H),2.81(dd,J=6,6Hz,2H),2.64–2.53(m,1H),2.09–1.99(m, 2H),1.82–1.73(m,2H),1.45–1.29(m,3H),1.38(s,9H),1.08–0.94(m,2H)

[0380] Preparation Example P12: tert-Butyl ({(1r,4r)-4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)carbamate (P12)

[0381]

[0382] Step 1: Synthesis of tert-butyl {[(1r,4r)-4-(6-bromo-2H-indazol-2-yl)cyclohexyl]methyl}carbamate (C20).

[0383] A suspension of tert-butyl {[(1r, 4r)-4-aminocyclohexyl] methyl}carbamate (5.00 g, 21.9 mmol) and 4-bromo-2-nitrobenzaldehyde (5.04 g, 21.9 mmol) in propan-2-ol (70 mL) was heated at 80 ° C for 4 hours. After the reaction mixture was cooled to room temperature, tributylphosphine (94%, 12 mL, 45 mmol) was added via a syringe within 5 minutes; then, the reaction mixture was heated at 80 ° C overnight. After cooling to room temperature, the reaction mixture was filtered and the filter cake was washed with heptane to give C20 as a tan solid. Yield: 6.52 g, 16.0 mmol, 73%. LCMS m / z 408.1 (bromine isotope pattern observed) [M + H] + . 1 H NMR (400MHz, DMSO-d6)8.45(s,1H),7.86–7.82(m,1H),7.67(d,J=8.8Hz,1H),7.12(dd,J=8.8,1.7Hz,1H),6.89(br t,J=6.0Hz,1H),4.50–4.38(m,1H),2.85(dd,J=6.3,6.3Hz,2H),2.17–2.07( m,2H),1.93–1.78(m,4H),1.54–1.41(m,1H),1.39(s,9H),1.20–1.04(m,2H)

[0384] Step 2: Synthesis of tert-butyl ({(1r,4r)-4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)carbamate (P12).

[0385] A mixture of C20 (6.52 g, 16.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-di-1,3,2-dioxaborolane (6.08 g, 23.9 mmol) and potassium acetate (95%, 4.95 g, 47.9 mmol) in 1,4-dioxane (200 mL) was degassed with nitrogen for 10 minutes, followed by the addition of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (Pd(dppf)Cl2; 652 mg, 0.798 mmol). After the reaction mixture was heated at 100° C. for 1 hour, it was allowed to cool and filtered through a pad of celite. The filter cake was rinsed with ethyl acetate, and the combined filtrates were concentrated in vacuo; silica gel chromatography (gradient: 30% to 70% ethyl acetate / heptane; loaded as a solution in dichloromethane) provided P12 as a colorless foam. Yield: 7.20 g, 15.8 mmol, 99%. LCMS m / z 456.4 [M+H]+ . 1 H NMR (400 MHz, DMSO-d6), characteristic peaks: d8.40 (s, 1H), 7.96–7.93 (m, 1H), 7.65 (br d, J=8.4 Hz, 1H), 7.25 (br d, J=8.4 Hz, 1H), 6.89 (br t, J=6.0 Hz, 1H), 4.53–4.39 (m, 1H), 2.85 (dd, J=6,6 Hz, 2H), 2.19–2.08 (m, 2H), 1.93–1.78 (m, 4H), 1.56–1.43 (m, 1H), 1.39 (s, 9H), 1.31 (s, 12H).

[0386] Preparation Example P13: 1-{(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride (P13)

[0387]

[0388] Step 1: Synthesis of tert-butyl ({(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methyl)carbamate (C21).

[0389] 4-Bromo-1-methyl-1H-pyrazole (233 mg, 1.45 mmol), P12 (600 mg, 1.32 mmol), aqueous potassium carbonate solution (2 M; 1.98 mL, 3.96 mmol), [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) [Pd(dtbpf)Cl2; 85.9 mg, 0.132 mmol], ethanol (5 mL), and water (1 mL) were combined in a reduced pressure vial, and the reaction mixture was heated at 85° C. for 1 hour. After the reaction mixture cooled, vacuum concentration was used to remove ethanol, and the resulting mixture was partitioned between ethyl acetate and water. The organic layer was washed with saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated in vacuo; silica gel chromatography (eluent: ethyl acetate, then 5% methanol / ethyl acetate) provided C21 as a colorless foam. Yield: 404 mg, 0.986 mmol, 75%. LCMS m / z 410.4 [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.31(br s,1H),8.15(s,1H),7.91–7.89(m,1H),7.76–7.72(m,1H),7.65(dd,J=8.6,0.9Hz,1H),7.25(dd,J=8.7,1.4Hz,1H),6.90(br t,J=5.9Hz,1H),4.47–4.34(m,1H),3.87(s,3H),2.85(dd,J=6,6Hz,2H),2.18–2. 07(m,2H),1.94–1.78(m,4H),1.54–1.42(m,1H),1.39(s,9H),1.20–1.05(m,2H).

[0390] Step 2: Synthesis of 1-{(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride (P13)

[0391] A solution of hydrogen chloride in 1,4-dioxane (4 M; 6 mL) was added to C21 (404 mg, 0.986 mmol). Propan-2-ol (3 mL) was added to aid solubility and stirring, and the reaction mixture was allowed to stir overnight before being diluted with diethyl ether (50 mL). The solid was collected by filtration and washed with diethyl ether to provide P13 as a solid. Yield: 362 mg, assumed quantitative. LCMS m / z 310.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6)d 8.37(br s,1H),8.17(s,1H),7.92(s,1H),7.74(br s,1H),7.66(br d,J=8.6Hz,1H),7.27(dd,J=8.7,1.4Hz,1H),4.50–4.39(m,1H),3.87(s,3H),2.78–2. 68(m,2H),2.22–2.12(m,2H),2.01–1.84(m,4H),1.78–1.65(m,1H),1.29–1.15(m,2H).

[0392] Preparation Example P14: 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (P14)

[0393]

[0394] Step 1: Synthesis of 1-[(1r,4r)-4-(6-bromo-2H-indazol-2-yl)cyclohexyl]methanamine hydrochloride (C22).

[0395] A solution of hydrogen chloride in 1,4-dioxane (4M; 25 mL, 100 mmol) was added to a solution of C20 (7.35 g, 18.0 mmol) in 1,4-dioxane (30 mL); the reaction mixture was stirred at room temperature for 3 hours and then stirred at 50 ° C overnight. After the reaction mixture was cooled, it was diluted with diethyl ether (100 mL). The solid was collected by filtration and washed with diethyl ether to give C22 as a solid. Yield: 6.20 g, 18.0 mmol, quantitative. LCMS m / z 308.5 (bromine isotope pattern observed) [M + H] + . 1 H NMR(400MHz,DMSO-d6)d8.49(br s,1H),8.06(br s,3H),7.86–7.83(m,1H),7.68(d,J=8.8Hz,1H),7.12(dd,J=8.8,1.7Hz,1H),4.47(tt,J=11.7,3.9Hz ,1H),2.78–2.66(m,2H),2.21–2.11(m,2H),2.01–1.82(m,4H),1.78–1.65(m,1H),1.29–1.14(m,2H).

[0396] Step 2: Synthesis of N-{[(1r,4r)-4-(6-bromo-2H-indazol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C23).

[0397] To a suspension of C22 (6.20 g, 18.0 mmol) and P1 (5.92 g, 20.1 mmol) in N,N-dimethylformamide (15 mL) was added N,N-diisopropylethylamine (14 mL, 80.4 mmol) followed by O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 95%; 9.66 g, 24.1 mmol). The reaction mixture was stirred at room temperature for 3 days before being poured into water (450 mL) with stirring. The resulting solid was separated by filtration and washed with water to provide C23 as a solid. Yield: 10.0 g, 17.1 mmol, 95%. LCMS m / z 584.2 (bromine isotope pattern observed) [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.57(br t,J=5.8Hz,1H),8.45(br s,1H),7.86–7.83(m,1H),7.67(d,J=8.8Hz,1H),7.65–7.56(m,2H),7.34(d,J=8.6Hz,2H),7.12(dd,J=8.8,1.7Hz,1H),6.92(d,J=8.6Hz,2H),5 .17(s,2H),4.53–4.41(m,1H),3.74(s,3H),3.17(dd,J=6,6Hz,2H),2.2 1–2.09(m,2H),1.96–1.80(m,4H),1.73–1.59(m,1H),1.28–1.13(m,2H).

[0398] Step 3: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (P14).

[0399] A mixture of C23 (10 g, 17.1 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-di-1,3,2-dioxaborolane (6.52 g, 25.7 mmol) and potassium acetate (95%, 5.30 g, 51.3 mmol) in 1,4-dioxane (250 mL) was degassed with nitrogen for 10 minutes. [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (700 mg, 0.857 mmol) was added and the reaction mixture was purged with nitrogen for another 5 minutes before being heated at 100°C for 2 hours. After the reaction mixture cooled, it was filtered through celite and the filter pad was rinsed with ethyl acetate. The combined filtrates were concentrated in vacuo, and the residue was purified by silica gel chromatography (gradient: 2% to 10% methanol / dichloromethane; loaded as a solution in dichloromethane) to afford P14 as a tan solid. Yield: 7.32 g, 11.6 mmol, 68%. LCMS m / z 632.3 [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.57(br t,J=5.8Hz,1H),8.40(br s,1H),7.96–7.94(m,1H),7.65(dd,J=8.4,1Hz,1H),7.66–7.56(m,2H),7.34(d,J=8.6Hz,2H),7.25(br d,J=8.4Hz,1H),6.92(d,J=8.7Hz,2H),5.17(s,2H),4.55–4.43(m,1H),3.74(s,3H),3.18(dd,J=6, 6Hz,2H),2.21–2.10(m,2H),1.96–1.81(m,4H),1.75–1.60(m,1H),1.31(s,12H),1.28–1.14(m,2H).

[0400] Preparation Example P15: N-{[(1r,4r)-4-(6-chloro-2H-pyrazolo[4,3-c]pyridin-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P15)

[0401]

[0402]

[0403] Step 1: Synthesis of tert-butyl {[(1r,4r)-4-(6-chloro-2H-pyrazolo[4,3-c]pyridin-2-yl)cyclohexyl]methyl}carbamate (C24).

[0404] A mixture of 6-chloro-4-nitropyridine-3-carboxaldehyde (2.00 g, 10.7 mmol) and tert-butyl {[(1r, 4r)-4-aminocyclohexyl] methyl} carbamate (2.45 g, 10.7 mmol) in propan-2-ol (50 mL) was heated at 80 ° C for 4 hours, after which the reaction mixture was cooled to room temperature. Tributylphosphine (6.51 g, 32.2 mmol) was then added, and the reaction mixture was heated at 80 ° C for another 6 hours. After the solvent was removed in vacuo, the residue was purified via reverse phase HPLC (column: Waters XBridge C18, 30 × 150 mm, 5 μm; mobile phase A: water containing 0.05% formic acid; mobile phase B: acetonitrile; gradient: 50% to 60% B; flow rate: 20 mL / min) to provide C24 as a white solid. Yield: 260 mg, 0.713 mmol, 7%. LCMS m / z 365.2 (chlorine isotope pattern observed) [M+H] + . 1H NMR (400MHz, DMSO-d6)d 9.01 (d, J=1.2Hz, 1H), 8.81 (brs, 1H), 7.68 (br s, 1H), 6.91 (br t,J=5.9Hz,1H),4.59–4.47(m,1H),2.85(dd,J=6,6Hz,2H),2.20–2.08(m, 2H),1.95–1.78(m,4H),1.54–1.40(m,1H),1.38(s,9H),1.20–1.04(m,2H).

[0405] Step 2: Synthesis of 1-[(1r,4r)-4-(6-chloro-2H-pyrazolo[4,3-c]pyridin-2-yl)cyclohexyl]methanamine hydrochloride (C25).

[0406] To a solution of C24 (260 mg, 0.713 mmol) in dichloromethane (4 mL) was added a 1,4-dioxane solution of hydrogen chloride (4 M; 1 mL, 4 mmol). After the reaction mixture was stirred at 15 ° C for 4 hours, it was concentrated in vacuo to afford C25 as a yellow oil. Yield: 170 mg, 0.564 mmol, 79%. LCMS m / z 265.1 (chlorine isotope pattern observed) [M + H] + .

[0407] Step 3: Synthesis of N-{[(1r,4r)-4-(6-chloro-2H-pyrazolo[4,3-c]pyridin-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P15).

[0408] To a solution of P1 (186 mg, 0.632 mmol), N,N-diisopropylethylamine (163 mg, 1.26 mmol) and O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 288 mg, 0.757 mmol) in dichloromethane (10 mL) was added C25 (167 mg, 0.554 mmol), and the reaction mixture was stirred at 15 ° C for 1 hour. It was then extracted with dichloromethane (3×30 mL), and the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Purification using silica gel chromatography (eluent: 4% methanol / dichloromethane) provided P15 as a yellow oil. Yield: 250 mg, 0.462 mmol, 83%. LCMS m / z 541.1 (chlorine isotope pattern observed) [M+H] + . 1H NMR (400 MHz, DMSO-d6), characteristic peaks: d 9.01 (d, J = 1.2 Hz, 1H), 8.80 (br s, 1H), 8.58 (br t, J = 5.8 Hz, 1H), 7.68 (br s, 1H), 7.66–7.56 (m, 2H), 7.34 (d, J = 8.6 Hz, 2H), 6.92 (d, J = 8.7 Hz, 2H), 5.17 (s, 2H), 4.63–4.49 (m, 1H), 3.74 (s, 3H), 2.21–2.10 (m, 2H).

[0409] Preparation Example P16: N-{[4-(6-bromo-2H-indazol-2-yl)bicyclo[2.2.2]oct-1-yl]methyl}-2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamide (P16)

[0410]

[0411] A solution of P8 (200 mg, 0.446 mmol) and 4-bromo-2-nitrobenzaldehyde (123 mg, 0.535 mmol) in propan-2-ol (10 mL) was stirred at 85 ° C for 4 hours, then cooled to room temperature and treated with tributylphosphine (2 mL, 8 mmol). After the reaction mixture was stirred at 85 ° C overnight, it was diluted with water (15 mL) and extracted with dichloromethane (3 × 10 mL). The combined organic layer was washed with saturated sodium chloride aqueous solution (100 mL), dried over sodium sulfate, filtered, concentrated in vacuo, and subjected to silica gel chromatography (gradient: 0% to 10% methanol / dichloromethane) to obtain P16 as a yellow solid. 1 H NMR data were obtained from a reaction carried out in a similar manner. Yield: 170 mg, 0.270 mmol, 61%. LCMS m / z 626.1 (bromine isotope pattern observed) [MH] - . 1 H NMR(400MHz, DMSO-d6)d 8.46(d,J=1.0Hz,1H),8.40(br t,J=6.3Hz,1H),7.86–7.83(m,1H),7.66(br d,J=8.8Hz,1H),7.38–7.32(m,1H),7.36(d,J=8.7Hz,2H),7.11(dd,J=8.8,1.7Hz,1H),6.94(d,J= 8.7Hz,2H),5.22(s,2H),3.75(s,3H),3.10(d,J=6.2Hz,2H),2.20–2.10(m,6H),1.71–1.60(m,6H).

[0412] Example 1: N-{[(1r,4r)-4-(1,3-benzoxazol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide (1)

[0413]

[0414] Step 1: Synthesis of N-{[(1r,4r)-4-(1,3-benzoxazol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C26).

[0415] 1,3,5-trichloro-1,3,5-triazinane-2,4,6-trione (161 mg, 0.693 mmol) and P4 (1.00 g, 2.31 mmol) are added to a mixture of triphenylphosphine (95%, 637 mg, 2.31 mmol) in 1,4-dioxane (40 mL) at 0 ° C. After the reaction mixture is stirred for 30 minutes, it is allowed to warm to room temperature, 2-aminophenol (378 mg, 3.46 mmol) is added, and the reaction mixture is stirred at 105 ° C overnight. Once it has cooled, the reaction mixture is filtered through a celite pad, and the pad is rinsed with 1,4-dioxane, ethyl acetate and dichloromethane in sequence. The combined filtrate is concentrated in vacuo to obtain C26 as an orange oil, which is directly entered into the next step. LCMSm / z 507.3[M+H] + Step 2: Synthesis of N-{[(1r,4r)-4-(1,3-benzoxazol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide (1).

[0416] A suspension of C26 (from the previous step; ≤ 2.31 mmol) in 1,4-dioxane (20 mL) at 0°C was treated with a solution of hydrogen chloride in 1,4-dioxane (4 M; 20 mL). After the reaction mixture was allowed to stir at room temperature for 2 hours, it was concentrated in vacuo and purified via silica gel chromatography (Gradient: 0% to 100% ethyl acetate / heptane; sample loaded into dichloromethane containing minimal methanol). The resulting material was partitioned between saturated aqueous sodium bicarbonate solution and ethyl acetate, and the organic layer was subsequently washed with water, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was subjected to silica gel chromatography (gradient: 0% to 100% ethyl acetate / heptane, followed by 0% to 10% methanol / dichloromethane; sample loaded into dichloromethane containing minimal methanol) to provide N-{[(1r,4r)-4-(1,3-benzoxazol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide (1) as a white solid. Yield: 46 mg, 0.12 mmol, 5% over 2 steps. LCMS m / z 387.3 [M+H] + .1 H NMR (400 MHz, methanol-d4) d 7.65–7.60 (m, 1H), 7.58–7.53 (m, 1H), 7.52–7.42 (m, 2H), 7.38–7.31 (m, 2H), 3.27 (d, J = 7.0 Hz, 2H), 2.98 (tt, J = 12.2, 3.6 Hz, 1H), 2.32–2.22 (m, 2H), 2.04–1.94 (m, 2H), 1.80–1.62 (m, 3H), 1.31–1.16 (m, 2H).

[0417] Example 2: 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (2)

[0418]

[0419]

[0420] Step 1: Synthesis of N-hydroxy-5-(trifluoromethyl)pyridine-2-carboximidamide (C27).

[0421] To a mixture of 5-(trifluoromethyl)pyridine-2-carbonitrile (200 mg, 1.16 mmol) and hydroxylamine hydrochloride (242 mg, 3.48 mmol) in ethanol (20 mL) was added sodium hydroxide (139 mg, 3.48 mmol). After the reaction mixture was stirred at room temperature for 3 hours, it was concentrated in vacuo to provide C27 as a white solid. Yield: 200 mg, 0.975 mmol, 84%. LCMS m / z 206.1 [M+H] + .

[0422] Step 2: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (C28)

[0423] To a mixture of P4 (400 mg, 0.923 mmol), N,N-diisopropylethylamine (358 mg, 2.77 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 526 mg, 1.38 mmol) in dichloromethane (20 mL) at 0 ° C was added C27 (227 mg, 1.11 mmol). The reaction mixture was stirred at room temperature for 6 hours, then diluted with water (20 mL) and extracted with dichloromethane (2×20 mL); the combined organic layers were washed with saturated aqueous sodium chloride solution (2×20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 7% methanol / dichloromethane) provided the acylated intermediate (100 mg, 0.161 mmol, 17%) as a white solid, LCMS m / z 621.3 [M+H] + .

[0424] The material was dissolved in a mixture of ethanol (4 mL) and water (1 mL), treated with sodium acetate (39.6 mg, 0.483 mmol), and stirred at 100 ° C for 1 hour under microwave irradiation. After the reaction mixture was concentrated in vacuo, it was purified using silica gel chromatography (gradient: 0% to 5% methanol / dichloromethane) to provide C28 as a white solid. Yield: 60 mg, 0.10 mmol, 11% (from P4). LCMS m / z 603.3 [M + H] + .

[0425] Step 3: Synthesis of 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (2)

[0426] To a solution of C28 (60 mg, 0.10 mmol) in dichloromethane (5 mL) was added a 1,4-dioxane solution of hydrogen chloride (4 M; 1 mL). After the reaction mixture was stirred at room temperature for 2 hours, it was concentrated in vacuo, diluted with dichloromethane (10 mL), treated with sodium bicarbonate (10 mg, 0.12 mmol) and concentrated under reduced pressure. Silica gel chromatography (gradient: 0% to 5% methanol / dichloromethane) gave 3,5-difluoro-4-hydroxy-N-{[(1r, 4r)-4-{3-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (2) as a white solid. Yield: 11.6 mg, 24.0 μmol, 24%. LCMSm / z 483.2[M+H] + . 1H NMR(400MHz,DMSO-d6)d 9.19–9.14(m,1H),8.48–8.39(m,2H),8.27(d,J=8.2Hz,1H),7.60–7.54(m,2H),3.19–3 .05(m,3H),2.25–2.13(m,2H),1.93–1.81(m,2H),1.67–1.51(m,3H),1.22–1.07(m,2H).

[0427] Example 3: 2,3,5-Trifluoro-4-hydroxy-N-[(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide (3)

[0428]

[0429] Step 1: Synthesis of 5-(trifluoromethyl)pyrimidine-2-carbonitrile (C29).

[0430] A solution of tetraethylammonium cyanide (1.88 g, 12.0 mmol) and 1,4-diazabicyclo[2.2.2]octane (1.47 g, 13.1 mmol) in acetonitrile (8 mL) was added to a mixture of 2-chloro-5-(trifluoromethyl)pyrimidine (2.00 g, 11.0 mmol) in acetonitrile (8 mL), and the reaction mixture was subsequently stirred at room temperature for 3 hours. The solvent was removed in vacuo to provide a residue containing C29; the pale yellow solid material was directly taken to the next step.

[0431] Step 2: Synthesis of N-hydroxy-5-(trifluoromethyl)pyrimidine-2-carboximidamide (C30).

[0432] A mixture of C29 (from the previous step; ≤ 11.0 mmol), hydroxylamine hydrochloride (1.52 g, 21.9 mmol) and N,N-diisopropylethylamine (4.26 g, 33.0 mmol) in methanol (20 mL) was stirred at 70° C. for 12 hours. The reaction mixture was concentrated in vacuo to afford C30 (1.70 g), which was directly carried to the next step. LCMS m / z 207.1 [M+H] + Step 3: Synthesis of tert-butyl [(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]carbamate (C31).

[0433] To a solution of C30 (from the previous step; 1.70 g, ≤8.25 mmol) and 4-{[(tert-butoxycarbonyl)amino]methyl}bicyclo[2.2.2]octane-1-carboxylic acid (2.57 g, 9.07 mmol) in N,N-dimethylformamide (10 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 4.70 g, 12.4 mmol) and N,N-diisopropylethylamine (3.20 g, 24.8 mmol). After the reaction mixture was stirred at 25 °C for 2 hours, it was diluted with water and filtered; the filtrate was concentrated in vacuo to provide the acyl intermediate as a yellow solid. Yield: 1.90 g, 4.03 mmol, 37% over 3 steps. LCMS m / z 472.2 [M+H] + .

[0434] To a solution of the acyl intermediate (2.00 g, 4.24 mmol) in a mixture of ethanol (6 mL) and water (3 mL) was added sodium acetate (1.04 g, 12.7 mmol). After the reaction mixture was stirred at 100 ° C. under microwave irradiation for 1 hour, it was concentrated in vacuo. Purification via silica gel chromatography (gradient: 0% to 6% methanol / dichloromethane) provided C31 as a white solid. Yield: 1.00 g, 2.21 mmol, 52% (from the acyl intermediate). LCMS m / z 454.2 [M + H] + . 1 H NMR (400 MHz, methanol-d4) d 9.35–9.33 (m, 2H), 2.87 (s, 2H), 2.15–2.03 (m, 6H), 1.64–1.53 (m, 6H), 1.45 (s, 9H).

[0435] Step 4: Synthesis of 1-(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methanamine (C32).

[0436] A solution of hydrogen chloride in 1,4-dioxane (4M; 5mL, 20mmol) was added to a solution of C31 (1.00g, 2.21mmol) in dichloromethane (15mL), and the reaction mixture was subsequently stirred at room temperature for 2 hours. It was then concentrated in vacuo, diluted with dichloromethane (10mL), treated with sodium bicarbonate, and concentrated under reduced pressure again. Silica gel chromatography (gradient: 0% to 7% methanol / dichloromethane) gave C32 as a white solid. Yield: 800mg, quantitative. LCMSm / z 354.2[M+H] + . 1HNMR(400MHz, methanol-d4)d 9.35(br s,2H),2.80(s,2H),2.22–2.10(m,6H),1.75–1.65(m,6H).

[0437] Step 5: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-[(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide (3).

[0438] To a solution of C32 (100 mg, 0.283 mmol) and 2,3,5-trifluoro-4-hydroxybenzoic acid (65.2 mg, 0.339 mmol) in N,N-dimethylformamide (5 mL) was added 1H-benzotriazol-1-ol (57.4 mg, 0.425 mmol), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (81.4 mg, 0.425 mmol) and N,N-diisopropylethylamine (110 mg, 0.851 mmol). After the reaction mixture was stirred at 25° C. for 4 hours, it was diluted with water (15 mL) and extracted with dichloromethane (3×10 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (100 mL), dried over sodium sulfate, filtered, concentrated in vacuo, and purified via reverse phase HPLC (column: Waters XBridge C18, 19×150 mm, 5 μm; mobile phase A: water containing 0.1% formic acid; mobile phase B: acetonitrile; gradient: 65% to 75% B; flow rate: 20 mL / min) to afford 2,3,5-trifluoro-4-hydroxy-N-[(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide (3) as a white solid. Yield: 105 mg, 0.199 mmol, 70%. LCMS m / z 528.0 [M+H] + . 1 H NMR(400MHz,DMSO-d6)d 11.36(br s,1H),9.49(s,2H),8.20(br t, J=6Hz, 1H), 7.28 (ddd, J=11.0, 6.2, 2.3Hz, 1H), 3.09 (d, J=6.3Hz, 2H), 2.06–1.93 (m, 6H), 1.61–1.50 (m, 6H).

[0439] Example 4: 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (4)

[0440]

[0441] Step 1: Synthesis of tert-butyl {[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}carbamate (C33)

[0442] To a solution of C30 (453 mg, 2.20 mmol) in N,N-dimethylformamide (8 mL) was added (1r,4r)-4-{[(tert-butoxycarbonyl)amino]methyl}cyclohexane-1-carboxylic acid (679 mg, 2.64 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 1.25 g, 3.29 mmol) and N,N-diisopropylethylamine (852 mg, 6.59 mmol). The reaction was stirred at 25 ° C for 2 hours, after which it was diluted with ice water (30 mL) and the solid was collected by filtration to provide the acyl intermediate as a brown solid. Yield: 510 mg, 1.14 mmol, 52%. LCMS m / z 446.1 [M + H] + . 1 H NMR (400 MHz, chloroform-d), characteristic peaks, integration is approximate: d 9.08 (br s, 2H), 3.05–2.96 (m, 2H), 2.48 (tt, J = 12.3, 3.6 Hz, 1H), 2.16–2.06 (m, 2H), 1.92–1.82 (m, 2H), 1.66–1.53 (m, 2H), 1.09–0.94 (m, 2H)

[0443] To a solution of the acyl intermediate (700 mg, 1.57 mmol) in dichloromethane (5 mL) was added a solution of tetrabutylammonium fluoride in tetrahydrofuran (1 M; 5 mL, 5 mmol), and the reaction mixture was stirred at 25 ° C for 4 hours. It was then concentrated in vacuo and subjected to silica gel chromatography (gradient: 0% to 50% ethyl acetate / petroleum ether) to afford C33 as a white solid. Yield: 340 mg, 0.795 mmol, 51%. LCMS m / z 450.1 [M + Na + ]. 1H NMR (400 MHz, chloroform-d) d 9.20–9.18 (m, 2H), 4.60 (br s, 1H), 3.11–2.99 (m, 3H), 2.34–2.24 (m, 2H), 2.00–1.91 (m, 2H), 1.84–1.69 (m, 2H), 1.6–1.50 (m, 1H, assumed; mostly obscured by water peak), 1.45 (s, 9H), 1.20–1.06 (m, 2H)

[0444] Step 2: Synthesis of 1-[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methanamine hydrochloride (C34).

[0445] A solution of hydrogen chloride in 1,4-dioxane (4 M; 2 mL, 8 mmol) was added to a solution of C33 (340 mg, 0.795 mmol) in dichloromethane (5 mL). After the reaction mixture was stirred at 25 ° C for 2 hours, it was concentrated in vacuo to provide C34 as a white solid. Yield: 200 mg, 0.550 mmol, 69%. LCMS m / z 328.1 [M + H] + . 1 H NMR(400MHz,DMSO-d6)d 9.51–9.49(m,2H),8.00(br s,3H),3.14(tt,J=12.1,3.6Hz,1H),2.75–2.65(m,2H),2.27–2.17(m,2H),1.97–1.87(m,2H),1.73–1.53(m,3H),1.23–1.09(m,2H).

[0446] Step 3: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (C35).

[0447] To a 0°C solution of P1 (27 mg, 91.8 μmol), C34 (30 mg, 82 μmol) and O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 52.3 mg, 0.138 mmol) in N,N-dimethylformamide (3 mL) was added N,N-diisopropylethylamine (35.5 mg, 0.275 mmol), and the reaction mixture was stirred at 25°C for 2 hours. It was then treated with ice water (30 mL) and the resulting solid was collected by filtration to provide C35 (60 mg) as a white solid. This material was taken directly to the next step. LCMS m / z 626.2 [M+Na + ]. 1 H NMR(400MHz,DMSO-d6)d 9.49(s,2H),8.56(br t,J=5.8Hz,1H),7.66–7.56(m,2H),7.34(d,J=8.5Hz,2H),6.92(d,J=8.4Hz,2H),5.17(s,2H),3.74(s ,3H),3.19–3.08(m,3H),2.24–2.15(m,2H),1.91–1.82(m,2H),1.67–1.51(m,3H),1.22–1.08(m,2H). Step 4: Synthesis of 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (4).

[0448] A solution of C35 (from the previous step; 60 mg, ≤82 μmol) in dichloromethane (4 mL) was treated with a solution of hydrogen chloride in 1,4-dioxane (4 M; 2 mL), and the reaction mixture was stirred at 25° C. for 2 hours. After removing the volatiles in vacuo, the residue was purified using silica gel chromatography (gradient: 0% to 10% methanol / dichloromethane) to afford 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide (4) as a white solid. Yield: 14.3 mg, 29.6 μmol, 36% over 2 steps. LCMS m / z 484.1 [M+H] + . 1H NMR(400MHz,DMSO-d6)d 10.84(br s,1H),9.49(s,2H),8.44(br t,J=5.8Hz,1H),7.64–7.52(m,2H),3.20–3.07(m,3H),2.25–2.15(m,2H),1.93–1.82(m,2H),1.67–1.51(m,3H),1.22–1.08(m,2H).

[0449] Example 5: 2,3,5-Trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide trifluoroacetate (5)

[0450]

[0451] A mixture of 2,3,5-trifluoro-4-hydroxybenzoic acid (50 mg, 0.26 mmol), P13 (75 mg, 0.22 mmol), 2-hydroxypyridine 1-oxide (26.5 mg, 0.239 mmol) and 1-methyl-1H-imidazole (52 μL, 0.65 mmol) in a mixture of water (0.32 mL) and N,N-dimethylformamide (1.3 mL) was stirred at room temperature for 5 minutes, followed by the addition of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (45.7 mg, 0.238 mmol) in a single portion, and the reaction mixture was stirred at room temperature overnight. After dilution with water, the reaction mixture was acidified by the addition of 1 M hydrochloric acid and extracted with ethyl acetate. The organic layer was washed with water, dried over magnesium sulfate, filtered, and concentrated in vacuo to give a solid that was purified via reverse phase HPLC (column: Waters Sunfire C18, 19×100 mm, 5 μm; mobile phase A: 0.05% trifluoroacetic acid in water (v / v); mobile phase B: 0.05% trifluoroacetic acid in acetonitrile (v / v); gradient: 5% to 95% B over 8.54 minutes, followed by 95% B over 1.46 minutes; flow rate: 25 mL / minute) to provide 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide trifluoroacetate (5). Yield: 46.3 mg, 95.8 μmol, 44%. LCMS m / z 484.6 [M+H] +. Retention time: 2.51 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0452] Example 6: 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-(5-methoxy-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}benzamide (6)

[0453]

[0454]

[0455] Step 1: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-{[(1r,4r)-4-(5-methoxy-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}benzamide (C36)

[0456] A mixture of P3 (134 mg, 0.331 mmol) and 5-bromo-2-methoxypyridine-4-carboxaldehyde (65 mg, 0.30 mmol) in toluene (8 mL) was stirred at 90° C. for 8 hours. The reaction mixture was then concentrated under reduced pressure and diluted with dimethyl sulfoxide (8 mL); copper (I) iodide (5.71 mg, 30.0 μmol), N 1 ,N 1 ,N 2 ,N 2 -tetramethylethane-1,2-diamine (3.49 mg, 30.0 μmol) and sodium azide (39.1 mg, 0.601 mmol). After the reaction mixture was stirred at 100 ° C for 8 hours, it was treated with water (20 mL) and extracted with ethyl acetate (2×20 mL). The combined organic layers were washed with saturated aqueous sodium chloride solution (2×20 mL), dried over sodium sulfate, filtered, concentrated in vacuo, and purified via silica gel chromatography (gradient: 0% to 8% methanol / dichloromethane) to provide C36 as a brown solid. Yield: 50 mg, 93 μmol, 31%. LCMSm / z 537.3 [M+H] + .

[0457] Step 2: Synthesis of 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-(5-methoxy-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}benzamide (6)

[0458] To a solution of C36 (50 mg, 93 μmol) in dichloromethane (5 mL) was added a 1,4-dioxane solution of hydrogen chloride (4 M; 1 mL). The reaction mixture was stirred at room temperature for 2 hours, then concentrated under reduced pressure, treated with dichloromethane (10 mL) and sodium bicarbonate (10 mg), and concentrated in vacuo again. Silica gel chromatography (gradient: 0% to 7% methanol / dichloromethane) provided 3,5-difluoro-4-hydroxy-N-{[(1r, 4r)-4-(5-methoxy-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}benzamide (6) as a white solid. Yield: 5.1 mg, 12 μmol, 13%. LCMSm / z417.2[M+H] + . 1 H NMR(400MHz,DMSO-d6)d 10.84(s,1H),8.92–8.87(m,1H),8.47(br t,J=5.8Hz,1H),8.37(s,1H),7.64–7.53(m,2H),6.88(d,J=1.2Hz,1H),4.59–4.49(m,1H),3.84(s,3H ),3.17(dd,J=6,6Hz,2H),2.21–2.10(m,2H),1.99–1.83(m,4H),1.74–1.60(m,1H),1.29–1.14(m,2H).

[0459] Example 7: 2,3,5-Trifluoro-4-hydroxy-N-({4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (7)

[0460]

[0461]

[0462] Step 1: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-({4-[6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (C37).

[0463] To a solution of P16 (100 mg, 0.159 mmol) and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-di-1,3,2-dioxaborolane (48.5 mg, 0.191 mmol) in 1,4-dioxane (5 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (11.6 mg, 15.9 μmol) and potassium acetate (46.8 mg, 0.477 mmol), and the reaction mixture was stirred at 90° C. for 12 hours. Concentration in vacuo provided C37, which was directly carried to the next step.

[0464] Step 2: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-({4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (C38).

[0465] To a solution of C37 (from the previous step; ≤0.159 mmol) and 2-bromopyrimidine (28.2 mg, 0.177 mmol) in 1,4-dioxane (5 mL) was added [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (10.8 mg, 14.8 μmol) and potassium carbonate (61.4 mg, 0.444 mmol). After the reaction mixture was stirred at 90 °C for 12 hours, it was concentrated in vacuo and purified via silica gel chromatography (gradient: 0% to 60% ethyl acetate / petroleum ether) to give C38 as a white solid. Yield: 40 mg, 64 μmol, 40% over 2 steps. LCMS m / z 628.2 [M+H] + .

[0466] Step 3: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-({4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (7).

[0467] To a solution of C38 (40 mg, 64 μmol) in dichloromethane (10 mL) was added a 1,4-dioxane solution of hydrogen chloride (4 M; 2 mL, 8 mmol), and the reaction mixture was stirred at 25 ° C for 2 hours. It was then concentrated in vacuo, treated with dichloromethane (10 mL) and sodium bicarbonate (1 g), and concentrated under reduced pressure. Silica gel chromatography (gradient: 0% to 7% methanol / dichloromethane) provided 2,3,5-trifluoro-4-hydroxy-N-({4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (7) as a white solid. Yield: 6.0 mg, 12 μmol, 19%. LCMSm / z 508.1[M+H] +. 1 H NMR(400MHz, DMSO-d6)d 11.38(br s,1H),8.91(d,J=4.8Hz,2H),8.67–8.65(m,1H),8.47(d,J=1.0Hz,1H),8.21(br t,J=6Hz,1H),8.08(dd,J=8.8,1.4Hz,1H),7.79(dd,J=8.8,0.9Hz,1H),7.43(t,J=4.8Hz,1H),7 .29(ddd,J=11.1,6.2,2.3Hz,1H),3.12(d,J=6.3Hz,2H),2.25–2.15(m,6H),1.74–1.63(m,6H).

[0468] Example 8: 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[6-(2-methoxypyrimidin-5-yl)-2H-pyrazolo[4,3-c]pyridin-2-yl]cyclohexyl}methyl)benzamide (8)

[0469]

[0470] To a solution of P15 (60 mg, 0.11 mmol), (2-methoxypyrimidin-5-yl)boronic acid (25.6 mg, 0.166 mmol), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (XPhos; 21.1 mg, 44.3 μmol), and potassium carbonate (46.0 mg, 0.333 mmol) in a mixture of 1,4-dioxane (2 mL) and water (0.4 mL) was added tris(dibenzylideneacetone)dipalladium(0) (20.3 mg, 22.2 μmol). The reaction mixture was stirred at 100° C. for 1 hour under microwave irradiation and then concentrated under reduced pressure. The residue was dissolved in dichloromethane (4 mL), treated with a solution of hydrogen chloride in 1,4-dioxane (4 M; 1 mL, 4 mmol), and stirred at 15° C. for 1 hour. After the solvent was removed in vacuo, the product was purified by reverse phase HPLC (column: Welch Xtimate C18, 30×250 mm, 10 μm; mobile phase A: water containing 0.05% formic acid; mobile phase B: acetonitrile; gradient: 43% to 95% B; flow rate: 50 mL / min) to give 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[6-(2-methoxypyrimidin-5-yl)-2H-pyrazolo[4,3-c]pyridin-2-yl]cyclohexyl}methyl)benzamide (8) as a solid. Yield: 5.2 mg, 10 μmol, 9%. LCMS m / z 495.1 [M+H] + . 1H NMR (400MHz, DMSO-d6)d 9.30 (s, 2H), 9.25 (d, J = 1.3Hz, 1H), 8.76 (br s, 1H), 8.43 (br s,1H),8.23–8.20(m,1H),7.62–7.49(m,2H),4.64–4.50(m,1H),3.98(s,3H),3.18(dd,J =6,6Hz,2H),2.24–2.12(m,2H),2.01–1.84(m,4H),1.75–1.60(m,1H),1.33–1.12(m,2H).

[0471] Example 9: 2,3,5-Trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (9)

[0472]

[0473]

[0474] Step 1: Synthesis of N-{[(1r,4r)-4-(6-bromo-2H-indazol-2-yl)cyclohexyl]methyl}-2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C39)

[0475] To a solution of C22 free base (4.67 g, 15.2 mmol) and P2 (4.97 g, 15.9 mmol) in N,N-dimethylformamide (30 mL) was added 4-methylmorpholine (9.99 mL, 90.9 mmol). The suspension was then treated in batches with chloro(dimethylamino)-N,N-dimethylformimine hexafluorophosphate (4.25 g, 15.1 mmol, divided into 4 equal portions), and the reaction mixture was stirred at room temperature for 5 hours before being added dropwise to ice-cold water (300 mL). The resulting suspension was stirred for 20 minutes and filtered; the filter cake was washed three times with water to give C39 as a light brown solid. Yield: 9.20 g, quantitative. LCMS m / z 602.3 (bromine isotope pattern observed) [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.53–8.43(m,2H),7.85(s,1H),7.67(d,J=8.8Hz,1H),7.40–7.30(m,3H),7.12(dd,J=8.8,1.7Hz,1H),6.94(d,J=8.6Hz,2H),5.22(s,2 H),4.53–4.40(m,1H),3.75(s,3H),3.17(dd,J=6,6Hz,2H),2.20–2.09(m,2H),1.97–1.81(m,4H),1.72–1.58(m,1H),1.29–1.13(m,2H).

[0476] Step 2: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (C40).

[0477] A mixture of C39 (700 mg, 1.16 mmol), pyrimidin-5-ylboronic acid (144 mg, 1.16 mmol), sodium carbonate (369 mg, 3.48 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex (94.2 mg, 0.115 mmol) in a mixture of 1,4-dioxane (20 mL) and water (5 mL) was stirred at 90 ° C for 16 hours. The reaction mixture was then concentrated in vacuo and purified via silica gel chromatography (gradient: 0% to 100% ethyl acetate / petroleum ether) to provide C40 as a white solid. Yield: 300 mg, 0.499 mmol, 43%. LCMS m / z 602.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)d 9.20(s,2H),9.18(s,1H),8.53–8.46(m,2H),8.08(br s,1H),7.85(d,J=8.7Hz,1H),7.45(dd,J=8.7,1.6Hz,1H),7.41–7.34(m,1H),7.35(d,J=8.6Hz,2H),6.94(d,J=8.6Hz,2H),5.22(s,2H ),4.59–4.45(m,1H),3.75(s,3H),3.18(dd,J=6,6Hz,2H),2.23–2.13(m,2H),2.01–1.86(m,4H),1.74–1.60(m,1H),1.32–1.16(m,2H).

[0478] Step 3: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (9).

[0479] To a suspension of C40 (300 mg, 0.499 mmol) in dichloromethane (4.0 mL) was added a solution of hydrogen chloride in 1,4-dioxane (4 M; 1 mL, 4 mmol). The reaction mixture was stirred at 25 ° C for 2 hours, then concentrated in vacuo and purified using silica gel chromatography (gradient: 0% to 100% ethyl acetate / petroleum ether) to afford 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (9) as a white solid. Yield: 200 mg, 0.415 mmol, 83%. LCMS m / z 482.2 [M + H] + . 1 H NMR(400MHz,DMSO-d6)d 11.41(br s,1H),9.21(s,2H),9.18(s,1H),8.49(s,1H),8.37–8.30(m,1H),8.09(br s,1H),7.85(d,J=8.7Hz,1H),7.45(dd,J=8.7,1.6Hz,1H),7.30(ddd,J=11.1,6.3,2.2Hz,1H),4.58–4.4 6(m,1H),3.21–3.14(m,2H),2.23–2.13(m,2H),2.02–1.86(m,4H),1.75–1.61(m,1H),1.33–1.16(m,2H).

[0480] Example 10: N-{[(1r,4r)-4-{6-[1-(2,2-difluoroethyl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide trifluoroacetate (10)

[0481]

[0482]

[0483] Step 1: Synthesis of tert-butyl {[(1r,4r)-4-{6-[1-(2,2-difluoroethyl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}carbamate (C41).

[0484] To a mixture of 4-bromo-1-(2,2-difluoroethyl)-1H-pyrazole (46.1 mg, 0.218 mmol) and P12 (100 mg, 0.220 mmol) was added 1,4-dioxane (1.8 mL) and water (0.6 mL), followed by potassium phosphate (140 mg, 0.660 mmol) and bis[di-tert-butyl(4-dimethylaminophenyl)phosphine]palladium(II) dichloride [Pd(amphos)2Cl2; 15.5 mg, 21.9 μmol]. The reaction mixture was heated at 85 ° C for 18 hours and then partitioned between water and ethyl acetate. After extracting the aqueous layer twice with ethyl acetate, the combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 7.5% methanol / dichloromethane) provided C41 as an oil. Yield: 80 mg, 0.17 mmol, 78%. LCMS m / z 460.3 [M+H] + . 1 H NMR (400 MHz, CHLOROFORM-d), characteristic peaks: d 7.91 (br s, 1H), 7.89 (s, 1H), 7.79 (br s, 1H), 7.74 (s, 1H), 7.65 (d, J = 8.7 Hz, 1H), 7.22 (dd, J = 8.7, 1.4 Hz, 1H), 6.13 (tt, J = 55.4, 4.3 Hz, 1H), 4.68–4.58 (m, 1H), 4.51 (td, J = 13.5, 4.3 Hz, 2H), 4.43–4.32 (m, 1H), 3.06 (dd, J = 6, 6 Hz, 2H), 2.39–2.28 (m, 2H), 1.46 (s, 9H).

[0485] Step 2: Synthesis of 1-[(1r,4r)-4-{6-[1-(2,2-difluoroethyl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methanamine trifluoroacetate (C42).

[0486] Trifluoroacetic acid (0.5 mL, 6 mmol) was added dropwise to a solution of C41 (80 mg, 0.17 mmol) in dichloromethane (2 mL). The reaction mixture was stirred at room temperature for 30 minutes and then concentrated to dryness in vacuo; the residue was azeotroped twice with dichloromethane to provide C42 (84 mg) as a colorless oil, most of which was directly used in the next step. LCMS m / z 360.3 [M+H] + . 1HNMR (400 MHz, methanol-d4), characteristic peaks: d 8.28–8.26 (m, 1H), 8.13 (s, 1H), 7.98 (s, 1H), 7.75 (br s, 1H), 7.72 (d, J = 8.7 Hz, 1H), 7.38–7.33 (m, 1H), 6.22 (tt, J = 55.2, 3.9 Hz, 1H), 4.61 (td, J = 14.4, 3.9 Hz, 2H), 4.56–4.44 (m, 1H), 2.90 (d, J = 7.0 Hz, 2H), 2.37–2.27 (m, 2H), 2.13–2.00 (m, 4H), 1.89–1.75 (m, 1H).

[0487] Step 3: Synthesis of N-{[(1r,4r)-4-{6-[1-(2,2-difluoroethyl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide trifluoroacetate (10).

[0488] A solution of C42 (from the previous step; 82 mg, ≤0.17 mmol) in N,N-dimethylformamide (1.8 mL) was treated with water (0.4 mL). 3,5-difluoro-4-hydroxybenzoic acid (36.2 mg, 0.208 mmol), 1-methyl-1H-imidazole (41.4 μL, 0.520 mmol) and 2-hydroxypyridine 1-oxide (64 mg, 5.76 mmol) were added sequentially, and the reaction mixture was stirred at room temperature for 20 minutes. 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (98%, 33.9 mg, 0.173 mmol) was then added, and stirring was continued at room temperature for 18 hours. The reaction mixture was diluted with water (10 mL) and acidified to pH 4 by adding 1 M hydrochloric acid, which was then extracted 3 times with ethyl acetate. The combined organic layers were washed five times with water, dried over magnesium sulfate, filtered, and concentrated in vacuo; reverse phase HPLC (column: Waters Sunfire C18, 19×100 mm, 5 μm; mobile phase A: 0.05% trifluoroacetic acid / water (v / v); mobile phase B: 0.05% trifluoroacetic acid / acetonitrile (v / v); gradient: 20% to 60% B over 8.5 minutes, then 60% to 95% B over 0.5 minutes; flow rate: 25 mL / minute) afforded N-{[(1r,4r)-4-{6-[1-(2,2-difluoroethyl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide trifluoroacetate (10). Yield: 29.3 mg, 46.5 μmol, 27% over 2 steps. LCMS m / z 516.5 [M+H] +. Retention time: 2.59 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0489] Example 11: 2,3,5-Trifluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[4-(trifluoromethyl)-1H-pyrazol-1-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide trifluoroacetate (11)

[0490]

[0491] Step 1: Synthesis of tert-butyl {[(1r,4r)-4-{6-[4-(trifluoromethyl)-1H-pyrazol-1-yl]-2H-indazol-2-yl}cyclohexyl]methyl}carbamate (C43).

[0492] A mixture of P12 (100 mg, 0.220 mmol), 4-(trifluoromethyl)-1H-pyrazole (120 mg, 0.882 mmol) and copper (II) acetate (53 mg, 0.29 mmol) in pyridine (1.3 mL) was heated at 90 ° C for 18 hours. For the first hour, the reaction mixture was open to the air; it was then capped, a needle was inserted through the cap into the atmosphere, and heating was continued for another 17 hours. The reaction mixture was concentrated to dryness in vacuo, and the residue was partitioned between dichloromethane and water. The organic layer was subjected to silica gel chromatography (gradient: 0% to 7.5% methanol / dichloromethane) to give C43 as a colorless oil. Yield: 80.0 mg, 0.173 mmol, 79%. LCMS m / z 464.3 [M+H] + . 1 H NMR (400 MHz, chloroform-d), characteristic peaks: d 8.21 (br s, 1H), 8.00 (br s, 1H), 7.92 (s, 1H), 7.91–7.89 (m, 1H), 7.77 (br d, J = 9.0 Hz, 1H), 7.49 (dd, J = 9.0, 1.9 Hz, 1H), 4.64 (br s, 1H), 4.42 (tt, J = 11.9, 3.7 Hz, 1H), 3.08 (dd, J = 6, 6 Hz, 2H), 2.40–2.28 (m, 2H), 2.07–1.89 (m, 4H), 1.70–1.55 (m, 1H), 1.46 (s, 9H).

[0493] Step 2: Synthesis of 1-[(1r,4r)-4-{6-[4-(trifluoromethyl)-1H-pyrazol-1-yl]-2H-indazol-2-yl}cyclohexyl]methanamine hydrochloride (C44).

[0494] To a solution of C43 (80.0 mg, 0.173 mmol) in 1,4-dioxane (2 mL) was added a solution of hydrogen chloride in 1,4-dioxane (4 M; 1 mL, 4 mmol), and the reaction mixture was stirred for 2 hours. The solvent was removed in vacuo to afford a residue that was azeotroped twice with dichloromethane to afford C44 (75 mg) as a white solid; most of this material was carried directly to the next step. LCMS m / z 364.3 [M+H] + .

[0495] Step 3: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[4-(trifluoromethyl)-1H-pyrazol-1-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide trifluoroacetate (11).

[0496] A solution of C44 (from the previous step; 69 mg, ≤0.16 mmol) in N,N-dimethylformamide (1.8 mL) was treated with water (0.4 mL) followed by the addition of the following reagents: 2,3,5-trifluoro-4-hydroxybenzoic acid (39.8 mg, 0.207 mmol), 1-methyl-1H-imidazole (55.0 μL, 0.690 mmol) and 2-hydroxypyridine 1-oxide (26.8 mg, 0.241 mmol). After the reaction mixture was stirred at room temperature for 20 minutes, 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (98%, 33.8 mg, 0.173 mmol) was added and stirring was continued for 18 hours. The reaction mixture was then diluted with water (10 mL), acidified to pH 4 by the addition of 1 M hydrochloric acid, and extracted three times with ethyl acetate. The combined organic layers were washed five times with water, dried over magnesium sulfate, filtered, concentrated in vacuo, and purified using reverse phase HPLC (column: Waters Sunfire C18, 19×100 mm, 5 μm; mobile phase A: 0.05% trifluoroacetic acid / water (v / v); mobile phase B: 0.05% trifluoroacetic acid / acetonitrile (v / v); gradient: 5% to 95% B over 8.54 minutes, followed by 95% B over 1.46 minutes; flow rate: 25 mL / minute) to provide 2,3,5-trifluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[4-(trifluoromethyl)-1H-pyrazol-1-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide trifluoroacetate (11). Yield: 41.1 mg, 63.1 μmol, 39% over 2 steps. LCMS m / z 538.5 [M+H] + Retention time: 3.11 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0497] Example 12: 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[1-(oxan-4-yl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide (12)

[0498]

[0499] Step 1: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-{[(1r,4r)-4-{6-[1-(oxan-4-yl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide (C45).

[0500] The experiment was performed in a library format.

[0501] A solution of P14 (60 mg, 100 μmol) in 1,4-dioxane (1 mL) was added to 4-bromo-1-(oxan-4-yl)-1H-pyrazole (150 μmol). Aqueous potassium phosphate (1.5 M; 0.20 mL, 300 μmol) was then added, followed by chloro[(di(1-adamantyl)-N-butylphosphine)-2-(2-aminobiphenyl)]palladium(II) (cata A Pd G2; 5 μmol), then the reaction vial was capped and shaken at 100 ° C for 16 hours. After removal of the solvent by concentrator, the residue was mixed with water (1 mL), extracted with ethyl acetate (3 x 1.5 mL), and concentrated again to provide C45; this material was directly carried forward to the next step.

[0502] Step 2: Synthesis of 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[1-(oxan-4-yl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide (12).

[0503] The experiment was performed in a library format.

[0504] A solution of trifluoroacetic acid (0.2 mL) in dichloromethane (0.8 mL) was added to C45 (from the previous step; ≤ 100 μmol), and the reaction vial was capped and shaken at 30 °C for 16 h. After removing the solvent using a concentrator, reverse phase HPLC (column: YMC-Actus Triart C18, 30×150 mm, 5 μm; mobile phase A: water containing 0.225% formic acid; mobile phase B: acetonitrile; gradient: 35% to 75% B; flow rate: 35 mL / min) afforded 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{6-[1-(oxan-4-yl)-1H-pyrazol-4-yl]-2H-indazol-2-yl}cyclohexyl]methyl}benzamide (12). Yield: 13.7 mg, 22.2 μmol, 22% over 2 steps. LCMS m / z 536 [M+H] +Retention time: 2.77 min (column: Waters XBridge C18, 2.1×50 mm, 5 μm; mobile phase A: water containing 0.0375% trifluoroacetic acid; mobile phase B: acetonitrile containing 0.01875% trifluoroacetic acid; gradient: 1% to 5% B over 0.6 min; 5% to 100% B over 3.4 min; flow rate: 0.8 mL / min).

[0505] Example 13: 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{5-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-3-yl}cyclohexyl]methyl}benzamide (13)

[0506]

[0507] Step 1: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-{[(1r,4r)-4-{5-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-3-yl}cyclohexyl]methyl}benzamide (C46).

[0508] To a mixture of 5-(trifluoromethyl)pyridine-2-carboxylic acid (47.0 mg, 0.246 mmol), N,N-diisopropylethylamine (86.6 mg, 0.670 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 127 mg, 0.334 mmol) in dichloromethane (20 mL) at 0°C was added P5 (100 mg, 0.223 mmol), and the reaction mixture was stirred at room temperature for 1 hour. It was then diluted with water (20 mL) and extracted with dichloromethane (2 x 20 mL); the combined organic layers were washed with saturated aqueous sodium chloride solution (2 x 20 mL), dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (gradient: 0% to 5% methanol / dichloromethane) provided the acyl intermediate as a white solid. Yield: 80 mg, 0.13 mmol, 58%. LCMS m / z 621.3 [M+H] + .

[0509] Sodium acetate (31.7 mg, 0.386 mmol) was added to a solution of the acyl intermediate (80 mg, 0.13 mmol) in a mixture of ethanol (4 mL) and water (1 mL). After the reaction mixture was stirred at 100 ° C for 1 hour under microwave irradiation, it was concentrated in vacuo. Silica gel chromatography (gradient: 0% to 7% methanol / dichloromethane) purification provided C46 as a white solid. Yield: 40 mg, 66 μ mol, 51% (from the acyl intermediate). LCMSm / z 625.3 [M + Na + ].

[0510] Step 2: Synthesis of 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{5-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-3-yl}cyclohexyl]methyl}benzamide (13).

[0511] A solution of hydrogen chloride in 1,4-dioxane (4M; 1 mL) was added to a solution of C46 (40 mg, 66 μmol) in dichloromethane (5 mL). The reaction mixture was stirred at room temperature for 2 hours, then concentrated in vacuo, diluted with dichloromethane (10 mL), and treated with sodium bicarbonate (10 mg, 0.12 mmol). After the solvent was removed under reduced pressure, the residue was subjected to silica gel chromatography (gradient: 0% to 6% methanol / dichloromethane) followed by reverse phase HPLC (column: Waters XBridge C18, 19×100 mm, 5 μm; mobile phase A: water containing 0.1% formic acid; mobile phase B: acetonitrile; gradient: 50% to 60% B; flow rate: 20 mL / min) to afford 3,5-difluoro-4-hydroxy-N-{[(1r,4r)-4-{5-[5-(trifluoromethyl)pyridin-2-yl]-1,2,4-oxadiazol-3-yl}cyclohexyl]methyl}benzamide (13) as a white solid. Yield: 9.0 mg, 19 μmol, 29%. LCMS m / z 483.2 [M+H] + . 1 H NMR (400 MHz, methanol-d4) d 9.09 (br s, 1H), 8.45 (d, half of the AB quartet, J = 8.3 Hz, 1H), 8.40 (dd, component of the ABX system, J = 8.4, 2.3 Hz, 1H), 7.51–7.41 (m, 2H), 3.27 (d, J = 6.9 Hz, 2H), 2.91 (tt, J = 12.2, 3.4 Hz, 1H), 2.24–2.14 (m, 2H), 2.03–1.92 (m, 2H), 1.80–1.59 (m, 3H), 1.29–1.15 (m, 2H).

[0512] Example 14: N-[(4-{5-[5-(difluoromethyl)pyrazin-2-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]-3,5-difluoro-4-hydroxybenzamide ammonium salt (14)

[0513]

[0514] The reaction was performed in a library format.

[0515] A stock solution of P7 (300 mg, 0.634 mmol) in ethyl acetate (6 mL) was used; 1 mL of this solution (0.106 mmol of P7) was treated with 5-(difluoromethyl)pyrazine-2-carboxylic acid (18.3 mg, 0.105 mmol), followed by triethylamine (42.2 μL, 0.303 mmol) and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphine 2,4,6-trioxide (50% by weight in ethyl acetate; 0.15 mL, 0.25 mmol). The reaction vial was heated at 100°C until oxadiazole formation occurred, then allowed to cool to room temperature, diluted with ethyl acetate (3 mL), and washed sequentially with water (2×3 mL) and saturated aqueous sodium chloride solution (3 mL). The organic layer was concentrated in vacuo, and the residue was dissolved in 1,1,1,3,3,3-hexafluoropropan-2-ol, treated with 1 equivalent of trifluoroacetic acid, and stirred until the deprotection of phenol was complete. The solvent was removed under reduced pressure and then subjected to reverse phase HPLC (column: Waters XBridge C18, 19×100 mm, 5 μm; mobile phase A: water containing 0.03% ammonium hydroxide; mobile phase B: acetonitrile containing 0.03% ammonium hydroxide; gradient: 5% to 95% B; flow rate: 25 mL / min) to obtain N-[(4-{5-[5-(difluoromethyl)pyrazin-2-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]-3,5-difluoro-4-hydroxybenzamide ammonium salt (14). Yield: 13.8 mg, 27.1 μmol, 26%. LCMS m / z 492.4 [M+H] + Retention time: 2.54 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0516] Example 15: 2,3,5-Trifluoro-4-hydroxy-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (15)

[0517]

[0518] Step 1: Synthesis of N-hydroxy-6-methoxypyridazine-3-carboxamidine (C47).

[0519] To a solution of 6-methoxypyridazine-3-carbonitrile (745 mg, 5.51 mmol) in methanol (3.7 mL) was added hydroxylamine hydrochloride (383 mg, 5.51 mmol) followed by triethylamine (0.776 mL, 5.57 mmol). The reaction mixture was stirred at room temperature for 4 days and then cooled in an ice bath for 15 minutes; the precipitated solid was collected by filtration to provide C47 as a purple solid. Yield: 690 mg, 4.10 mmol, 74%. LCMS m / z 169.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6)d 10.13 (s, 1H), 7.94 (d, J = 9.3Hz, 1H), 7.22 (d, J = 9.3Hz, 1H), 5.98 (br s, 2H), 4.05 (s, 3H).

[0520] Step 2: Synthesis of tert-butyl ({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]oct-1-yl}methyl)carbamate (C48).

[0521] O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU; 312 mg, 0.821 mmol) was added to a solution of C47 (155 mg, 0.547 mmol) in N,N-dimethylformamide (3 mL). After the reaction mixture was stirred for 20 minutes, 4-{[(tert-butoxycarbonyl)amino]methyl}bicyclo[2.2.2]octane-1-carboxylic acid (101 mg, 0.601 mmol) and N,N-diisopropylethylamine (0.286 mL, 1.64 mmol) were added and stirring was continued at room temperature for 18 hours. The reaction mixture was then diluted with water; the solid was collected by filtration and washed with water to give the acyl intermediate as a white solid. Yield: 134 mg, 0.309 mmol, 56%. LCMSm / z 434.4[M+H] + . 1 H NMR (400 MHz, methanol-d4) d 8.22 (d, J = 9.3 Hz, 1H), 7.21 (d, J = 9.3 Hz, 1H), 6.61–6.52 (m, 1H; assumed to be amide protons, slow change), 4.14 (s, 3H), 2.83 (d, J = 6.5 Hz, 2H), 2.00–1.91 (m, 6H), 1.54–1.45 (m, 6H), 1.44 (s, 9H).

[0522] The acyl intermediate (134 mg, 0.309 mmol) and sodium acetate (51.2 mg, 0.624 mmol) were taken up in a mixture of water (0.1 mL) and ethanol (1 mL), and the reaction vial was heated at 120° C. under microwave irradiation for 2.5 hours. The reaction mixture was then diluted with water (approximately 0.5 mL) and filtered; the filter cake was washed with ethanol to afford C48 as an off-white solid. Yield: 75 mg, 0.18 mmol, 58% (from the acyl intermediate). LCMS m / z 416.4 [M+H] + . 1 H NMR (400 MHz, methanol-d4) d 8.20 (d, J = 9.2 Hz, 1H), 7.33 (d, J = 9.2 Hz, 1H), 6.68–6.58 (m, 1H), 4.19 (s, 3H), 2.88 (d, J = 6.4 Hz, 2H), 2.13–2.02 (m, 6H), 1.63–1.53 (m, 6H), 1.45 (s, 9H).

[0523] Step 3: Synthesis of 1-{4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]octan-1-yl}methanamine trifluoroacetate (C49).

[0524] Trifluoroacetic acid (0.15 mL, 1.9 mmol) was added dropwise to a solution of C48 (75 mg, 0.18 mmol) in dichloromethane (2 mL) at 0 ° C. After the reaction mixture was stirred for 30 minutes, trifluoroacetic acid (0.15 mL, 1.9 mmol) was added again; after 30 minutes, the reaction mixture was treated with trifluoroacetic acid (20 μL, 0.26 mmol) again and stirred for another 5 minutes. It was then concentrated in vacuo, and the residue was azeotroped once with toluene and once with dichloromethane to give C49 (84 mg) as an oil. Most of this material was used in the following steps. LCMS m / z 316.2 [M+H] + Step 4: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (15).

[0525] A solution of C49 (from the previous step; 84 mg, ≤0.18 mmol) in a mixture of N,N-dimethylformamide (1.8 mL) and water (0.41 mL) was treated sequentially with 2,3,5-trifluoro-4-hydroxybenzoic acid (41.9 mg, 0.218 mmol), 1-methyl-1H-imidazole (43.4 μL, 0.544 mmol), and 2-hydroxypyridine 1-oxide (20.2 mg, 0.182 mmol). After the reaction mixture was stirred at room temperature for 20 minutes, 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (35.5 mg, 0.185 mmol) was added; stirring was continued at room temperature for 18 hours, after which the reaction mixture was diluted with water (10 mL), acidified to pH 4 by the addition of methanesulfonic acid, and extracted three times with ethyl acetate. The combined organic layers were washed five times with water, dried over magnesium sulfate, filtered, and concentrated in vacuo. Purification via reverse phase HPLC (column: Waters Sunfire C18, 19×100 mm, 5 μm; mobile phase A: 0.05% trifluoroacetic acid in water (v / v); mobile phase B: 0.05% trifluoroacetic acid in acetonitrile (v / v); gradient: 5% to 95% B over 8.54 minutes, followed by 95% B over 1.46 minutes; flow rate: 25 mL / minute) afforded 2,3,5-trifluoro-4-hydroxy-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide (15). Yield: 26.6 mg, 54.3 μmol, 30% over 2 steps. LCMS m / z 490.4 [M+H] + . Retention time: 2.57 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0526] Example 16: 3,5-difluoro-N-{[(1r,4r)-4-(6-fluoro-2H-indazol-2-yl)cyclohexyl]methyl}-4-hydroxybenzamide ammonium salt (16)

[0527]

[0528] Step 1: Synthesis of 3,5-difluoro-N-{[(1r,4r)-4-(6-fluoro-2H-indazol-2-yl)cyclohexyl]methyl}-4-[(4-methoxyphenyl)methoxy]benzamide (C50).

[0529] The reaction was performed in a library format.

[0530] A solution of P3 (60.7 mg, 0.150 mmol) in propan-2-ol (0.6 mL) was added to 4-fluoro-2-nitrobenzaldehyde (0.15 mmol). The reaction vial was capped, then evacuated and filled with nitrogen. This evacuation cycle was repeated twice, and the reaction mixture was subsequently shaken at 80° C. for 4 hours and then cooled to room temperature. After the addition of tributylphosphine (0.1 mL, 0.4 mmol), the reaction mixture was shaken at 80° C. for 18 hours. It was then partitioned between half-saturated aqueous sodium bicarbonate solution (1.5 mL) and ethyl acetate (2.4 mL) and vortexed. The organic layer was eluted through a solid phase extraction column (6 mL) containing sodium sulfate (about 1 g); this extraction procedure was repeated twice, and the combined eluents were concentrated in vacuo to provide C50, which was directly taken to the next step.

[0531] Step 2: Synthesis of 3,5-difluoro-N-{[(1r,4r)-4-(6-fluoro-2H-indazol-2-yl)cyclohexyl]methyl}-4-hydroxybenzamide ammonium salt (16).

[0532] The reaction was performed in a library format.

[0533] A solution of p-toluenesulfonic acid (57.1 mg, 0.300 mmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (0.6 mL) was added to C50 (from the previous step; ≤ 0.150 mmol), and the reaction mixture was shaken at room temperature for 3 days. After removing the solvent using a Genevac concentrator, it was purified by reverse phase HPLC (column: Waters XBridge C18, 19×100 mm, 5 μm; mobile phase A: water containing 0.03% ammonium hydroxide; mobile phase B: acetonitrile containing 0.03% ammonium hydroxide; gradient: 5% to 95% B over 8.54 minutes, then 95% B over 1.46 minutes; flow rate: 25 mL / min) to give 3,5-difluoro-N-{[(1r,4r)-4-(6-fluoro-2H-indazol-2-yl)cyclohexyl]methyl}-4-hydroxybenzamide ammonium salt (16). Yield: 11.4 mg, 27.1 μmol, 18% over 2 steps. LCMS m / z 404.4 [M+H] + Retention time: 2.61 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0534] Example 17: N-{[(1r,4r)-4-{3-[6-(2,2-dimethylpropionamido)pyridazin-3-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide ammonium salt (17)

[0535]

[0536] Step 1: Synthesis of 6-chloro-N-hydroxypyridazine-3-carboxamidine (C51)

[0537] To a solution of 6-chloropyridazine-3-carbonitrile (698 mg, 5.00 mmol) in methanol (15 mL) was added hydroxylamine hydrochloride (382 mg, 5.50 mmol) followed by triethylamine (0.775 mL, 5.56 mmol). The reaction mixture was stirred for 2 hours, after which the solid was collected by filtration to provide C51 as a brown solid. Yield: 465 mg, 2.69 mmol, 54%. LCMS m / z 173.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6)d 10.43 (s, 1H), 8.08 (d, J = 9.1Hz, 1H), 7.88 (d, J = 9.0Hz, 1H), 6.15 (br s, 2H).

[0538] Step 2: Synthesis of N-({(1r,4r)-4-[3-(6-chloropyridazin-3-yl)-1,2,4-oxadiazol-5-yl]cyclohexyl}methyl)-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C52).

[0539] A solution of P4 (595 mg, 1.37 mmol) in N, N-dimethylformamide (9 mL) was treated with O-(7-azabenzotriazole-1-yl)-N, N, N', N'-tetramethyluronium hexafluorophosphate (HATU; 783 mg, 2.06 mmol). After 30 minutes, C51 (261 mg, 1.51 mmol) and N, N-diisopropylethylamine (0.717 mL, 4.12 mmol) were added, and the reaction mixture was subsequently stirred at room temperature for 18 hours. The precipitate was collected by filtration and washed with dichloromethane to obtain an acyl intermediate as an off-white solid. Yield: 358 mg, 0.609 mmol, 44%. LCMSm / z 588.3[M+H] + . 1H NMR (400MHz, DMSO-d6)d8.52(br t,J=5.8Hz,1H),8.15(d,J=9.0Hz,1H),8.01(d,J=9.0Hz,1H),7.64–7.55(m,2H),7.34(d,J=8.6Hz,2H),7.29(br s,2H),6.92(d,J=8.6Hz,2H),5.17(s,2H),3.74(s,3H),3.12(dd,J=6,6Hz,2H),2.57–2.44(m,1H,assumed;almost entirely obscured bysolvent peak),2.05–1.96(m,2H),1.85–1.75(m,2H),1.61–1.48(m,1H),1.47–1.32(m,2H),1.06–0.92(m,2H).

[0540] Under microwave irradiation, a portion of the acyl intermediate (219 mg, 0.372 mmol) and sodium acetate (61.7 mg, 0.752 mmol) in a mixture of ethanol (4.5 mL) and water (0.45 mL) was heated at 120 ° C for 1 hour. The resulting solid was isolated by filtration and washed with a 10:1 mixture of ethanol and water to provide C52 as a white solid. Yield: 172 mg, 0.302 mmol, 81% (from the acyl intermediate). LCMS m / z 570.3 (chlorine isotope pattern observed) [M + H] + . 1 H NMR(400MHz,DMSO-d6)d 8.55(br t,J=5.8Hz,1H),8.32(d,J=9.0Hz,1H),8.14(d,J=8.9Hz,1H),7.65–7.56(m,2H),7.34(d,J=8.6Hz,2H),6.92(d,J=8.6Hz,2H ),5.17(s,2H),3.74(s,3H),3.20–3.09(m,3H),2.25–2.14(m,2H),1.92–1.83(m,2H),1.69–1.51(m,3H),1.23–1.08(m,2H).

[0541] Step 3: Synthesis of N-{[(1r,4r)-4-{3-[6-(2,2-dimethylpropionamido)pyridazin-3-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C53).

[0542] A mixture of C52 (46 mg, 81 μmol), 2,2-dimethylpropionamide (9.8 mg, 97 μmol), palladium(II) acetate (0.906 mg, 4.04 μmol), ([1,1'-binaphthyl]-2,2'-diyl)bis(diphenylphosphine) (BINAP; 5.03 mg, 8.08 μmol) and cesium carbonate (65.7 mg, 0.202 mmol) in 1,4-dioxane (1 mL) was degassed in vacuo and filled with nitrogen. This evacuation cycle was repeated twice before the reaction vial was heated at 100 °C for 18 hours. After the reaction mixture was partitioned between water and ethyl acetate, the aqueous layer was extracted twice with ethyl acetate, and the combined organic layers were dried over magnesium sulfate, filtered, and concentrated in vacuo to provide C53 (64 mg) as a brown oil. This material was taken directly to the next step. LCMS m / z 635.4 [M+H] + .

[0543] Step 4: Synthesis of N-{[(1r,4r)-4-{3-[6-(2,2-dimethylpropionamido)pyridazin-3-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide ammonium salt (17).

[0544] Trifluoroacetic acid (0.3 mL, 4 mmol) was added to a solution of C53 (from the previous step; 64 mg, ≤81 μmol) in dichloromethane (1 mL). The reaction mixture was stirred at room temperature for 1 hour before being concentrated in vacuo and azeotroped twice with dichloromethane. Reverse phase HPLC (column: Waters XBridge C18, 19×100 mm, 5 μm; mobile phase A: water containing 0.03% ammonium hydroxide; mobile phase B: acetonitrile containing 0.03% ammonium hydroxide; gradient: 5% to 50% B over 8.5 minutes, then 50% to 95% B over 0.5 minutes; then 95% B over 1.0 minutes; flow rate: 25 mL / min) afforded N-{[(1r,4r)-4-{3-[6-(2,2-dimethylpropionamido)pyridazin-3-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}-3,5-difluoro-4-hydroxybenzamide ammonium salt (17). Yield: 4.2 mg, 7.9 μmol, 10% over 2 steps. LCMS m / z 515.3 [M+H] + Retention time: 2.83 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0545] Example 18: 3,5-Difluoro-4-hydroxy-N-({(1r,4r)-4-[4-(quinoxalin-6-yl)-1H-1,2,3-triazol-1-yl]cyclohexyl}methyl)benzamide (18)

[0546]

[0547] Step 1: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[4-(quinoxalin-6-yl)-1H-1,2,3-triazol-1-yl]cyclohexyl}methyl)benzamide (C54).

[0548] The reaction was performed in a library format.

[0549] A solution of P6 (100 μmol) in N,N-dimethylformamide (0.50 mL) was treated with an aqueous solution of sodium azide (2.0 M; 0.20 mL, 400 μmol) followed by an aqueous solution of sodium carbonate (0.2 M; 0.10 mL, 20 μmol). The reaction vial was capped and the reaction mixture was heated at 125 ° C for 10 minutes under microwave irradiation. After the reaction mixture was cooled to room temperature, 6-ethynylquinoxaline (100 μmol) and copper (I) iodide (2.0 mg, 10 μmol) were added and microwave irradiation was continued at 125 ° C for 40 minutes. When the reaction mixture returned to room temperature, it was treated with an aqueous solution of sodium hypochlorite (8% to 10%; 1.0 mL) and the vial was shaken at 30 ° C for 5 minutes; the reaction mixture was heated to 40 minutes under microwave irradiation. A concentrator removed the solvent to provide C54. This material was carried directly to the next step.

[0550] Step 2: Synthesis of 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[4-(quinoxalin-6-yl)-1H-1,2,3-triazol-1-yl]cyclohexyl}methyl)benzamide (18).

[0551] The reaction was performed in a library format.

[0552] To a solution of C54 (from the previous step; ≤100 μmol) in dichloromethane (0.8 mL) was added a solution of hydrogen chloride in 1,4-dioxane (4 M; 0.2 mL, 800 μmol), and the reaction vial was capped and shaken at 30° C. for 16 h. After the solvent was removed by concentrator, the residue was purified by reverse phase HPLC (column: YMC-Actus Triart C18, 30×150 mm, 5 μm; mobile phase A: water containing ammonium hydroxide (pH 10); mobile phase B: acetonitrile; gradient: 10% to 50% B; flow rate: 35 mL / min) to obtain 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[4-(quinoxalin-6-yl)-1H-1,2,3-triazol-1-yl]cyclohexyl}methyl)benzamide (18). Yield: 9.1 mg, 20 μmol, 20%. LCMS m / z 465 [M+H] + Retention time: 2.46 minutes (analytical conditions, column: Waters XBridge C18, 2.1×50 mm, 5 μm; mobile phase A: water containing 0.0375% trifluoroacetic acid; mobile phase B: acetonitrile containing 0.01875% trifluoroacetic acid; gradient: 1% to 5% B over 0.6 minutes; 5% to 100% B over 3.4 minutes; flow rate: 0.8 mL / minute).

[0553] Example 19: 2,3,5-Trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(4-methylpiperazin-1-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide trifluoroacetate (19)

[0554]

[0555] Step 1: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[6-(4-methylpiperazin-1-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide (C55).

[0556] In a glove box under nitrogen, a scintillation vial was charged with C39 (100 mg, 0.166 mmol), cesium carbonate (162 mg, 0.497 mmol), and methanesulfonic acid (2-dicyclohexylphosphino-2′,6′-isopropoxy-1,1′-biphenyl) [2-(2′-amino-1,1′-biphenyl)] palladium (II) (RuPhos Pd G3; 13.9 mg, 16.6 μmol). The contents of the vial were stirred for 2 minutes, followed by the addition of toluene (1.7 mL). To the resulting solution was added 1-methylpiperazine (27.6 μL, 0.249 mmol), and the vial was transferred to a heating block. After the reaction mixture was slowly heated to 90°C with vigorous stirring, it was maintained at 90°C overnight. It was then allowed to cool to room temperature, concentrated in vacuo, taken up in ethyl acetate (50 mL), and washed sequentially with water (3×50 mL) and saturated aqueous sodium chloride solution (25 mL). The organic layer was concentrated under reduced pressure to give an oil (106 mg). LCMS analysis indicated that both C55 and 19 were present in the material, most of which was carried directly to the next step. LCMS m / z 622.5 and 502.4 [M+H] + .

[0557] Step 2: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(4-methylpiperazin-1-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide trifluoroacetate (19).

[0558] Trifluoroacetic acid (50 μL, 0.65 mmol) was added to a solution of C55 and 19 (from the previous step; 103 mg, ≤0.161 mmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (1.5 mL). After the reaction mixture was stirred at room temperature overnight, it was concentrated in vacuo and purified via reverse phase HPLC [column: Waters Sunfire C18, 19×100 mm, 5 μm; mobile phase A: 0.05% trifluoroacetic acid / water (v / v); mobile phase B: 0.05% trifluoroacetic acid / acetonitrile (v / v); gradient: 5% to 35% B over 8.5 minutes, then 35% to 95% B over 0.5 minutes; flow rate: 25 mL / min] to give 2,3,5-trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(4-methylpiperazin-1-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide trifluoroacetate (19). Yield: 40 mg, 65 μmol, 40% over 2 steps. LCMS m / z 502.3 [M+H] +Retention time: 1.91 minutes (analytical conditions, column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; mobile phase A: water containing 0.05% trifluoroacetic acid (v / v); mobile phase B: acetonitrile containing 0.05% trifluoroacetic acid (v / v); gradient: 5.0% to 95% B over 4.0 minutes, then 95% B over 1.0 minute; flow rate: 2 mL / minute).

[0559] Example 20: 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[5-(1-methyl-1H-pyrazol-3-yl)-1-oxo-1,3-dihydro-2H-isoindol-2-yl]cyclohexyl}methyl)benzamide (20)

[0560]

[0561] Step 1: Synthesis of N-{[(1r,4r)-4-(5-bromo-1-oxo-1,3-dihydro-2H-isoindol-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (C56).

[0562] To a solution of P3 (400 mg, 0.989 mmol) and triethylamine (150 mg, 1.48 mmol) in toluene (10 mL) was added methyl 4-bromo-2-(bromomethyl)benzoate (305 mg, 0.990 mmol). After the reaction mixture was stirred at 100 ° C for 16 hours, it was concentrated in vacuo; silica gel chromatography (eluent: 5% methanol / dichloromethane) gave C56 as a white solid. Yield: 332 mg, 0.554 mmol, 56%. LCMS m / z 599.0 (bromine isotope pattern observed) [M + H] + . 1 H NMR(400MHz,DMSO-d6)d 8.55(br t,J=5.6Hz,1H),7.84(s,1H),7.69–7.55(m,4H),7.33(d,J=8.5Hz,2H),6.92(d,J=8.4Hz,2H),5.17(s,2H),4.43(s,2 H),4.04–3.92(m,1H),3.74(s,3H),3.12(dd,J=6,6Hz,2H),1.89–1.70(m,4H),1.63–1.46(m,3H),1.19–1.04(m,2H).

[0563] Step 2: Synthesis of 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({(1r,4r)-4-[5-(1-methyl-1H-pyrazol-3-yl)-1-oxo-1,3-dihydro-2H-isoindol-2-yl]cyclohexyl}methyl)benzamide (C57).

[0564] To a mixture of C56 (100 mg, 0.167 mmol), (1-methyl-1H-pyrazol-3-yl)boronic acid (25.2 mg, 0.200 mmol) and potassium carbonate (69.2 mg, 0.501 mmol) in 1,4-dioxane (10 mL) was added tetrakis(triphenylphosphine)palladium(0) (19.3 mg, 16.7 μmol), and the reaction mixture was stirred at 100 ° C for 16 hours. After removing the solvent via vacuum concentration, silica gel chromatography (eluent: 5% methanol / dichloromethane) provided C57 as an oil. Yield: 42 mg, 70 μmol, 42%. LCMS m / z 601.2 [M + H] + .

[0565] Step 3: Synthesis of 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[5-(1-methyl-1H-pyrazol-3-yl)-1-oxo-1,3-dihydro-2H-isoindol-2-yl]cyclohexyl}methyl)benzamide (20).

[0566] To a solution of C57 (37 mg, 62 μmol) in dichloromethane (5 mL) was added a 1,4-dioxane solution of hydrogen chloride (4 M; 1 mL). The reaction mixture was stirred at 25°C for 1 hour and then concentrated in vacuo; purified via reverse phase HPLC (column: Waters XBridge C18, 19×100 mm, 5 μm; mobile phase A: water containing 0.1% formic acid; mobile phase B: acetonitrile; gradient: 25% to 45% B; flow rate: 20 mL / min) to provide 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[5-(1-methyl-1H-pyrazol-3-yl)-1-oxo-1,3-dihydro-2H-isoindol-2-yl]cyclohexyl}methyl)benzamide (20). Yield: 15.8 mg, 32.9 μmol, 53%. LCMS m / z 481.2 [M+H] + . 1H NMR(400MHz,DMSO-d6)d 8.37(br t,J=5.8Hz,1H),7.97(br s,1H),7.89(dd,J=7.9,1.4Hz,1H),7.77(d,J=2.3Hz,1H),7.66(d,J=7.9Hz,1H),7.59–7.47(m,2H),6.79(d,J=2.3Hz,1H),4.46(s ,2H),4.00(tt,J=12.2,3.8Hz,1H),3.90(s,3H),3.12(dd,J=6,6Hz,2H),1.90–1.73(m,4H),1.65–1.49(m,3H),1.20–1.05(m,2H).

[0567] Example 21: 2,3,5-Trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide hydrochloride (21)

[0568]

[0569]

[0570] Step 1: Synthesis of methyl 2-[4-(tert-butoxycarbonyl)piperazin-1-yl]pyrimidine-4-carboxylate (C58).

[0571] Potassium carbonate (2.18 g, 15.8 mmol) was added to a solution of methyl 2-chloropyrimidine-4-carboxylate (95%, 956 mg, 5.26 mmol) and tert-butyl piperazine-1-carboxylate (1.00 g, 5.37 mmol) in acetonitrile (26 mL), and the reaction mixture was stirred at 65° C. After 1.5 hours, LCMS analysis indicated the presence of C58: LCMS m / z 267.2 [(M-2-methylprop-1-ene)+H] + The reaction mixture was allowed to stir at 65° C. for another hour, then diluted with water and extracted three times with dichloromethane. The combined organic layers were concentrated in vacuo to afford C58 (1.75 g) as a yellow solid, most of which was carried forward to the next step. 1 HNMR (400MHz, chloroform-d) δ8.51(d,J=4.8Hz,1H),7.14(d,J=4.8Hz,1H),3.96(s,3H),3.92–3.84(m,4H),3.55–3.47(m,4H),1.49(s,9H).

[0572] Step 2: Synthesis of 2-[4-(tert-Butyloxycarbonyl)piperazin-1-yl]pyrimidine-4-carboxylic acid (C59).

[0573] A solution of lithium hydroxide (1.26 g, 52.6 mmol) in a mixture of tetrahydrofuran (10 mL), water (10 mL) and methanol (5 mL) was added to C58 (from the previous step; 1.70 g, ≤ 5.11 mmol). The reaction mixture was heated at 50 ° C for 1 hour, allowed to cool to room temperature, and concentrated in vacuo to remove most of the solvent. After the residue was acidified to pH 2 to 3 by adding 1M hydrochloric acid, the mixture was extracted three times with ethyl acetate. At this time, the aqueous layer was acidified again to bring the pH to 2 and extracted twice with ethyl acetate. All organic layers were combined, dried over magnesium sulfate, filtered, and concentrated under reduced pressure to provide C59 as a light yellow solid. Yield: 1.42 g, 4.60 mmol, 90% over 2 steps. LCMSm / z 307.2[MH] - . 1 H NMR (400 MHz, CHLOROFORM-d) δ 8.62 (d, J = 4.7 Hz, 1H), 7.31 (d, J = 4.7 Hz, 1H), 3.90–3.82 (m, 4H), 3.58–3.51 (m, 4H), 1.50 (s, 9H).

[0574] Step 3: Synthesis of tert-butyl 4-(4-{3-[4-({2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octan-1-yl]-1,2,4-oxadiazol-5-yl}pyrimidin-2-yl)piperazine-1-carboxylate (C60).

[0575] N, N-diisopropylethylamine (0.532mL, 3.05mmol) is added dropwise to a solution of C59 (345mg, 1.12mmol) and bis(pentafluorophenyl) carbonate (98%, 450mg, 1.12mmol) in tetrahydrofuran (5mL). After the reaction mixture was stirred at room temperature for 30 minutes, additional bis(pentafluorophenyl) carbonate (98%, 20mg, 51μmol) was added and stirring was continued for 10 minutes, followed by the addition of P10 (500mg, 1.02mmol), followed by further stirring at room temperature for 30 minutes. The reaction mixture was then treated with a solution of tetrabutylammonium fluoride in tetrahydrofuran (1.0M; 5.09mL, 5.09mmol) and heated at 50°C overnight. After cooling to room temperature, the reaction mixture was treated with a small amount of aqueous sodium bicarbonate solution, diluted with water, and extracted three times with ethyl acetate. The combined organic layers were dried over magnesium sulfate, filtered, concentrated in vacuo, and purified via silica gel chromatography (gradient: 30% to 100% ethyl acetate / heptane) to afford C60 as a yellow solid. Yield: 474 mg (corrected for residual dichloromethane), 0.621 mmol, 61%. LCMS m / z 764.5 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.67(d,J=4.8Hz,1H),8.35(br t,J=6.3Hz,1H),7.38–7.31(m,1H),7.36(br d,J=8.6Hz,2H),7.30(d,J=4.8Hz,1H),6.94(br d,J=8.7Hz,2H),5.22(s,2H),3.83–3.77(m,4H),3.75(s,3H),3.47–3.40(m,4 H), 3.07 (d, J = 6.2Hz, 2H), 1.94–1.84 (m, 6H), 1.57–1.48 (m, 6H), 1.43 (s, 9H).

[0576] Step 4: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-[(4-{5-[2-(piperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide (C61).

[0577] A solution of C60 (840 mg, 1.10 mmol) and pyridine (0.711 mL, 8.79 mmol) in dichloromethane (36 mL) was cooled to approximately -15 ° C and treated dropwise with trimethylsilyl trifluoromethanesulfonate (0.796 mL, 4.40 mmol). The reaction mixture was stirred at -15 ° C overnight, but the temperature of the cooling bath had reached 12 ° C by morning. The reaction mixture was then cooled in an ice bath, followed by the slow addition of aqueous sodium bicarbonate solution (20 mL), and the resulting mixture was stirred for 10 minutes. The aqueous layer was adjusted to pH 10 and extracted three times with dichloromethane; the combined organic layers were washed successively with saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was co-evaporated three times with dichloromethane to provide C61 as a yellow solid. Yield: 673 mg, 1.01 mmol, 92%. LCMS m / z 664.4 [M+H] + . 1 H NMR (400MHz, chloroform-d) δ8.51(d,J=4.8Hz,1H),7.59(ddd,J=11.7,6.8,2.3Hz,1H),7.34(d,J=8.6Hz,2H),7.22(d, J=4.8Hz,1H),6.88(d,J=8.6Hz,2H),6.62–6.50(m,1H),5.24(s,2H),3.96–3.86(m,4H),3.80(s,3H),3.30(br d,J=6Hz,2H),3.04–2.91(m,4H),2.07–1.96(m,6H),1.66–1.56(m,6H).

[0578] Step 5: Synthesis of 2,3,5-trifluoro-4-[(4-methoxyphenyl)methoxy]-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide (C62).

[0579] To a solution of C61 (100 mg, 0.151 mmol) and formaldehyde (43 mg, 1.43 mmol) in 1,2-dichloroethane (8 mL) was added sodium triacetoxyborohydride (91 mg, 0.43 mmol). After the reaction mixture was stirred at 25 ° C for 1 hour, it was subjected to aqueous post-treatment and extracted with dichloromethane (2×30 mL); the combined organic layers were washed with saturated aqueous sodium chloride solution, dried over sodium sulfate, filtered, and concentrated in vacuo. Silica gel chromatography (eluent: 5% methanol / dichloromethane) provided C62 as a yellow solid. Yield: 72.0 mg, 0.106 mmol, 70%. LCMS m / z 678.2 [M + H]+ . 1 H NMR(400MHz,DMSO-d6)δ8.65(d,J=4.9Hz,1H),8.36(br t,J=6.2Hz,1H),7.37–7.30(m,1H),7.36(d,J=8.5Hz,2H),7.27(d,J=4.8Hz,1H),6.94(d,J=8.4Hz,2H),5.22(s,2H),3.8 4–3.76(m,4H),3.75(s,3H),3.06(d,J=6.2Hz,2H),2.45–2.34(m,4H),2.23(s,3H),1.92–1.84(m,6H),1.58–1.47(m,6H).

[0580] Step 6: Synthesis of 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide hydrochloride (21).

[0581] A solution of hydrogen chloride in 1,4-dioxane (4M: 1 mL, 4 mmol) was added to a solution of C62 (72.0 mg, 0.106 mmol) in dichloromethane (4 mL). After the reaction mixture was stirred at 25° C. for 1 hour, it was concentrated in vacuo and purified via reverse phase HPLC (column: Waters XBridge C18, 19×150 mm, 5 μm, mobile phase A: water containing 0.05% formic acid; mobile phase B: acetonitrile; gradient: 15% to 45% B; flow rate: 20 mL / min) to provide 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide hydrochloride (21) as a white solid. Yield: 30.0 mg, 50.5 μmol, 48%. LCMS m / z 558.3 ​​[M+H] + . 1 H NMR (400 MHz, DMSO-d6), characteristic peaks: δ8.75 (d, J = 4.9 Hz, 1H), 8.19 (br t, J = 6 Hz, 1H), 7.42 (d, J = 4.9 Hz, 1H), 7.27 (ddd, J = 11.0, 6.2, 2.4 Hz, 1H), 3.66–3.2 (m, 8H, hypothetical; completely obscured by water peak), 3.07 (d, J = 6.2 Hz, 2H), 2.84 (s, 3H), 1.96–1.83 (m, 6H), 1.60–1.47 (m, 6H).

[0582] Using similar procedures, the compounds of Examples 22-214 were synthesized as described in Tables 1 and 2.

[0583] Table 1. Structures and IUPAC names of Examples 1-214.

[0584]

[0585]

[0586]

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600]

[0601]

[0602]

[0603]

[0604]

[0605]

[0606]

[0607]

[0608]

[0609]

[0610]

[0611]

[0612] Table 2. Synthesis methods and physicochemical data of Examples 22-214.

[0613]

[0614]

[0615]

[0616]

[0617]

[0618]

[0619]

[0620]

[0621]

[0622]

[0623]

[0624]

[0625] 1. P3 was reacted with 2-nitrobenzaldehyde, and the resulting imine was ring-closed with triethyl phosphite and deprotected using hydrogen chloride in 1,4-dioxane to give Example 22.

[0626] 2. Use trifluoroacetic acid instead of hydrogen chloride for the final deprotection.

[0627] 3. Using the method described for the synthesis of C25 in Preparation P15, the desired 1-[(1r,4r)-4-(6-methoxy-2H-indazol-2-yl)cyclohexyl]methanamine hydrochloride was prepared.

[0628] 4. In this case, the boronate coupling is catalyzed by tetrakis(triphenylphosphino)palladium(0) rather than [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II).

[0629] 5. Using the method described for the synthesis of C25 in Preparation P15, the desired chloro-substituted 1-[(1r,4r)-4-(2H-indazol-2-yl)cyclohexyl]methanamine hydrochloride was prepared.

[0630] 6. Using the method described for the synthesis of P13 in Preparation P13, the desired 1-{(1r,4r)-4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride was prepared.

[0631] 7. Using the method described for the synthesis of P13 in Preparation P13, the desired 1-{(1r,4r)-4-[6-(pyrazin-2-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride was prepared.

[0632] 8. The synthesis of tert-butyl {[(1r,4r)-4-(6-bromoimidazo[1,2-a]pyridin-2-yl)cyclohexyl]methyl}carbamate was carried out from C19 using the method described for the synthesis of P11 in Preparation P11. This material was converted to the desired 1-{(1r,4r)-4-[6-(1-methyl-1H-pyrazol-4-yl)imidazo[1,2-a]pyridin-2-yl]cyclohexyl}methanamine hydrochloride by reaction with 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole in the presence of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and potassium carbonate, followed by deprotection with hydrogen chloride.

[0633] 9. According to the method described for the preparation of P15 in Preparation Example P15, the desired N-{[(1r,4r)-4-(5-chloro-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide was synthesized.

[0634] 10. Cyclopropylmethanol was deprotected with sodium hydride in tetrahydrofuran at 0° C., followed by addition of N-{[(1r,4r)-4-(5-chloro-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (see footnote 9), and the reaction mixture was heated at 80° C. for 16 hours. Subsequent deprotection using trifluoroacetic acid provided Example 32.

[0635] 11. Analytical HPLC conditions. Column: Waters XBridge C18, 2.1×50 mm, 5 μm; Mobile phase A: 0.0375% trifluoroacetic acid in water; Mobile phase B: 0.01875% trifluoroacetic acid in acetonitrile; Gradient: 10% B over 0.50 min; 10% to 100% B over 3.5 min; Flow rate: 0.8 mL / min.

[0636] 12. A mixture of N-{[(1r,4r)-4-(5-chloro-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide (see footnote 9), [6-(trifluoromethyl)pyridin-2-yl]methanol, tris(dibenzylideneacetone)dipalladium(0), 5-(di-tert-butylphosphine)-1',3',5'-triphenyl-1'H-1,4'-bipyrazole (BippyPhos), and sodium hydroxide was heated in a 4:1 mixture of 2-methylbutan-2-ol and dichloromethane at 105° C. for 16 hours. Subsequent deprotection using trifluoroacetic acid provided Example 33.

[0637] 13. Analytical HPLC conditions. Column: Waters XBridge C18, 2.1×50 mm, 5 μm; Mobile phase A: 0.0375% trifluoroacetic acid in water; Mobile phase B: 0.01875% trifluoroacetic acid in acetonitrile; Gradient: 1% to 5% B over 0.6 min; 5% to 100% B over 3.4 min; Flow rate: 0.8 mL / min.

[0638] 14. Intermediate P14 is reacted with an appropriate aromatic bromide in the presence of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and tripotassium phosphate, followed by deprotection with hydrogen chloride.

[0639] 15. Column: Waters Atlantis dC18, 4.6×50 mm, 5 μm; Mobile phase A: Water containing 0.05% trifluoroacetic acid (v / v); Mobile phase B: Acetonitrile containing 0.05% trifluoroacetic acid (v / v); Gradient: 5.0% to 95% B, linear over 4.0 minutes, then 95% B over 1.0 minute; Flow rate: 2 mL / minute.

[0640] 16. In this case, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) was used instead of chloro[(di(1-adamantyl)-N-butylphosphine)-2-(2-aminobiphenyl)]palladium(II) (cata A Pd G2).

[0641] 17. Coupling was performed using the conditions described in step 1 of Example 10; deprotection of the product was then achieved using trifluoroacetic acid.

[0642] 18. In this case, a chlorine reactant is used instead of a bromide.

[0643] 19. In this case, 5-bromo-1H-pyrrolo[2,3-b]pyridine is reacted with p-toluenesulfonyl chloride in the presence of N,N-diisopropylethylamine, and the resulting 5-bromo-1-(4-methylbenzene-1-sulfonyl)-1H-pyrrolo[2,3-b]pyridine is used in the coupling reaction.

[0644] 20. In this case, the acyl intermediate was cyclized by treatment with sodium acetate rather than tetrabutylammonium fluoride.

[0645] 21. Using the method described for the preparation of P9 in Preparation P9, the desired 4-({3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamido}methyl)bicyclo[2.2.2]octane-1-carboxylic acid was synthesized, but using P1 as the starting material.

[0646] 22. Use 1,1,1,3,3,3-hexafluoropropan-2-ol with methanesulfonic acid instead of hydrogen chloride for the final deprotection.

[0647] 23. Conversion of P1 to the desired N-[(4-aminobicyclo[2.2.2]octan-1-yl)methyl]-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide was carried out using the method described in Preparation P3. This intermediate was condensed with 2-nitro-5-(trifluoromethyl)benzaldehyde, followed by ring closure of the resulting imine with triethyl phosphite and deprotection using hydrogen chloride to afford Example 125.

[0648] 24.P7 was reacted with 4-[(fluorosulfonyl)oxy]benzoic acid (A. Baranczak et al., J. Am. Chem. Soc. 2015, 137, 7404–7414), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride, 1H-benzotriazol-1-ol, and N,N-diisopropylethylamine to give the acyl intermediate; this was subjected to tetrabutylammonium fluoride to provide 3,5-difluoro-N-({4-[5-(4-hydroxyphenyl)-1,2,4-oxadiazol-3-yl]bicyclo[2.2.2]octan-1-yl}methyl)-4-[(4-methoxyphenyl)methoxy]benzamide. Reaction with (4-acetamidophenyl)imidodithiodifluoride (AISF) and cesium carbonate, followed by deprotection via treatment with hydrogen chloride, gave Example 126.

[0649] 25. Use p-toluenesulfonic acid instead of hydrogen chloride for the final deprotection.

[0650] 26. In this case, the acyl intermediate was cyclized by heating in 1-methylpyrrolidin-2-one rather than by treatment with sodium acetate.

[0651] 27. Methyl 4-(aminomethyl)bicyclo[2.2.2]octane-1-carboxylate was protected by reaction with benzyl chloroformate and triethylamine, followed by ester cleavage using sodium hydroxide. The resulting 4-({[(benzyloxy)carbonyl]amino}methyl)bicyclo[2.2.2]octane-1-carboxylic acid was converted to benzyl {[4-(1,3-benzoxazol-2-yl)bicyclo[2.2.2]oct-1-yl]methyl}carbamate using the method described for the synthesis of C26 from P4 in Example 1; subsequent hydrogenation over palladium on carbon afforded the desired 1-[4-(1,3-benzoxazol-2-yl)bicyclo[2.2.2]oct-1-yl]methylamine.

[0652] 28. Using the method described for the synthesis of C32 in Example 3, the desired 1-(4-{3-[6-(trifluoromethyl)pyridazin-3-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methanamine was prepared.

[0653] 29. Using the method employed for the synthesis of C25 in Preparation P15, the desired chloro-substituted 1-[(1r,4r)-4-(2H-indazol-2-yl)cyclohexyl]methanamine hydrochloride was prepared.

[0654] 30. Reaction of C49 with P1 mediated by O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine provides 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide. This material was demethylated using trimethylsilyl chloride and potassium iodide to give 3,5-difluoro-4-[(4-methoxyphenyl)methoxy]-N-({4-[3-(6-oxo-1,6-dihydropyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]oct-1-yl}methyl)benzamide, which was N-methylated using methyl 4-nitrobenzene-1-sulfonate and cesium carbonate; deprotection with trifluoroacetic acid provided Example 146.

[0655] 31. Using the procedure described for the synthesis of 6 from P3 in Example 6, tert-butyl {[(1r,4r)-4-aminocyclohexyl]methyl}carbamate was converted to the desired 1-{(1r,4r)-4-[5-(trifluoromethyl)-2H-pyrazolo[3,4-c]pyridin-2-yl]cyclohexyl}methanamine hydrochloride.

[0656] 32. 1-{(1r,4r)-4-[6-(1-ethyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride was prepared from P12 using the method described for the synthesis of P13 in Preparation P13.

[0657] 33. 1-{(1r,4r)-4-[6-(difluoromethyl)-2H-indazol-2-yl]cyclohexyl}methanamine trifluoroacetate was prepared from C20 by reaction with 2-(difluoromethanesulfonyl)pyridine and zinc in the presence of nickel(II) chloride ethylene glycol dimethyl ether complex, 4-methylpyridine-2,6-dicarboximidamide (see JME Hughes and PSFier, Org. Lett. 2019, 21, 5650–5654) and tetraethylammonium iodide; subsequent deprotection was performed with trifluoroacetic acid.

[0658] 34. tert-Butyl ({(1r,4r)-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)carbamate was prepared in the same manner as in Preparation P12. Using the method described in Preparation P13, it was subsequently converted to the desired 1-{(1r,4r)-4-[5-(pyrimidin-2-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride.

[0659] 35. tert-Butyl ({(1r,4r)-4-[5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2H-indazol-2-yl]cyclohexyl}methyl)carbamate (see footnote 34) was converted to the desired 1-{(1r,4r)-4-[5-(1-methyl-1H-pyrazol-4-yl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride via the method described for the synthesis of C41 from P12 in Example 10, followed by deprotection with hydrogen chloride.

[0660] 36. Conversion of P12 to the desired 2-(4-{2-[(1r,4r)-4-(aminomethyl)cyclohexyl]-2H-indazol-6-yl}-1H-pyrazol-1-yl)ethan-1-ol was carried out using the method described for the synthesis of C41 from P12 in Example 10, followed by deprotection with hydrogen chloride.

[0661] 37. The reaction of P12 with 4-bromo-1-(oxetan-3-yl)-1H-pyrazole was carried out using the method described for the synthesis of C41 from P12 in Example 10. Subsequent deprotection with hydrogen chloride also cleaved the oxetane ring to provide 2-(4-{2-[(1r,4r)-4-(aminomethyl)cyclohexyl]-2H-indazol-6-yl}-1H-pyrazol-1-yl)-3-chloropropan-1-ol.

[0662] 38. The synthesis of tert-butyl {[(1r,4r)-4-(6-bromoimidazo[1,2-a]pyridin-2-yl)cyclohexyl]methyl}carbamate was carried out from C19 using the method described for the synthesis of P11 in Preparation P11. This material was coupled with 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole in the presence of [1,1′-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) and potassium carbonate, followed by deprotection with hydrogen chloride to give the desired 1-[(1r,4r)-4-{6-[1-(difluoromethyl)-1H-pyrazol-4-yl]imidazo[1,2-a]pyridin-2-yl}cyclohexyl]methanamine hydrochloride.

[0663] 39. In this case, 2,3-difluoro-4-hydroxybenzoic acid rather than P1 was used for the final amide formation.

[0664] 40. The desired 1-(4-{3-[5-(trifluoromethyl)pyrazin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methanamine hydrochloride was prepared as follows: 4-{[(tert-Butyloxycarbonyl)amino]methyl}bicyclo[2.2.2]octane-1-carboxylic acid was reacted with N'-hydroxy-5-(trifluoromethyl)pyrazine-2-carboximidamide using 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and 1-methyl-1H-imidazole; the resulting acyl intermediate was cyclized by heating at 120° C. in 1-methylpyrrolidin-2-one and then deprotected with hydrogen chloride.

[0665] 41. In this case, hydrogen chloride was used instead of trifluoroacetic acid for the intermediate deprotection. In addition, the final coupling was performed with O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine instead of the reagents described in Example 15.

[0666] 42. Tert-butyl {[(1r,4r)-4-aminocyclohexyl]methyl}carbamate is condensed with 2-nitro-5-(trifluoromethyl)benzaldehyde, followed by ring closure of the resulting imine with triethyl phosphite and deprotection with hydrogen chloride to afford the desired 1-{(1r,4r)-4-[5-(trifluoromethyl)-2H-indazol-2-yl]cyclohexyl}methanamine hydrochloride.

[0667] 43. In this case, hydrogen chloride was used instead of trifluoroacetic acid for the deprotection of the intermediate.

[0668] 44. Reaction of tert-butyl {[(1r,4r)-4-aminocyclohexyl]methyl}carbamate with methyl 2-(bromomethyl)-4-chlorobenzoate and triethylamine followed by deprotection with hydrogen chloride provides the desired 2-[(1r,4r)-4-(aminomethyl)cyclohexyl]-5-chloro-2,3-dihydro-1H-isoindol-1-one hydrochloride.

[0669] 45. Using the method described for the synthesis of C25 in Preparation P15, the desired 1-[(1r,4r)-4-(5-chloro-2H-pyrazolo[3,4-c]pyridin-2-yl)cyclohexyl]methanamine hydrochloride was prepared.

[0670] 46. ​​In this case, 3-hydroxybenzoic acid is used instead of 4-[(fluorosulfonyl)oxy]benzoic acid.

[0671] 47. Treatment of tert-butyl [4-(hydroxymethyl)bicyclo[2.2.2]octan-1-yl]carbamate with methanesulfonyl chloride and triethylamine, followed by displacement of the resulting mesylate group using sodium azide and potassium carbonate, provides tert-butyl [4-(azidomethyl)bicyclo[2.2.2]octan-1-yl]carbamate. This material is hydrogenated over palladium on carbon, and the resulting primary amine is acylated with P1 by reaction with 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and 1H-benzotriazol-1-ol. The product is deprotected via hydrogenation over palladium on carbon to provide Example 192.

[0672] 48. In this case, the acyl intermediate was cyclized by treatment with tetrabutylammonium fluoride rather than sodium acetate.

[0673] 49. 4-{[(tert-Butoxycarbonyl)amino]methyl}bicyclo[2.2.2]octane-1-carboxylic acid reacts with N'-hydroxy-6-(trifluoromethyl)pyridine-3-carboximidamide and N'-hydroxy-4-(trifluoromethyl)benzene-1-carboximidamide in the presence of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and 1-methyl-1H-imidazole, followed by heating the acyl intermediate in 1-methylpyrrolidin-2-one. The resulting cyclized material is deprotected with hydrogen chloride to provide the desired 1-(4-{3-[6-(trifluoromethyl)pyridin-3-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methanamine hydrochloride.

[0674] 50. In this case, hydrogen chloride was used for the final deprotection step.

[0675] 51. Using the methods described for the synthesis of Preparations P8 and P16, the desired N-{[4-(6-bromo-2H-indazol-2-yl)bicyclo[2.2.2]octan-1-yl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide was prepared.

[0676] 52. N-{[4-(6-Bromo-2H-indazol-2-yl)bicyclo[2.2.2]octan-1-yl]methyl}-3,5-difluoro-4-[(4-methoxyphenyl)methoxy]benzamide described in footnote 51 was deprotected using hydrogen chloride to give Example 205.

[0677] 53. The final coupling was performed with 2,3,5-trifluoro-4-hydroxybenzoic acid instead of P1.

[0678] 54. The desired 1-{(1r,4r)-4-[3-(6-methylpyridazin-3-yl)-1,2,4-oxadiazol-5-yl]cyclohexyl}methanamine was prepared using the procedure described for the conversion of C29 to C32 in Example 3, except that the cyclization to form the oxadiazole was performed using heat rather than microwave irradiation and NaOAc.

[0679] The coupling of 2,3,5-trifluoro-4-hydroxybenzoic acid with 1-{(1r,4r)-4-[3-(6-methylpyridazin-3-yl)-1,2,4-oxadiazol-5-yl]cyclohexyl}methanamine was carried out using O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropylethylamine.

[0680] 56. Reaction of (1r,4r)-4-{[(tert-Butoxycarbonyl)amino]methyl}cyclohexane-1-carboxylic acid with N-hydroxyacetamidine in the presence of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and triethylamine provides the coupling product, which cyclizes to the oxadiazole via refluxing in toluene. Removal of the protecting group by treatment with methanol containing hydrogen chloride provides the desired 1-[(1r,4r)-4-(3-methyl-1,2,4-oxadiazol-5-yl)cyclohexyl]methanamine hydrochloride.

[0681] The following scheme can of course be modified by those skilled in the art.

[0682] hHSD17B13 IC 50 Determination (FACβ estradiol)

[0683] The HSD17B13 enzyme inhibition potency of the test compounds was determined using a purified protein biochemical enzyme activity assay using the NAD(P)H-Glo luciferase readout (Promega). During the metabolism of beta-estradiol (substrate) to estradiol (product), the HSD17B13 enzyme uses the oxidized form of nicotinamide adenine dinucleotide (NAD+) as a cofactor, while simultaneously converting NAD+ to its reduced form (NADH). Promega NAD(P)H-Glo TM The assay is a homogeneous bioluminescent assay that generates a light signal from a biochemical reaction involving NADH (or nicotinamide adenine dinucleotide phosphate, NADPH). In the presence of NADH (or NADPH), the reductase enzyme reduces the pre-luciferin reductase substrate to form luciferin. The Ultra-Glo TMRecombinant luciferase (rLuciferase) quantifies luciferin, and the light signal generated is proportional to the amount of NAD(P)H in the sample. A substrate mixture consisting of 12 μM final assay concentration (FAC)β-estradiol (Sigma, E8875) and 500 μM FAC NAD+ (Sigma, N8285) in assay buffer (25 mM Tris-HCl and 0.02% Triton, pH 7.6, Sigma T2444 and X-100) was added (2 μL / well) to a 384-well assay plate (Corning 3824) containing 80 nL of 50x FAC compound, serially diluted 1 to 3.162 in 100% DMSO to generate an 11-point concentration-response curve (80 μM top concentration) with replicate points at each concentration. The reaction was initiated by adding 2 μL / well of purified HSD17B13 protein (30 nM FAC in assay buffer). The compound, substrate mixture and HSD17B13 protein were incubated in the dark at room temperature for 2 hours, and then 3 μL / well NAD (P) H-Glo detection reagent (made from luciferase detection reagent and reductase / reductase substrate according to the manufacturer's instructions, Promega, G9061) was added. The detection reagent was incubated in the dark at room temperature for 1 hour, and then the plate was read on an Envision microplate reader (Perkin Elmer) using a luminescence protocol. The data expressed in relative luminescence units (RLU) were then normalized relative to the control wells using Activity Base (IDBS). Zero percent effect (ZPE) is defined as the RLU generated by uninhibited HSD17B13 protein (vehicle control). 100% effect (HPE) is defined as the RLU generated in the wells of a Pfizer proprietary compound known to cause 100% inhibition of HSD17B13 protein containing 40 μM FAC. The concentration and % effect of each compound were plotted using a four-parameter logistic dose-response equation using Activity Base, and the concentration required for 50% inhibition (IC 50 ).

[0684] HEK_Beta-Est LCMS_IC 50 _Tekcel

[0685] The inhibitory potency of the test compounds was determined using a whole cell HEK-HSD17B13 inhibition assay with an LCMS readout.

[0686] During the metabolism of β-estradiol (substrate), HSD17B13 uses the oxidized form of nicotinamide adenine dinucleotide (NAD+) as a cofactor, converting NAD+ to its reduced form (NADH) and converting β-estradiol into its product (estrone). Estrone production is quantified by LCMS and used as a measure of HSD17B13 enzyme activity.

[0687] HEK cells stably expressing wild-type human HSD17B13 were plated at 10,000 cells / well in 50 μL of growth medium (DMEM containing 10% heat-inactivated FBS, 400 μg / ml geneticin, 1× L-glutamine, and 1× non-essential amino acids, Invitrogen 11965-092, 16140-071, 10131-027, 25030-081, 11140-050) in poly-D-lysine-coated 384-well plates (Corning Biocoat, 354663) and incubated overnight at 37°C (95% O2: 5% CO2) (covered). After overnight incubation, an intermediate compound plate (Greiner, 781280) containing 160 nL of 375x FAC test compound was diluted 1 to 187.5 with 30 μL of warm assay medium (DMEM, 1x L-glutamine and 1x non-essential amino acids) to give a 2x FAC compound (80 μM highest concentration) in 0.53% DMSO. The test compound had been serially diluted 1 to 3.162 in 100% DMSO to give an 11-point concentration response curve with replicates at each concentration. The growth medium was then removed from the cell plate and replaced with 10 μL of 2x FAC test compound and incubated at 37°C (95% O2: 5% CO2) for 1 hour (covered), followed by the addition of assay medium / 0.2% DMSO containing 25 μM FACβ estradiol. The reaction was incubated at 37°C (95% O2: 5% CO2) for 2 hours (covered), and then 10 μL of the reaction was transferred from the assay plate to a new 384-well deep-well plate (Matrix4325) and 90 μL of stop reagent (containing the internal standard 17b estradiol-2,3,4- 13C3 in 50% methanol in water) was diluted 1 to 10. The amount of product (estrone) was then quantified by LCMS. The data, expressed as product area ratio (PAR), was then normalized relative to the control wells using Activity Base (IDBS). Zero percent effect (ZPE) was defined as the PAR generated by uninhibited HSD17B13 (vehicle control). 100% effect (HPE) was defined as the PAR generated from wells containing 20 μM FAC, a Pfizer proprietary compound known to cause 100% inhibition of HSD17B13. Using a four-parameter logistic dose-response equation, the concentration and % effect of each compound were plotted using Activity Base, and the concentration required for 50% inhibition (IC) was determined. 50 ).

[0688] In Table 3, the measurement data of the example according to the above measurement are presented below (IC 50 )(Based on the number of replicates tested (No.), take two (2) significant figures as the geometric mean).

[0689] Table 3: Bioassay data

[0690]

[0691]

[0692]

[0693]

[0694]

[0695]

[0696]

[0697]

[0698]

[0699]

[0700]

[0701] Throughout this application, various publications are cited. The disclosures of these publications are hereby incorporated by reference into this patent application in their entirety for all purposes.

[0702] It will be apparent to those skilled in the art that various modifications and variations may be made without departing from the scope or spirit of the invention. In view of the description and implementation of the invention disclosed herein, other embodiments of the invention will be apparent to those skilled in the art. The description and examples are intended to be considered as exemplary only, and the true scope and spirit of the invention are indicated by the appended claims.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, in A is -NH-C(O)- or heteroaryl having 1, 2, 3 or 4 heteroatoms selected from O, N and S, wherein A is optionally substituted by one or two R 4 replace; B is absent or is H, aryl, heteroaryl, heterocyclyl, fluorine, chlorine, bromine, oxo, cyano, hydroxy, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) fluoroalkyl, (C1-C6) alkoxy, or (C1-C6) fluoroalkoxy, wherein the heteroaryl or heterocyclyl has 1, 2 or 3 heteroatoms selected from O, N and S, and wherein B is optionally replaced by one or two R 5 replace; C does not exist or is H, -NH-C(O)-R 7 、-S(O)2-R 7 、-OS(O)2-R 7 , fluorine, chlorine, bromine, oxo, cyano, hydroxy, (C1-C6) alkyl, (C3-C6) cycloalkyl, (C1-C6) alkoxy, (C3-C6) cyclic ether, (C1-C6) fluoroalkyl, (C1-C6) fluoroalkoxy, aryl, heteroaryl or heterocyclic group, wherein the heteroaryl or heterocyclic group has 1, 2 or 3 heteroatoms selected from O, N and S, and wherein C is optionally replaced by one, two or three R 6 replace; R 1 、R 2 and R 3 are each independently selected from H and fluorine; Each R 4 、R 5 and R 6 independently selected from oxo, hydroxy, chlorine, fluorine, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)fluoroalkyl, (C3-C6)cycloalkyl, and heterocyclyl having 1, 2 or 3 heteroatoms selected from O and N; R 7 is hydroxy, chloro, fluoro, (C1-C6)alkyl, (C1-C6)alkoxy, (C1-C6)fluoroalkyl, or (C3-C6)cycloalkyl; and n is 0, 1 or 2.

2. The compound according to claim 1 or a pharmaceutically acceptable salt of the compound, wherein the compound has Formula IA:

3. The compound according to claim 1 or a pharmaceutically acceptable salt of the compound, wherein the compound has Formula IB:

4. A compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein R 1 、R 2 and R 3 At least one of them is fluorine.

5. The compound or pharmaceutically acceptable salt of any one of the preceding claims, wherein A is thiazolyl, pyrazolyl, oxazolyl, imidazolyl, isoxazolyl, isothiazolyl, imidazotriazinyl, imidazopyridazinyl, imidazopyridinyl, benzimidazolyl, benzothiazolyl, purinyl, pyridopyridazinyl, quinazolinyl, indazolyl, imidazopyridinyl, benzoxazolyl, pyrazolopyridinyl, isoindolinonyl, triazolyl, or oxadiazolyl.

6. A compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein A is 7. The compound or pharmaceutically acceptable salt of any one of the preceding claims, wherein B is absent or is H, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, pyrazolyl, piperazinyl, quinoxalinyl, phenyl, triazolyl, thiazolyl, thiadiazolyl, oxazolyl, imidazolyl, indazolyl, (Ci-C6)alkyl, (Ci-C6)fluoroalkyl, (Ci-C6)alkoxy, bromo, chloro, fluoro, or oxo, and wherein B is optionally substituted with one or two fluoro, oxo, hydroxy, (Ci-C6)alkyl, (C3-C6)cycloalkyl, (Ci-C6)fluoroalkyl, (Ci-C6)alkoxy, or (C3-C6)cyclic ether.

8. The compound according to any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein B is pyrimidinyl, (C1-C3)fluoroalkyl-substituted pyrimidinyl, (C1-C3)alkyl-substituted pyrazolyl, methoxy-substituted pyridazinyl, difluoromethyl-substituted pyrazinyl, trifluoromethyl-substituted pyrimidinyl, or methoxy-substituted pyrimidinyl.

9. The compound or pharmaceutically acceptable salt of any one of the preceding claims, wherein C is absent or is H, pyridyl, piperazinyl, oxolanyl, (C3-C6)cycloalkyl, (C1-C6)alkyl, (C1-C6)fluoroalkyl, (C1-C6)alkoxy, cyano, bromo, chloro, fluoro, or oxo, and wherein C is optionally substituted with one, two, or three fluoro, oxo, hydroxy, (C1-C6)alkyl, (C3-C6)cycloalkyl, (C1-C6)fluoroalkyl, or (C1-C6)alkoxy.

10. The compound or pharmaceutically acceptable salt of any one of the preceding claims, wherein C is absent or is pyridyl, piperazinyl, (C3-C6)cycloalkyl, (C1-C6)alkyl, (C1-C6)fluoroalkyl; and wherein C is optionally substituted with one, two or three fluoro, oxo, hydroxy, or (C1-C6)alkyl.

11. A compound, wherein the compound is: 2,3,5-Trifluoro-4-hydroxy-N-[(4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide; 2,3,5-Trifluoro-4-hydroxy-N-({4-[6-(pyrimidin-2-yl)-2H-indazol-2-yl]bicyclo[2.2.2]octan-1-yl}methyl)benzamide; 2,3,5-Trifluoro-4-hydroxy-N-({(1r,4r)-4-[6-(pyrimidin-5-yl)-2H-indazol-2-yl]cyclohexyl}methyl)benzamide; 2,3,5-Trifluoro-4-hydroxy-N-({4-[3-(6-methoxypyridazin-3-yl)-1,2,4-oxadiazol-5-yl]bicyclo[2.2.2]oct-1-yl}methyl)benzamide; N-[(4-{5-[5-(difluoromethyl)pyrazin-2-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]oct-1-yl)methyl]-3,5-difluoro-4-hydroxybenzamide; 3,5-Difluoro-4-hydroxy-N-{[(1r,4r)-4-{3-[5-(trifluoromethyl)pyrimidin-2-yl]-1,2,4-oxadiazol-5-yl}cyclohexyl]methyl}benzamide; 3,5-difluoro-4-hydroxy-N-({(1r,4r)-4-[6-(2-methoxypyrimidin-5-yl)-2H-pyrazolo[4,3-c]pyridin-2-yl]cyclohexyl}methyl)benzamide, 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(piperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide, or 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]oct-1-yl)methyl]benzamide, or a pharmaceutically acceptable salt of the compound.

12. A compound, wherein the compound is 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide, or a pharmaceutically acceptable salt of the compound.

13. A compound, wherein the compound is 2,3,5-trifluoro-4-hydroxy-N-[(4-{5-[2-(4-methylpiperazin-1-yl)pyrimidin-4-yl]-1,2,4-oxadiazol-3-yl}bicyclo[2.2.2]octan-1-yl)methyl]benzamide hydrochloride.

14. A compound, wherein the compound is 15. A method for treating the following diseases by administering a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt of the compound to a human in need of treatment: fatty liver, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, non-alcoholic steatohepatitis with liver fibrosis, non-alcoholic steatohepatitis with cirrhosis, non-alcoholic steatohepatitis with cirrhosis, hepatocellular carcinoma, alcoholic fatty liver disease, alcoholic steatohepatitis, hepatitis B, hepatitis C, biliary cirrhosis, renal clear cell carcinoma, head and neck squamous cell carcinoma, colorectal adenocarcinoma, mesothelioma, gastric adenocarcinoma, adrenocortical carcinoma, renal papillary cell carcinoma, cervical and endocervical cancer, bladder urothelial carcinoma, lung adenocarcinoma, type I diabetes, idiopathic type I diabetes diabetes mellitus (type Ib), latent autoimmune diabetes in adults (LADA), early-onset type 2 diabetes (EOD), atypical diabetes of the young (YOAD), maturity-onset diabetes of the young (MODY), malnutrition-related diabetes, gestational diabetes, restenosis after angioplasty, peripheral vascular disease, intermittent claudication, postprandial dyslipidemia, metabolic acidosis, ketosis, arthritis, diabetic retinopathy, macular degeneration, cataracts, diabetic nephropathy, glomerulosclerosis, chronic renal failure, diabetic neuropathy, skin and connective tissue disorders, foot ulcers and ulcerative colitis, endothelial dysfunction and impaired vascular compliance, kidney disease, end-stage renal disease, chronic kidney disease at risk of progression, and maple syrup urine disease.

16. The method of claim 15, wherein alcoholic fatty liver disease, alcoholic steatohepatitis, hepatitis B, hepatitis C, or biliary cirrhosis is treated.

17. The method of claim 15, wherein fatty liver, nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, nonalcoholic steatohepatitis with liver fibrosis, nonalcoholic steatohepatitis with cirrhosis, or nonalcoholic steatohepatitis with cirrhosis or hepatocellular carcinoma is treated.

18. The method of claim 15, wherein nonalcoholic steatohepatitis is treated.

19. A method of reducing cirrhosis, decompensated cirrhosis, progression to Model for End-Stage Liver Disease (MELD), liver transplantation, liver-related death, or hepatocellular carcinoma in a human by administering to a human a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt of said compound.

20. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt of the compound and a pharmaceutically acceptable carrier, vehicle or diluent.

21. A pharmaceutical combination composition comprising: a therapeutically effective amount of a composition comprising: a first compound, wherein the first compound is a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt of the compound; a second compound which is an antidiabetic agent, a therapeutic agent for nonalcoholic steatohepatitis, a therapeutic agent for nonalcoholic fatty liver disease, or an anti-heart failure therapeutic agent, and Pharmaceutically acceptable carrier, vehicle or diluent.

22. The pharmaceutical combination composition according to claim 21, wherein the therapeutic agent for non-alcoholic steatohepatitis or non-alcoholic fatty liver disease is an ACC inhibitor, a KHK inhibitor, a DGAT-2 inhibitor, a FXR agonist, metformin, an incretin analog, or an incretin receptor modulator.

23. The pharmaceutical combination composition according to claim 21, wherein the antidiabetic agent is an SGLT-2 inhibitor, metformin, an incretin mimetic, an incretin receptor modulator, a DPP-4 inhibitor, or a PPAR agonist.

Images