RU2807545C2 - Tetrazole-containing inhibitors of apoptotic signal-regulating kinase 1 and methods of their use - Google Patents

Abstract

FIELD: organic chemistry; pharmaceuticals.

SUBSTANCE: group of inventions relates to new compounds that inhibit apoptotic signal-regulating kinase 1 (ASK-1). The following is disclosed: the compounds of formula I or a pharmaceutically acceptable salt thereof, wherein selected from the group consisting of represents or ; represents; R¹ is selected from the group consisting of the following: 1) hydrogen; 2) -C₁-C₆ alkyl; 3) halogen -C₁-C₆ alkyl; and 4) -C₃-C₆ cycloalkyl; R² represents X represents either N or CR³; R³ is selected from the group consisting of the following: 1) hydrogen; 2) halogen; 3) -C₁-C₆ alkyl; and 4) -C₁-C₆ alkoxy; R⁴ is selected from the group consisting of the following: 1) hydrogen; 2) halogen; 3) -C₁-C₆ alkyl; 4) halogen-C₁-C₆ alkyl; and 5) -N(R⁵)(R⁶); where R⁵ and R⁶ are combined with the nitrogen atom to which they are attached to form a 3–8 membered heterocycloalkyl containing 1 nitrogen atom, 2 nitrogen atoms or a nitrogen atom and an oxygen atom, optionally substituted with a C₁-C₄-alkyl. In addition, a pharmaceutical composition for the treatment of an ASK-1-mediated disease or condition based on the above compounds is disclosed, as well as the use of the said compounds for the treatment of an ASK-1-mediated disease or condition.

EFFECT: group of inventions provides effective inhibition of apoptotic signal-regulating kinase 1 (ASK-1).

11 cl, 1 tbl, 84 ex

Description

Field of technology to which the invention relates

The present invention generally relates to compounds and pharmaceutical compositions that can be used as ASK-1 inhibitors. In particular, the present invention relates to compounds that can be used as ASK-1 inhibitors, methods for their preparation and their use.

Prior Art

Apoptotic signal-regulating kinase 1 (ASK-1) is a member of the cilantro mitogen-activated protein kinase (MAPKKK, MAP3K) family of kinases that, when activated, phosphorylates MAP kinase kinases (MAPKK, MAP2K), which in turn activate MAP kinases (MAPK). MAP kinases elicit responses by phosphorylating cellular substrates, thereby controlling the activity of transcription factors that ultimately control gene expression. Specifically, ASK-1, also known as MAPKKK5, phosphorylates MAPKK4/MAPKK7 or MAPKK3/MAPKK6, which then phosphorylates and activates c-Jun N-terminal protein kinase (JNK) and p38 MAPK, respectively (H. Ichijo, et al., Cell Comm. Signal 2009, 7, 1-10; K. Takeda, et al., Annu. Rev. Pharmacol. Toxicol. 2008, 48, 199-225; H. Nagai, et al., J. Biochem. Mol. Biol. 2007, 40, 1-6). Activation of the JNK and p38 pathways triggers further stress responses such as apoptosis, inflammation or differentiation (H. Ichijo, et al., Science 1997, 275, 90-94; K. Takeda, et al., J. Biol. Chem. 2000 , 275, 9805-9813; K. Tobiume, et al., EMBO Rep. 2001, 2, 222-228; K. Sayama, et al., J. Biol. Chem. 2001, 276, 999-1004).

ASK-1 activity is controlled by thioredoxin (Trx), which binds to the N-terminal end of ASK-1 (M. Saitoh, et al., EMBO J. 1998, 17, 2596-2606). ASK-1 is activated by autophosphorylation at Thr838 in response to external stimuli, including oxidative stress, lipopolysaccharide (LPS), reactive oxygen species (ROS), endoplasmic reticulum (ER) stress, increased intracellular calcium ion concentration, Fas ligand and various cytokines such as tumor necrosis factor (TNF) (H. Nishitoh, et al., Genes Dev. 2002, 16, 1345-1355; K. Takeda, et al., EMBO Rep. 2004, 5, 161-166; A. Matsuzawa, et al., Nat. Immunol. 2005, 6, 587-592).

ASK-1 has been associated with autoimmune diseases, neurodegenerative diseases, inflammatory diseases, chronic kidney disease, cardiovascular diseases, metabolic diseases, and acute and chronic liver diseases (R. Hayakawa, et al., Proc. Jpn. Acad., Ser. B 2012, 88, 434-453).

More specifically, ASK-1 has been associated with hepatic steatosis, including non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH). In a mouse model, a high-fat diet caused hepatic steatosis by causing fat accumulation and fatty acid oxidation. This led to the appearance of reactive oxygen species (ROS), which caused hepatocyte dysfunction and death (S. K. Mantena, et al., Free Radic. Biol. Med. 2008, 44, 1259-1272; S. K. Mantena, et al., Biochem. J. 2009, 417, 183-193). Moreover, TNF has been shown to play a critical role in hepatocyte apoptosis through the ASK-1-JNK pathway, and reduced hepatic steatosis and fibrosis were observed in TNF-deficient mice (W. Zhang, et al., Biochem. Biophys. Res Commun 2010, 391, 1731-1736).

Small molecule compounds that act as ASK-1 inhibitors have been disclosed in the following publications: WO 2008/016131, WO 2009/027283, WO 2009/0318425, WO 2009/123986, US 2009/0318425, WO 2011/041293, WO 201 1/ 097079, US 2011/0009410, G.P. Volynets, et al., J. Med. Chem. 2011, 54, 2680-2686, WO 2012/003387, WO 2012/011548, WO 2012/080735, Y. Terao, et al., Bioorg. Med. Chem. Lett. 2012, 22, 7326-7329, WO 2013/112741, G.P. Volynets, et al., Eur. J. Med. Chem. 2013, 16, 104-115, US 2014/0018370, WO 2014/100541, WO 2015/095059, WO 2016/049069, WO 2016/049070, WO 2018/090869, WO 2018/13386 5, WO 2018/133866, WO 2018 /148204, WO 2018/149284, WO 2018/151830, WO/2018/157856, WO 2018/157857, WO 2018/160406, WO 2018/169742, WO 2018/183122, WO 2018/1 87506, WO 2018/209354, WO 2018/218042, WO 2018/218044, WO 2018/218051, WO 2018/233553, US 2019/0062310, WO 2019/070742, WO 2019/050794, WO 2019/051265 and WO 2 019/034096.

There is a need to develop ASK-1 inhibitors for the treatment and prevention of diseases.

Brief description of the invention

The present invention relates to compounds and pharmaceutical compositions that can be used as ASK-1 inhibitors. In particular, the present invention relates to compounds that can be used as ASK-1 inhibitors, methods of their preparation and their use. In addition, the present invention includes a method for preparing these compounds.

The main aspect of the present invention describes a compound having formula (I), or a pharmaceutically acceptable salt or ester thereof:

,

Where

is an optionally substituted aryl or an optionally substituted heteroaryl containing 1, 2 or 3 heteroatoms selected from N, O and S, provided that is not ;

In some embodiments, selected from the following:

,

represents or ;

represents or ;

R ¹ and R ² are each independently selected from the group consisting of the following:

1) Hydrogen atom;

2) Optionally substituted -C ₁ -C ₈ alkyl;

3) Optionally substituted -C ₂ -C ₈ alkenyl;

4) Optionally substituted -C ₂ -C ₈ alkynyl;

5) Optionally substituted -C ₃ -C ₈ cycloalkyl;

6) Optionally substituted aryl;

7) Optionally substituted arylalkyl;

8) Optionally substituted 3-8 membered heterocycloalkyl;

9) Optionally substituted heteroaryl; And

10) Optionally substituted heteroarylalkyl;

X represents N or CR ³ ;

R ³ is selected from the group consisting of the following:

1) Hydrogen atom;

2) Halogen;

3) Optionally substituted -C ₁ -C ₈ alkyl; And

4) Optionally substituted -C ₁ -C ₈ alkoxy;

R ⁴ is selected from the group consisting of the following:

1) Hydrogen atom;

2) Halogen;

3) Optionally substituted -C ₁ -C ₈ alkyl;

4) Optionally substituted -C ₂ -C ₈ alkenyl;

5) Optionally substituted -C ₂ -C ₈ alkynyl;

6) Optionally substituted -C ₃ -C ₈ cycloalkyl;

7) Optionally substituted aryl;

8) Optionally substituted arylalkyl;

9) Optionally substituted 3-8 membered heterocycloalkyl;

10) Optionally substituted heteroaryl;

11) Optionally substituted heteroarylalkyl; And

12) −N(R ⁵ )(R ⁶ );

wherein R ⁵ and R ⁶ are independently selected from the group consisting of hydrogen, -C ₁ -C ₈ alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, each of which is optionally substituted with 1-3 substituents selected from halogen, alkyl, mono- or dialkylamino groups, alkyl or aryl or heteroaryl amide, -CN, alkoxy groups, -CF ₃ , aryl and heteroaryl. Alternatively, R ⁵ and R ⁶ are combined with the nitrogen atom to which they are attached to form an optionally substituted heterocyclic moiety.

In another embodiment, the present invention provides a pharmaceutical composition comprising a therapeutically effective amount of a compound or combination of compounds of the present invention, or a pharmaceutically acceptable salt, ester, or combination thereof, in combination with a pharmaceutically acceptable carrier or excipient.

In another embodiment, the present invention describes a method of preventing or treating a disease or condition involving ASK-1. This method involves administering a therapeutically effective amount of a compound having formula (I). The present invention also describes the use of a compound having formula (I) for the preparation of a medicament for the prevention or treatment of a disease or pathological condition involving ASK-1. Such diseases include autoimmune diseases, neurodegenerative diseases, inflammatory diseases, chronic kidney disease, cardiovascular diseases, metabolic disorders, and acute and chronic liver diseases.

Detailed Description of the Invention

In a first embodiment of the present invention, a compound having the above formula (I) or a pharmaceutically acceptable salt or ester thereof is described.

In some embodiments of compounds having formula (I), X is CH.

In some embodiments of compounds having formula (I), X is CF.

In some embodiments of compounds having formula (I), X is N.

In some embodiments of compounds having formula (I), R ¹ is selected from the groups depicted below

,

where each of the listed groups optionally has additional substituents.

In some embodiments of compounds having formula (I), R ⁴ is selected from the groups shown below:

,

where each of the listed groups optionally has additional substituents.

In some embodiments of compounds having formula (I), R ² is selected from the groups depicted below:

.

In some embodiments, the present invention describes compounds having formula (II-a ~ II-h), and pharmaceutically acceptable salts and esters thereof:

Where , R ² and R ⁴ have the meanings indicated above.

In some embodiments, the present invention describes compounds having formula (III-a ~ III-n), and pharmaceutically acceptable salts and esters thereof,

where R ¹ , R ² and R ⁴ have the meanings indicated above.

In some embodiments, the present invention describes compounds having formula (IV-a through IV-n), and pharmaceutically acceptable salts and esters thereof,

where R ¹ , R ² and R ⁴ have the meanings indicated above.

In some embodiments, the present invention describes a method of preventing or treating a disease or condition involving ASK-1. This method involves administering a therapeutically effective amount of a compound having formula (I), or a pharmaceutically acceptable salt thereof, to a subject in need thereof. The present invention also describes the use of a compound having formula (I), or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a disease or condition involving ASK-1.

In some embodiments, the disease or condition involving ASK-1 is an autoimmune disease, neurodegenerative disease, inflammatory disease, chronic kidney disease, renal disease, cardiovascular disease, metabolic disease, or acute or chronic liver disease.

In some embodiments, the chronic liver disease is primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug-induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition-induced cholestasis (PNAC), cholestasis induced by bacterial overgrowth or sepsis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), graft-versus-host disease caused by liver transplantation, liver regeneration in a living liver donor , congenital liver fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjögren's syndrome, sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis or alpha 1-antitrypsin deficiency. In some embodiments, the gastrointestinal disease is inflammatory bowel disease (IBD) (including ulcerative colitis and Crohn's disease), irritable bowel syndrome, microflora overgrowth, malabsorption, post-radiation colitis, or microscopic colitis.

In some embodiments, the kidney disease is diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic post-transplant glomerulopathy, chronic interstitial nephritis, or polycystic kidney disease.

In some embodiments, the cardiovascular disease is atherosclerosis, arteriosclerosis, ischemia-reperfusion injury in stroke, cardiac hypertrophy, respiratory disease, heart attack, myocardial ischemia.

In some embodiments, the metabolic disease is insulin resistance, type I and type II diabetes, or obesity.

In some embodiments, the chronic kidney disease is polycystic kidney disease, pyelonephritis, renal fibrosis, and glomerulonephritis.

Another aspect of the present invention is a method for preparing the compounds described in the present invention using the synthetic methods described herein.

Definitions

The following are definitions of terms used to describe the present invention. These definitions apply to terms used in the specification and claims unless otherwise specifically stated, individually or as part of a larger group.

The term “alkyl” as used herein means saturated linear or branched hydrocarbon radicals. Suitable alkyl groups include "C ₁ -C ₃ alkyl", "C ₁ -C ₆ alkyl", "C ₁ -C ₁₀ alkyl", "C ₂ -C ₄ alkyl" or "C ₃ -C ₆ alkyl", which means alkyl groups containing 1 - 3, 1 - 6, 1 - 10 carbon atoms, 2 - 4 and 3 - 6 carbon atoms, respectively. Examples of C ₁ -C ₈ alkyl radicals include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, tert-butyl, neopentyl, n-hexyl, heptyl and octyl radicals.

The term “alkenyl” as used herein means linear or branched hydrocarbon radicals containing at least one carbon-carbon double bond formed by the removal of one carbon atom. Suitable alkenyl groups include “C ₂ -C ₁₀ alkenyl”, “C ₂ -C ₈ alkenyl”, “C ₂ -C ₄ alkenyl” or “C ₃ -C ₆ alkenyl”, which mean alkenyl groups containing 2 - 10, 2 – 8, 2 – 4 or 3 – 6 carbon atoms, respectively. Alkenyl groups include, but are not limited to, for example, ethenyl, propenyl, butenyl, 1-methyl-2-buten-1-yl, heptenyl, octenyl, and the like.

The term “alkynyl” as used herein means linear or branched hydrocarbon radicals containing at least one carbon-carbon triple bond formed by the removal of one carbon atom. Suitable alkynyl groups include “C ₂ -C ₁₀ alkynyl”, “C ₂ -C ₈ alkynyl”, “C ₂ -C ₄ alkynyl” or “C ₃ -C ₆ alkynyl,” which mean alkynyl groups containing 2 to 10, 2 – 8, 2 – 4 or 3 – 6 carbon atoms, respectively. Representative alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 1-butynyl, heptynyl, octinyl, and the like.

The term “cycloalkyl” as used herein means a monocyclic or polycyclic saturated carbocyclic ring or bi- or tricyclic group of fused, bridged or spiro structure, and the carbon atoms may optionally be oxo-substituted or optionally substituted by an exocyclic olefin, imine or oxime double bond . Preferred cycloalkyl groups include C ₃ -C ₁₂ cycloalkyl, C ₃ -C ₆ cycloalkyl, C ₃ -C ₈ cycloalkyl and C ₄ -C ₇ cycloalkyl. Examples of C ₃ -C ₁₂ cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopentyl, cyclooctyl, 4-methylene-cyclohexyl, bicyclo[2.2.1]heptyl, bicyclo[3.1.0]hexyl, spiro [2.5]octyl, 3-methylenebicyclo[3.2.1]octyl, spiro[4.4]nonanyl, etc.

The term “cycloalkenyl” as used herein means a monocyclic or polycyclic saturated carbocyclic ring or bi- or tricyclic group of fused, bridged or spiro structure containing at least one carbon-carbon double bond, and the carbon atoms may optionally be oxo-substituted or optionally substituted with an exocyclic olefin, imine or oxime double bond. Preferred cycloalkenyl groups include C ₃ -C ₁₂ cycloalkenyl, C ₃ -C ₈ cycloalkenyl or C ₅ -C ₇ cycloalkenyl groups. Examples of C ₃ -C ₁₂ cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, bicyclo[2.2.1]hept-2-enyl, bicyclo[3.1.0]hex-2-enyl , spiro[2.5]oct-4-enyl, spiro[4.4]non-1-enyl, bicyclo[4.2.1]non-3-en-9-yl, etc.

The term "aryl" as used herein means a mono- or polycyclic carbocyclic ring system containing at least one aromatic ring, including but not limited to phenyl, naphthyl, tetrahydronaphthyl, indanyl and indenyl. Polycyclic aryl is a polycyclic ring system that contains at least one aromatic ring. Polycyclic aryls may contain fused rings, covalently linked rings, or a combination thereof.

The term "heteroaryl" as used herein means a mono- or polycyclic aromatic radical containing on the ring one or more atoms selected from S, O and N; and the remaining ring atoms are carbon atoms, wherein any N or S atom included in the ring may optionally be oxidized. Heteroaryl includes, but is not limited to, pyridinyl, pyrazinyl, pyrimidinyl, pyrazolyl, imidazolyl, thiazolyl, oxazolyl. isoxazolyl, thiadiazolyl, oxadiazolyl, thiophenyl, furanyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolyl, quinoxalinyl. The polycyclic heteroaryl may contain fused rings, covalently linked rings, or a combination thereof.

According to the present invention, aromatic groups may be substituted or unsubstituted.

The term “bicyclic aryl” or “bicyclic heteroaryl” means a ring system consisting of two rings, where at least one ring is aromatic; and the two rings may be fused or covalently attached to each other.

As used herein, the term “arylalkyl” means a functional group in which an alkylene chain is attached to an aryl group, for example, -CH ₂ CH ₂ -phenyl. The term “substituted arylalkyl” means an arylalkyl functional group in which the aryl group is substituted. Likewise, the term “heteroarylalkyl” means a functional group in which an alkylene chain is attached to a heteroaryl group. The term “substituted heteroarylalkyl” means a heteroarylalkyl functional group in which the heteroaryl group is substituted.

The term “alkylene” as used herein means a diradical formed from a branched or straight-chain hydrocarbon chain typically containing from 1 to 20 carbon atoms (e.g., 1 to 10 carbon atoms, or 1, 2, 3, 4, 5, or 6 carbon atoms). This term can be illustrated by such groups as methylene (−CH ₂ −), ethylene (−CH ₂ CH ₂ −), propylene isomers (for example, −CH ₂ CH ₂ CH ₂ − and -CH(CH ₃ )CH ₂ −) and etc.

The term “substituted” as used herein means the independent substitution of one, two, three or more hydrogen atoms with substituents including, but not limited to, deuterium, -F, -Cl, -Br, -I, -OH, protected hydroxy -group, -NO ₂ , -CN, -NH ₂ , N ₃ , protected amino group, alkoxy group, thioalkoxy group, oxo group, C ₁ -C ₁₂ -alkyl, C ₂ -C ₁₂ -alkenyl, C ₂ -C ₁₂ -alkynyl, -halogen-C ₁ -C ₁₂ -alkyl, -halogen-C ₂ -C ₁₂ -alkenyl, -halogen-C ₂ -C ₁₂ -alkynyl, -halogen-C ₃ -C ₁₂ - cycloalkyl, -NH-C ₁ -C ₁₂ -alkyl, -NH-C ₂ -C ₁₂ -alkenyl, -NH-C ₂ -C ₁₂ -alkynyl, -NH-C ₃ -C ₁₂ -cycloalkyl, -NH-aryl , -NH-heteroaryl, -NH-heterocycloalkyl, -dialkylamino, -diarylamino group, -diheteroarylamino group, -OC ₁ -C ₁₂ -alkyl, -OC ₂ -C ₁₂ -alkenyl, -OC ₂ -C ₁₂ -alkynyl , -OC ₃ -C ₁₂ -cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocycloalkyl, -C(O)- C ₁ -C ₁₂ -alkyl, -C(O)- C ₂ -C ₁₂ -alkenyl, -C(O)-C ₂ -C ₁₂ -alkynyl, -C(O)-C ₃ -C ₁₂ -cycloalkyl, -C(O)-aryl, -C(O)-heteroaryl, -C( O)-heterocycloalkyl, -CONH ₂ , -CONH-C ₁ -C ₁₂ -alkyl, -CONH-C ₂ -C ₁₂ -alkenyl, -CONH-C ₂ -C ₁₂ -alkynyl, -CONH-C ₃ -C ₁₂ -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -CO ₂ -C ₁ -C ₁₂ -alkyl, -CO ₂ -C ₂ -C ₁₂ -alkenyl, -CO ₂ -C ₂ -C ₁₂ -alkynyl, -CO ₂ -C ₃ -C ₁₂ -cycloalkyl, -CO ₂ -aryl, -CO ₂ -heteroaryl, -CO ₂ -heterocycloalkyl, -CONH ₂ , -CONH-C ₁ -C ₁₂ -alkyl, - CONH-C ₂ -C ₁₂ -alkenyl, -CONH-C ₂ -C ₁₂ -alkynyl, -CONH-C ₃ -C ₁₂ -cycloalkyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocycloalkyl, -NHC( O)-C ₁ -C ₁₂ -alkyl, -NHC(O)-C ₂ -C ₁₂ -alkenyl, -NHC(O)-C ₂ -C ₁₂ -alkynyl, -NHC(O)-C ₃ -C ₁₂ -cycloalkyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocycloalkyl, -NHCO ₂ -C ₁ -C ₁₂ -alkyl, -NHCO ₂ -C ₂ -C ₁₂ -alkenyl , -NHCO ₂ -C ₂ -C ₁₂ -alkynyl, -NHCO ₂ -C ₃ -C ₁₂ -cycloalkyl, -NHCO ₂ - aryl, -NHCO ₂ -heteroaryl, -NHCO ₂ -heterocycloalkyl, -NHC(O)NH ₂ , -NHC(O)NH-C ₁ -C ₁₂ -alkyl, -NHC(O)NH-C ₂ -C ₁₂ -alkenyl, -NHC(O)NH-C ₂ -C ₁₂ -alkynyl, -NHC(O )NH-C ₃ -C ₁₂ -cycloalkyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocycloalkyl, NHC(S)NH ₂ , -NHC(S) NH-C ₁ -C ₁₂ -alkyl, -NHC(S)NH-C ₂ -C ₁₂ -alkenyl, -NHC(S)NH-C ₂ -C ₁₂ -alkynyl, -NHC(S)NH-C ₃ - C ₁₂ -cycloalkyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocycloalkyl, -NHC(NH)NH ₂ , -NHC(NH)NH-C ₁ - C ₁₂ -alkyl, -NHC(NH)NH-C ₂ -C ₁₂ -alkenyl, -NHC(NH)NH-C ₂ -C ₁₂ -alkynyl, -NHC(NH)NH-C ₃ -C ₁₂ -cycloalkyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocycloalkyl, -NHC(NH)-C ₁ -C ₁₂ -alkyl, -NHC(NH)-C ₂ - C ₁₂ -alkenyl, -NHC(NH)-C ₂ -C ₁₂ -alkynyl, -NHC(NH)-C ₃ -C ₁₂ -cycloalkyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, - NHC(NH)-heterocycloalkyl, -C(NH)NH-C ₁ -C ₁₂ -alkyl, -C(NH)NH-C ₂ -C ₁₂ -alkenyl, -C(NH)NH-C ₂ -C ₁₂ - alkynyl, -C(NH)NH-C ₃ -C ₁₂ -cycloalkyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocycloalkyl, -S(O)- C ₁ -C ₁₂ -alkyl, -S(O)-C ₂ -C ₁₂ -alkenyl, -S(O)-C ₂ -C ₁₂ -alkynyl, -S(O)-C ₃ -C ₁₂ -cycloalkyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocycloalkyl-SO ₂ NH ₂ , -SO ₂ NH- C ₁ -C ₁₂ -alkyl, -SO ₂ NH- C ₂ - C ₁₂ -alkenyl, -SO ₂ NH-C ₂ -C ₁₂ -alkynyl, -SO ₂ NH-C ₃ -C ₁₂ -cycloalkyl, -SO ₂ NH- aryl, -SO ₂ NH-heteroaryl, -SO ₂ NH- heterocycloalkyl, -NHSO ₂ -C ₁ -C ₁₂ -alkyl, -NHSO ₂ -C ₂ -C ₁₂ -alkenyl, -NHSO ₂ -C ₂ -C ₁₂ -alkynyl, -NHSO ₂ -C ₃ -C ₁₂ -cycloalkyl, -NHSO ₂ -aryl, -NHSO ₂ -heteroaryl, -NHSO ₂ -heterocycloalkyl, -CH ₂ NH ₂ , -CH ₂ SO ₂ CH ₃ , -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -C ₃ -C ₁₂ -cycloalkyl, polyalkoxyalkyl, polyalkoxy group, -methoxymethoxy group, -methoxyethoxy group, -SH, -SC ₁ -C ₁₂ -alkyl, -SC ₂ -C ₁₂ -alkenyl, -SC ₂ -C ₁₂ - alkynyl, -SC ₃ -C ₁₂ -cycloalkyl, -S-aryl, -S-heteroaryl, -S-heterocycloalkyl, methylthiomethyl or –L'–R', where L' represents C ₁ -C ₆ alkylene, C ₂ - C ₆ alkenylene or C ₂ -C ₆ alkynylene, and R' represents aryl, heteroaryl, heterocycle, C ₃ -C ₁₂ cycloalkyl or C ₃ -C ₁₂ cycloalkenyl. In some embodiments, the substituents are independently selected from halogen, preferably Cl and F; C _1- C ₄ alkyl, preferably methyl and ethyl; halogen-C _1- C ₄ -alkyl, such as fluoromethyl, difluoromethyl and trifluoromethyl; C ₂ -C ₄ alkenyl; halogen-C ₂ -C ₄ -alkenyl; C ₃ -C ₆ -cycloalkyl, such as cyclopropyl; C _1- C ₄ alkoxy groups such as methoxy group and ethoxy group; halogen-C _1- C ₄ -alkoxy groups such as fluoromethoxy group, difluoromethoxy group and trifluoromethoxy group, -CN; -OH; _NH2 ; C _1- C ₄ -alkylamino groups; di(C _1- C ₄ -alkyl)amino groups; and _NO2 . It should be understood that aryls, heteroaryls, alkyls, etc. may be further substituted. In some cases, each substituent on a substituted moiety is optionally further substituted with one or more groups, where each group is independently selected from C ₁ -C ₆ -alkyl, -F, -Cl, -Br, -I, -OH, -NO ₂ . -CN or _-NH2 . Preferably, the substituted alkyl group, such as a substituted methyl group, is substituted with one or more halogen atoms, more preferably one or more fluorine or chlorine atoms.

According to the present invention, any of the aryls, substituted aryls, heteroaryls and substituted heteroaryls described herein can be any aromatic group. Aromatic groups may be substituted or unsubstituted.

It should be understood that any alkyl, alkenyl, alkynyl, cycloalkyl and cycloalkenyl moiety described herein may also be an aliphatic group, an alicyclic group or a heterocyclic group. An “aliphatic group” is a non-aromatic moiety that may contain any combination of carbon atoms, hydrogen atoms, halogen atoms, oxygen, nitrogen or other atoms, and optionally contains one or more unsaturation elements, such as double and/or triple bonds. The aliphatic group may be straight, branched or cyclic, and preferably contains from about 1 to about 24 carbon atoms, more often from about 1 to about 12 carbon atoms. In addition to aliphatic hydrocarbon groups, aliphatic groups include, for example, polyalkoxyalkyls such as polyalkylene glycols, polyamines and polyimines. Such aliphatic groups may be further substituted. It should be understood that aliphatic groups can be used in place of the alkyl, alkenyl, alkynyl, alkylene, alkenylene and alkynylene groups described herein.

The term “alicyclic” as used herein means a monovalent group formed from a monocyclic or polycyclic saturated carbocyclic compound by the removal of one hydrogen atom. Examples thereof include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[2.2.1]heptyl and bicyclo[2.2.2]octyl. Such alicyclic groups may be further substituted.

As used herein, the term “alkoxy,” used alone or in combination with other terms, means, unless otherwise specified, an alkyl group containing the stated number of carbon atoms attached to the remainder of the molecule through an oxygen atom, for example, methoxy-, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) group, and higher homologues and isomers. Preferred alkoxy groups are (C ₁ -C ₃ ) alkoxy groups.

The term “aryloxy” means an aryl-O- group, where aryl group is as defined above, and includes optionally substituted aryl groups, also defined above. The term “arylthio” means the group R−S−, where R corresponds to the definition of aryl given above.

The terms “heterocyclic” or “heterocycloalkyl” can be used interchangeably and mean a non-aromatic ring or bi- or tricyclic group of fused, bridged or spiro structure, wherein (i) each ring system contains at least one heteroatom independently selected from oxygen, sulfur and nitrogen, (ii) each ring system may be saturated or unsaturated, (iii) the nitrogen and sulfur heteroatoms may optionally be oxidized, (iv) the nitrogen heteroatom may optionally be quaternized, (v) any of the rings described above may be fused to an aromatic ring and (vi) the remaining atoms in the ring are carbon atoms, which may optionally be oxo-substituted or optionally substituted by an exocyclic olefin, imine or oxime double bond. Representative heterocycloalkyl groups include, but are not limited to, 1,3-dioxolane, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, piperazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiazolidinyl, isothiazolidinyl, quinoxalinyl, pyridazinonyl, 2- azabicyclo[2.2 .1]-heptyl, 8-azabicyclo[3.2.1]octyl, 5-azaspiro[2.5]octyl, 1-oxa-7-azaspiro[4.4]nonanyl, 7-oxooxepan-4-yl and tetrahydrofuryl. Such heterocyclic groups may be further substituted. Heteroaryl or heterocyclic groups may be C-linked or N-linked (where appropriate).

It should be understood that any alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclic and cycloalkenyl moiety described herein may also be an aliphatic group or an alicyclic group.

In various embodiments of the present invention, the substituted or unsubstituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, arylalkyl, heteroarylalkyl and heterocycloalkyl may be monovalent or divalent. Thus, alkylene, alkylene and alkynylene, cycloalkylene, cycloalkenylene, cycloalkynylene, arylalkylene, heteroarylalkylene and heterocycloalkylene groups are included in the above definitions and can be used in the formulas given in this text, subject to the rules of valency.

The term "halogen" as used herein means an atom selected from fluorine, chlorine, bromine and iodine.

The term “optionally substituted” as used herein means that the corresponding group may be substituted or unsubstituted. In one embodiment, the corresponding group is optionally substituted with zero substituent, i.e. the corresponding group is unsubstituted. In another embodiment, the corresponding group is optionally replaced by one or more additional groups individually and independently selected from the groups described herein.

The term “hydrogen” includes hydrogen and deuterium. In addition, the indication of an atom includes other isotopes of that atom, provided that the resulting compound is pharmaceutically acceptable.

In some embodiments, compounds meeting the given formulas include isotopically labeled compounds. An “isotopically labeled compound” is a compound in which at least one atomic position is enriched in a specific isotope of the specified element to a degree substantially greater than the natural abundance of that isotope. For example, one or more hydrogen atom positions in a compound may be enriched in deuterium to a degree substantially greater than the natural abundance of deuterium, such as enrichment to at least 1%, preferably at least 20%, or at least 50%. Such a deuterated compound may, for example, be metabolized more slowly than its non-deuterated counterpart and therefore exhibit a longer half-life when administered to a subject. Such compounds can be synthesized using methods known in the art, for example, using deuterated starting compounds. Unless otherwise indicated, isotopically labeled compounds are pharmaceutically acceptable.

The compounds described herein may contain one or more asymmetric centers, thereby allowing the formation of enantiomers, diastereomers and other stereoisomeric forms that can be defined in terms of absolute stereochemistry, for example (R)- or (S)-, or (D) - or (L)- for amino acids. The present invention includes all possible isomers, as well as their racemic and optically pure forms. Optical isomers can be prepared from their respective optically active precursors according to the methods described above or by resolution of racemic mixtures. The cleavage can be carried out in the presence of a cleavage agent, chromatographically or by repeated crystallizations, or using a combination of these methods known to those skilled in the art. Additional details regarding cleavage can be found in Jacques, et al., Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). When compounds described herein contain an olefinic double bond, other unsaturations, or other centers of geometric asymmetry, and unless otherwise indicated, the compounds are meant to include both types of geometric isomers, i.e. E and Z geometric isomers or cis and trans isomers. Likewise, all tautomeric forms are also included within the scope of the present invention. Tautomers can be cyclic or acyclic. The configuration of all carbon-carbon double bonds depicted in this text is given for convenience only and does not indicate a specific configuration unless otherwise indicated in the text; thus, a carbon-carbon double bond or a carbon-heteroatom double bond, depicted herein as trans, may be cis, trans, or a mixture of both in any proportion.

The term “subject” as used herein means a mammal. Thus, the subject may be, for example, dogs, cats, horses, cows, pigs, guinea pigs, etc. Preferably, the subject is a human. When the subject is a person, the subject can be called a patient.

As used herein, the term "pharmaceutically acceptable salt" means salts of the compounds prepared by the process of the present invention that are, within the meaning of current medical knowledge, suitable for use in contact with human and lower animal tissues without undesirable toxicity, irritation, allergic reactions, etc. etc., and which correspond to a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art.

Berge et al. described pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). Such salts can be prepared in situ during the final isolation and purification of the compounds of the present invention, or separately by reaction of the free base with a suitable organic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, for example, salts formed by the amino group with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids, such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorasulfonate, citrate, cyclopentane propionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalene sulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Representative alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts include, where available, non-toxic ammonium salts, quaternary ammonium salts and amine salts formed with counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl, sulfonate and arylsulfonate.

As used herein, the term "pharmaceutically acceptable ester" means esters that hydrolyze in vivo, and includes esters that readily degrade in the human body to release the parent compound or a salt thereof. Suitable ester groups include, for example, those formed from pharmaceutically acceptable carboxylic acids, in particular alkanoic, alkene, cycloalkanoic and alkanedioic acids, in which each alkyl or alkenyl moiety preferably contains more than 6 carbon atoms. Examples of suitable esters include, but are not limited to, C ₁ -C ₆ alkanoic acid esters such as acetate, propionate, butyrate and pivalate esters.

The term “hydroxyl-activating group” as used herein means a labile chemical moiety that is known in the art to activate a hydroxyl group such that it is removed by synthetic operations such as substitution or elimination reactions. Examples of hydroxyl activating groups include, but are not limited to, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate, and the like.

The term “activated hydroxyl” as used herein means a hydroxy group activated by the hydroxyl activating group described above, including, for example, mesylate, tosylate, triflate, p-nitrobenzoate, phosphonate groups.

The term “hydroxy protecting group” as used herein means a labile chemical moiety that is known in the art to protect the hydroxyl group from unwanted reactions during synthetic transformations. After such synthetic transformations, the hydroxy protecting group can be selectively removed. Hydroxy protecting groups known in the art are generally described in T.H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of hydroxyl protecting groups include benzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, tert-butoxycarbonyl, isopropoxycarbonyl, diphenylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl, allyloxycarbonyl, acetyl, formyl, chloroacetyl, trifluoroacetyl, methoxyacetyl, phenoxyacetyl, benzoyl, methyl, t - butyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, allyl, benzyl, triphenylmethyl (trityl), methoxymethyl, methylthiomethyl, benzyloxymethyl, 2-(trimethylsilyl)ethoxymethyl, methanesulfonyl, trimethylsilyl, triisopropylsilyl, etc.

The term "substituted hydroxy" as used herein means a hydroxy group protected by the hydroxyl protecting group described above, including, for example, a benzoyl, acetyl, trimethylsilyl, triethylsilyl, methoxymethyl group.

The term “hydroxy-prodrug group” as used herein means a pro-group that is known in the art to temporarily alter the physicochemical and consequent biological properties of the parent drug molecule by capping or masking the hydroxy group. After synthetic manipulation, the hydroxy prodrug group described above should be able to convert back to a hydroxy group in vivo. Hydroxy prodrug groups known in the art are generally described in Kenneth B. Sloan, Prodrugs, Topical an Ocular Drug Delivery, (Drugs and the Pharmaceutical Sciences; Volume 53), Marcel Dekker, Inc., New York (1992) and “Prodrugs of Alcohols and Phenols” by S. S. Dhareshwar and V. J. Stella, in Prodrugs Challenges and Rewards Part-2, (Biotechnology: Pharmaceutical Aspects), edited by V. J. Stella, et al, Springer and AAPSPress, 2007, pp. 31-99.

The term “amino” as used herein means a –NH ₂ group.

The term “substituted amino group” as used herein means a –NRR group, wherein each R is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl and heterocycloalkyl, provided that both R groups cannot represent a hydrogen, or a −Y−Z group in which Y is an optionally substituted alkylene and Z is alkenyl, cycloalkenyl or alkynyl.

The term “amine protecting group” as used herein means a labile chemical moiety that is known in the art to protect the amino group from unwanted reactions during synthetic manipulation. After completion of the synthetic manipulations, the amino protecting group described herein can be selectively removed. Amine protecting groups known in the art are generally described in T.H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, New York (1999). Examples of amino protecting groups include, but are not limited to, t-butoxycarbonyl, 9-fluorenylmethoxycarbonyl, benzyloxycarbonyl, and the like.

The term "leaving group" means a functional group or atom that can be replaced by another functional group or atom by a substitution reaction, such as a nucleophilic substitution reaction. For example, representative leaving groups include chlorine, bromine and iodine; sulfoester groups such as mesylate, tosylate, brosylate, nosylate and the like; and acyloxy groups such as acetoxy, trifluoroacetoxy and the like.

As used herein, the Aztec term "pharmaceutically acceptable ester" means esters of compounds produced by the process of the present invention that are hydrolyzed in vivo, and includes esters that readily degrade in the human body to release the parent compound or a salt thereof. Suitable ester groups include, for example, those formed from pharmaceutically acceptable aliphatic carboxylic acids, in particular alkanoic, alkenic, cycloalkanic and alkanedioic acids, in which each alkyl or alkenyl moiety preferably contains no more than 6 carbon atoms. Examples of suitable esters include, but are not limited to, formates, acetates, propionates, butyrates, acrylates and ethyl succinates.

The term “pharmaceutically acceptable prodrugs” as used herein means prodrugs of compounds prepared by the method of the present invention that are, according to accepted medical knowledge, suitable for use in contact with human and lower animal tissues without unnecessary toxicity, irritation, allergic reactions, etc. etc., which correspond to a reasonable benefit/risk ratio and are effective in the areas of their intended application, as well as, if possible, zwitterionic forms of the compounds of the present invention. The term “prodrug” as used herein means a compound that can be converted in vivo by metabolic mechanisms (eg, hydrolysis) to produce a compound corresponding to the formulas of the present invention. Various forms of prodrugs are known in the art, for example, they are described in Bundgaard, (ed.), Design of Prodrugs, Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, Vol. 4, Academic Press (1985); Krogsgaard-Larsen, et al., (ed). "Design and Application of Drugs, Textbook of Drug Design and Development, Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug Delivery Reviews, 8:1-38(1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285 et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel Drug Delivery Systems, American Chemical Society (1975); and Bernard Testa & Joachim Mayer, “Hydrolysis In Drug and Prodrug Metabolism: Chemistry, Biochemistry and Enzymology,” John Wiley and Sons, Ltd. (2002).

The term "treatment" as used herein means alleviation, amelioration, reduction, removal or modulation, i.e. causing regression of a disease or painful condition. Treatment may also include inhibition, i.e. stopping the development of an existing disease or pathological condition, and reducing or alleviating, i.e. causing regression of an existing disease or pathological condition, for example, in cases where the disease or pathological condition already exists.

The term “preventing” as used herein means completely or nearly completely stopping the onset of a disease or condition in a patient or subject, especially when the patient or subject is predisposed thereto or is at risk of acquiring the disease or condition.

In addition, the compounds of the present invention, for example, salts of these compounds, may exist in hydrated or non-hydrated (anhydrous) form or as solvates with other solvent molecules. Non-limiting examples of hydrates include monohydrates, dihydrates, etc. Non-limiting examples of solvates include ethanol solvates, acetone solvates, etc.

The term "solvates" means solvent-added forms that contain stoichiometric or non-stoichiometric amounts of solvent. Some compounds tend to trap a fixed molar proportion of solvent molecules in the crystalline state, forming a solvate. If the solvent is water, the resulting solvate is a hydrate. When the solvent is an alcohol, the solvate is an alcoholate. Hydrates are formed by the combination of one or more molecules of water with one of the substances, where water retains its molecular state as an H ₂ O molecule, and such a combination is capable of forming one or more hydrates.

As used herein, the term "analogue" means a chemical compound that is structurally similar to another but slightly different in composition (for example, the substitution of an atom for an atom of a different element, or the presence of a particular functional group, or the substitution of one functional group for another functional group ). Thus, an analogue is a compound that is similar or comparable in function and appearance to a reference compound.

The term "aprotic solvent" as used herein means a solvent that is relatively inert in terms of proton activity, i.e. does not act as a proton donor. Examples include, but are not limited to, hydrocarbons such as, for example, hexane and toluene, halogenated hydrocarbons, such as, for example, methylene chloride, ethylene chloride, chloroform, and the like, heterocyclic compounds, such as, for example, tetrahydrofuran and N -methylpyrrolidinone, and ethers such as diethyl ether, bis-methoxymethyl ether. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for certain compounds and reaction conditions, depending on factors such as, for example, the solubility of the reagents, the reactivity of the reagents, and preferred temperature ranges. Additional discussion of aprotic solvents can be found in organic chemistry textbooks or specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

The terms “protogenic organic solvent” or “protic solvent” as used herein mean solvents that are prone to abstraction of a proton, such as alcohols, for example, methanol, ethanol, propanol, isopropanol, butanol, t-butanol, etc. Such solvents are well known to those skilled in the art, and individual solvents or mixtures thereof may be preferred for certain compounds and reaction conditions, depending on factors such as, for example, the solubility of the reagents, the reactivity of the reagents, and preferred temperature ranges. Additional discussion of protic solvents can be found in organic chemistry textbooks or specialized monographs, for example: Organic Solvents Physical Properties and Methods of Purification, 4th ed., edited by John A. Riddick et al., Vol. II, in the Techniques of Chemistry Series, John Wiley & Sons, NY, 1986.

Combinations of substituents and variables covered by the present invention can only be those that result in stable compounds. The term "stable" as used herein means compounds that are stable enough to be produced and that maintain their integrity for a sufficient period of time to be used for the purposes specified herein (e.g., therapeutic or prophylactic administration subject).

The synthesized compounds can be isolated from the reaction mixture and further purified by methods such as column chromatography, high performance liquid chromatography or recrystallization. In addition, the various chemical steps can be carried out in a different order or sequence to obtain the target compounds. In addition, solvents, temperatures, reaction times, etc. are indicated in this text for illustrative purposes only, and varying the reaction conditions allows you to obtain the desired isoxazole products of the present invention. Synthetic chemistries and protecting group methodologies (protection and deprotection) that are used in the synthesis of the compounds described herein include, for example, those described in R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); T.W. Greene and P.G.M. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995).

The compounds of the present invention can be modified by changing various functional groups using the synthetic methods described herein in order to enhance specific biological properties. Such modifications include modifications that improve biological penetration into the desired biological system (eg, blood, lymphatic system, central nervous system), increase oral bioavailability, increase solubility allowing administration by injection, alter metabolism and elimination rate.

Pharmaceutical compositions

The pharmaceutical compositions of the present invention contain a therapeutically effective amount of a compound of the present invention together with one or more pharmaceutically acceptable carriers. As used herein, the term "pharmaceutically acceptable carrier" means a non-toxic, inert solid or semi-solid or liquid excipient, diluent, encapsulating material or excipient of any type. Some examples of substances that can act as pharmaceutically acceptable carriers are sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; powdered gum tragacanth; malt; gelatin; talc; excipients such as cocoa butter and wax for suppositories; oils such as peanut oil, cottonseed oil; safflower oil; Sesame oil; olive oil; corn oil and soybean oil; glycols such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffer additives such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic sodium chloride solution; Ringer's solution; ethyl alcohol and phosphate-buffered saline, and may be formulated with non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as colors, disintegrants, coating materials, sweeteners, flavors and fragrances, preservatives and antioxidants. The pharmaceutical compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (eg, in the form of powders, ointments, or drops), buccally, or as an oral or nasal spray.

The pharmaceutical compositions of the present invention can be administered orally, parenterally, as inhalation sprays, topically, rectally, nasally, buccally, intravaginally or as an implantable reservoir, preferably orally or by injection. The pharmaceutical compositions of the present invention may contain any conventional non-toxic pharmaceutically acceptable carriers or adjuvants. In some cases, the pH of the formulation may be adjusted using pharmaceutically acceptable acids, bases, or buffering agents to enhance the stability of the formulation of the compound or formulation thereof. The term “parenteral” as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasternal, intrathecal, intralesional, or intracranial administration by injection or infusion techniques.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredients, liquid dosage forms may contain inert diluents commonly used in the art, for example, water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butylene glycol, dimethylformamide, oils (especially cottonseed, peanut, corn, olive, castor or sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters, and mixtures thereof. In addition to inert diluents, compositions for oral administration may also include adjuvants such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents and flavoring agents.

Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be prepared according to techniques known to those skilled in the art, using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic diluent or solvent suitable for parenteral administration, for example a solution in 1,3-butanediol. Suitable carriers and solvents also include water, Ringer's solution, USP solution, and isotonic sodium chloride solution. In addition, sterile fatty oils are also widely used as a solvent or suspending medium. Any brand of fatty oil can be used for this purpose, including synthetic mono- and diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectable drugs.

Injectable preparations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents into sterile solid compositions that must be dissolved or dispersed in sterile water or other sterile environment immediately prior to use.

To increase the duration of action of a drug, it is often desirable to slow the absorption of the drug by subcutaneous or intramuscular injection. This can be achieved by using liquid suspensions of crystalline or amorphous substances with low solubility in water. The rate of absorption of the drug then depends on the rate of its dissolution, which in turn may depend on the crystal size and crystalline form. Alternatively, slowing the absorption of a parenterally administered drug can be achieved by dissolving or suspending the drug in an oily vehicle. Injectable depot formulations are prepared by forming microencapsulated drug matrices in biodegradable polymers such as polylactide-polyglycolide. The rate of drug release can be controlled by the ratio of drug to polymer as well as the nature of the polymer used. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Deposited injectable drugs are also prepared by incorporating the drug into liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or intravaginal use are preferably suppositories, which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or suppository wax that is solid at room temperature but liquid at body, and therefore melt in the anus or vaginal cavity, releasing the active substance.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms the active substance is mixed with at least one inert pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate, and/or: a) excipients such as starches, lactose, sucrose, glucose, mannitol and silica acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and acacia gum, c) humectants such as glycerin, d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch , alginic acid, some silicates and sodium carbonate, e) solution retarders such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate, h) adsorbents such as kaolin and bentonite, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid compositions of the same type can also be used as fillers in soft and hard gelatin capsules, using excipients such as lactose or milk sugar, high molecular weight polyethylene glycols, etc.

The active substances can also be in microencapsulated form with one or more of the excipients listed above. Solid dosage forms such as tablets, dragees, capsules, pills and granules can also be formulated with coatings and coatings such as enteric coatings, sustained release coatings and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active ingredient may be mixed with at least one inert diluent such as sucrose, lactose or starch. Such solid dosage forms may also contain, as is customary in pharmaceutical practice, additional substances other than inert diluents, for example, tableting lubricants and other tabletting auxiliaries such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, dosage forms may also contain buffering agents. They may optionally contain opacifiers and may also be formulated to release the active substance(s) exclusively or preferentially in a specific part of the gastrointestinal tract, optionally in a delayed manner. Examples of bases for such compositions include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active substance is mixed under sterile conditions with a pharmaceutically acceptable carrier and, if necessary, with suitable preservatives or buffer additives. Ophthalmic preparations, eye drops, ophthalmic ointments, powders and solutions are also included within the scope of the present invention.

Ointments, pastes, creams and gels may contain, in addition to the compounds of the present invention, excipients such as animal and vegetable oils and fats, waxes, paraffins, starch, gum tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

Powders and sprays may contain, in addition to the compound of the present invention, excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays may additionally contain commonly known propellants such as chlorofluorocarbons.

Transdermal patches have the added benefit of... they allow controlled administration of compounds into the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption accelerators can also be used to enhance the delivery of the compound through the skin. The rate of entry into the body can be controlled by selecting a rate-determining membrane or by dispersing the compound in a polymer matrix or gel.

Unless otherwise specified, all technical and scientific terms used in this text have the meanings commonly known to those skilled in the art. All publications, patents, published patent applications and other references mentioned in this text are incorporated herein in their entirety by reference.

Abbreviations

In this text, in the description, diagrams and Examples, the following abbreviations and abbreviations are used:

BOP-Cl = bis(2-oxo-3-oxazolidinyl)phosphine chloride;

CDI = carbonyldiimidazole;

DBU = 1,8-diazabicycloundec-7-ene;

DCC = N,N'-dicyclohexylcarbodiimide;

DCM = dichloromethane;

DIPEA = N,N-diisopropylethylamine;

DMAP = N,N-dimethylaminopyridine;

DME = 1,2-dimethoxyethane;

DMF = N,N-dimethylformamide;

DMPU = 1,3-Dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone;

EDC = 1-(3-diethylaminopropyl)-3-ethylcarbodiimide hydrochloride;

Et ₃ N = triethylamine;

EtOAc = ethyl acetate;

HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate;

HCl = hydrochloric acid;

mCPBA = meta-chloroperoxybenzoic acid;

NMO = N-methylmorpholine-N-oxide;

PE = petroleum ether

PhMe = toluene;

PyAOP = 7-azabenzotriazol-1-yloxy)tripyrrolidinophosphonium hexafluorophosphate;

PyBOP = benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate;

THF = tetrahydrofuran;

TEA = triethylamine;

RT = room temperature.

Synthesis methods

The compounds and methods of the present invention will be better understood in conjunction with the following synthetic schemes, which illustrate the methods by which the compounds of the present invention can be prepared. They are provided for illustrative purposes only and do not limit the scope of the present invention. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art, and such changes and modifications, including but not limited to those related to the chemical structures, substituents, derivatives and/or methods of the present invention, may be made without departure beyond the scope of the present invention and the scope of the claims.

As shown in Scheme 1, an ester compound having the formula (S-4), where R ⁴ , X and have the above meanings, can be prepared by a Suzuki-Miyaura coupling reaction or from a compound having formula (S-1) and the corresponding boronic acids and their esters; or from a compound having formula (S-2) and a corresponding halide. Alternatively, the compound having formula (S-3), which can be prepared by a Suzuki-Miyaura coupling reaction, can be converted into an ester compound having formula (S-4) by a Pd-catalyzed CO-insertion reaction into alcohols. Hydrolysis of the compound having formula (S-4) under basic conditions gives a carboxylic acid, which is the compound having formula (S-5).

Scheme 1

As shown in Scheme 2, the reaction of a carboxylic acid compound having formula (S-5) and an amine compound having formula (S-6) in an aprotic solvent with a suitable capping agent in the presence of an organic base gives the amide compound, having formula (I), where , R ⁴ , X, And have the meanings indicated above. A suitable capping agent may be, but is not limited to, BOP-Cl, CDI, DCC, EDC, HATU, PyAOP or PyBOP. The organic base may be, but is not limited to, Et ₃ N, DIPEA, pyridine or N-methylmorpholine. The aprotic solvent may include, but is not limited to, THF, DCM, and DMF. The reaction temperature ranges from -20°C to 80°C.

Scheme 2

As shown in Scheme 3, a carboxylic acid compound having formula (S-5) can first be converted to an acyl chloride having formula (S-7) using reagents such as, but not limited to, oxalyl chloride or Gosez's reagent . Reaction of acyl chloride (S-7) with an amine compound having formula (S-6) in the presence of an organic base such as pyridine gives a compound having formula (I), where , R ⁴ , X, And have the meanings indicated above.

Scheme 3.

As shown in Scheme 4, an amide compound having formula (I), where , R ⁴ , X, And have the above meanings, can also be prepared from an ester compound having formula (S-4) and an amine compound having formula (S-6) under ester-amide exchange conditions. These conditions include, but are not limited to: Cs ₂ CO ₃ or K ₂ CO ₃ in DMF at elevated temperature; or AlMe ₃ in DCM at room or elevated temperature.

Scheme 4

Examples

Example 1

Stage 1-1

Triethylamine (242.1 mg, 2.39 mmol) and _Tf2O (506.3 mg, 1.79 mmol) were added dropwise to a solution of ethyl 4-hydroxypyridine-2-carboxylate (200 mg, 1.20 mmol) in DCM (5 ml) at 0°C. The mixture was warmed to room temperature, stirred for 1 hour and evaporated in vacuum. Purification of the residue by silica gel column chromatography using 0 - 20% EtOAc in petroleum ether as the eluent gave compound 1-a (170 mg, 47.49%) as a yellow oil.

Stage 1-2

A mixture of compound 1-a (150 mg, 0.50 mmol), 1-cyclopropyl-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (234.7 mg, 1.00 mmol), Pd(PPh ₃ ) ₄ (57.9 mg, 0.05 mmol) and K ₂ CO ₃ (138.6 mg, 1.00 mmol) in dioxane (5 ml) were stirred at 100°C in a nitrogen atmosphere for 1 hour. The solvent was removed in vacuo. Purification of the residue by silica gel chromatography using EtOAc in petroleum ether (0-50%) as eluent gave compound 1-b (60 mg, 46.52%) as a yellow oil.

Stage 1-3

A mixture of compound 1-b (60 mg, 0.23 mmol), compound 1-c (71.4 mg, 0.35 mmol) and Cs ₂ CO ₃ (152.0 mg, 0.47 mmol) in DMF was stirred at 120°C for 2 hours. The solvent was removed in vacuo. Purification of the residue on a C-18 column using acetonitrile in H ₂ O (60-70%) as eluent gave Example 1 (7.8 mg) as a white solid. [M+H] ⁺ , 416. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.61 (s, 1H), 8.65 - 8.61 (m, 2H), 8.40 (s, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.04 - 8.01 (m, 3H), 7.62 (dd, J = 5.3, 1.6 Hz, 1H), 5.89 (m, 1H), 3.72 (m, 1H), 1.76 (d, J = 6.6 Hz, 6H), 1.23 (m, 2H), 1.14 (m, 2H).

Example 2

Stage 2-1

At 0°C, Tf ₂ O (1.86 g, 6.60 mmol) was added dropwise to a solution of ethyl 4-hydroxy-5-methylpyridine-2-carboxylate (800 mg, 4.42 mmol) and Et ₃ N (1.33 g, 13.20 mmol) in DXM (5 ml). The resulting mixture was stirred at room temperature for 1 hour and evaporated in vacuum. Purification of the residue by silica gel column chromatography using EtOAc in petroleum ether (0 - 24%) as eluent gave compound 2-a (550 mg, 39.8%) as a yellow oil.

Stage 2-2

A mixture of compound 2-a (550 mg, 1.76 mmol), 1-cyclopropyl-4-(tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (822.1 mg, 3.51 mmol), Pd(PPh ₃ ) ₄ (202.9 mg, 0.18 mmol) and K ₂ CO ₃ (485.3 mg, 3.51 mmol) in dioxane (10 ml) were stirred at 100°C for 2 hours in a nitrogen atmosphere. The solvent was removed in vacuo. The residue was purified by silica gel column chromatography using EtOAc in petroleum ether (0 - 35%) as eluent to give 2-b (400 mg, 83.97%) as a yellow oil.

Stage 2-3

A mixture of compound 2-b (80 mg, 0.29 mmol), compound 1-c (90.3 mg, 0.44 mmol) and Cs ₂ CO ₃ (192.1 mg, 0.59 mmol) in DMF (2 ml) was stirred at 100°C for 1 hours. The solvent was removed in vacuo. The resulting crude product was purified on a C18 column using acetonitrile in H ₂ O (50-70%) as eluent to give Example 2 (15.3 mg) as a white solid. [M+H] ⁺ , 430; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.54 (s, 1H), 8.61 (d, J = 8.3 Hz, 1H), 8.52 (s, 1H), 8.31 (s, 1H), 8.09 (d, J = 8.3 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.87 (s, 2H), 5.88 (m, 1H), 3.72 (m, 1H), 2.57 (s, 3H), 1.75 ( d, J = 6.7 Hz, 6H), 1.24 (m, 2H), 1.16 (m, 2H).

Example 3

Stage 3-1

In a mixture of 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (2.0 g, 8.55 mmol), 2-chloro-5-fluoro -4-iodopyridine (2 g, 7.77 mmol), and K ₂ CO ₃ (3.2 g, 23.31 mmol) in dioxane (10 ml) and H ₂ O (2 ml) were added Pd(dppf)Cl ₂ (568.5 mg, 0.78 mmol) at room temperature in a nitrogen atmosphere. The resulting mixture was stirred at 80°C for 3 hours under a nitrogen atmosphere. After cooling to room temperature, the mixture was diluted with ethyl acetate (20 ml) and washed with water (10 ml *2) and saturated sodium chloride solution (10 ml). The organic layer was dried over anhydrous Na ₂ SO ₄ , filtered and evaporated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/EtOAc (5:1) as the eluent to give 3-a (1.3 g, 70.41%) as a white solid.

Stage 3-2

In a 50 mL autoclave, a solution of compound 3-a (1.2 g, 5.0 mmol), Pd(dppf)Cl ₂ (385 mg, 0.5 mmol) and Et ₃ N (1.52 g, 15 mmol) in BuOH (20 mL) was saturated with CO and stirred under a pressure of 10 atm CO at 70 ° ^C for 16 hours. The resulting mixture was evaporated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/EtOAc (3:1) as the eluent to give 3-b (1.2 g, 78.35%) as a yellow oil.

Stage 3-3

To a solution of compound 3-b (600 mg, 1.98 mmol) in acetonitrile (5 ml) was added 1N. HCl solution (5 ml), and the resulting mixture was stirred at 100°C for 14 hours under a nitrogen atmosphere. The solvent was removed under reduced pressure. The residue was purified by reverse phase flash chromatography with 0–30% MeCN in water to give compound 3-c (420 mg, 85.79%) as a white solid.

Stage 3-4

Compound 1-c (227.2 mg, 1.11 mmol) was added to a solution of compound 3-c (250 mg, 1.01 mmol), HATU (499.8 mg, 1.31 mmol) and DIPEA (392.1 mg, 3.03 mmol) in DMF (5 ml). The resulting mixture was stirred at room temperature for 4 hours under a nitrogen atmosphere. The residue was purified by reverse phase flash chromatography with 40 - 70% MeCN in water to give Example 3 (150 mg, 34.22%) as an off-white solid. [M+H] ⁺ , 434; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.39 (s, 1H), 8.60 (dd, J = 8.4, 1.0 Hz, 1H), 8.53 – 8.51 (m, 2H), 8.12 – 8.09 (m, 3H ), 8.02 (t, J = 8.0 Hz, 1H), 5.85 (m, 1H), 3.73 (m, 1H), 1.76 (d, J = 6.7 Hz, 6H), 1.26 (m, 2H), 1.12 (m , 2H).

Example 4

Stage 4-1

DBU (0.2 ml) was added to a solution of Example 3 (30 mg, 0.07 mmol) in pyrrolidine (1 ml). The resulting mixture was stirred at 80°C for 4 hours under a nitrogen atmosphere. The reaction mixture was purified by reverse phase flash chromatography with 40-70% MeCN in water to give Example 4 (17.9 mg, 53.37%) as a white solid. [M+H] ⁺ , 434; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.39 (s, 1H), 8.60 (dd, J = 8.3, 1.0 Hz, 1H), 8.20 (s, 1H), 8.04 – 8.06 (m, 2H), 7.97 (t, J = 8.0 Hz, 1H), 7.68 (m, 2H), 5.90 (m, 1H), 3.69 (m, 1H), 3.20 (m, 4H), 1.95 (m, 4H), 1.75 (d , J = 6.7 Hz, 6H), 1.26 (m, 2H), 1.12 (m, 2H).

Example 5

Example 5 was prepared using a procedure similar to that described in Example 4. [M+H] ⁺ , 499; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.47 (s, 1H), 8.61 (d, J = 8.3 Hz, 1H), 8.38 (s, 1H), 8.25 (s, 1H), 8.20 (s, 1H), 8.16 (s, 1H), 8.08 (d, J = 7.4 Hz, 1H), 7.99 (t, J = 8.0 Hz, 1H), 5.88 (m, 1H), 3.71 (m, 1H), 3.03 ( m, 4H), 1.76 – 1.70 (m, 10H), 1.70 (m, 2H), 1.26 (m, 2H), 1.14 (m, 2H).

Example 6

Example 6 was prepared using a procedure similar to that described in Example 4. [M+H] ⁺ , 501; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.45 (s, 1H), 8.60 (dd, J = 8.3, 1.0 Hz, 1H), 8.39 (s, 1H), 8.25 (s, 1H), 8.18 – 8.15 (m, 2H), 8.09 (dd, J = 7.7, 1.0 Hz, 1H), 8.00 (t, J = 8.3 Hz, 1H), 5.87 (m, 1H), 3.88 (t, J = 4.4 Hz, 4H ), 3.70 (m, 1H), 3.11 (t, J = 4.4 Hz, 4H), 1.75 (d, J = 6.7 Hz, 6H), 1.70 (m, 2H), 1.26 (m, 2H), 1.14 (m , 2H).

Example 7

Example 7 was prepared using a procedure similar to that described in Example 4. [M+H] ⁺ , 501; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.45 (s, 1H), 8.59 (dd, J = 8.3, 1.0 Hz, 1H), 8.40 (s, 1H), 8.24 (s, 1H), 8.12 ( s, 1H), 8.09 (dd, J =7.6, 0.8 Hz, 1H), 7.99 (t, J = 8.0 Hz, 1H), 5.88 (m, 1H), 3.71 (m, 1H), 3.19 (t, J = 4.4 Hz, 4H), 2.71 (t, J = 4.4 Hz, 4H), 2.47 (s, 3H), 1.75 (d, J = 6.7 Hz, 6H), 1.26 (m, 2H), 1.14 (m, 2H ).

Example 8

Stage 8-1

To a solution of compound 3-c (150 mg, 0.61 mmol), HATU (348 mg, 0.91 mmol) and DIPEA (239 mg, 1.87 mmol) in DMF (5 ml) was added compound 8-a (141 mg, 0.67 mmol) at room temperature in a nitrogen atmosphere. The reaction mixture was stirred at room temperature for 2 hours and purified by reverse phase flash chromatography with 50–80% MeCN in water to give Example 8 (130 mg, 48.8%) as a white solid. [M+H] ⁺ 440; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.58 (s, 1H), 8.51 – 8.50 (m, 2H), 8.17 (s, 1H), 8.11 – 8.08 (m, 2H), 5.88 (m, 1H ), 3.73 (m, 1H), 1.74 (d, J = 6.7 Hz, 6H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 9

Example 9 was prepared using a procedure similar to that described for Step 4-1. [M+H] ⁺ 491; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.58 (s, 1H), 8.18 (s, 1H), 8.13 (s, 1H), 8.05 (s, 1H), 7.69 – 7.67 (m, 2H), 5.91 (m, 1H), 3.69 (m, 1H), 3.20 (t, J = 6.4 Hz, 4H), 1.95 (t, J = 6.4 Hz, 4H), 1.74 (d, J = 6.7 Hz, 6H), 1.25(m, 2H), 1.12(m, 2H).

Example 10

Example 10 was prepared using a procedure similar to that described for Step 4-1. [M+H] ⁺ 507; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.62 (s, 1H), 8.37 (s, 1H), 8.24 (s, 1H), 8.18 (s, 1H), 8.17 – 8.15 (m, 2H), 5.88 (m, 1H), 3.88 (t, J = 4.4 Hz, 4H), 3.70 (m, 1H), 3.11 (t, J = 4.4 Hz, 4H), 1.74 (d, J = 6.6 Hz, 6H), 1.22 (m, 2H), 1.15 (m, 2H).

Example 11

Example 11 was prepared using a procedure similar to that described for Step 4-1. [M+H] ⁺ 505; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.63 (s, 1H), 8.36 (s, 1H), 8.24 (s, 1H), 8.22 – 8.12 (m, 3H), 5.89 (m, 1H), 3.71 (m, 1H), 3.02 (t, J = 5.2 Hz, 4H), 1.74 – 1.63 (m, 12H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 12

Example 12 was prepared using a procedure similar to that described for Step 4-1. [M+H] ⁺ 520; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.62 (s, 1H), 8.39 (s, 1H), 8.22 (s, 1H), 8.18 (s, 1H), 8.15 (s, 1H), 8.10 ( s, 1H), 5.88 (m, 1H), 3.71 (m, 1H), 3.22 (t, J = 4.9 Hz, 4H), 2.79 (s, 4H), 2.51 (s, 3H), 1.74 (d, J = 6.7 Hz, 6H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 13

Stage 13-1

Mixture of methyl 4-chloro-5-methylpyridin-2-carboxylate (200 mg, 1.08 mmol), 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-pyrazole (302.7 mg, 1.29 mmol), Pd(dppf)Cl ₂ -CH ₂ Cl ₂ (88.0 mg, 0.11 mmol) and K ₂ CO ₃ (446.8 mg, 3.23 mmol) in dioxane (4 ml) and water (1 ml) was stirred at 80°C for 2 hours under nitrogen atmosphere. After cooling to room temperature, the residue was purified by reverse phase flash chromatography with 40-70% MeCN in H ₂ O to give 13-a (60 mg, 21.64%) as a yellow solid.

Stage 13-2

NaOH (18.7 mg, 0.47 mmol) was added to a stirred mixture of compound 13-a (60 mg, 0.23 mmol) in MeOH (2 ml) and H ₂ O (1 ml) at 0°C. The resulting mixture was stirred for 1 hour at 50°C, cooled to room temperature, acidified to pH 3 with an aqueous solution of HCl and evaporated in vacuum. The residue was purified by reverse phase flash chromatography with 10-50% MeCN/H ₂ O to give compound 13-b (40 mg, 70.51%) as a white solid.

Stage 13-3

Compound 13-c (52 mg, 0.20 mmol) was added to a solution of compound 13-b (40 mg, 0.16 mmol), HATU (91 mg, 0.24 mmol) and DIPEA (62 mg, 0.48 mmol) in DMF (2 ml). The reaction mixture was stirred at room temperature for 16 hours, and then purified by reverse phase flash chromatography with 10-80% MeCN/H ₂ O, obtaining compound 13-d (30 mg, 37.4%) as a white solid.

Stage 13-4

K2CO3 (42 mg, 0.3 mmol) was added to a solution of compound 13-d (30 mg, 0.06 mmol) in _MeOH ( ₂ ml). The reaction mixture was stirred at room temperature for 2 hours. The solid precipitate was separated by filtration. The filtrate was purified by reverse phase flash chromatography with 10-60% MeCN/H ₂ O to give Example 13 (30 mg) as a white solid. [M+H] ⁺ 446; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.56 (s, 1H), 8.51 (s, 1H), 8.40 (dd, J = 8.3, 1.0 Hz, 1H), 8.30 (s, 1H), 8.12 ( dd, J = 8.3, 1.0 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.87 (s, 2H), 5.91 (m, 1H), 4.19 (d, J = 5.6 Hz, 2H), 3.72 (m, 1H), 3.27 (s, 1H), 2.57 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 14

Stage 14-1

A mixture of 2-chloro-4-iodo-5-(trifluoromethyl)pyridine (1 g, 3.25 mmol), 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- yl)-1H-pyrazole (990 mg, 4.23 mmol), Pd(dppf)Cl ₂ (375 mg, 0.3 mmol) and potassium carbonate (1.35 g, 9.75 mmol) in ethylene glycol dimethyl ether (5 ml) and H ₂ O ( 5 ml) was stirred at 100°C for 3 hours. The reaction mixture was evaporated and purified by preparative HPLC using 0-100% MeCN/ _H2O as eluent to give 14-a (750 mg, 80%) as a yellow solid.

Stage 14-2

In a 50 mL autoclave, a solution of compound 14-a (750 mg, 2.61 mmol), Pd(dppf)Cl ₂ (425 mg, 0.52 mmol) and TEA (957 mg, 7.83 mmol) in BuOH (10 mL) was stirred under a pressure of 10 atm CO at 70°C overnight. The reaction mixture was evaporated in vacuum. Purification of the residue by preparative HPLC using 0-100% MeCN/H ₂ O as eluent gave 14-b (660 mg, 72%) as a yellow solid.

Stage 14-3

In a 50 ml autoclave, 1N was added. an aqueous solution of HCl (5 ml) to a solution of compound 14-b (660 mg, 1.87 mmol) in MeCN (10 ml). The resulting solution was stirred at 100°C overnight. The reaction mixture was evaporated and the residue was purified by preparative HPLC using 0-40% MeCN/H ₂ O as eluent to give compound 14-c (420 mg, 76%) as a white solid.

Stage 14-4

To a solution of compound 14-c (30 mg, 0.10 mmol), HATU (58 mg, 0.15 mmol) and 4-methylmorpholine (33 mg, 0.30 mmol) in DMF (2 ml) was added compound 1-c (24.8 mg, 0.12 mmol ). The resulting mixture was stirred at room temperature overnight and purified by preparative HPLC using 0-80% MeCN/H ₂ O as eluent to give Example 14 (16.5 mg) as a white solid. [M+H] ⁺ 484; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.48 (s, 1H), 9.01 (s, 1H), 8.60 (d, J = 8.4, 1.0 Hz, 1H), 8.43 (s, 1H), 8.15 ( dd, J = 8.4, 1.0 Hz, 1H), 8.04 (t, J = 8.4 Hz, 1H), 7.89 (d, J = 13.1 Hz, 2H), 5.86 (m, 1H), 3.73 (m, 1H), 1.76 (d, J = 6.7 Hz, 6H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 15

Stage 15-1

Example 15 was prepared using a procedure similar to that described for Step 14-4. [M+H] ⁺ 490; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.69 (s, 1H), 8.98 (s, 1H), 8.42 (s, 1H), 8.20 (s, 1H), 7.89 (d, J = 14.2 Hz, 2H), 5.87 (m, 1H), 3.73 (m, 1H), 1.75 (d, J = 6.7 Hz, 6H), 1.25 (m, 2H), 1.16 (m, 2H).

Example 16

Example 16 was prepared from compound 14-c using a procedure similar to that described for Step 13-3 and Step 13-4. [M+H] ⁺ 500; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.51 (s, 1H), 9.00 (s, 1H), 8.44 (m, 2H), 8.17 (dd, J = 8.0, 1.0 Hz, 1H), 8.05 ( t, J = 8.0 Hz, 1H), 7.90 (d, J = 14.3 Hz, 2H), 5.88 (m, 1H), 4.20 (m, 2H), 3.73 (m, 1H), 1.74 (d, J = 6.8 Hz, 3H), 1.23 (m, 2H), 1.14 (m, 2H).

Example 17

Example 17 was prepared from compound 13-b and compound 17-a using a procedure similar to that described for Step 14-4. [M+H] ⁺ 430; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.60 (d, J = 8.3 Hz, 1H), 8.52 (s, 1H), 8.30 (s, 1H), 8.06 (t, J = 8.1 Hz, 1H), 7.87 (s, 2H), 7.68 (d, J = 7.8 Hz, 1H), 3.98 (m, 1H), 3.72 (m, 1H), 2.56 (s, 3H), 1.53 ( d, J = 6.9 Hz, 6H), 1.25 (m, 2H), 1.15 (m, 2H).

Example 18

Stage 18-1

Mixture of compound 18-a (US 2017/0121308) (100 mg, 0.45 mmol), 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1H-pyrazole (211 mg, 0.90 mmol), Pd(PPh ₃ ) ₄ (104 mg, 0.09 mmol) and K ₂ CO ₃ (187 mg, 1.35 mmol) in dioxane (5 ml) were stirred at 100°C for overnight in a nitrogen atmosphere. The solvent was evaporated in vacuo and the residue was purified by 0-65% MeCN/H ₂ O reverse phase column chromatography to give compound 18-b (95 mg, 71.8%) as a yellow solid.

Stage 18-2

Compound 18-b (30 mg, 0.10 mmol), compound 1-c (43.0 mg, 0.20 mmol), Cs ₂ CO ₃ (100 mg, 0.31 mmol) and DMF (5 ml) were placed in an 8 ml hermetically sealed tube. The resulting mixture was stirred at 100°C for 3 hours under nitrogen, and purified by reverse phase flash chromatography with 0-65% MeCN/H ₂ O, obtaining Example 18 (8 mg) as an off-white solid. [M+H] ⁺ 466; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.54 (s, 1H), 9.03 (s, 1H), 8.61 (dd, J = 8.4, 0.9 Hz, 1H), 8.39 (s, 1H), 8.14 ( dd, J = 8.4, 0.9 Hz, 1H), 8.03 (t, J = 8.4 Hz, 1H), 7.91 – 7.82 (m, 2H), 6.84 (t, J = 54.0 Hz, 1H), 5.87 (m, 1H ), 3.74 (m, 1H), 1.77 (d, J = 6.7 Hz, 6H), 1.26 (m, 2H), 1.19 (m, 2H).

Example 19

Example 19 was prepared from compound 18-b and compound 8-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 472; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.73 (s, 1H), 9.00 (s, 1H), 8.38 (s, 1H), 8.20 (s, 1H), 7.86 (d, J = 17.1 Hz, 2H), 6.84 (t, J = 54.0 Hz, 1H), 5.88 (m, 1H), 3.74 (m, 1H), 1.75 (d, J = 6.7 Hz, 6H), 1.26 (m, 2H), 1.17 ( m, 2H).

Example 20

Stage 20-1

A mixture of methyl 4-chloro-5-methylpyridine-2-carboxylate (300 mg, 1.62 mmol), 1-tert-butyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- yl)-1H-pyrazole (606.5 mg, 2.42 mmol), Pd(PPh ₃ ) ₄ (373.5 mg, 0.32 mmol), K ₂ CO ₃ (670.1 mg, 4.85 mmol) in dioxane (5 ml) was stirred at 100°C in a nitrogen atmosphere overnight. The mixture was cooled to room temperature and purified by reverse phase flash chromatography with 0-40% MeCN/H ₂ O to give compound 20-a (140 mg, 31.69%) as a yellow oil.

Stage 20-2

Example 20 was prepared from compound 20-a and compound 1-c using a procedure similar to that described for Step-18-2. [M+H] ⁺ 446; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.57 (s, 1H), 8.62 (d, J = 8.2 Hz, 1H), 8.53 (s, 1H), 8.34 (s, 1H), 8.10 (d, J = 7.5 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.94 (s, 2H), 5.89 (m, 1H), 2.59 (s, 3H), 1.76 (d, J = 6.6 Hz, 6H), 1.70 (s, 9H).

Example 21

Example 21 was prepared from compounds 20-a and 17-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 446; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.58 (s, 1H), 8.61 (d, J = 8.2 Hz, 1H), 8.53 (s, 1H), 8.33 (s, 1H), 8.07 (t, J = 8.0 Hz, 1H), 7.97 (s, 1H), 7.94 (s, 1H), 7.69 (d, J = 7.7 Hz, 1H), 4.00 (m, 1H), 2.59 (s, 3H), 1.71 ( s, 9 H), 1.54 (d, J = 6.9 Hz, 6H).

Example 22

Example 22 was prepared from compound 20-a and compound 22-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 462; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.60 (s, 1H), 8.51 (s, 1H), 8.41 (d, J = 8.3 Hz, 1H), 8.33 (s, 1H), 8.13 (d, J = 7.6 Hz, 1H), 8.02 (t, J = 8.0 Hz, 1H), 7.93 (s, 1H), 7.92 (s, 1H), 5.90 (m, 1H), 4.19 (d, J = 6.0 Hz, 2H), 2.59 (s, 3H), 1.74 (d, J = 7.6 Hz, 3H), 1.69 (s, 9H).

Example 23

Stage 23-1

Mixture of methyl 5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-carboxylate (300 mg, 1.08 mmol), 4-bromo-1- (propan-2-yl)-1H-imidazole (307.0 mg, 1.62 mmol), Pd(PPh ₃ ) ₄ (250.2 mg, 0.22 mmol), K ₂ CO ₃ (448.8 mg, 3.25 mmol) in dioxane (5 ml) stirred at 100°C under nitrogen atmosphere overnight. The resulting mixture was evaporated in vacuo and purified by reverse phase flash chromatography with 0-37% MeCN/H ₂ O to give compound 23-a (180 mg, 64.12%) as a yellow oil.

Stage 23-2

Example 23 was prepared from compound 23-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 432; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.57 (s, 1H), 8.68 (s, 1H), 8.63 (d, J = 8.4 Hz, 1H), 8.51 (s, 1H), 8.08 (d, J = 7.5 Hz, 1H), 7.99 (t, J = 7.9 Hz, 1H), 7.71 (s, 1H), 7.44 (s, 1H), 5.90 (m, 1H), 4.46 (m, 1H), 2.68 ( s, 3H), 1.75 (d, J = 6.6 Hz, 6H), 1.60 (d, J = 6.7 Hz, 6H).

Example 24

Example 24 was prepared from compound 23-a and compound 17-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 432; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.58 (s, 1H), 8.69 (s, 1H), 8.62 (d, J = 8.2 Hz, 1H), 8.51 (s, 1H), 8.05 (t, J = 8.1 Hz, 1H), 7.74 (s, 1H), 7.67 (d, J = 7.6 Hz, 1H), 7.44 (d, J = 1.3 Hz, 1H), 4.47 (m, 1H), 4.00 (m, 1H), 2.68 (s, 3H), 1.61 (d, J = 6.7 Hz, 6H), 1.53 (d, J = 6.9 Hz, 6H).

Example 25

Example 25 was prepared from compound 23-a and compound 22-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 448; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.61 (s, 1H), 8.68 (s, 1H), 8.51 (s, 1H), 8.38 (d, J = 8.3 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.76 (s, 1H), 7.45 (s, 1H), 5.94 (m, 1H), 4.48 (m, 1H), 4.17 ( m, 2H), 2.68 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H), 1.62 (d, J = 6.7 Hz, 6H).

Example 26

Example 26 was prepared using a procedure similar to that described in Example 20. [M+H] ⁺ 432; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.62 (d, J = 8.3 Hz, 1H), 8.53 (s, 1H), 8.33 (s, 1H), 8.10 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.91 (s, 1H), 7.85 (s, 1H), 5.88 (m, 1H), 4.62 (m, 1H), 2.58 ( s, 3H), 1.76 (d, J = 6.6 Hz, 6H), 1.62 (d, J = 6.7 Hz, 6H).

Example 27

Example 27 was prepared using a procedure similar to that described in Example 21. [M+H] ⁺ 432; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.63 (s, 1H), 8.60 (d, J = 8.3 Hz, 1H), 8.52 (s, 1H), 8.34 (s, 1H), 8.07 (t, J = 8.1 Hz, 1H), 7.92 (s, 1H), 7.86 (s, 1H), 7.70 (d, J = 7.8 Hz, 1H), 4.64 (m, 1H), 4.02 (m, 1H), 2.59 ( s, 3H), 1.63 (d, J = 6.7 Hz, 6H), 1.54 (d, J = 6.9 Hz, 6H).

Example 28

Example 28 was prepared using a procedure similar to that described in Example 22. [M+H] ⁺ 448; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.69 (s, 1H), 8.52 (s, 1H), 8.41 (d, J = 8.0 Hz, 1H), 8.33 (s, 1H), 8.15 (d, J = 7.1 Hz, 1H), 8.02 (t, J = 8.0 Hz, 1H), 7.93 (s, 1H), 7.87 (s, 1H), 5.94 (m, 1H), 4.63 (m, 1H), 4.19 ( m, 2H), 2.60 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H), 1.63 (d, J = 6.7 Hz, 6H).

Example 29

Stage 29-1

To a solution of compound 22-a (60.1 mg, 0.27 mmol) in DCM (5 ml) at 0°C was added _Me3Al in toluene (0.4 ml, 0.568 mmol) under a nitrogen atmosphere. The mixture was stirred for 1 hour and a solution of compound 18-b (80 mg, 0.27 mmol) in DCM (1 ml) was added. The resulting mixture was stirred at 35°C for 16 hours, diluted with DCM, and washed with aqueous Rochelle salt and saturated sodium chloride solution. The organic layer was dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure. The resulting crude product was purified by preparative HPLC using 35-60% MeCN/H ₂ O as eluent to give Example 29 (11.8 mg) as a white solid. [M+H] ⁺ 482; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.57 (s, 1H), 9.02 (s, 1H), 8.46 – 8.37 (m, 2H), 8.17 (d, J = 7.6 Hz, 1H), 8.05 ( t, J = 8.1 Hz, 1H), 7.90 (s, 1H), 7.85 (s, 1H), 6.84 (t, J = 53.2 Hz, 1H), 5.90 (m, 1H), 4.27 – 4.17 (m, 2H), 3.75 (m, 1H), 1.75 (d, J = 6.8 Hz, 3H), 1.27 (m, 2H), 1.17 (m, 2H).

Example 30

Stage 30-1

Mixture of methyl 5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)picolinate (300 mg, 1.08 mmol), 4-bromo-1-(trifluoromethyl)- 1H-pyrazole (256 mg, 1.19 mmol), Pd(PPh ₃ ) ₄ (104 mg, 0.1 mmol) and K ₂ CO ₃ (447 mg, 3.24 mmol) in dioxane (10 ml) were stirred at 100°C in a nitrogen atmosphere 1 hour. The solvent was removed under reduced pressure. The residue was purified by reverse phase flash chromatography with 0-60% CH ₃ CN in water to give compound 30-a (175 mg, 56.7%) as a white solid.

Stage 30-2

Example 30 was prepared from compound 30-a and compound 1-c using a procedure similar to that described for Step 29-1. [M+1] ⁺ 458; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.50 (s, 1H), 8.62 – 8.60 (m, 2H), 8.33 (s, 1H), 8.18 (s, 1H), 8.15 – 8.06 (m, 2H ), 8.01 (t, J = 8.0 Hz, 1H), 5.87 (m, 1H), 2.58 (s, 3H), 1.76 (d, J = 6.7 Hz, 6H).

Example 31

Example 31 was prepared from compound 30-a and compound 17-a using a procedure similar to that described for Step 29-1. [M+1] ⁺ 458; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.51 (s, 1H), 8.61 – 8.59 (m, 2H), 8.32 (s, 1H), 8.17 (s, 1H), 8.13 (s, 1H), 8.08 (t, J = 8.1 Hz, 1H), 7.70 (d, J = 7.8 Hz, 1H), 3.97 (m, 1H), 2.58 (s, 3H), 1.53 (d, J = 6.9 Hz, 7H).

Example 32

Example 32 was prepared from compound 30-a and compound 22-a using a procedure similar to that described for Step 29-1. [M+1] ⁺ 474; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.53 (s, 1H), 8.61 (s, 1H), 8.42 (d, J = 8.3 Hz, 1H), 8.33 (s, 1H), 8.18 (s, 1H), 8.16 – 8.13 (m, 2H), 8.02 (t, J = 8.0 Hz, 1H), 5.90 (m, 1H), 4.21 (m, 2H), 2.59 (s, 3H), 1.74 (d, J = 6.8 Hz, 3H).

Example 33

Stage 33-1

A mixture of methyl 5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-carboxylate (250 mg, 0.90 mmol), 3-bromo-1- (propan-2-yl)-1H-pyrazole (341.1 mg, 1.80 mmol), Pd(PPh ₃ ) ₄ (208.5 mg, 0.18 mmol) and K ₂ CO ₃ (374.0 mg, 2.71 mmol) in dioxane (5 ml) stirred at 100°C under nitrogen atmosphere overnight. The reaction mixture was evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography with 0-40% MeCN/H ₂ O to give compound 33-a (200 mg, 85.50%) as a yellow oil.

Stage 33-2

Example 33 was prepared from compound 33-a and compound 1-c using a procedure similar to that described for Step 29-1. [M+H] ⁺ 432. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.60 (s, 1H), 8.67 – 8.59 (m, 2H), 8.56 (s, 1H), 8.10 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.56 (d, J = 2.3 Hz, 1H), 6.70 (d, J = 2.4 Hz, 1H), 5.92 (m, J = 6.7 Hz , 1H), 4.61 (m, J = 6.7 Hz, 1H), 2.70 (s, 3H), 1.76 (d, J = 6.6 Hz, 6H), 1.61 (d, J = 6.6 Hz, 6H).

Example 34

Example 34 was prepared from compound 33-a and compound 17-a using a procedure similar to that described for Step 29-1. [M+H] ⁺ 432. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.58 (s, 1H), 8.65 – 8.57 (m, 2H), 8.54 (s, 1H), 8.05 (t, J = 8.0 Hz, 1H), 7.68 (d, J = 7.8 Hz, 1H), 7.56 (d, J = 2.3 Hz, 1H), 6.69 (d, J = 2.3 Hz, 1H), 4.60 (m, J = 6.8 Hz , 1H), 4.00 (m, J = 6.9 Hz, 1H), 2.69 (s, 3H), 1.61 (d, J = 6.7 Hz, 6H), 1.54 (d, J = 6.9 Hz, 6H).

Example 35

Example 35 was prepared from compound 33-a and compound 22-a using a procedure similar to that described for Step 29-1. [M+H] ⁺ 448. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.63 (s, 1H), 8.59 (s, 1H), 8.53 (s, 1H), 8.42 (d, J = 8.3 Hz , 1H), 8.11 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.56 (d, J = 2.3 Hz, 1H), 6.70 (d, J = 2.3 Hz, 1H ), 5.94 (m, J = 6.5 Hz, 1H), 4.60 (m, J = 6.7 Hz, 1H), 4.19 (d, J = 6.0 Hz, 2H), 3.30 (s, 1H), 2.70 (s, 3H ), 1.74 (d, J = 6.8 Hz, 3H), 1.61 (d, J = 6.7 Hz, 6H)

Example 36

Stage 36-1

In a reaction test tube, a mixture of methyl 4-chloro-5-methylpyridine-2-carboxylate (100 mg, 0.54 mmol), 2-(tributylstannyl)pyridine (190 mg, 0.52 mmol) and Pd(PPh ₃ ) ₄ was heated in a microwave oven. (49 mg, 0.04 mmol) in DMF (2 ml) under microwave conditions at 160°C for 1 hour. The solvent was removed in vacuo and the residue was purified by preparative HPLC using 0-90% MeCN/H ₂ O as eluent to give 60 mg (49%) of compound 36-a.

Stage 36-2

Example 36 was prepared from compound 36-a and compound 1-c using a procedure similar to that described for Step 18-2. [M+H] ⁺ 401; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.79 (d, J = 4.4 Hz, 1H), 8.67 – 8.59 (m, 2H), 8.39 (s, 1H), 8.10 ( d, J = 7.2 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.90 (td, J = 7.8, 1.8 Hz, 1H), 7.57 (d, J = 7.8 Hz, 1H), 7.41 ( dd, J = 7.6, 4.9 Hz, 1H), 5.90 (m, 1H), 2.55 (s, 3H), 1.77 (d, J = 6.7 Hz, 6H).

Example 37

Stage 37-1

A mixture of methyl 4-chloro-5-methylpyridin-2-carboxylate (100 mg, 0.54 mmol), (pyridin-3-yl)boronic acid (100 mg, 0.80 mmol), Pd(PPh ₃ ) ₄ (62 mg, 0.05 mmol ) and K ₂ CO ₃ (224 mg, 1.62 mmol) in dioxane (5 ml) were stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified by preparative HPLC using 0-80% MeCN/ _H2O as eluent to give 60 mg (48.8%) of compound 37-a as a yellow solid.

Stage 37-2

Example 37 was prepared from compound 37-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 401; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.52 (s, 1H), 8.75 (d, J = 4.8 Hz, 1H), 8.71 (s, 1H), 8.66 (s, 1H), 8.61 (d, J = 8.0 Hz, 1H), 8.24 (s, 1H), 8.10 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.77 (d, J = 8.0 Hz, 1H) , 7.49 (dd, J = 7.8, 4.8 Hz, 1H), 5.88 (m, 1H), 2.45 (s, 3H), 1.77 (d, J = 6.7 Hz, 6H).

Example 38

Stage 38-1

A mixture of methyl 4-chloro-5-methylpyridin-2-carboxylate (100 mg, 0.54 mmol), (pyridin-4-yl)boronic acid (100 mg, 0.80 mmol), Pd(PPh ₃ ) ₄ (62 mg, 0.05 mmol ) and K ₂ CO ₃ (223 mg, 1.62 mmol) in ethylene glycol dimethyl ether (5 ml) and H ₂ O (1 ml) were stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified by preparative HPLC using 0-60% MeCN/H ₂ O as eluent to give 70 mg (61%) of compound 38-a as a yellow solid.

Stage 38-2

Compound 1-c (57 mg, 0.28 mmol) was added to a mixture of compound 38-a (50 mg, 0.23 mmol), HATU (266 mg, 0.46 mmol) and 4-methylmorpholine (118 mg, 1.17 mmol) in DMF (2 ml ). The resulting solution was stirred at room temperature overnight. The resulting solution was purified by preparative HPLC using 0-80% MeCN/H ₂ O as eluent to obtain 8.3 mg of Example 38 as a white solid. [M+H] ⁺ 401; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.51 (s, 1H), 8.80 (d, J = 5.0 Hz, 2H), 8.66 (s, 1H), 8.59 (d, J = 8.4 Hz, 1H) , 8.22 (s, 1H), 8.11 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.35 (d, J = 5.2 Hz, 2H), 5.88 (m, 1H) , 2.44 (s, 3H), 1.77 (d, J = 6.7 Hz, 6H).

Example 39

Stage 39-1

Mixture of methyl 4-chloro-5-methylpyridin-2-carboxylate (100 mg, 0.54 mmol), 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1,3-thiazole (242.6 mg, 1.08 mmol), Pd(PPh ₃ ) ₄ (124.5 mg, 0.11 mmol) and K ₂ CO ₃ (223.4 mg, 1.62 mmol) in dioxane (5 ml) were stirred at 100°C in a nitrogen atmosphere overnight. The reaction mixture was evaporated in vacuo and purified by reverse phase flash chromatography with 0-33% MeCN/H ₂ O to give compound 39-a (40 mg, 29.90%) as a yellow solid.

Stage 39-2

Example 39 was prepared from compound 39-a and compound 1-c using a procedure similar to that described for Step 18-2. [M+H] ⁺ 421; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.49 (s, 1H), 8.61 (t, J = 4.2 Hz, 2H), 8.34 (s, 1H), 8.10 (d, J = 7.5 Hz, 1H) , 8.01 (t, J = 8.0 Hz, 1H), 7.91 (s, 1H), 5.87 (m, 1H), 2.84 (s, 3H), 2.60 (s, 3H), 1.76 (d, J = 6.7 Hz, 6H).

Example 40

Stage 40-1

A mixture of methyl 5-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-carboxylate (100 mg, 0.36 mmol), 4-bromo-2- methyl 1,3-thiazole (128.5 mg, 0.72 mmol), Pd(PPh ₃ ) ₄ (83.4 mg, 0.07 mmol) and K ₂ CO ₃ (149.6 mg, 1.08 mmol) in dioxane (5 ml) were stirred at 100° C in a nitrogen atmosphere overnight. The reaction mixture was evaporated under reduced pressure and the residue was purified by reverse phase flash chromatography with 0-36% MeCN/H ₂ O to give compound 40-a (80 mg, 89.29%) as a brown oil.

Stage 40-2

Example 40 was prepared from compound 40-a and compound 1-c using a procedure similar to that described for Step 18-2. [M+H] ⁺ 421; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.54 (s, 1H), 8.66 – 8.57 (m, 3H), 8.09 (d, J = 7.5 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.48 (s, 1H), 5.89 (m, 1H), 2.83 (s, 3H), 2.64 (s, 3H), 1.76 (d, J = 6.6 Hz, 6H).

Example 41

Compound 41-a was prepared using a procedure similar to that described for Step 37-1. Example 41 was prepared from compound 41-a and compound 22-a using a procedure similar to that described for Step 29-1. [M+H] ⁺ 417. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.76 (s, 1H), 8.72 (s, 1H), 8.64 (s, 1H), 8.42 (d, J = 8.3 Hz, 1H), 8.23 (s, 1H), 8.13 (d, J = 7.2 Hz, 1H), 8.02 (t, J = 8.1 Hz, 1H), 7.78 (d, J = 7.7 Hz , 1H), 7.51 (s, 1H), 5.92 (m, 1H), 4.20 (d, J = 5.3 Hz, 2H), 2.46 (s, 3H), 1.75 (d, J = 6.9 Hz, 3H).

Example 42

Compound 42-a was prepared using a procedure similar to that described for Step 40-1. Example 42 was prepared from compound 42-a and 1-c using a procedure similar to that described for Step 29-1. [M+H] ⁺ 447. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.49 (s, 1H), 8.69 – 8.60 (m, 2H), 8.33 (s, 1H), 8.11 (d, J = 7.6 Hz, 1H), 8.02 (t, J = 7.9 Hz, 1H), 7.88 (s, 1H), 5.87 (m, 1H), 2.60 (s, 3H), 2.43 (s, 1H), 1.76 (d, J = 6.7 Hz, 6H), 1.28 (d, J = 8.1 Hz, 2H), 1.21 (d, J = 8.1 Hz, 2H).

Example 43

Example 43 was prepared from compounds 42-a and 22-a using a procedure similar to that described for Step 29-1. [M+H] ⁺ 463. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.53 (s, 1H), 8.58 (s, 1H), 8.42 (d, J = 8.3 Hz, 1H), 8.32 (s , 1H), 8.14 (d, J = 7.6 Hz, 1H), 8.02 (t, J = 8.0 Hz, 1H), 7.85 (s, 1H), 5.90 (m, 1H), 4.19 (d, J = 6.0 Hz , 2H), 3.15 (s, 1H), 2.60 (s, 3H), 2.41 (m, 1H), 1.74 (d, J = 6.8 Hz, 3H), 1.30 – 1.16 (m, 4H).

Example 44

Compound 44-a was prepared using a procedure similar to that described for Step 40-1. Example 44 was prepared from compounds 44-a and 1-c using a procedure similar to that described for Step 29-1. [M+H] ⁺ 446. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.52 (s, 1H), 8.62 (d, J = 8.3 Hz, 1H), 8.53 (s, 1H), 8.37 (s , 1H), 8.10 (m, 1H), 8.00 (t, J = 7.9 Hz, 1H), 7.26 (d, J = 3.7 Hz, 1H), 6.87 (d, J = 3.7 Hz, 1H), 5.89 (m , 1H), 2.63 (s, 3H), 2.17 (m, 1H), 1.76 (d, J = 6.6 Hz, 6H), 1.16 – 1.07 (m, 2H), 0.87 – 0.85 (m, 2H).

Example 45

Example 45 was prepared from compounds 44-a and 22-a using a procedure similar to that described for Step 29-1. [M+H] ⁺ 462. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.52 (s, 1H), 8.41 (d, J = 8.3 Hz, 1H), 8.36 (s , 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.26 (d, J = 3.8 Hz, 1H), 6.87 (d, J = 3.7 Hz, 1H ), 5.92 (m, 1H), 4.19 (t, J = 6.3 Hz, 2H), 3.51 (s, 1H), 3.22 (t, J = 6.4 Hz, 1H), 2.62 (s, 3H), 2.17 (m , 1H), 1.74 (d, J = 6.8 Hz, 3H), 1.16 – 1.07 (m, 2H), 0.87 – 0.85 (m, 2H).

Example 46

Stage 46-1

Mixture of 5-bromo-2-fluoro-4-methylbenzoic acid (150 mg, 0.64 mmol), 1-cyclopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2- yl)-1H-pyrazole (226.0 mg, 0.97 mmol), Pd(PPh ₃ ) ₄ (74.4 mg, 0.06 mmol) and K ₂ CO ₃ (266.9 mg, 1.93 mmol) in DME (5 ml) and H ₂ O ( 1 ml) was stirred at 100°C in a nitrogen atmosphere for 1 hour. The mixture was left to cool to room temperature and acidified to pH 3~4 with concentrated HCl. The solvent was removed in vacuo and the residue was purified by reverse phase flash chromatography with 0-40% MeCN/H ₂ O to give 46-a (150 mg, 89.54%) as a light brown solid.

Stage 46-2

A mixture of compound 46-a (50 mg, 0.19 mmol) and 1-Chloro-N,N,2-trimethyl-1-propenylamine (30.8 mg, 0.23 mmol) in DCM (2 ml) was stirred at room temperature for 1 hour. Pyridine (45.6 mg, 0.58 mmol) and compound 1-c (58.9 mg, 0.29 mmol) were added to the resulting solution. The mixture was stirred at room temperature for 1 hour. The solvent was removed in vacuo and the residue was purified by reverse phase flash chromatography with 0-60% CH ₃ CN/H ₂ O to give Example 46 (12.5 mg) as a white solid. [M+H] ⁺ 447; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.6 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.14 – 8.08 (m, 2H), 7.99 (t, J = 8.0 Hz, 1H), 7.65 – 7.63 (m, 2H), 7.13 (d, J = 13.1 Hz, 1H), 5.75 (m, 1H), 3.68 (m, 1H), 2.49 (s, 3H), 1.74 (d, J = 6.7 Hz, 6H), 1.19 (m, 2H), 1.12 (m, 2H).

Example 47

Stage 47-1

A mixture of compound 46-a (50 mg, 190 mmol) and 1-Chloro-N,N,2-trimethyl-1-propenylamine (30.8 mg, 230 mmol) in DCM (2 ml) was stirred at room temperature for 1 hour. Pyridine (45.6 mg, 0.58 mmol) and compound 47-a (75.6 mg, 0.29 mmol) were added and the resulting mixture was stirred at room temperature for 1 hour. The reaction was quenched by adding MeOH (2 ml), and K ₂ CO ₃ (53.1 mg, 0.38 mmol) was added. The resulting mixture was stirred at room temperature for 5 hours. The solvent was removed in vacuo and the residue was purified by reverse phase flash chromatography with 0-60% MeCN/H ₂ O to give Example 47 (9.5 mg) as a white solid. [M+H] ⁺ 463; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.6 Hz, 1H), 8.49 (d, J = 8.3 Hz, 1H), 8.14 – 8.11 (m, 2H), 8.02 (t, J = 8.0 Hz, 1H), 7.65 – 7.60 (m, 2H), 7.13 (d, J = 13.1 Hz, 1H), 5.76 (m, 1H), 4.23 – 4.12 (m, 2H), 3.69 (m, 1H ), 2.50 (s, 3H), 1.73 (d, J = 6.8 Hz, 3H), 1.22 (m, 2H), 1.09 (m, 2H).

Example 48

Example 48 was prepared from compound 46-a and compound 17-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 463; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 16.7 Hz, 1H), 8.54 (d, J = 8.3 Hz, 1H), 8.13 (d, J = 8.2 Hz, 1H), 8.05 (t, J = 8.1 Hz, 1H), 7.72 (d, J = 7.9 Hz, 1H), 7.68 – 7.65 (m, 1H), 7.14 (d, J = 13.1 Hz, 1H), 3.90 (m, 1H) , 3.68 (m, 1H), 2.50 (s, 3H), 1.53 (d, J = 6.9 Hz, 6H), 1.22 (m, 2H), 1.14 (m, 2H).

Example 49

Example 49 was prepared using a procedure similar to that described for Example 46. [M+H] ⁺ 449; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.20 (d, J = 16.6 Hz, 1H), 8.55 (d, J = 8.4 Hz, 1H), 8.15 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.68 (s, 1H), 7.60 (s, 1H), 7.13 (d, J = 13.2 Hz, 1H), 5.76 (m, 1H), 4.58 (m, 1H), 2.51 (s, 3H), 1.74 (d, J = 6.7 Hz, 6H), 1.59 (d, J = 6.7 Hz, 6H).

Example 50

Example 50 was prepared using a procedure similar to that described for Example 47. [M+H] ⁺ 465; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.22 (d, J = 16.5 Hz, 1H), 8.49 (d, J = 8.3 Hz, 1H), 8.19 – 8.09 (m, 2H), 8.02 (t, J = 7.9 Hz, 1H), 7.68 (s, 1H), 7.61 (s, 1H), 7.14 (d, J = 13.2 Hz, 1H), 5.71 (m, 1H), 4.59 (m, 1H), 4.18 ( m, 2H), 2.51 (s, 3H), 1.73 (d, J = 6.8 Hz, 3H), 1.60 (d, J = 6.7 Hz, 6H).

Example 51

Example 51 was prepared using a procedure similar to that described for Example 48. [M+H] ⁺ 449; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.17 (d, J = 16.8 Hz, 1H), 8.54 (d, J = 8.3 Hz, 1H), 8.15 (d, J = 8.3 Hz, 1H), 8.06 (t, J = 8.1 Hz, 1H), 7.72 (d, J = 8.3 Hz, 1H), 7.69 (s, 1H), 7.61 (s, 1H), 7.14 (d, J = 13.1 Hz, 1H), 4.58 (m, 1H), 3.90 (m, 1H), 2.51 (s, 3H), 1.60 (d, J = 6.7 Hz, 6H), 1.53 (d, J = 6.9 Hz, 6H).

Example 52

Example 52 was prepared using a procedure similar to that described for Example 46. [M+H] ⁺ 463; ¹ HNMR (400 MHz, Chloroform-d) δ 9.20 (d, J = 16.7 Hz, 1H), 8.56 (dd, J = 8.3, 0.9 Hz, 1H), 8.16 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 8.0 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.73 – 7.67 (m, 2H), 7.14 (d, J = 13.2 Hz, 1H), 5.77 (m, 1H) , 2.52 (s, 3H), 1.75 (d, J = 6.7 Hz, 6H), 1.68 (s, 9H).

Example 53

Example 53 was prepared using a procedure similar to that described for Example 48. [M+H] ⁺ 463; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.17 (d, J = 16.8 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.15 (d, J = 8.3 Hz, 1H), 8.06 (t, J = 8.1 Hz, 1H), 7.74 – 7.69 (m, 3H), 7.14 (d, J = 13.1 Hz, 1H), 3.91 (m, 1H), 2.51 (s, 3H), 1.68 (s, 9H), 1.53 (d, J = 6.9 Hz, 6H).

Example 54

Example 54 was prepared using a procedure similar to that described for Example 47. [M+H] ⁺ 479; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.20 (d, J = 16.8 Hz, 1H), 8.50 (d, J = 8.3 Hz, 1H), 8.16 – 8.11 (m, 2H), 8.02 (t, J = 8.0 Hz, 1H), 7.71 – 7.69 (m, 2H), 7.14 (d, J = 13.2 Hz, 1H), 5.76 (m, 1H), 4.24 – 4.13 (m, 2H), 2.52 (s, 3H ), 1.73(d, J = 6.9 Hz, 3H), 1.66 (s, 9H).

Example 55

Stage 55-1

A mixture of methyl 5-bromo-2-fluoro-4-methylbenzoate (1.7 g), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi (1,3 ,2-dioxaborolane) (2.3 g, 7.7 mmol), Pd(dppf)Cl ₂ CH ₂ Cl ₂ (24 mg, 0.03 mmol) and KOAc (1.6 g, 19.2 mmol) in dioxane (20 ml) were stirred at 85°C during the night. The solvent was removed in vacuo. The residue was dissolved in EtOAc, washed with saturated NaCl solution, dried over Na ₂ SO ₄ and evaporated in vacuum. Purification of the residue on a silica gel column using petroleum ether/EtOAc (10:1 to 5:1) as eluent gave 1.5 g of 55-a (79.3%, 2 steps) as an off-white solid.

Stage 55-2

A mixture of compound 55-a (300 mg, 1.0 mmol), 4-bromo-1-(trifluoromethyl)-1H-pyrazole (258 mg, 1.2 mmol), Pd(PPh ₃ ) ₄ (115 mg, 0.10 mmol) and K ₂ CO ₃ (414 mg, 3.0 mmol) in DME (8 ml) and H ₂ O (2 ml) were stirred at 100°C under nitrogen atmosphere for 1 hour. The reaction mixture was cooled to room temperature and acidified to pH 3~4 with concentrated HCl. The solvent was removed in vacuo and the residue was purified by reverse phase flash chromatography with 0-60% CH ₃ CN in water to give compound 55-b (170 mg, 57.8%) as a light brown solid.

Stage 55-3

Example 55 was prepared from compound 55-b using a procedure similar to that described for Step 46-2. [M+H] ⁺ 475; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.5 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.18 (d, J = 8.1 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.02 (t, J = 8.0 Hz, 1H), 7.80 – 7.97 (m, 2H), 5.75 (m, 1H), 2.50 (s, 3H), 1.75 (d, J = 6.6 Hz, 6H).

Example 56

Example 56 was prepared from compound 55-b and compound 17-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 475; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 16.6 Hz, 1H), 8.54 (d, J = 8.3 Hz, 1H), 8.17 (d, J = 8.1 Hz, 1H), 8.07 (t, J = 8.1 Hz, 1H), 7.97 – 7.94 (m, 2H), 7.74 (d, J = 7.9 Hz, 1H), 7.21 (d, J = 13.0 Hz, 1H), 3.89 (m, 1H) , 2.50 (s, 3H), 1.54 (d, J = 6.9 Hz, 6H).

Example 57

Example 57 was prepared from compound 55-b using a procedure similar to that described for Step 47-1. [M+1] ⁺ 491; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.3 Hz, 1H), 8.48 (d, J = 8.3 Hz, 1H), 8.17 – 8.11 (m, 2H), 8.03 (t, J = 8.0 Hz, 1H), 7.97 – 7.94 (m, 2H), 7.20 (d, J = 12.9 Hz, 1H), 5.74 (m, 1H), 4.18 (m, 2H), 2.50 (s, 3H), 1.73 (d, J = 6.8 Hz, 3H).

Example 58

Stage 58-1

Mixture of compound 55-a (300 mg, 1.02 mmol), 3-bromo-1-(propan-2-yl)-1H-pyrazole (267.6 mg, 1.42 mmol), Pd(PPh ₃ ) ₄ (118 mg, 0.10 mmol ), K ₂ CO ₃ (422 mg, 3.00 mmol) in 1,4-dioxane (10 ml) was stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified by silica gel column chromatography using 0-50% EtOAc/petroleum ether as eluent to give 210 mg (74.6%) of compound 58-a as a white solid.

Stage 58-2

Example 58 was prepared from compound 58-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 449. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.17 (d, J = 16.2 Hz, 1H), 8.57 (d, J = 8.3 Hz, 1H), 8.39 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 7.5 Hz, 1H), 7.99 (t, J = 8.0 Hz, 1H), 7.51 (d, J = 2.3 Hz, 1H), 7.13 (d, J = 13.2 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 5.78 (m, 1H), 4.57 (m, 1H), 2.60 (s, 3H), 1.75 (d, J = 6.7 Hz, 6H), 1.59 (d, J = 6.7 Hz, 6H).

Example 59

Example 59 was prepared from compound 58-a and compound 17-a using a procedure similar to that described for Step 18-2. [M+H] ⁺ 449. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.14 (d, J = 16.5 Hz, 1H), 8.56 (d, J = 8.3 Hz, 1H), 8.38 (d, J = 8.4 Hz, 1H), 8.05 (t, J = 8.0 Hz, 1H), 7.72 (d, J = 7.8 Hz, 1H), 7.51 (d, J = 2.3 Hz, 1H), 7.14 (d, J = 13.2 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 4.58 (m, 1H), 3.92 (m, 1H), 2.60 (s, 3H), 1.59 (d, J = 6.8 Hz, 6H), 1.54 (d, J = 6.8 Hz, 6H).

Example 60

Stage 60-1

LiOH (52 mg, 2.17 mmol) was added to a solution of compound 58-a (60 mg, 0.217 mmol) in MeOH (6 ml) and H ₂ O (2 ml). The resulting solution was stirred at room temperature for 2 hours. The solvent was removed in vacuo and the residue was purified by preparative flash HPLC using 0-100% MeCN/H ₂ O as eluent, yielding 40 mg (70%) of compound 60-a as a white solid.

Stage 60-2

Example 60 was prepared from compound 60-a using a procedure similar to that described for Step 47-1. [M+H] ⁺ 465. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.19 (d, J = 16.1 Hz, 1H), 8.50 (d, J = 8.3 Hz, 1H), 8.37 (d, J = 8.4 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.51 (d, J = 2.2 Hz, 1H), 7.13 (d, J = 13.2 Hz, 1H), 6.46 (d, J = 2.3 Hz, 1H), 5.77 (m, 1H), 4.58 (m, 1H), 4.25 – 4.10 (m, 2H), 2.99 (s, 1H), 2.60 (s , 3H), 1.73 (d, J = 7.0 Hz, 3H), 1.59 (d, J = 6.7 Hz, 6H).

Example 61

Stage 61-1

Mixture of compound 55-a (300 mg, 1.02 mmol), 4-bromo-1-(propan-2-yl)-1H-imidazole (230 mg, 1.22 mmol), Pd(PPh ₃ ) ₄ (117.7 mg, 0.10 mmol ) and K ₂ CO ₃ (422 mg, 3.06 mmol) in 1,4-dioxane (2 ml) were stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified on a silica gel column using 30% EtOAc in petroleum ether as eluent to give 130 mg (46%) of compound 61-a as a yellow solid.

Stage 61-2

Compound 61-a (130 mg, 0.47 mmol) was added to a mixture of sodium hydroxide (94 mg, 2.35 mmol) in methanol (6 ml) and H ₂ O (2 ml). The resulting solution was stirred at room temperature for 2 hours. The solvent was removed in vacuo and the resulting crude product was purified by preparative flash HPLC using 0-100% MeCN/H ₂ O as eluent to give 100 mg (81%) of compound 61-b as a white solid.

Stage 61-3

Example 61 was prepared from compound 61-b using a procedure similar to that described for Step 46-2. [M+H] ⁺ 449; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.19 (d, J = 15.6 Hz, 1H), 8.57 (d, J = 8.3 Hz, 1H), 8.42 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 7.6 Hz, 1H), 7.99 (t, J = 8.0 Hz, 1H), 7.67 (s, 1H), 7.18 (s, 1H), 7.15 (d, J = 4.8 Hz, 1H), 5.80 (m, 1H), 4.43 (m, 1H), 2.63 (s, 3H), 1.75 (d, J = 6.6 Hz, 6H), 1.59 (d, J = 6.7 Hz, 6H).

Example 62

Example 62 was prepared from compound 61-b and compound 17-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 449; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 15.9 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.42 (d, J = 8.4 Hz, 1H), 8.04 (t, J = 8.1 Hz, 1H), 7.71 (d, J = 7.6 Hz, 1H), 7.67 (s, 1H), 7.28 (s, 1H), 7.12 (d, J = 13.2 Hz, 1H), 4.43 (m, 1H), 3.92 (m, 1H), 2.62 (s, 3H), 1.59 (d, J = 6.8 Hz, 2H), 1.53 (d, J = 6.8 Hz, 2H).

Example 63

Example 63 was prepared from compound 61-b using a procedure similar to that described for Step 47-1. [M+H] ⁺ 465; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.36 (d, J = 13.4 Hz, 1H), 8.47 (d, J = 8.3 Hz, 1H), 8.38 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 7.6 Hz, 1H), 7.99 (t, J = 7.9 Hz, 1H), 7.69 (s, 1H), 7.17 (s, 1H), 7.10 (d, J = 12.8 Hz, 1H), 5.83 (m, 1H), 4.44 (m, 1H), 4.16 (m, 2H), 2.59 (s, 3H), 1.72 (d, J = 6.8 Hz, 3H), 1.60 (d, J = 6.7 Hz, 6H) .

Example 64

Stage 64-1

A mixture of 5-bromo-2-fluoro-4-methylbenzoic acid (100 mg, 0.43 mmol), 2-(tributylstannyl)pyridine (190 mg, 0.52 mmol), Pd(PPh ₃ ) ₄ (49 mg, 0.04 mmol) in DMF (2 ml) in the microwave at 160°C for 1 hour. The solvent was removed in vacuo and the residue was purified by preparative flash HPLC using 0-100% MeCN/H ₂ O as the eluent, yielding 40 mg (37%) of compound 64-a as a yellow solid.

Stage 64-2

Example 64 was prepared from compound 64-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 418; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.19 (d, J = 16.1 Hz, 1H), 8.74 (d, J = 4.4 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.25 (d, J = 8.3 Hz, 1H), 8.11 (d, J = 7.5 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.84 (td, J = 7.7, 1.8 Hz, 1H), 7.48 (d, J = 7.8 Hz, 1H), 7.34 (dd, J = 7.5, 5.0 Hz, 1H), 7.19 (d, J = 13.1 Hz, 1H), 5.78 (m, 1H), 2.49 (s, 3H) , 1.75 (d, J = 6.7 Hz, 6H).

Example 65

Stage 65-1

Mixture of 5-bromo-2-fluoro-4-methylbenzoic acid (100 mg, 0.40 mmol), (pyridin-3-yl)boronic acid (64 mg, 0.52 mmol), Pd(PPh ₃ ) ₄ (99 mg, 0.08 mmol ) and K ₂ CO ₃ (179 mg, 1.3 mmol) in ethylene glycol dimethyl ether (4 ml) and H ₂ O (2 ml) were stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified by preparative flash HPLC using 0-70% MeCN/H ₂ O as eluent, yielding 40 mg (37%) of compound 65-a as a yellow solid.

Stage 65-2

Example 65 was prepared from compound 65-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 418; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.19 (d, J = 16.2 Hz, 1H), 8.69 – 8.64 (m, 2H), 8.55 (d, J = 8.3 Hz, 1H), 8.12 – 8.09 ( m, 2H), 8.01 (t, J = 8.0 Hz, 1H), 7.71 (m, 1H), 7.45 (dd, J = 7.5, 5.1 Hz, 1H), 7.22 (d, J = 13.0 Hz, 1H), 5.75 (m, 1H), 3.51 (s, 1H), 2.39 (s, 3H), 1.75 (d, J = 6.7 Hz, 6H).

Example 66

Stage 66-1

Mixture of 5-bromo-2-fluoro-4-methylbenzoic acid (100 mg, 0.43 mmol), (pyridin-4-yl)boronic acid (64 mg, 0.52 mmol), Pd(PPh ₃ ) ₄ (99 mg, 0.08 mmol ) and K ₂ CO ₃ (179 mg, 1.3 mmol) in ethylene glycol dimethyl ether (4 ml) and H ₂ O (2 ml) were stirred at 100°C overnight. The solvent was removed in vacuo and the resulting crude product was purified by preparative flash HPLC using 0-100% MeCN/H ₂ O as eluent to give 70 mg (75%) of compound 66-a as a white solid.

Stage 66-2

Example 66 was prepared from compound 66-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 418; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.2 Hz, 1H), 8.75 (s, 2H), 8.55 (d, J = 8.3 Hz, 1H), 8.14 – 8.09 (m, 2H), 8.01 (t, J = 7.9 Hz, 1H), 7.35 (d, J = 5.0 Hz, 2H), 7.23 (d, J = 13.0 Hz, 1H), 5.75 (m, 1H), 2.40 (s, 3H), 1.75 (d, J = 6.7 Hz, 6H).

Example 67

Stage 67-1

Mixture of 5-bromo-2-fluoro-4-methylbenzoic acid (100 mg, 0.43 mmol), 2-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )-1,3-thiazole (193.2 mg, 0.86 mmol), Pd(PPh ₃ ) ₄ (99.2 mg, 0.09 mmol) and K ₂ CO ₃ (177.9 mg, 1.29 mmol) in ethylene glycol dimethyl ether (5 ml) and H ₂ O (1 ml) was stirred at 100°C under nitrogen atmosphere overnight. The mixture was left to cool to room temperature and acidified to pH 3~4 by adding 1N. HCl solution. The solvent was removed in vacuo and the residue was purified by reverse phase chromatography using 0-38% MeCN/H ₂ O=38:62 as eluent, yielding compound 67-a (40 mg, 37.10%) as a yellow solid .

Stage 67-2

Example 67 was prepared from compound 67-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 438; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 16.1 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.20 (d, J = 8.1 Hz, 1H), 8.11 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 7.9 Hz, 1H), 7.63 (s, 1H), 7.18 (d, J = 12.9 Hz, 1H), 5.75 (m, 1H), 2.79 (s, 3H), 2.49 (s, 3H), 1.74 (d, J = 6.7 Hz, 6H).

Example 68

Stage 68-1

Mixture of methyl 2-fluoro-4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (100 mg, 0.34 mmol), 4-bromo-2- methyl 1,3-thiazole (72 mg, 0.40 mmol), Pd(PPh ₃ ) ₄ (30 mg, 0.03 mmol), K ₂ CO ₃ (141 mg, 1.02 mmol) in ethylene glycol dimethyl ether (5 ml) and H ₂ O (1 ml) was stirred at 100°C overnight. The solvent was removed in vacuo and the residue was purified by preparative flash HPLC using 0-80% MeCN/H ₂ O as the eluent, yielding 55 mg (65%) of compound 68-a as a yellow solid.

Stage 68-2

Example 68 was prepared from compound 68-a using a procedure similar to that described for Step 46-2. [M+H] ⁺ 438; ¹ H-NMR (400 MHz, Chloroform-d) δ 9.17 (d, J = 16.1 Hz, 1H), 8.56 (d, J = 8.3 Hz, 1H), 8.37 (d, J = 8.4 Hz, 1H), 8.10 (d, J = 7.6 Hz, 1H), 8.00 (t, J = 8.0 Hz, 1H), 7.20 (s, 1H), 7.15 (d, J = 13.2 Hz, 1H), 5.77 (m, 1H), 2.81 (s, 3H), 2.54 (s, 3H), 1.75 (d, J = 6.8 Hz, 6H).

Example 69

Example 69 was prepared using a procedure similar to that described for Example 61. [M+H] ⁺ 463. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.15 (d, J = 16.1 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.20 (d, J = 8.2 Hz, 1H), 8.10 (d, J = 7.6 Hz, 1H), 7.99 (t, J = 8.0 Hz, 1H), 7.13 (d, J = 13.1 Hz, 1H), 6.90 (d, J = 3.5 Hz, 1H), 6.79 (d, J = 3.6 Hz, 1H), 5.77 (m, 1H), 2.52 (s, 3H), 2.13 (m, J = 8.7 , 5.0 Hz, 1H), 1.74 (d, J = 6.7 Hz, 6H), 1.15 (m, 2H), 0.80 (m, 2H).

Example 70

Example 70 was prepared using a procedure similar to that described for Example 63. [M+H] ⁺ 479. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.13 (d, J = 16.3 Hz, 1H), 8.48 (d, J = 8.3 Hz, 1H), 8.19 (d, J = 8.2 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 7.13 (d, J = 13.1 Hz, 1H), 6.90 (d, J = 3.6 Hz, 1H), 6.79 (d, J = 3.6 Hz, 1H), 5.75 (m, 1H), 4.17 (m, J = 11.9, 5.9 Hz, 2H), 2.52 (s, 3H), 2.15 (m, 1H), 1.73 (d, J = 6.8 Hz, 3H), 1.05 (m, 2H), 0.80 (m, 2H).

Example 71

Stage 71-1

A solution of 5-bromo-4-fluoro-2-methylbenzoic acid (900 mg, 3.9 mmol), methoxy(methyl)amine hydrochloride (573 mg, 5.9 mmol), HATU (2.3 g, 6.0 mmol) and DIPEA (2 g, 15.5 mmol) in DCM (20 ml) was stirred at room temperature overnight. The resulting solution was then diluted with saturated NaCl solution and extracted with ethyl acetate (30 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/EtOAc (2:1) as eluant to give 5-bromo-4-fluoro-N-methoxy-N,2-dimethylbenzamide (900 mg, 84.4%) as light yellow oil.

Stage 71-2

In a round-bottomed 3-neck flask, a solution of 5-bromo-4-fluoro-N-methoxy-N,2-dimethylbenzamide (900 mg, 3.3 mmol) in THF was cooled to -30°C under a nitrogen atmosphere. Add CH ₃ MgCl in THF (3M) (14.6 ml, 43.8 mmol) dropwise over 10 minutes, maintaining the temperature below 0°C. The reaction mixture was stirred at 0°C for 0.5 hour, quenched with saturated NH ₄ Cl solution at 0°C and extracted with ethyl acetate (30 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/EtOAc (2:1) as eluent to give 1-(5-bromo-4-fluoro-2-methylphenyl)ethan-1-one (600 mg, 79.6% ) as a light yellow solid.

Stage 71-3

Mixture of 1-(5-bromo-4-fluoro-2-methylphenyl)ethan-1-one (600 mg, 2.6 mmol), 1-cyclopropane-carbonyl-1H-1,2,3-benzotriazole (885 mg, 4.8 mmol ), magnesium bromide diethyl ether complex (4.9 g, 19.2 mmol) and DIPEA (1.2 g, 9.3 mmol) in DCM were stirred at room temperature for 3 days. The resulting mixture was diluted with saturated NaCl solution (10 ml) and then extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions (column: silica gel C18; mobile phase: acetonitrile in a mixture of formic acid (0.1%) and water, gradient from 0% to 100% in 25 minutes; detector: UV 254 nm) , giving 1-(5-bromo-4-fluoro-2-methylphenyl)-3-cyclopropyl propane-1,3-dione (400 mg, 51.4%) as a yellow oil.

Stage 71-4

A solution of 1-(5-bromo-4-fluoro-2-methylphenyl)-3-cyclopropylpropane-1,3-dione (400 mg, 1.3 mmol) and hydroxylamine hydrochloride (318 mg, 4.6 mmol) in MeOH (10 ml) was stirred at 45°C overnight. The resulting solution was diluted with saturated NaCl solution (20 ml) and extracted with ethyl acetate (20 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions (column: silica gel C18; mobile phase: acetonitrile in water, gradient from 0 to 100% in 25 minutes; detector: UV 254 nm), obtaining 3-(5-bromo- 4-fluoro-2-methylphenyl)-5-cyclopropyl-1,2-oxazole (170 mg, 42.9%) as a yellow oil.

Stage 71-5

A mixture of 3-(5-bromo-4-fluoro-2-methylphenyl)-5-cyclopropyl-1,2-oxazole (170 mg, 0.5 mmol), Pd(dppf)Cl ₂ CH ₂ Cl ₂ (258 mg, 0.3 mmol ) and TEA (478 mg, 4.8 mmol) in DMF/MeOH (4/1 ml) were stirred at 75°C under CO overnight. The resulting mixture was diluted with saturated NaCl solution and then extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions (column: C18 silica gel; mobile phase: acetonitrile in water, gradient from 0 to 80% in 20 minutes; detector: UV 254 nm) to obtain methyl 5-(5-cyclopropyl -1,2-oxazol-3-yl)-2-fluoro-4-methylbenzoate (80 mg, 50.6%) as a yellow oil.

Stage 71-6

A mixture of methyl 5-(5-cyclopropyl-1,2-oxazol-3-yl)-2-fluoro-4-methylbenzoate (80 mg, 0.3 mmol) and LiOH (24 mg, 0.9 mmol) in THF/H ₂ O ( 1/0.3 ml) was stirred at room temperature for 2 hours. The mixture was then acidified to pH 4~5 with 1M aqueous HCl and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure to give 5-(5-cyclopropyl-1,2-oxazol-3-yl)-2-fluoro-4-methylbenzoic acid (50 mg, 65.8%) in the form of a yellow oil.

Stage 71-7

To a stirred solution of 5-(5-cyclopropyl-1,2-oxazol-3-yl)-2-fluoro-4-methylbenzoic acid (50 mg, 0.2 mmol) in DCM (1 ml), (1-chloro- 2-methylprop-1-en-1-yl)dimethylamine (33 mg, 0.2 mmol) at room temperature under nitrogen atmosphere. After stirring for 1 hour, a solution of pyridine (48 mg, 0.6 mmol) and 6-(1-isopropyl-1H-tetrazol-5-yl)pyridin-2-amine (86 mg, 0.4 mmol) in DCM (1 ml) was added ). After stirring for a further 1 hour, the reaction was quenched with water (10 ml) and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ , filtered and evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions (column: C18 silica gel; mobile phase: acetonitrile in water, gradient from 0% to 90% in 20 minutes; detector: UV 254 nm), obtaining 14 mg of Example 71 as off-white solid. MS (m/z): [M + H] ⁺ = 448.20, t = 1.928 min. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.13 (d, J = 15.8 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.47 (d, J = 8.0 Hz, 1H), 8.12 (d, J = 7.6 Hz, 1H), 8.02 (t, J = 8.0 Hz, 1H), 7.20 (d, J = 12.9 Hz, 1H), 6.20 (s, 1H), 5.75 (m, 1H), 2.62 (s, 3H), 2.10 (m, 1H), 1.74 (d, J = 6.7 Hz, 6H), 1.18 – 1.08 (m, 2H), 0.97 – 0.89 (m, 2H).

Example 72-A and 72-B

Stage 72-1

A mixture of ethyl 2-chloro-5-fluoropyridine-4-carboxylate (1.0 g, 4.91 mmol) and LiOH (1.5 g, 6.26 mmol) in THF/H ₂ O (20/5 ml) was stirred at room temperature for 2 hours. The mixture was acidified with aqueous HCl and extracted with ethyl acetate (30 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ . The resulting filtrate was evaporated under reduced pressure to obtain 760 mg (88.4%) of 2-chloro-5-methylisonicotinic acid as an off-white solid.

Stage 72-2

2-Chloro-N-methoxy-N,5-dimethylisonicotinamide was prepared according to the procedure described for Step 71-1. 700 mg (73.6%) was obtained as an off-white solid.

Stage 72-3

1-(2-Chloro-5-methylpyridin-4-yl)ethan-1-one was prepared according to the procedure described for Step 71-2. 400 mg (84.4%) was obtained as a light yellow oil.

Stage 72-4

1-(2-Chloro-5-methylpyridin-4-yl)-3-cyclopropylpropane-1,3-dione was prepared according to the procedure described for Step 71-3. 220 mg (41.3%) was obtained as a white solid.

Stage 72-5

A mixture of 3-(2-Chloro-5-methylpyridin-4-yl)-5-cyclopropylisoxazole and 5-(2-chloro-5-methylpyridin-4-yl)-3-cyclopropylisoxazole (190 mg, 96.2%) was prepared according to the method , described for Step 71-4 as a light yellow solid.

Stage 72-6

Methyl 4-(5-cyclopropylisoxazol-3-yl)-5-methylpicolinate (20 mg) and methyl 4-(3-cyclopropylisoxazol-5-yl)-5-methylpicolinate (18 mg) were prepared according to the procedure described for Step 71- 5, as a light yellow solid.

Stage 72-7

Example 72-A was prepared according to the procedure described for Step 29-1. 10 mg was obtained as a light yellow solid. MS (m/z): [M + H] ⁺ = 431.20; ¹ H-NMR (400 MHz, Chloroform-d) δ 10.50 (s, 1H), 8.66 (s, 1H), 8.61 (d, J = 8.3 Hz, 1H), 8.42 (s, 1H), 8.11 (d, J = 7.5 Hz, 1H), 8.01 (t, J = 8.0 Hz, 1H), 6.33 (s, 1H), 5.88 (m, 1H), 2.68 (s, 3H), 2.18 (m, 1H), 1.76 ( d, J = 6.7 Hz, 6H), 1.20 – 1.17 (m, 2H), 1.13 – 1.10 (m, 2H).

Example 72-B was prepared according to the procedure described for Step 29-1. 4.8 mg was obtained as an off-white solid. MS (m/z): [M + H] ⁺ = 431.20. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.46 (s, 1H), 8.68 – 8.59 (m, 2H), 8.57 (s, 1H), 8.11 (d, J = 7.5 Hz, 1H), 8.02 ( t, J = 8.0 Hz, 1H), 6.46 (s, 1H), 5.86 (m, 1H), 2.69 (s, 3H), 2.12 (m, 1H), 1.76 (d, J = 6.7 Hz, 6H), 1.21 – 1.13 (m, 2H), 1.01 – 0.92 (m, 2H).

Example 73

Stage 73-1

Mixture of 1-methylpyrazol-4-ylboronic acid (113 mg, 0.89 mmol), 5-bromo-2-methoxy-4-methylbenzoic acid (200 mg, 0.82 mmol), Pd(PPh ₃ ) ₄ (94 mg, 0.08 mmol) and K ₂ CO ₃ (451 mg, 3.26 mmol) in DME/H ₂ O (5/1 ml) were stirred at 100°C under nitrogen atmosphere overnight. The mixture was diluted with water (10 ml), acidified to pH 4~5 by adding 1M aqueous HCl and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated. The residue was purified by silica gel column chromatography using DCM/MeOH (30:1) as eluent to give 2-methoxy-4-methyl-5-(1-methyl-1H-pyrazol-4-yl)benzoic acid (140 mg, Y = 69.9%) as a yellow oil.

Stage 73-2.

Example 73 was prepared according to the procedure described for Step 71-7. 12 mg was obtained as an off-white solid. MS (m/z): [M+H] ⁺ = 433.20. ¹ H-NMR (400 MHz, DMSO-d ₆ ) δ 10.73 (s, 1H), 8.45 (m, 1H), 8.14 (t, J = 8.0 Hz, 1H), 8.01 – 7.94 (m, 2H), 7.87 (s, 1H), 7.67 (s, 1H), 7.22 (s, 1H), 5.85 (m, 1H), 4.01 (s, 3H), 3.90 (s, 3H), 2.46 (s, 3H), 1.63 ( d, J = 6.6 Hz, 6H).

Example 74

Example 74 was prepared according to the procedure described for Step 71-7. 12 mg was obtained as an off-white solid. MS (m/z) 432.20, 433.20 [M+H] ⁺ ; ¹ H-NMR (400 MHz, DMSO-d6) δ 10.69 (s, 1H), 8.43 (d, J = 8.3 Hz, 1H), 8.21 (t, J = 8.1 Hz, 1H), 7.97 (s, 1H) , 7.86 (s, 1H), 7.70 (d, J = 7.8 Hz, 1H), 7.66 (s, 1H), 7.20 (s, 1H), 3.99 (s, 3H), 3.89 – 3.80 (m, 4H), 2.45 (s, 3H), 1.40 (d, J = 6.9 Hz, 6H).

Example 75

Stage 75-1

A solution of 5-(1-cyclopropyl-1H-pyrazol-4-yl)-2-fluoro-4-methylbenzoic acid (300 mg, 1.2 mmol) and sodium methoxide (15 ml, 30%) was stirred at 80°C overnight . The reaction was quenched with water, acidified to pH 4~5 with 1M aqueous HCl and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure. The residue was purified by silica gel column chromatography using DCM/MeOH (30:1) as eluent to give 5-(1-cyclopropyl-1H-pyrazol-4-yl)-2-methoxy-4-methylbenzoic acid (310 mg , 98.77%) as a light pink solid.

Stage 75-2

Example 75 was prepared according to the procedure described for Step 71-7. 15.3 mg was obtained as an off-white solid. MS (m/z): [M + H] ⁺ = 459.40. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.61 (dd, J = 8.3, 1.1 Hz, 1H), 8.25 (s, 1H), 8.04 (dd, J = 7.6, 1.1 Hz, 1H), 7.97 (t, J = 7.9 Hz, 1H), 7.65 (d, J = 12.6 Hz, 2H), 6.96 (s, 1H), 5.81 (m, 1H), 4.11 (s, 3H), 3.68 (m, 1H), 2.51 (s, 3H), 1.77 (d, J = 6.7 Hz, 6H), 1.24 - 1.21 (m, 2H), 1.18 – 1.05 (m, 2H).

Example 76

Example 76 was prepared according to the procedure described for Step 71-7. 12.6 mg was obtained as an off-white solid. MS (m/z): [M + H] ⁺ = 459.25. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.55 (s, 1H), 8.58 (dd, J = 8.3, 0.7 Hz, 1H), 8.24 (s, 1H), 8.02 (t, J = 8.1 Hz, 1H), 7.69 – 7.60 (m, 3H), 6.96 (s, 1H), 4.10 (s, 3H), 3.92 (m, 1H), 3.68 (m, 1H), 2.51 (s, 3H), 1.57 (d , J = 6.9 Hz, 6H), 1.25 – 1.13 (m, 2H), 1.17 – 1.04 (m, 2H).

Example 77

(1-Chloro-2- methylprop-1-en-1-yl)dimethylamine (30 mg, 0.2 mmol) at room temperature under nitrogen atmosphere. After 2 hours, a solution of 6-[1-[(2R)-1-[(tert-butyldimethylsilyl)oxy]propan-2-yl]-1H-1,2,3,4-tetrazol-5-yl]pyridin- 2-amine (93 mg, 0.3 mmol) and pyridine (44 mg, 0.6 mmol) in DCM (1 ml). The reaction mixture was stirred for 2 hours and conc. HCl (0.4 ml) at room temperature. After 1 hour, the resulting mixture was evaporated under reduced pressure. The residue was purified by reverse phase flash chromatography under the following conditions (column: silica gel C18; mobile phase: acetonitrile in water, gradient from 0 to 100% in 25 minutes; detector: UV 254 nm) to obtain 5-(1-cyclopropyl-1H - pyrazol-4-yl)-N-(6-[1-[(2R)-1-hydroxypropan-2-yl]-1H-1,2,3,4-tetrazol-5-yl]pyridin-2- yl)-2-methoxy-4-methylbenzamide (13 mg, 14.8%) as an off-white solid. MS (m/z): [M + H] ⁺ = 475.25. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.65 (s, 1H), 8.57 (dd, J = 7.1, 2.2 Hz, 1H), 8.23 (s, 1H), 8.04 – 7.94 (m, 2H), 7.66 – 7.59 (m, 2H), 6.95 (s, 1H), 5.68 (m, 1H), 4.16 (d, J = 6.4 Hz, 2H), 4.14 (s, 3H), 3.93 (s, 1H), 3.67 (m, 1H), 2.50 (s, 3H), 1.74 (d, J = 6.9 Hz, 3H), 1.22 – 1.19 (m, 2H), 1.17 – 1.02 (m, 2H).

Example 78

Stage 78-1

Mixture of methyl 2-fluoro-4-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (400 mg, 1.4 mmol), 3-bromo-1- (propan-2-yl)-1H-pyrazole (312 mg, 1.8 mmol), K ₂ CO ₃ (560 mg, 4.1 mmol) and Pd(PPh ₃ ) ₄ (80 mg, 0.1 mmol) in DME/H ₂ O (/5/1 ml) was stirred at 100°C in a nitrogen atmosphere for 1 hour. The resulting mixture was cooled to room temperature, diluted with water (20 ml) and extracted with ethyl acetate (20 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure. The residue was purified by silica gel column chromatography using petroleum ether/EtOAc (20:1) as the eluent to give methyl 2-fluoro-4-methyl-5-[1-(propan-2-yl)-1H-pyrazol- 3-yl] benzoate (173 mg, 46%) as a yellow oil.

Stage 78-2

Into a stirred solution of methyl 2-fluoro-4-methyl-5-[1-(propan-2-yl)-1H-pyrazol-3-yl]benzoate (173 mg, 0.6 mmol) in THF/H ₂ O (3/ 1 ml) LiOH (200 mg, 8.4 mmol) was added and the resulting mixture was stirred at room temperature overnight. The mixture was then acidified to pH 4~5 by adding 1M HCl solution and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure to give 2-fluoro-5-(1-isopropylpyrazol-3-yl)-4-methylbenzoic acid (160 mg) as a gray solid.

Stage 78-3

A mixture of 2-fluoro-5-(1-isopropylpyrazol-3-yl)-4-methylbenzoic acid (160 mg, 0.6 mmol) and 30% sodium methoxide solution in methanol (12 ml) was stirred at 80°C overnight . The mixture was acidified to pH 4~5 by adding 1M HCl solution and extracted with ethyl acetate (10 ml x 3). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and evaporated under reduced pressure. The residue was purified by silica gel column chromatography using DCM/MeOH (20:1) as eluent to give 5-(1-isopropylpyrazol-3-yl)-2-methoxy-4-methylbenzoic acid (162 mg, 96.8%) as a gray solid.

Stage 78-4

Example 78 was prepared according to the procedure described for Step 71-7. MS (m/z): [M + H] ⁺ = 461.35. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.54 (s, 1H), 8.62 (d, J = 8.3 Hz, 1H), 8.49 (s, 1H), 8.03 (d, J = 7.4 Hz, 1H) , 7.96 (t, J = 7.9 Hz, 1H), 7.49 (d, J = 2.3 Hz, 1H), 6.96 (s, 1H), 6.45 (d, J = 2.2 Hz, 1H), 5.83 (m, 1H) , 4.57 (m, 1H), 4.11 (s, 3H), 2.62 (s, 3H), 1.78 (d, J = 6.7 Hz, 6H), 1.59 (d, J = 6.7 Hz, 6H).

Example 79

Example 79 was prepared according to the procedure described for Step 71-7. MS (m/z): [M + H] ⁺ = 461.25. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.53 (s, 1H), 8.60 (d, J = 8.3 Hz, 1H), 8.48 (s, 1H), 8.01 (t, J = 8.1 Hz, 1H) , 7.65 (d, J = 7.7 Hz, 1H), 7.49 (d, J = 2.2 Hz, 1H), 6.95 (s, 1H), 6.44 (d, J = 2.2 Hz, 1H), 4.57 (m, 1H) , 4.10 (s, 3H), 3.93 (m, 1H), 2.62 (s, 3H), 1.59 – 1.56 (m, 12H).

Example 80

Example 80 was obtained as a white solid using a procedure similar to that described in Example 77. MS (m/z): [M + H] ⁺ = 477.25. ¹ H-NMR (400 MHz, Chloroform-d) δ 10.64 (s, 1H), 8.59 (d, J = 7.8 Hz, 1H), 8.47 (s, 1H), 8.02 – 7.93 (m, 2H), 7.49 ( d, J = 2.2 Hz, 1H), 6.94 (s, 1H), 6.44 (d, J = 2.2 Hz, 1H), 5.70 (m, 1H), 4.58 (m, 1H), 4.17 – 4.14 (m, 5H ), 2.61 (s, 3H), 1.75 (d, J = 6.8 Hz, 3H), 1.59 (d, J = 6.6 Hz, 6H).

Example 81

Example 81 was prepared according to the synthesis scheme shown above. [M+H] ⁺ 475.2. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.19 (d, J = 16.9 Hz, 1H), 8.56 (d, J = 8.1 Hz, 1H), 8.15 (d, J = 3.7 Hz, 1H), 8.13 (d, J = 5.0 Hz, 1H), 8.02 (dd, J = 8.0, 8.0 Hz, 1H), 7.65 (s, 1H), 7.64 (s, 1H), 7.14 (d, J = 13.3 Hz, 1H) , 6.09 (ddt, J = 9.6, 6.6, 3.3 Hz, 1H), 4.40-4.34 (m, 2H), 4.22 (dd, J = 9.9, 3.6 Hz, 1H), 4.20-4.13 (m, 1H), 3.72 -3.66 (m, 1H), 2.83-2.76 (m, 1H), 2.68 – 2.54 (m, 1H), 2.50 (s, 3H), 1.25 – 1.18 (m, 2H), 1.16 – 1.05 (m, 2H) .

Example 82

Example 82 was prepared according to the synthesis scheme shown above. [M+H] ⁺ 475.2. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.18 (d, J = 16.9 Hz, 1H), 8.55 (d, J = 8.1 Hz, 1H), 8.14 (d, J = 3.7 Hz, 1H), 8.12 (d, J = 5.0 Hz, 1H), 8.01 (dd, J = 8.0, 8.0 Hz, 1H), 7.65 (s, 1H), 7.64 (s, 1H), 7.14 (d, J = 13.3 Hz, 1H) , 6.09 (ddt, J = 9.6, 6.6, 3.3 Hz, 1H), 4.40-4.34 (m, 2H), 4.22 (dd, J = 9.9, 3.6 Hz, 1H), 4.20-4.13 (m, 1H), 3.72 -3.66 (m, 1H), 2.83-2.76 (m, 1H), 2.68 – 2.54 (m, 1H), 2.50 (s, 3H), 1.25 – 1.18 (m, 2H), 1.16 – 1.05 (m, 2H) .

Example 83

Example 83 was prepared according to the synthesis scheme shown above. [M+H] ⁺ 489.2. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.26 (d, J = 16.4 Hz, 1H), 8.51 (d, J = 8.3 Hz, 1H), 8.17 – 8.08 (m, 2H), 8.03 (dd, J = 8.0, 8.0 Hz, 1H), 7.65 (s, 1H), 7.63 (s, 1H), 7.14 (d, J = 13.1 Hz, 1H), 5.59 (td, J = 7.8, 5.4 Hz, 1H), 4.71 (kv, J = 6.2 Hz, 1H), 3.77-3.69 (m, 1H), 2.50 (s, 3H), 2.48 – 2.40 (m, 2H), 2.38-2.30 (m, 1H), 2.19 – 1.97 ( m, 2H), 1.96 – 1.80 (m, 1H), 1.27 – 1.17 (m, 2H), 1.15 – 1.06 (m, 2H).

Example 84

Example 84 was prepared according to the synthesis scheme shown above. [M+H] ⁺ 489.2. ¹ H-NMR (400 MHz, Chloroform-d) δ 9.17 (d, J = 17.4 Hz, 1H), 8.52 (d, J = 8.3 Hz, 1H), 8.14 (d, J = 8.3 Hz, 1H), 8.12 – 8.08 (m, 1H), 8.03 (dd, J = 8.0, 8.0 Hz, 1H), 7.65 (d, J = 0.8 Hz, 1H), 7.63 (s, 1H), 7.14 (d, J = 13.2 Hz, 1H), 5.64 – 5.49 (m, 1H), 4.64 (kv, J = 5.9 Hz, 1H), 3.73-3.69 (m, 1H), 2.67-2.57 (m, 1H), 2.50 (s, 2H), 2.40 -2.34 (m, 1H), 2.32-2.26 (m, 1H), 2.22 – 2.11 (m, 1H), 2.10-1.96 (m, 1H), 1.90 – 1.78 (m, 1H), 1.26 – 1.18 (m, 2H), 1.14 – 1.06 (m, 2H).

Tests

The ability (IC ₅₀ ) of compounds to inhibit ASK1 kinase activity was determined using the HTRF® KinEASE™ Assay System.

ASK1 was purchased from Thermofisher (Catalogue #PV4011), ATP was purchased from Sigma (Catalogue #A7699), HTRF® KinEASE™ Assay System was obtained from Cisbio (Bedford, Mass). ½ area plates were purchased from Perkin Elmer (Catalogue ##6005560). HTRF® KinEASE™-STK is a general method for measuring serine/threonine kinase activity using a time-resolved fluorescence inductive resonance energy transfer (TR-FRET) immunoassay. The IC ₅₀ value for each compound was determined in the presence of the compound (various concentrations ranging from 0 to 10 μM) and a fixed amount of ATP and peptide substrates. The test compound, 1 μM STK3 peptide substrate and 5 nM ASK1 kinase were incubated with a kinase reaction buffer containing 50 mM HEPES pH 7.5, 0.01% BRIJ-35, 10 mM MgCl ₂ and 1 mM EGTA for 30 minutes. 100 μM ATP was added to initiate the kinase reaction, and the mixture was incubated for 3 hours. Eu ³⁺ -Cryptate-labeled STK3 antibodies and 125 nM streptavidin-XL665 were added in one dose along with a stop reagent from the Cisbio kit to stop the kinase reaction. Fluorescence was measured using an Envision Multilabeled 2014 instrument from PerkinElmer. Fluorescence levels were measured at 615 nm (Cryptat) and 665 nm (XL665) and the 665 nm/615 nm ratio was calculated for each well. The TR-FRET result is proportional to the level of phosphorylation. Staurosporine was used as a positive control. The IC ₅₀ value was determined using the XLfit 5.3 program.

The ASK1 inhibition of compounds having formula (I) was determined by the method described above. The IC ₅₀ value ranges are designated as follows: A < 1 nM; 1 nM < B < 10 nM; 10 nM < C < 100 nM; 100 nM < D < 1 µM; E > 1 µM.

Table 1

Example IC ₅₀ Example IC ₅₀ Example IC ₅₀ 1 C 2 C 3 C 4 C 5 C 6 B 7 B 8 C 9 B 10 B eleven C 12 B 13 B 14 C 15 C 16 B 17 C 18 B 19 B 20 B 21 C 22 B 23 C 24 E 25 B 26 B 27 B 28 B 29 B thirty C 31 E 32 B 33 C 34 E 35 B 36 C 37 B 38 C 39 B 40 C 41 B 42 B 43 B 44 C 45 B 46 B 47 B 48 B 49 B 50 B 51 B 52 B 53 B 54 B 55 C 56 D 57 B 58 C 59 E 60 C 61 C 62 E 63 C 64 D 65 B 66 C 67 C 68 D 69 C 70 B 71 C 72-A/B C 73 B 74 B 75 A 76 A 77 A 78 B 79 C 80 A 81 B 82 B 83 B 84 B

While the present invention has been described and illustrated by reference to particular preferred embodiments thereof, those skilled in the art will appreciate that various changes in form and detail may be made without departing from the spirit and scope of the present invention as described in the Claims. .

Claims (44)

1. Compound of formula I: or a pharmaceutically acceptable salt thereof, wherein selected from the group consisting of represents or ; represents ; R ¹ is selected from the group consisting of the following: 1) hydrogen; 2) -C ₁ -C ₆ alkyl; 3) halogen -C ₁ -C ₆ alkyl; And 4) -C ₃ -C ₆ cycloalkyl; R ² represents X represents N or CR ³ ; R ³ is selected from the group consisting of the following: 1) hydrogen; 2) halogen; 3) -C ₁ -C ₆ alkyl; And 4) -C ₁ -C ₆ alkoxy; R ⁴ is selected from the group consisting of the following: 1) hydrogen; 2) halogen; 3) -C ₁ -C ₆ alkyl; 4) halogen-C ₁ -C ₆ alkyl; And 5) -N(R ⁵ )(R ⁶ ); where R ⁵ and R ⁶ are combined with the nitrogen atom to which they are attached to form a 3-8 membered heterocycloalkyl containing 1 nitrogen atom, 2 nitrogen atoms or a nitrogen atom and an oxygen atom, optionally substituted with a C ₁ -C ₄ -alkyl. 2. A compound according to claim 1, having one of formulas (II-a), (II-c), (II-e) and (IIg), or a pharmaceutically acceptable salt thereof: Where , R ² and R ⁴ correspond to the definitions given in paragraph 1. 3. A compound according to claim 1, having one of the formulas (III-a), (III-b), (III-f), (III-g), (III-h), (III-i), (III -j), (III-m) and (III-n), or a pharmaceutically acceptable salt thereof: where R ¹ , R ² and R ⁴ correspond to the definitions given in paragraph 1. 4. A compound according to claim 1, having one of the formulas (IV-a), (IV-b), (IV-c), (IV-f), (IV-g), (IV-h), (IV -i), (IV-j), (IV-m) and (IV-n), or a pharmaceutically acceptable salt thereof: where R ¹ , R ² and R ⁴ correspond to the definitions given in paragraph 1. 5. A compound according to claim 1, selected from the following compounds, or a pharmaceutically acceptable salt thereof: 6. A pharmaceutical composition for the treatment of an ASK-1 mediated disease or condition selected from the group consisting of an autoimmune disease, a neurodegenerative disease, an inflammatory disease, a chronic kidney disease, a renal disease, a cardiovascular disease, a metabolic disorder, or an acute, or chronic liver disease, containing an effective amount of a compound according to any one of paragraphs. 1-5 and a pharmaceutically acceptable carrier or excipient. 7. Use of a compound according to any one of paragraphs. 1-5 for treating an ASK-1 mediated disease or condition in a subject in need thereof, selected from the group consisting of an autoimmune disease, a neurodegenerative disease, an inflammatory disease, a chronic kidney disease, a kidney disease, a cardiovascular disease, a disorder metabolism or acute or chronic liver disease. 8. The use of claim 7, wherein the ASK-1 mediated disease or condition is a chronic liver disease selected from the group consisting of primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (PSC) , drug-induced cholestasis, intrahepatic cholestasis of pregnancy, parenteral nutrition-induced cholestasis (PNAC), cholestasis caused by microflora overgrowth or sepsis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH ), graft-versus-host disease caused by liver transplantation, liver regeneration in a living liver donor, congenital liver fibrosis, choledocholithiasis, granulomatous liver disease, intra- or extrahepatic malignancy, Sjögren's syndrome, sarcoidosis, Wilson's disease, Gaucher's disease, hemochromatosis or alpha 1-antitrypsin deficiency. 9. The use of claim 7, wherein the ASK-1 mediated disease or condition is a kidney disease selected from the group consisting of diabetic nephropathy, focal segmental glomerulosclerosis (FSGS), hypertensive nephrosclerosis, chronic glomerulonephritis, chronic post-transplant glomerulopathy, chronic interstitial nephritis, renal fibrosis and polycystic kidney disease. 10. Use according to claim 7, wherein the disease or pathological condition mediated by ASK-1 is a cardiovascular disease selected from the group consisting of atherosclerosis, arteriosclerosis, ischemia-reperfusion injury in stroke, cardiac hypertrophy, respiratory diseases, cardiac attacks, myocardial ischemia. 11. Use of the compound according to any one of paragraphs. 1-5 for the treatment of an ASK-1 mediated disease selected from diabetic nephropathy, type II diabetes, rheumatoid arthritis, osteoarthritis, multiple sclerosis, myocardial ischemia, cerebral ischemia, Alzheimer's disease and Parkinson's disease, in a subject in need thereof.