PHD Inhibitor Compounds, Compositions, and Their Uses

Abstract

The present invention provides in part novel small molecule PHD inhibitors having a structure according to formula (A) or a sub-formula thereof: or a pharmaceutically acceptable salt thereof. The compounds provided herein can be used to treat diseases including heart diseases (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease).

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application No. 62 / 992,616, filed on March 20, 2020, which is hereby incorporated by reference in its entirety. Background Art

[0003] Hypoxia is a condition or state in which the supply of oxygen is insufficient for normal life functions, such as where there is a low arterial oxygen supply. Hypoxia can lead to functional damage and structural tissue damage of cells. The activation of cellular defense mechanisms during hypoxia is mediated by HIF (hypoxia - inducible factor) proteins. In response to hypoxic conditions, HIFα levels increase in most cells due to reduced prolyl hydroxylation of HIFα. The prolyl hydroxylation of HIFα is accomplished by a family of proteins variously called proteins containing prolyl hydroxylase domains (PHD1, 2, and 3), also known as HIF prolyl hydroxylases (HPH - 3, 2, and 1) or EGLN - 2, 1, and 3. PHD proteins are oxygen sensors and regulate the stability of HIF in an oxygen - dependent manner. The three PHD isoforms act differently in the regulation of HIF and can have other non - HIF - related regulatory roles.

[0004] In fact, many studies have shown that the stabilization of HIF can attenuate tissue inflammation and promote its repair. Thus, compounds that can inhibit the activity of PHD proteins can be particularly beneficial in new therapies (Lee et al. (2019) Exp. Mol. Med. 51:68).

[0005] Described herein are novel small - molecule PHD inhibitors for treating diseases including heart diseases (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease). Summary of the Invention

[0006] The present invention provides, among other things, novel small - molecule PHD inhibitors and for treating diseases including but not limited to heart diseases (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease).

[0007] In one aspect, provided herein are compounds having a structure according to formula (A),

[0008]

[0009] or a pharmaceutically acceptable salt thereof, wherein:

[0010] Ar 1 is an aryl or heteroaryl optionally substituted with one or more groups selected from: halogen, CN, OH, C 1-3 alkyl optionally substituted with CN or one or more halogens and C 1-3 alkoxy; and

[0011] Ar 2 is pyridin-2-yl optionally substituted with one or more groups selected from: halogen; amino; amide; OH; sulfonyl; sulfinyl; carbonyl; phosphoryl; C 3-6 cycloalkyl; C 3-6 heterocycloalkyl optionally substituted with sulfonyl or ═O; C 1-3 alkyl optionally substituted with carbonyl or one or more halogens; and heteroaryl optionally substituted with C 1-3 alkyl or phenyl.

[0012] In an embodiment, Ar 1 is

[0013] wherein

[0014] X is N or CR 1a ;

[0015] Y and Z are independently CH or N;

[0016] R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted with CN;

[0017] Each occurrence, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens and C 1-3 alkoxy; and

[0018] m is 1, 2, 3 or 4.

[0019] In an embodiment, Ar 1 is

[0020]

[0021] In an embodiment, Ar 1 is

[0022] wherein R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C optionally substituted with CN 1-3 alkyl.

[0023] In the examples, R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C optionally substituted with CN 1-3 alkyl.

[0024] In the examples, each time it is taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 alkyl and C 1-3 alkoxy.

[0025] In the examples, Ar 2 is

[0026] wherein

[0027] each time it is taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl and C 3-6 cycloalkyl;

[0028] R 3 is SO 2 R 6 、SOR 7 R 8 、SOR 9 、COR 10 、(CH 2 ) p COOH、NHR 11 、POR 12 R 13 、halogen, cycloalkyl, heterocycloalkyl optionally substituted with SO 2 R 14 or =O, heteroaryl optionally substituted with C 1-3 alkyl or phenyl, or C optionally substituted with one or more halogens 1-3 alkyl;

[0029] R 6 is C 1-3 alkyl, NHCOR 15 、NR 16 R 17 or phenyl;

[0030] R7 It is C 1-3 Alkyl, C 3-5 Cycloalkyl, phenyl or NR 18 R 19 ;

[0031] R 8 is NH or NCH 3 ;

[0032] R 10 It is C 1-3 Alkyl or NHSO 2 R 20 ;

[0033] R 11 It is COR 21 or SO 2 R 22 ;

[0034] R 9 , R 12 , R 13、 R 14 , R 15 and R 20 Each is independently C 1-3 alkyl;

[0035] R 21 is a heterocycloalkyl, cycloalkyl or C 1-3 alkyl;

[0036] R 22 YesNR 23 R 24 or C optionally substituted by carboxyl 1-3 alkyl;

[0037] R 4 , R 5 , R 16 , R 17 , R 18 , R 19 , R 23 and R 24 Each independently is H or C 1-3 alkyl;

[0038] p is 1, 2, or 3; and

[0039] n is 0, 1, 2 or 3.

[0040] In the embodiment, Ar 2 yes Where R 3 Selected from the group consisting of: F, Cl, Br and I.

[0041] In the embodiment, Ar2 is wherein R 11 is COR 21 or SO 2 R 22 ; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with carboxyl; and R 23 and R 24 are independently H or C 1-3 alkyl.

[0042] In an embodiment, Ar 2 is wherein R 3 is cycloalkyl or heterocycloalkyl optionally substituted with SO 2 R 14 or =O; and R 14 is C 1-3 alkyl.

[0043] In an embodiment, Ar 2 is wherein R 3 is heteroaryl optionally substituted with C 1-3 alkyl or phenyl.

[0044] In an embodiment, the cycloalkyl or optionally substituted heterocycloalkyl is selected from the group consisting of:

[0045] In an embodiment, the optionally substituted heteroaryl is selected from the group consisting of:

[0046] In an embodiment, each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl and C 3-6 cycloalkyl, wherein R 4 and R 5 are each independently H or C 1-3 alkyl.

[0047] In an embodiment, R 3 is SO 2 R 6 , SOR 7 R 8 , SOR 9, COR 10 , (CH 2 ) p COOH, NHR 11 , POR 12 R 13 , halogen, cycloalkyl, heterocycloalkyl optionally substituted by SO 2 R 14 or =O, heteroaryl optionally substituted by C 1-3 alkyl or phenyl, or C 1-3 alkyl optionally substituted by one or more halogens, wherein R 6 is C 1-3 alkyl, NHCOR 15 , NR 16 R 17 or phenyl; R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ; R 8 is NH or NCH 3 ; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; R 11 is COR 21 or SO 2 R 22 ; R 9 , R 12 , R 13、 R 14 , R 15 and R 20 each independently is C 1-3 alkyl; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted by carboxyl; R 4 , R 5 , R 16 , R 17 , R 18 , R 19 , R 23 and R 24 each independently is H or C 1-3 alkyl; and p is 1, 2 or 3.

[0048] In the examples, the compound of formula (A) has the following structure,

[0049] or a pharmaceutically acceptable salt thereof.

[0050] In embodiments of formula (I), X is N or CR 1a ; Y and Z are independently CH or N; each occurrence, R 1 is independently selected from the group consisting of hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens, and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH, or C 1-3 alkyl optionally substituted with CN; each occurrence, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl, and C 3-6 cycloalkyl; R 3 is SO 2 R 6 , SOR 7 R 8 , SOR 9 , COR 10 , (CH 2 ) p COOH, NHR 11 , POR 12 R 13 , halogen, cycloalkyl, heterocycloalkyl optionally substituted with SO 2 R 14 or =O, heteroaryl optionally substituted with C 1-3 alkyl or phenyl, or C 1-3 alkyl optionally substituted with one or more halogens; R 4 and R 5 are each independently H or C 1-3 alkyl; R 6 is C 1-3 alkyl, NHCOR 15 , NR 16 R 17 , or phenyl; R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl, or NR 18 R 19 ; R 8 is NH or NCH 3 ; R 9 is C 1-3 alkyl; R 10 is C 1-3 alkyl or NHSO 2R 20 ; R 11 is COR 21 or SO 2 R 22 ; R 12 and R 13 are each independently C 1-3 alkyl; R 14 is C 1-3 alkyl; R 15 is C 1-3 alkyl; R 16 and R 17 are each independently H or C 1-3 alkyl; R 18 and R 19 are each independently H or C 1-3 alkyl; R 20 is C 1-3 alkyl; R 21 is heteroalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or optionally carboxyl-substituted C 1-3 alkyl; R 23 and R 24 are each independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; n is 0, 1, 2 or 3; and p is 1, 2 or 3.

[0051] In an embodiment, the compound of formula (A) or formula (I) has the following structure,

[0052] or a pharmaceutically acceptable salt thereof.

[0053] In an embodiment, X is N or CR 1a ; Z is CH or N; each time taken, R 1 is independently selected from the group consisting of hydrogen, halogen, CN, OH, optionally one or more halogen-substituted C 1-3 alkyl and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or optionally CN-substituted C 1-3 alkyl; each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 、OH、C 1-3 alkyl and C 3-6 cycloalkyl; R 3 is SO 2 R 6, SOR 7 R 8 , SOR 9 , COR 10 , (CH 2 ) p COOH, NHR 11 , POR 12 R 13 , halogen, cycloalkyl, heterocycloalkyl optionally substituted by SO 2 R 14 or =O, heteroaryl optionally substituted by C 1-3 alkyl or phenyl, or C 1-3 alkyl optionally substituted by one or more halogens; R 4 and R 5 are each independently H or C 1-3 alkyl; R 6 is C 1-3 alkyl, NHCOR 15 , NR 16 R 17 or phenyl; R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ; R 8 is NH or NCH 3 ; R 9 is C 1-3 alkyl; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; R 11 is COR 21 or SO 2 R 22 ; R 12 and R 13 are each independently C 1-3 alkyl; R 14 is C 1-3 alkyl; R 15 is C 1-3 alkyl; R 16 and R 17 are each independently H or C 1-3 alkyl; R 18 and R 19 are independently H or C 1-3 alkyl; R 20 is C 1-3 alkyl; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R24 or a C alkyl optionally substituted with a carboxyl group; R 1-3 alkyl; R 23 and R 24 are independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; n is 0, 1, 2 or 3; and p is 1, 2 or 3.

[0054] In the examples, the compounds of formula (A), formula (I) or formula (II) have the following structure,

[0055] or a pharmaceutically acceptable salt thereof.

[0056] In the examples of formula (III), each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, a C alkyl optionally substituted with one or more halogens and C 1-3 alkoxy; R 1-3 is H, CN, halogen, C 1a alkoxy, OH or a C alkyl optionally substituted with CN; each time taken, R 1-3 is independently selected from the group consisting of: hydrogen, halogen, NR 1-3 R 2 , OH, C 4 alkyl and C 5 cycloalkyl; R 1-3 is SO 3-6 R 3 is SO 2 R 6 is SOR 7 R 8 is SOR 9 is COR 10 is (CH 2 ) p COOH, NHR 11 is POR 12 R 13 is halogen, cycloalkyl, a heterocycloalkyl optionally substituted with SO 2 R 14 or =O, a heteroaryl optionally substituted with C 1-3 alkyl or phenyl or a C alkyl optionally substituted with one or more halogens; R 1-3 alkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 6 is C 1-3 alkyl, NHCOR 15 is NR 16 R 17 or phenyl; R 7 is C1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ; R 8 is NH or NCH 3 ; R 9 is C 1-3 alkyl; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; R 11 is COR 21 or SO 2 R 22 ; R 12 and R 13 are each independently C 1-3 alkyl; R 14 is C 1-3 alkyl; R 15 is C 1-3 alkyl; R 16 and R 17 are independently H or C 1-3 alkyl; R 18 and R 19 are independently H or C 1-3 alkyl; R 20 is C 1-3 alkyl; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or optionally carboxyl-substituted C 1-3 alkyl; R 23 and R 24 are independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; n is 0, 1, 2 or 3; and p is 1, 2 or 3.

[0057] In the examples, the compounds of formula (A), formula (I), formula (II) or formula (III) have the following structure,

[0058] or a pharmaceutically acceptable salt thereof.

[0059] In the examples of formula (IV), each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens and C 1-3 alkoxy; R 1a is H, CN, halogen, C1-3 alkoxy, OH or C1-3 Alkyl; each time taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ; R 8 is NH or NCH 3 ; R 18 and R 19 are each independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0060] In an embodiment. R 1 is C 1-3 alkyl. In an embodiment, R 1 is CH 3 .

[0061] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (IV) has the following structure,

[0062] or a pharmaceutically acceptable salt thereof.

[0063] In an embodiment, R 1a is CN or halogen; R 2 is selected from the group consisting of hydrogen or C 1-3 alkyl; R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ; R 8 is NH or NCH 3 ; and R 18 and R 19 are each independently H or C 1-3 alkyl.

[0064] In an embodiment, R 1a is CN.

[0065] In an embodiment, R 1a is halogen. In an embodiment, R 1a is Cl.

[0066] In an embodiment, R 2 is C 1-3 alkyl.

[0067] In an embodiment, R 2 is CH 3 .

[0068] In an embodiment, R 7 is C 1-3 alkyl. In an embodiment, R 7 is CH 3 . In an embodiment, R 7 is CH 2 CH 3 . In an embodiment, R 7 is CH(CH 3 ) 2 . In an embodiment, R 7 is C 3-5 cycloalkyl. In an embodiment, R 7 is cyclopropyl. In an embodiment, R 7 is cyclopentyl. In an embodiment, R 7 is phenyl. In an embodiment, R 7 is NR 18 R 19 , and wherein R 18 and R 19 are each independently H or C 1-3 alkyl.

[0069] In an embodiment, R 18 and R 19 are independently H. In an embodiment, R 18 is H and R 19 is C 1-3 alkyl. In an embodiment, R 19 is CH 3 . In an embodiment, R 18 and R 19 are independently CH 3 .

[0070] In an embodiment, R 8 is NH. In an embodiment, R 8 is NCH 3 .

[0071] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0072] or a pharmaceutically acceptable salt thereof.

[0073] In an embodiment, X is N or CR1a ; Z is N or CH; each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted by one or more halogens and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted by CN; each time taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 6 is C 1-3 alkyl, NHCOR 15 , NR 16 R 17 or phenyl; R 15 is C 1-3 alkyl; R 16 and R 17 are each independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0074] In an embodiment, X is N. In an embodiment, X is CR 1a .

[0075] In an embodiment, R 1a is CN. In an embodiment, R 1a is halogen. In an embodiment, R 1a is Cl. In an embodiment, R 1a is F. In an embodiment, R 1a is Br. In an embodiment, R 1a is C 1-3 alkoxy.

[0076] In an embodiment, R 1a is methoxy. In an embodiment, R 1a is H. In an embodiment, R 1a is C 1-3 alkyl optionally substituted by CN. In an embodiment, R 1a is CH 2 CN. In an embodiment, R 1a is OH.

[0077] In an embodiment, Z is CH. In an embodiment, Z is N.

[0078] In an embodiment, R 1 is H. In an embodiment, R 1 is C 1-3 alkyl. In an embodiment, R 1 is CH 3 . In an embodiment, R 1 is C 1-3 alkoxy. In an embodiment, R 1 is methoxy. In an embodiment, R 1 is CN.

[0079] In an embodiment, R 2 is H. In an embodiment, R 2 is C 1-3 alkyl. In an embodiment, R 2 is CH 3 .

[0080] In an embodiment, R 6 is C 1-3 alkyl. In an embodiment, R 6 is CH 3 . In an embodiment, R 6 is NHCOR 15 , and wherein R 15 is C 1-3 alkyl. In an embodiment, R 15 is CH 3 . In an embodiment, R 6 is NR 16 R 17 , and wherein R 16 and R 17 are each independently H or C 1-3 alkyl. In an embodiment, R 6 is NH 2 . In an embodiment, R 6 is phenyl.

[0081] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0082] or a pharmaceutically acceptable salt thereof, wherein R 3 is cycloalkyl or a heterocycloalkyl optionally substituted with SO 2 R 14 or ═O.

[0083] In an embodiment, each time taken, R 1Independently selected from the group consisting of: hydrogen, halogen, CN, OH, C alkyl optionally substituted with one or more halogens and C alkoxy; R is H, CN, halogen, C alkoxy, OH or C alkyl optionally substituted with CN; each time taken, R is independently selected from the group consisting of: hydrogen, halogen, NR R, OH, C alkyl and C cycloalkyl; R and R are each independently H or C alkyl; R is C alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3. 1-3 alkyl and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted with CN; each time taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 14 is C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0084] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VI) has the following structure,

[0085] or a pharmaceutically acceptable salt thereof, wherein R 3 is cycloalkyl or heterocycloalkyl optionally substituted with SO 2 R 14 or =O.

[0086] In an embodiment, R 2 is hydrogen or C 1-3 alkyl; and R 14 is C 1-3 alkyl.

[0087] In an embodiment, R 2 is H. In an embodiment, R 2 is C 1-3 alkyl. In an embodiment, R 2 is CH 3 .

[0088] In an embodiment, R 3 is cycloalkyl.

[0089] In an embodiment, R 3 is cyclopropyl.

[0090] In an embodiment, R 3 is heterocycloalkyl optionally substituted with SO 2 R 14 or =O, and wherein R 14 is C 1-3 alkyl.

[0091] In an embodiment, R 3 is

[0092] In an embodiment, R 3 is

[0093] In an embodiment, R 3 is

[0094] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0095] or a pharmaceutically acceptable salt thereof.

[0096] In an embodiment, each time taken, R 1 is independently selected from the group consisting of hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens, and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted with CN; each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 、OH、C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 11 is COR 21 or SO 2 R 22 ; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with carboxyl; R 23 and R 24 are independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0097] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VII) has the following structure,

[0098] or a pharmaceutically acceptable salt thereof.

[0099] In the examples, R 2 is hydrogen or C 3-6 cycloalkyl; R 11 is COR 21 or SO 2 R 22 ; R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl; and R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with carboxyl, wherein R 23 and R 24 are independently H or C 1-3 alkyl.

[0100] In the examples, R 2 is H. In the examples, R 2 is C 1-3 alkyl. In the examples, R 2 is CH 3 .

[0101] In the examples, R 11 is COR 21 , and wherein R 21 is heterocycloalkyl, cycloalkyl or C 1-3 alkyl.

[0102] In the examples, R 21 is heterocycloalkyl. In the examples, R 21 is In the examples, R 21 is In the examples, R 21 is cycloalkyl. In the examples, R 21 is cyclopropyl. In the examples, R 21 is C 1-3 alkyl. In the examples, R 21 is CH 2 CH 3 .

[0103] In the examples, R 11 is SO 2 R 22 , wherein R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with carboxyl, and wherein R 23 and R 24Independently is H or C 1-3 alkyl group.

[0104] In an embodiment, R 22 is C 1-3 alkyl group optionally substituted by carboxyl group. In an embodiment, R 22 is CH 3 . In an embodiment, R 22 is CH 2 CH 3 . In an embodiment, R 22 is CH 2 COOH. In an embodiment, R 22 is NR 23 R 24 , and wherein R 23 and R 24 are each independently H or C 1-3 alkyl group. In an embodiment, R 22 is NHCH 3 . In an embodiment, R 22 is N(CH 3 ) 2 .

[0105] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0106] or a pharmaceutically acceptable salt thereof, wherein R 3 is a heteroaryl group optionally substituted by C 1-3 alkyl group or phenyl group.

[0107] In an embodiment, each time taken, R 1 is independently selected from the group consisting of hydrogen, halogen, CN, OH, C 1-3 alkyl group optionally substituted by one or more halogen atoms and C 1-3 alkoxy group; R 1a is H, CN, halogen, C 1-3 alkoxy group, OH or C 1-3 alkyl group optionally substituted by CN; each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl group and C 3-6 cycloalkyl group; R 4 and R 5 are each independently H or C 1-3 alkyl group; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0108] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VIII) has the following structure,

[0109] or a pharmaceutically acceptable salt thereof, wherein R 3 is a heteroaryl optionally substituted by C 1-3 alkyl or phenyl.

[0110] In an embodiment, R 2 is H.

[0111] In an embodiment, R 3 is a heteroaryl. In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is a heteroaryl optionally substituted by C 1-3 alkyl or phenyl. In an embodiment, R 3 is In an embodiment, R 3 is In an embodiment, R 3 is

[0112] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0113] or a pharmaceutically acceptable salt thereof.

[0114] In an embodiment, each time taken, R 1 is independently selected from the group consisting of hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted by one or more halogens, and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH, or C 1-3 alkyl optionally substituted by CN; each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 、OH、C 1-3 alkyl and C3-6 Naphthenyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; R 20 is C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0115] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (IX) has the following structure,

[0116] or a pharmaceutically acceptable salt thereof.

[0117] In an embodiment, R 1a is CN or a halogen; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; and R 20 is C 1-3 alkyl.

[0118] In an embodiment, R 1a is CN. In an embodiment, R 1a is a halogen. In an embodiment, R 1a is Cl.

[0119] In an embodiment, R 10 is C 1-3 alkyl. In an embodiment, R 10 is CH 3 . In an embodiment, R 10 is CH(CH 3 ) 2 . In an embodiment, R 10 is CH 2 CH 3 . In an embodiment, R 10 is NHSO 2 R 20 , and wherein R 20 is C 1-3 alkyl. In an embodiment, R 20 is CH 3 .

[0120] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0121] or a pharmaceutically acceptable salt thereof.

[0122] In an embodiment, each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens, and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH, or C 1-3 alkyl optionally substituted with CN; each time taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl, and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 9 is C 1-3 alkyl; m is 1, 2, 3, or 4; and n is 0, 1, 2, or 3.

[0123] In an embodiment, R 1a is CN.

[0124] In an embodiment, R 1 is H.

[0125] In an embodiment, R 2 is H.

[0126] In an embodiment, R 9 is C 1-3 alkyl. In an embodiment, R 9 is CH 3 .

[0127] In an embodiment, the compound of formula (A), formula (I), formula (II), or formula (III) has the following structure,

[0128] or a pharmaceutically acceptable salt thereof.

[0129] In an embodiment, each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens, and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH, or C 1-3 alkyl optionally substituted with CN; each time taken, R 2Independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; n is 0, 1, 2 or 3; and p is 1, 2 or 3.

[0130] In an embodiment, R 1a is CN.

[0131] In an embodiment, R 1 is H.

[0132] In an embodiment, R 2 is H.

[0133] In an embodiment, p is 1.

[0134] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0135] or a pharmaceutically acceptable salt thereof, wherein R 3 is halogen.

[0136] In an embodiment, each time taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted with CN; each time taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; m is 1, 2, 3 or 4; and n is 0, 1, 2 or 3.

[0137] In an embodiment, R 1a is CN.

[0138] In an embodiment, R 1 is H.

[0139] In an embodiment, R2 It is H.

[0140] In the embodiment, R 3 is Cl. In the embodiment, R 3 is Br. In the embodiment, R 3 is F.

[0141] In the embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0142] or a pharmaceutically acceptable salt thereof.

[0143] In the embodiment, each time it is taken, R 1 is independently selected from the group consisting of: hydrogen, halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens and C 1-3 alkoxy; R 1a is H, CN, halogen, C 1-3 alkoxy, OH or C 1-3 alkyl optionally substituted with CN; and each time it is taken, R 2 is independently selected from the group consisting of: hydrogen, halogen, NR 4 R 5 , OH, C 1-3 alkyl and C 3-6 cycloalkyl; R 4 and R 5 are each independently H or C 1-3 alkyl; R 12 is C 1-3 alkyl; R 13 is C 1-3 alkyl; and m is 1, 2, 3 or 4.

[0144] In the embodiment, R 1a is CN.

[0145] In the embodiment, R 1 is H.

[0146] In the embodiment, R 2 is C 1-3 alkyl. In the embodiment, R 2 is CH 3 .

[0147] In the embodiment, R 12 is C 1-3 alkyl. In the embodiment, R 12 is CH 3 .

[0148] In the embodiment, R13 is C 1-3 alkyl. In an embodiment, R 13 is CH 3 .

[0149] In some embodiments, the compound is any one of Compounds 1 to 33:

[0150]

[0151]

[0152]

[0153] In an embodiment, in Compounds of Formula (A) and (I) to (XIII), at least one hydrogen atom is replaced by a deuterium atom, such as any one of Compounds 1 to 33.

[0154] On the other hand, the present invention features a pharmaceutical composition comprising any of the compounds described herein (e.g., Compounds of Formula (A) and (I) to (XIII), such as any one of Compounds 1 to 33) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0155] On the other hand, the present invention features a method for treating a disease mediated by PHD activity, the method comprising administering to a subject any of the compounds described herein (e.g., Compounds of Formula (A) and (I) to (XIII), such as any one of Compounds 1 to 33) or a pharmaceutically acceptable salt thereof.

[0156] In an embodiment, the disease mediated by PHD activity is ischemia-reperfusion injury (e.g., stroke, myocardial infarction, or acute kidney injury).

[0157] In an embodiment, the disease mediated by PHD activity is inflammatory bowel disease (e.g., ulcerative colitis or Crohn's disease).

[0158] In an embodiment, the disease mediated by PHD activity is cancer (e.g., colorectal cancer).

[0159] In an embodiment, the disease mediated by PHD activity is liver disease.

[0160] In an embodiment, the disease mediated by PHD activity is atherosclerosis.

[0161] In an embodiment, the disease mediated by PHD activity is cardiovascular disease.

[0162] In embodiments, diseases mediated by PHD activity are diseases or conditions of the eye (e.g., radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia).

[0163] In embodiments, diseases mediated by PHD activity are anemia (e.g., anemia associated with chronic kidney disease).

[0164] In embodiments, diseases mediated by PHD activity are hyperoxia.

[0165] In embodiments, diseases mediated by PHD activity are retinopathy of prematurity.

[0166] In embodiments, diseases mediated by PHD activity are bronchopulmonary dysplasia (BPD).

[0167] In embodiments, diseases mediated by PHD activity are ischemic heart disease, valvular heart disease, congestive heart failure, acute lung injury, pulmonary fibrosis, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), acute liver failure, liver fibrosis, or cirrhosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0168] Figure 1 is an exemplary illustration showing the principle of the TR-FRET assay for PHD enzymes (PHD1, PHD2, and PHD3). In the presence of 2-oxoglutarate and O 2 the PHD enzyme hydroxylates proline 564 of the biotinylated HIF-1α peptide, resulting in biotinylated HIF-1α-hydroxyproline, succinate, and CO 2 . The resulting proximity of the donor fluorophore complex (i.e., monoclonal antibody anti-6His-terbium (Tb)-cryptate Gold) bound to the His-tagged VHL protein / EloB / EloC complex (His-VBC) and the acceptor fluorophore (i.e., SA-D2 complex) bound to HIF-1α-hydroxyproline generates a fluorescence resonance energy transfer signal that can be detected and quantified. DETAILED DESCRIPTION

[0169] Definitions

[0170] To facilitate a better understanding of the present invention, certain terms are first defined below. Additional definitions of the following terms and other terms are set forth throughout the specification. Publications and other reference materials cited herein to describe the background art of the present invention and to provide additional details regarding its implementation are incorporated herein by reference.

[0171] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to a human at any stage of development. In some embodiments, "animal" refers to a non-human animal at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, and / or pig). In some embodiments, animals include but are not limited to mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, the animal can be a transgenic animal, a genetically engineered animal, and / or a clone.

[0172] About or approximately: As used herein, when applied to one or more target values, the term "about" or "approximately" refers to a value similar to the reference value. In certain embodiments, the term "about" or "approximately" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater than or less than) of the stated value, unless otherwise specified or otherwise apparent from the context (unless the number exceeds 100% of the possible value).

[0173] As used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly indicates otherwise. Thus, for example, reference to "a composition" includes a mixture of two or more such compositions.

[0174] Throughout the specification and claims of this specification, the word "comprise" and other forms thereof, such as "comprising" and "comprises", mean "including but not limited to", and are not intended to exclude, for example, other additives, components, integers, or steps.

[0175] "Optional" or "optionally" means that the subsequent described event or circumstance may or may not occur, and the description includes instances where the event or circumstance occurs and instances where the event or circumstance does not occur.

[0176] Improve, increase, or decrease: As used herein, the terms “improve,” “increase,” or “decrease” or their grammatical equivalents refer to a value relative to a baseline measurement, which is a measurement in the same individual before initiation of the treatment described herein, or in a control subject (or control subjects) in the absence of the treatment described herein. A “control subject” is a subject having the same form of disease as the subject being treated, and the subject is approximately the same age as the subject being treated.

[0177] In vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, such as in a test tube or reaction vessel, in cell culture, etc., and not within a multicellular organism.

[0178] In vivo: As used herein, the term “in vivo” refers to events that occur within a multicellular organism such as a human and non-human animals. In the context of a cell-based system, the term can be used to refer to events that occur within a living cell (e.g., as opposed to an in vitro system).

[0179] Patient: As used herein, the term “patient” or “subject” refers to any organism to which the provided composition can be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is a human. Humans include prenatal and postnatal forms.

[0180] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” refers to a substance that, within the scope of sound medical judgment, is commensurate with a reasonable benefit / risk ratio and is suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications.

[0181] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, S.M. Berge et al. described pharmaceutically acceptable salts in detail in Journal of Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipates, alginates, ascorbates, aspartates, benzenesulfonates, benzoates, bisulfates, borates, butyrates, camphorates, camphorsulfonates, citrates, cyclopentanepropionates, digluconates, dodecyl sulfates, ethanesulfonates, formates, fumarates, glucoheptanoates, glycerophosphates, gluconates, hemisulfates, heptanoates, hexanoates, hydroiodides, 2-hydroxyethanesulfonates, lactates, lactobionates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, pamoates, pectates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts and N+(C1-4 alkyl)4 salts. Representative alkali metal salts or alkaline earth metal salts include sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, etc. In appropriate cases, additional pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates and arylsulfonates. Additional pharmaceutically acceptable salts include salts formed by quaternization of amines, which quaternization is carried out using appropriate electrophiles (e.g., alkyl halides) to form quaternized alkylated amino salts.

[0182] Subject: As used herein, the term "subject" refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cow, pig, sheep, horse or primate). Humans include prenatal and postnatal forms. In many embodiments, the subject is a human. A subject can be a patient, which refers to a human presenting to a healthcare provider for diagnosis or treatment of a disease. The term "subject" is used interchangeably herein with "individual" or "patient". A subject can have or be predisposed to a disease or disorder, but may or may not exhibit symptoms of the disease or disorder.

[0183] Basically: As used herein, the term "basically" refers to a qualitative condition that exhibits all or nearly all of the range or degree of a characteristic or property of interest. Those of ordinary skill in the biological arts will appreciate that biological and chemical phenomena rarely, if ever, achieve complete and / or proceed to completion or realization or avoid an absolute result. Thus, the term "basically" is used herein to obtain the inherent completeness that is potentially lacking in many biological and chemical phenomena.

[0184] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" of a therapeutic agent refers to an amount that is sufficient to treat, diagnose, prevent, and / or delay the onset of symptoms of a disease, disorder, and / or condition when administered to a subject suffering from or susceptible to the disease, disorder, and / or condition. Those of ordinary skill in the art will recognize that a therapeutically effective amount is typically administered via a dosing regimen that includes at least one unit dose.

[0185] Treatment: As used herein, the term "treatment" refers to any method used to partially or completely relieve, ameliorate, mitigate, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a particular disease, disorder, and / or condition. To reduce the risk of developing a pathology associated with a disease, treatment may be administered to a subject who does not exhibit signs of the disease and / or who exhibits only early signs of the disease.

[0186] Aliphatic: As used herein, the term aliphatic refers to C 1 -C 40 hydrocarbons and includes saturated and unsaturated hydrocarbons. Aliphatic can be straight-chain, branched-chain, or cyclic. For example, C 1 -C 20 Aliphatic can contain C 1 -C 20 alkyl (e.g., straight-chain or branched-chain C 1 -C 20 saturated alkyl), C 2 -C 20 alkenyl (e.g., straight-chain or branched-chain C 4 -C 20 dienyl, straight-chain or branched-chain C 6 -C 20 trienyl, etc.) and C 2 -C 20 alkynyl (e.g., straight-chain or branched-chain C 2 -C 20 alkynyl). C 1 -C 20 Aliphatic can contain C 3 -C 20 cycloaliphatic (e.g., C 3 -C 20Naphthenyl, C 4 -C 20 cycloalkenyl or C 8 -C 20 cycloalkynyl). In certain embodiments, the aliphatic may include one or more cycloaliphatic and / or one or more heteroatoms such as oxygen, nitrogen or sulfur, and may optionally be substituted with one or more substituents such as alkyl, halogen, alkoxy, hydroxy, amino, aryl, ether, ester or amide. The aliphatic group is unsubstituted or substituted with one or more substituents as described herein. For example, the aliphatic may be substituted with one or more of the following: (e.g., 1, 2, 3, 4, 5 or 6 independently selected substituents) halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 , -NHR', -N(R') 2 , -SR' or -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted alkyl (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted C 1 -C 3 alkyl. In some embodiments, the aliphatic is unsubstituted. In some embodiments, the aliphatic does not contain any heteroatoms.

[0187] Alkyl: As used herein, the term "alkyl" means acyclic straight-chain and branched hydrocarbon groups, e.g., "C 1 -C 20"Alkyl" refers to an alkyl group having from 1 to 20 carbons. The alkyl group can be straight-chain or branched-chain. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl, hexyl, isohexyl, and the like. The term "lower alkyl" means a straight-chain or branched-chain alkyl group having from 1 to 6 carbon atoms. Considering the beneficial effects of the present disclosure, other alkyl groups will be apparent to those skilled in the art. The alkyl group can be unsubstituted or substituted with one or more substituents as described herein. For example, the alkyl group can be substituted with one or more of the following: (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 , -NHR', -N(R') 2 , -SR' or -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently an unsubstituted alkyl group (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted C 1 -C 3 alkyl. In some embodiments, the alkyl group is substituted (e.g., 1, 2, 3, 4, 5, or 6 substituents as described herein). In some embodiments, the alkyl group is substituted with an -OH group and can also be referred to herein as "hydroxyalkyl", where the prefix represents the -OH group and "alkyl" is as described herein. In some embodiments, the alkyl group is substituted with an -OR' group and can also be referred to herein as "alkoxy".

[0188] Attaching the suffix "-ene" to a group indicates that the group is a divalent moiety. For example, arylene is the divalent moiety of an aryl group, and heteroarylene is the divalent moiety of a heteroaryl group.

[0189] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent straight-chain or branched-chain hydrocarbon group, and examples thereof include methylene, ethylene, isopropylidene, and the like. Similarly, as used herein, the term "alkenylene" refers to an unsaturated divalent straight-chain or branched-chain hydrocarbon group having one or more unsaturated carbon-carbon double bonds, which may be present at any stable point along the chain, and the term "alkynylene" as used herein refers to an unsaturated divalent straight-chain or branched-chain hydrocarbon group having one or more unsaturated carbon-carbon triple bonds, which may be present at any stable point along the chain. In certain embodiments, the alkylene, alkenylene, or alkynylene may include one or more cycloaliphatic groups and / or one or more heteroatoms such as oxygen, nitrogen, or sulfur, and may optionally be substituted by one or more substituents such as alkyl, halogen, alkoxy, hydroxy, amino, aryl, ether, ester, or amide. For example, the alkylene, alkenylene, or alkynylene may be substituted by one or more of the following: (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 2, -NHR', -N(R') 2 2, -SR' or -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently an unsubstituted alkyl (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted C 1 -C 3 alkyl. In certain embodiments, the alkylene, alkenylene, or alkynylene is unsubstituted. In certain embodiments, the alkylene, alkenylene, or alkynylene does not contain any heteroatoms.

[0190] Alkenyl: As used herein, "alkenyl" means any straight or branched hydrocarbon chain having one or more unsaturated carbon-carbon double bonds, which can be present at any stable point along the chain. For example, "C 2 -C 20 alkenyl" refers to an alkenyl group having 2 to 20 carbons. For example, alkenyl includes prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, hex-5-enyl, 2,3-dimethylbut-2-enyl, etc. In some embodiments, alkenyl includes 1, 2 or 3 carbon-carbon double bonds. In some embodiments, alkenyl includes a single carbon-carbon double bond. In some embodiments, multiple double bonds (e.g., 2 or 3) are conjugated. Alkenyl can be unsubstituted or substituted with one or more of the substituents described herein. For example, alkenyl can be substituted with one or more of the following: (e.g., 1, 2, 3, 4, 5 or 6 independently selected substituents) halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 , -NHR', -N(R') 2 , -SR' or -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted alkyl (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted C 1 -C 3 alkyl. In some embodiments, alkenyl is unsubstituted. In some embodiments, alkenyl is substituted (e.g., 1, 2, 3, 4, 5 or 6 substituents as described herein). In some embodiments, alkenyl is substituted with an -OH group and can also be referred to herein as "hydroxyalkenyl", where the prefix indicates the -OH group and "alkenyl" is as described herein.

[0191] Alkynyl: As used herein, "alkynyl" means any hydrocarbon chain in a straight-chain configuration or a branched-chain configuration that has one or more carbon-carbon triple bonds at any stable point along the chain. For example, "C 2 -C 20 alkynyl" refers to an alkynyl group having 2 to 20 carbons. Examples of alkynyls include prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl, hex-5-ynyl, etc. In some embodiments, the alkynyl includes one carbon-carbon triple bond. The alkynyl can be unsubstituted or substituted with one or more substituents as described herein. For example, the alkynyl can be substituted with one or more of the following: (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 , -NHR', -N(R') 2 , -SR' or -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted alkyl (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some embodiments, R' is independently unsubstituted C 1 -C 3 alkyl. In some embodiments, the alkynyl is unsubstituted. In some embodiments, the alkynyl is substituted (e.g., 1, 2, 3, 4, 5, or 6 substituents as described herein).

[0192] Aryl: The term "aryl", used alone or as part of a larger moiety such as "aralkyl", refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system having a total of six to fourteen ring members, wherein the ring system has a single point of attachment to the remainder of the molecule, at least one of the rings in the system is aromatic, and each ring in the system contains 4 to 7 ring members. In some embodiments, aryl has 6 ring carbon atoms ("C 6 aryl", e.g., phenyl). In some embodiments, aryl has 10 ring carbon atoms ("C 10 aryl", e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, aryl has 14 ring carbon atoms ("C 14 aryl", e.g., anthracenyl). "Aryl" also encompasses ring systems in which the aromatic ring as defined above is fused to one or more carbocyclic or heterocyclic groups, where the linking group or point of attachment is on the aromatic ring, and in such cases, the number of carbon atoms continues to represent the number of carbon atoms in the aromatic ring system. Exemplary aryls include phenyl, naphthyl, and anthracene.

[0193] Arylene: As used herein, the term "arylene" refers to a divalent aryl (i.e., having two points of attachment to the molecule). Exemplary arylenes include phenylene (e.g., unsubstituted phenylene or substituted phenylene).

[0194] Halogen or halo group: As used herein, the term "halogen" or "halo group" means fluorine, chlorine, bromine, or iodine.

[0195] Amide: The term "amide" or "amide group" refers to a chemical moiety having the formula -C(O)N(R') 2 , -C(O)N(R')-, -NR'C(O)R', -NR'C(O)N(R') 2 -, or -NR'C(O)-, wherein each R' is independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, aralkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, or heterocycloalkyl (bonded through a ring carbon), and each moiety as such can be optionally substituted as described herein, unless otherwise specified in the specification, or two R's can combine with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring.

[0196] Amino: The term "amino" or "amine" refers to -N(R') 2a group, wherein each R' is independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, aralkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), sulfonyl, amide or carbonyl, and unless otherwise specified in the specification, each moiety in said moieties may itself be optionally substituted as described herein, or two R's may combine with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring. In an embodiment, the amino group is -NHR', where R' is aryl ("arylamino"), heteroaryl ("heteroarylamino"), amide or alkyl ("alkylamino").

[0197] Sulfonyl: The term "sulfonyl" refers to -S(=O) 2 R' or -S(=O) 2 -group, where R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), amino, cycloalkyl, aryl, aralkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), and unless otherwise specified in the specification, each moiety in said moieties may itself be optionally substituted as described herein. For example, in one embodiment, the sulfonyl is -SO 2 R', where R' is an alkyl substituted by a carbonyl.

[0198] Sulfinyl: The term "sulfinyl" refers to a chemical moiety having the formula -S(=O)R', -S(=O)- or -S(=O)(=NR')-, where R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, aralkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), and unless otherwise specified in the specification, each moiety in said moieties may itself be optionally substituted as described herein.

[0199] Carbonyl: The term "carbonyl" refers to a -C(=O)R' or -C(=O)- group, where R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a carbon chain), cycloalkyl, aryl, aralkyl, amino, hydroxy, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), and unless otherwise specified in the specification, each moiety in said moieties may itself be optionally substituted as described herein.

[0200] Phosphoryl: The term "phosphoryl" refers to -P(=O)(R') 2or a -P(=O)(R')- group, wherein each R' is independently selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon or heteroatom), cycloalkyl, aryl, aralkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl or heterocycloalkyl (bonded through a ring carbon) groups, and unless otherwise specified in the specification, each moiety in said moieties may be optionally substituted as described herein, or two R's may combine with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring.

[0201] Heteroalkyl: The term "heteroalkyl" means a branched or unbranched alkyl, alkenyl or alkynyl group having from 1 to 14 carbon atoms in addition to 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O, S and P. Heteroalkyl includes tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, aminophosphates, sulfonamides and disulfides. The heteroalkyl group may optionally contain a monocyclic, bicyclic or tricyclic ring, wherein each ring desirably has three to six members. Examples of heteroalkyl include polyethers such as methoxymethyl and ethoxyethyl.

[0202] Subheteroalkyl: As used herein, the term "subheteroalkyl" represents the divalent form of a heteroalkyl group as described herein.

[0203] Heteroaryl: As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic or tricyclic carbocyclic system having a total of six to fourteen ring members, wherein said ring system has a single point of attachment to the remainder of the molecule, wherein at least one ring in the system is aromatic, wherein each ring in the system contains 4 to 7 ring members, and wherein at least one ring atom is a heteroatom such as, but not limited to, nitrogen and oxygen.

[0204] Heterocycloalkyl: As used herein, the term "heterocycloalkyl" is a non-aromatic ring wherein at least one atom is a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus, and the remaining atoms are carbon. Heterocycloalkyl may be substituted or unsubstituted.

[0205] Deuterium: The term "deuterium" ("D" or " 2 H") is also known as heavy hydrogen. Deuterium is an isotope of hydrogen wherein the atomic nucleus consists of one proton and one neutron and is twice the mass of the ordinary hydrogen atomic nucleus (one proton).

[0206] Isotope: The term "isotope" refers to variants of a particular chemical element that differ in the number of neutrons and thus in the number of nucleons. All isotopes of a given element have the same number of protons but different numbers of neutrons in each atom.

[0207] The term "substituted" means that a particular group or moiety bears one or more substituents. The term "unsubstituted" means that the designated group bears no substituents. The term "optionally substituted" means that the designated group is either unsubstituted or substituted with one or more substituents. In cases where the term "substituted" is used to describe a structural system, substitution is understood to occur at any position permitted by the valences on the system, such that substitution results in a stable compound (e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reactions). In cases where a particular moiety or group is not explicitly noted as being optionally substituted or substituted with any designated substituents, it is understood that such moiety or group is intended to be unsubstituted.

[0208] When a ring system (e.g., cycloalkyl, heterocyclic, aryl, or heteroaryl) is substituted with multiple substituents that vary within a defined range, it is understood that the total number of substituents does not exceed the normal available valences under the existing conditions. It is also understood that hydrogen atoms are assumed to be present to fill the remaining valences of the ring system. Substituted groups only encompass combinations of substituents and variables that result in stable or chemically viable compounds. A stable compound or a chemically viable compound is a compound that has, among other things, sufficient stability to permit its preparation and detection.

[0209] A variety of substituents are well known, and methods for the formation of substituents and their introduction into a variety of parent groups are also well known. Representative substituents include, but are not limited to, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aralkyl, alkaryl, aryl, aryloxyalkyl, arylamino, heteroarylamino, heteroaryl, heteroaryloxyalkyl, heterocycloalkyl, hydroxyalkyl, aminoalkyl, haloalkyl, thioalkyl, alkylthioalkyl, carboxyalkyl, imidazolidinyl, indolylalkyl, mono-, di- and trihaloalkyl, mono-, di- and trihaloalkyl, amino, alkylamino, dialkylamino, amide, cyano, alkoxy, hydroxy, sulfonamide, halo (e.g., -Cl and -Br), nitro, oxime, -COOR 50 、-COR 50 、-SO 0-2 R 50 、-SO 2 NR 50 R 51 、NR 52 SO 2 R 50 、═C(R 50 R 51 )、═N-OR 50 、═N-CN、═C(halo) 2 、═S、═O、-CON(R 50 R 51 )、-OCOR 50 、-OCON(R50 R 51 )、 -N(R 52 )CO(R 50 )、 -N(R 52 )COOR 50 and -N(R 52 )CON(R 50 (R 51 ), where, with or without substituents, R 50 , R 51 and R 52 can independently be selected from the following: a hydrogen atom and branched or straight-chain C 1-6 -alkyl, C 3-6 -cycloalkyl, C 4-6 -heterocycloalkyl, heteroaryl, and aryl. Where permitted, R 50 and R 51 can be joined together to form a carbocyclic or heterocyclic system.

[0210] In a preferred embodiment, the substituents are selected from halogen, -COR', -CO 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 , -NHR', -N(R') 2 , -SR' and -SO 2 R', where each instance of R' is independently C 1 -C 20 aliphatic (e.g., C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). In some of its embodiments, R' is independently unsubstituted alkyl (e.g., unsubstituted C 1 -C 20 alkyl, C 1 -C 15 alkyl, C 1 -C 10 alkyl or C 1 -C 3 alkyl). Preferably, R' is independently unsubstituted C 1 -C 3 alkyl.

[0211] Any formula given herein is intended to represent compounds having the structures depicted by the structural formulas and certain variants or forms. Specifically, compounds of any formula given herein may have asymmetric centers and thus exist in different enantiomeric forms. All optical isomers and stereoisomers of the compounds of the general formula and mixtures thereof are considered to be within the scope of the formula. Thus, any formula given herein is intended to represent racemates, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. In addition, certain structures may exist as geometric isomers (i.e., cis and trans isomers), as tautomers, or as atropisomers. Additionally, any formula given herein is intended to include hydrates, solvates, and polymorphs of such compounds and mixtures thereof.

[0212] Compounds of the Invention

[0213] Compounds that are potent inhibitors of PHD are disclosed herein. In some embodiments, the compounds of the invention have an enzymatic half-maximal inhibitory concentration (IC 50 ) value of less than 100 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 50 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 25 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 20 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 15 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 10 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 5 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of less than 1 μM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50 value of about 3 nM to about 5 nM against any one of PHD1, PHD2, and PHD3. In some embodiments, the compounds of the invention have an IC 50Value. In some embodiments, the compounds of the present invention have an IC of about 10 nM to about 20 nM against any one of PHD1, PHD2, and PHD3 50 Value. In some embodiments, the compounds of the present invention have an IC of about 20 nM to about 50 nM against any one of PHD1, PHD2, and PHD3 50 Value. In some embodiments, the compounds of the present invention have an IC of about 50 nM to about 100 nM against any one of PHD1, PHD2, and PHD3 50 Value. In some embodiments, the compounds of the present invention have an IC of about 100 nM to about 200 nM against any one of PHD1, PHD2, and PHD3 50 Value. In some embodiments, the compounds of the present invention have an IC of about 200 nM to about 500 nM against any one of PHD1, PHD2, and PHD3 50 Value. In some embodiments, the compounds of the present invention have an IC of about 500 nM to about 1000 nM against any one of PHD1, PHD2, and PHD3 50 Value.

[0214] Representative examples of this category show inhibitory activity against PHD1, PHD2, and PHD3 in vitro.

[0215] Exemplary compounds are described herein. Specifically, these selective inhibitors can have a pyrazole moiety (e.g., 5-hydroxy-substituted pyrazole) connecting two aromatic moieties.

[0216] Compounds of formula (A) and (I)-(XIII)

[0217] On the one hand, the present invention provides compounds having a structure according to formula (A):

[0218]

[0219] Or a pharmaceutically acceptable salt thereof, wherein:

[0220] Ar 1 Is an aryl or heteroaryl optionally substituted with one or more groups selected from: halogen, CN, OH, C optionally substituted with CN or one or more halogens 1-3 Alkyl and C 1-3 Alkoxy; and

[0221] Ar 2 Is a pyridin-2-yl optionally substituted with one or more groups selected from: halogen; amino; amide; OH; sulfonyl; sulfinyl; carbonyl; phosphoryl; C 3-6 Cycloalkyl; C optionally substituted with sulfonyl or =O 3-6Heterocycloalkyl; C optionally substituted with carbonyl or one or more halogens 1-3 alkyl; and heteroaryl optionally substituted with C 1-3 alkyl or phenyl.

[0222] In embodiments, Ar 1 is unsubstituted aryl. In embodiments, Ar 1 is substituted aryl. In embodiments, Ar 1 is substituted phenyl.

[0223] In embodiments, Ar 1 is unsubstituted 6-membered heteroaryl. In embodiments, Ar 1 is substituted 6-membered heteroaryl.

[0224] In embodiments, Ar 1 is substituted with one or more groups selected from: halogen, CN, OH, C optionally substituted with CN or one or more halogens 1-3 alkyl and C 1-3 alkoxy. In some embodiments, Ar 1 is substituted with 1 substituent. In some embodiments, Ar 1 is substituted with 2 substituents. In some embodiments, Ar 1 is substituted with 3 substituents. In some embodiments, Ar 1 is substituted with 4 substituents.

[0225] In embodiments, Ar 1 includes one or more R 1 groups, where each R 1 is independently selected from hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 alkyl and C 1-3 alkoxy. In embodiments, Ar 1 includes a number of R 1 groups represented by m, where m is 1, 2, 3, or 4. When R 1 is present, R 1 can replace hydrogen in the parent molecular structure. In embodiments, when R 1 is present and is a non-hydrogen moiety, R 1 represents a substituent. In embodiments, R 1 is independently selected from halogen, CN, OH, C optionally substituted with one or more halogens 1-3 alkyl and C 1-3 alkoxy.

[0226] Thus, it should also be understood that for any value of m described herein, hydrogen is present where appropriate to complete the valence of Ar 1The valence requirements are met at the constituent atoms such that the molecule is a stable compound (e.g., the molecule is a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reactions). Ar is described herein 1 、R 1 and exemplary embodiments of m are provided.

[0227] In an embodiment, Ar 1 is

[0228] wherein

[0229] X is N or CR 1a ;

[0230] Y and Z are independently CH or N; and

[0231] m is 1, 2, 3, or 4.

[0232] In an embodiment, R 1 is not hydrogen. In an embodiment, when R 1 is present and is a non-hydrogen moiety, R 1 represents a substituent.

[0233] In an embodiment, the value of m is based on the number of nitrogen atoms present in the ring. In an embodiment, when only one of Y and Z is N, m is 1, 2, or 3. In an embodiment, when each of Y and Z is N, m is 1 or 2.

[0234] In an embodiment, X is N. In an embodiment, X is CR 1a .

[0235] In an embodiment, Y is CH. In an embodiment, Z is N.

[0236] In an embodiment, m is 1. In an embodiment, m is 2. In an embodiment, m is 3. In an embodiment, m is 4.

[0237] In an embodiment, both Y and Z are N and m is 1 or 2. In an embodiment, m is 1 and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valence. In an embodiment, m is 2.

[0238] In an embodiment, both Y and Z are CH and m is 1, 2, 3, or 4. In an embodiment, m is 1 and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valence. In an embodiment, m is 2 and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valence. In an embodiment, m is 3 and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valence. In an embodiment, m is 4.

[0239] In an embodiment, one of Y and Z is CH and the other is N, and m is 1, 2, or 3. In an embodiment, m is 1, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 2, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3.

[0240] In an embodiment, Ar 1 is

[0241] where

[0242] X is N or CR 1a ;

[0243] Z is CH or N; and

[0244] m is 1, 2, 3, or 4.

[0245] In an embodiment, Z is N, and m is 1, 2, or 3. In an embodiment, m is 1, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 2, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3.

[0246] In an embodiment, Z is CH, and m is 1, 2, 3, or 4. In an embodiment, m is 1, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 2, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 4.

[0247] In an embodiment, X is N. In an embodiment, X is CR 1a .

[0248] In an embodiment, Ar 1 is

[0249] where

[0250] m is 1, 2, 3, or 4.

[0251] In an embodiment, m is 1, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 2, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3, and any remaining unsubstituted carbocyclic atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 4.

[0252] In an embodiment, R 1a is H.

[0253] In an embodiment, R 1a is CN.

[0254] In an embodiment, R 1a is OH.

[0255] In an embodiment, R 1a is a halogen. In an embodiment, R 1a is F. In an embodiment, R 1a is Cl. In an embodiment, R 1a is Br. In an embodiment, R 1a is I.

[0256] In an embodiment, R 1a is C 1-3 alkoxy. In an embodiment, R 1a is methoxy. In an embodiment, R 1a is ethoxy. In an embodiment, R 1a is propoxy.

[0257] In an embodiment, R 1a is C 1-3 alkyl.

[0258] In an embodiment, R 1a is unsubstituted C 1-3 alkyl. In an embodiment, R 1a is CH 3 .

[0259] In an embodiment, R 1a is substituted C 1-3 alkyl. In an embodiment, R 1a is C 1-3 alkyl substituted by a CN group. In an embodiment, R 1a is CH 2 CN.

[0260] In an embodiment, each time it is taken, R 1 is hydrogen.

[0261] In an embodiment, each time it is taken, R 1 is CN.

[0262] In an embodiment, each time it is taken, R 1 is OH.

[0263] In an embodiment, each time it is taken, R 1 is a halogen. In an embodiment, the halogen is Cl. In an embodiment, the halogen is Br. In an embodiment, the halogen is I.

[0264] In an embodiment, each time it is taken, R 1 is C 1-3 alkyl.

[0265] In an embodiment, each time it is taken, R 1 is unsubstituted C 1-3 alkyl. In an embodiment, each time it is taken, R 1 is CH 3 .

[0266] In an embodiment, each time it is taken, R 1 is substituted C 1-3 alkyl. In an embodiment, each time it is taken, R 1 is C alkyl substituted by one or more halogens 1-3 alkyl. In an embodiment, the halogen is F. In an embodiment, the halogen is Cl. In an embodiment, the halogen is Br. In an embodiment, the halogen is I.

[0267] In an embodiment, each time it is taken, R 1 is CF 3 .

[0268] In an embodiment, each time it is taken, R 1 is C 1-3 alkoxy. In an embodiment, each time it is taken, R 1 is OMe.

[0269] In an embodiment, Ar 2 is pyridin-2-yl optionally substituted by one or more groups selected from the following: halogen; amino; amide; OH; sulfonyl (e.g., SO 2 R 6 ); sulfinyl; (e.g., SOR 7 R 8 or SOR 9 ); carbonyl; (e.g., COR 10 ); phosphoryl; (e.g., POR 12 R 13 ); C 3-6 cycloalkyl; C 3-6 heterocycloalkyl optionally substituted by sulfonyl or =O or C 1-3 alkyl optionally substituted by carbonyl or one or more halogens; and heteroaryl optionally substituted by C 1-3 alkyl or phenyl. In an embodiment, Ar 2 is unsubstituted pyridin-2-yl. In an embodiment, Ar 2 is substituted pyridin-2-yl. In an embodiment, Ar 2is pyridin-2-yl substituted with 1 or 2 substituents as described herein. In an embodiment, Ar 2 is pyridin-2-yl substituted with 3 substituents as described herein.

[0270] In an embodiment, Ar 2 is

[0271] wherein

[0272] each time taken, R 2 is independently selected from the group consisting of hydrogen, halogen, NR 4 R 5 、OH, C 1-3 alkyl, and C 3-6 cycloalkyl;

[0273] R 3 is SO 2 R 6 、SOR 7 R 8 、SOR 9 、COR 10 、(CH 2 ) p COOH, NHR 11 、POR 12 R 13 、halogen, cycloalkyl, heterocycloalkyl optionally substituted with SO 2 R 14 or =O, heteroaryl optionally substituted with C 1-3 alkyl or phenyl, or C 1-3 alkyl optionally substituted with one or more halogens;

[0274] R 6 is C 1-3 alkyl, NHCOR 15 、NR 16 R 17 or phenyl;

[0275] R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 ;

[0276] R 8 is NH or NCH 3 ;

[0277] R 10 is C 1-3 alkyl or NHSO 2 R 20 ;

[0278] R 11 is COR 21 or SO 2 R 22 ;

[0279] R 9 、R 12 、R 13、 R 14 、R 15 and R 20 are each independently C 1-3 alkyl;

[0280] R 21 is heteroalkyl, cycloalkyl or C 1-3 alkyl;

[0281] R 22 is NR 23 R 24 or optionally C 1-3 alkyl substituted with carboxyl;

[0282] R 4 、R 5 、R 16 、R 17 、R 18 、R 19 、R 23 and R 24 are each independently H or C 1-3 alkyl;

[0283] p is 1, 2 or 3; and

[0284] n is 0, 1, 2 or 3.

[0285] In an embodiment, n is 0. In an embodiment, n is 1. In an embodiment, n is 2. In an embodiment, n is 3.

[0286] In an embodiment, n is 0 and any remaining unsubstituted carbocyclic atoms are assumed to be hydrogen bonded to fill valences.

[0287] In an embodiment, n is 1 and any remaining unsubstituted carbocyclic atoms are assumed to be hydrogen bonded to fill valences. In an embodiment, n is 2 and any remaining unsubstituted carbocyclic atoms are assumed to be hydrogen bonded to fill valences. In an embodiment, n is 3.

[0288] In an embodiment, each time taken, R 2 is hydrogen.

[0289] In an embodiment, each time taken, R 2 is OH.

[0290] In an embodiment, each time it is taken, R 2 is a halogen. In an embodiment, the halogen is Cl. In an embodiment, the halogen is Br. In an embodiment, the halogen is I.

[0291] In an embodiment, each time it is taken, R 2 is NR 4 R 5 where R 4 and R 5 are each independently H or C 1-3 alkyl.

[0292] In an embodiment, R 4 and R 5 are both H.

[0293] In an embodiment, one of R 4 and R 5 is H and the other is C 1-3 alkyl. In an embodiment, C 1-3 alkyl is CH 3 .

[0294] In an embodiment, each time it is taken, R 2 is C 1-3 alkyl.

[0295] In an embodiment, each time it is taken, R 2 is C 3-6 cycloalkyl.

[0296] In an embodiment, R 3 is SO 2 R 6 where R 6 is C 1-3 alkyl, NHCOR 15 , NR 16 R 17 or phenyl.

[0297] In an embodiment, R 3 is SOR 7 R 8 where R 7 is C 1-3 alkyl, C 3-5 cycloalkyl, phenyl or NR 18 R 19 and where R 8 is NH or NCH 3 ;

[0298] In an embodiment, R 3 is SOR 9 where R 9 is C1-3 Alkyl

[0299] In an embodiment, R 3 is COR 10 wherein R 10 is C 1-3 alkyl or NHSO 2 R 20 and wherein R 20 is C 1-3 alkyl

[0300] In an embodiment, R 3 is (CH 2 ) p COOH

[0301] In an embodiment, p is 1, 2 or 3. In an embodiment, p is 1. In an embodiment, p is 2. In an embodiment, p is 3.

[0302] In an embodiment, R 3 is NHR 11 wherein R 11 is COR 21 or SO 2 R 22 and wherein R 21 is heteroalkyl, cycloalkyl or C 1-3 alkyl; R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with carboxyl; and wherein R 23 and R 24 are each independently H or C 1-3 alkyl

[0303] In an embodiment, R 3 is POR 12 R 13 wherein R 12 and R 13 are C 1-3 alkyl

[0304] In an embodiment, R 3 is a halogen

[0305] In an embodiment, R 3 is cycloalkyl or heteroalkyl. In an embodiment, the cycloalkyl or heteroalkyl is unsubstituted. In an embodiment, the cycloalkyl or heteroalkyl is substituted.

[0306] In an embodiment, R 3 is heteroaryl. In an embodiment, the heteroaryl is unsubstituted. In an embodiment, the heteroaryl is substituted.

[0307] In an embodiment, R 3 is C 1-3 alkyl. In an embodiment, the C 1-3 alkyl is unsubstituted. In an embodiment, the C 1-3 alkyl is substituted with one or more halogens.

[0308] In an embodiment, the compound of formula (A) has the following structure,

[0309] or a pharmaceutically acceptable salt thereof, wherein X, Y, Z, R 1 , R 2 and R 3 are as defined anywhere herein.

[0310] In an embodiment, the compound of formula (A) or formula (I) has the following structure,

[0311] or a pharmaceutically acceptable salt thereof, wherein X, Z, R 1 , R 2 and R 3 are as defined anywhere herein.

[0312] In an embodiment, the compound of formula (A), formula (I) or formula (II) has the following structure,

[0313] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 , R 2 and R 3 are as defined anywhere herein.

[0314] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0315] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 are as defined anywhere herein.

[0316] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (IV) has the following structure,

[0317] or a pharmaceutically acceptable salt thereof, wherein A, R 1a and R 2 are as defined anywhere herein.

[0318] In an embodiment, R 7is C 1-3 alkyl

[0319] In an embodiment, R 7 is C 3-5 cycloalkyl

[0320] In an embodiment, R 7 is phenyl

[0321] In an embodiment, R 7 is NR 18 R 19 wherein R 18 and R 19 are each independently H or C 1-3 alkyl

[0322] In an embodiment, R 18 and R 19 are both H

[0323] In an embodiment, R 18 and R 19 are both C 1-3 alkyl. In an embodiment, R 18 and R 19 are both CH 3 .

[0324] In an embodiment, R 18 is H and R 19 is C 1-3 alkyl. In an embodiment, R 19 is CH 3 .

[0325] In an embodiment, R 8 is NH

[0326] In an embodiment, R 8 is NCH 3 .

[0327] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0328] or a pharmaceutically acceptable salt thereof, wherein X, Z, R 1 and R 2 are as defined anywhere herein

[0329] In an embodiment, R 6 is C 1-3 alkyl. In an embodiment, R 6 is CH 3 .

[0330] In an embodiment, R6 is NHCOR 15 , where R 15 is C 1-3 alkyl. In an embodiment, R 6 is NHCOCH 3 .

[0331] In an embodiment, R 6 is NR 16 R 17 , where R 16 and R 17 are each independently H or C 1-3 alkyl.

[0332] In an embodiment, R 16 and R 17 are both H.

[0333] In an embodiment, R 16 and R 17 are both C 1-3 alkyl. In an embodiment, R 16 and R 17 are both CH 3 .

[0334] In an embodiment, R 16 is H and R 17 is C 1-3 alkyl. In an embodiment, R 17 is CH 3 .

[0335] In an embodiment, R 6 is phenyl.

[0336] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0337] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 are as defined anywhere herein.

[0338] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VI) has the following structure,

[0339] or a pharmaceutically acceptable salt thereof, wherein R 2 is as defined anywhere herein.

[0340] In an embodiment, R 3 is cycloalkyl.

[0341] In an embodiment, R 3 is an unsubstituted cycloalkyl. In an embodiment, R 3 is

[0342] In an embodiment, R 3 is a substituted cycloalkyl. In an embodiment, R 3 is a cycloalkyl substituted with SO 2 R 14 or =O, where R 14 is C 1-3 alkyl.

[0343] In an embodiment, R 3 is a heterocycloalkyl.

[0344] In an embodiment, R 3 is an unsubstituted heterocycloalkyl. In an embodiment, R 3 is

[0345] In an embodiment, R 3 is a substituted heterocycloalkyl. In an embodiment, R 3 is a heterocycloalkyl substituted with SO 2 R 14 or =O, where R 14 is C 1-3 alkyl. In an embodiment, R 3 is

[0346] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0347] or a pharmaceutically acceptable salt thereof, where R 1a , R 1 and R 2 are as defined anywhere herein.

[0348] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VII) has the following structure,

[0349] or a pharmaceutically acceptable salt thereof, where R 2 is as defined anywhere herein.

[0350] In an embodiment, R 11 is COR 21 , where R 21 is a heterocycloalkyl, cycloalkyl or C 1-3 alkyl.

[0351] In an embodiment, R 21 is a cycloalkyl group. In an embodiment, R 21 is

[0352] In an embodiment, R 21 is a heterocycloalkyl group. In an embodiment, R 21 is

[0353] In an embodiment, R 21 is C 1-3 alkyl. In an embodiment, R 21 is CH 2 CH 3 .

[0354] In an embodiment, R 11 is SO 2 R 22 , where R 22 is NR 23 R 24 or C 1-3 alkyl optionally substituted with a carboxyl group, and where R 23 and R 24 are each independently H or C 1-3 alkyl.

[0355] In an embodiment, R 22 is C 1-3 alkyl. In an embodiment, R 22 is unsubstituted C 1-3 alkyl. In an embodiment, R 22 is C 1-3 alkyl substituted with a carboxyl group. In an embodiment, R 22 is CH 2 COOH.

[0356] In an embodiment, R 22 is NR 23 R 24 , where R 23 and R 24 are each independently H or C 1-3 alkyl.

[0357] In an embodiment, R 23 and R 24 are both H.

[0358] In an embodiment, R 23 and R 24 are both C 1-3 alkyl. In an embodiment, R 23 and R 24All are CH 3 .

[0359] In an embodiment, R 23 is H and R 24 is C 1-3 alkyl. In an embodiment, R 24 is CH 3 .

[0360] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0361] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 are as defined anywhere herein.

[0362] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (VIII) has the following structure,

[0363] or a pharmaceutically acceptable salt thereof.

[0364] In an embodiment, R 3 is heteroaryl. In an embodiment, the heteroaryl is thiazole, oxazole, pyridine, triazole, tetrazole or pyrazole.

[0365] In an embodiment, R 3 is unsubstituted heteroaryl. In an embodiment, R 3 is

[0366] In an embodiment, R 3 is heteroaryl substituted with C 1-3 alkyl or phenyl. In an embodiment, R 3 is

[0367] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0368] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 are as defined anywhere herein.

[0369] In an embodiment, the compound of formula (A), formula (I), formula (II), formula (III) or formula (IX) has the following structure,

[0370] or a pharmaceutically acceptable salt thereof, wherein R 1a as defined anywhere herein.

[0371] In an embodiment, R 10 is C 1-3 alkyl.

[0372] In an embodiment, R 10 is NHSO 2 R 20 wherein R 20 is C 1-3 alkyl. In an embodiment, R 10 is NHSO 2 CH 3 .

[0373] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0374] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 as defined anywhere herein.

[0375] In an embodiment, R 9 is C 1-3 alkyl.

[0376] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0377] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 as defined anywhere herein.

[0378] In an embodiment, p is 1. In an embodiment, p is 2. In an embodiment, p is 3.

[0379] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0380] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 as defined anywhere herein.

[0381] In an embodiment, R 3 is halogen. In an embodiment, R 3 is F. In an embodiment, R3 is Cl. In an embodiment, R 3 is Br. In an embodiment, R 3 is I.

[0382] In an embodiment, the compound of formula (A), formula (I), formula (II) or formula (III) has the following structure,

[0383] or a pharmaceutically acceptable salt thereof, wherein R 1a , R 1 and R 2 are as defined anywhere herein.

[0384] In an embodiment, R 12 and R 13 are both C 1-3 alkyl. In an embodiment, R 12 and R 13 are both CH 3 .

[0385] Exemplary compounds

[0386] In some embodiments, the PHD inhibitor compound is any one of Compounds 1 to 33 or a pharmaceutically acceptable salt thereof.

[0387]

[0388]

[0389]

[0390] Isotopologues

[0391] It should be understood that in the compounds described herein (e.g., the compounds of any one of formula (A) and (I) to (XIII), such as any one of Compounds 1 to 33), atoms can exhibit their natural isotope abundances, or one or more of the atoms can be artificially enriched with specific isotopes having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number predominantly found in nature. The present invention is intended to encompass all suitable isotopic variants of the compounds described herein (e.g., the compounds of any one of formula (A) and (I) to (XIII), such as any one of Compounds 1 to 33). For example, the different isotopic forms of hydrogen (H) include protium ( 1 H), deuterium ( 2 H), and tritium ( 3 H). Protium is the major hydrogen isotope found in nature.

[0392] In some embodiments, one or more hydrogens in the hydrogen of the compounds described herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) are replaced by deuterium. Enriching deuterium can provide certain therapeutic advantages, such as increasing the in vivo half-life or reducing the dosage requirement, or can provide compounds that can be used as standards for characterizing biological samples. In some embodiments, one or more hydrogens in the hydrogen of the compounds described herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) are replaced by tritium. Tritium is radioactive and can therefore provide radiolabeled compounds that can be used as tracers in metabolic or kinetic studies.

[0393] Isotope enrichment of the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) can be achieved without undue experimentation by conventional techniques well known to those skilled in the art or by methods similar to those described in the protocols and examples herein, using appropriate isotopically enriched reagents and / or intermediates.

[0394] The term "isotopologue" refers to a species having the same chemical structure and formula as a particular compound provided herein, except for the isotopic substitution sites and / or isotopic enrichment levels at one or more positions, such as hydrogen to deuterium. Thus, as used herein, the term "compound" encompasses a collection of molecules having the same chemical structure but also having isotopic variations among the constituent atoms of the molecule. Thus, it will be apparent to those skilled in the art that a compound represented by a particular chemical structure containing the indicated deuterium atoms will also contain a lesser amount of isotopologues having hydrogen atoms at one or more of the designated deuterium positions in the structure. The relative amounts of such isotopologues in the provided compounds depend on many factors, including but not limited to the isotopic purity of the deuterated reagents used to prepare the compounds and the efficiency of deuterium incorporation in the individual synthetic steps used to prepare the compounds.

[0395] When a position is designated as "H" or "hydrogen", the position is understood to have hydrogen with its isotopic composition at natural abundance. When a position is designated as "D" or "deuterium", the position is understood to have deuterium with an abundance greater than 3340 times the natural abundance of deuterium, the natural abundance of deuterium being 0.015% (i.e., the term "D" or "deuterium" indicates at least 50.1% deuterium incorporation).

[0396] In an embodiment, the isotopic enrichment factor for each deuterium at the site of a potential deuteration site designated on the compound provided herein can be at least 3500 (52.5% deuterium incorporation), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0397] Synthesis of the compounds of the present invention

[0398] The compounds described herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) can be prepared according to methods known in the art, including the exemplary syntheses provided herein as examples.

[0399] The abbreviations and acronyms used herein include the following:

[0400]

[0401]

[0402] Compositions and Methods

[0403] The present invention provides the use of a compound of any one of formulas (A) and (I) to (XIII) or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating various conditions or disorders as described herein. In one embodiment, a pharmaceutical composition is provided that comprises at least one compound of any one of formulas (A) and (I) to (XIII) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient or carrier. In various embodiments, the medicament or pharmaceutical composition can further comprise at least one additional therapeutic agent or be used in combination with at least one additional therapeutic agent.

[0404] The compounds or medicaments of the present invention or compositions comprising said compounds can be used to inhibit the activity of PHD. Inhibiting PHD can be particularly beneficial for treating diseases including heart diseases (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease).

[0405] In one embodiment, the method of the present invention comprises administering to a patient in need thereof a therapeutically effective amount of a compound of any one of formulas (A) and (I) to (XIII) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition comprising one or more compounds of any one of formulas (A) and (I) to (XIII).

[0406] The present invention also relates to a method of inhibiting PHD activity. In one embodiment, the method comprises contacting PHD with an effective amount of one or more compounds selected from the group consisting of: a compound of any one of formulas (A) and (I) to (XIII) or a pharmaceutically acceptable salt thereof.

[0407] In still other embodiments, the compounds disclosed herein (e.g., a compound of any one of formulas (A) and (I) to (XIII), such as any one of Compounds 1 to 33) or a pharmaceutically acceptable salt thereof can be used to treat or prevent anemia, including treating anemia conditions associated with: chronic kidney disease, polycystic kidney disease, aplastic anemia, autoimmune hemolytic anemia, myelodysplastic syndrome related anemia, Churg-Strauss syndrome, Diamond Blackfan anemia, Fanconi's anemia, Felty syndrome, graft-versus-host disease, hematopoietic stem cell transplantation, hemolytic uremic syndrome, myelodysplastic syndrome, paroxysmal nocturnal hemoglobinuria, myelofibrosis, pancytopenia, pure red cell aplasia, Henoch-Schönlein purpura, refractory anemia with excess blasts, rheumatoid arthritis, Shwachman syndrome, sickle cell disease, thalassemia major, thalassemia minor, thrombocytopenic purpura, anemic or non-anemic patients undergoing surgery, anemia associated with or secondary to trauma, sideroblastic anemia, anemia secondary to other treatments, including: reverse transcriptase inhibitors for treating HIV, corticosteroids, cisplatin or non-cisplatin containing chemotherapeutic drugs, vinca alkaloids, mitotic inhibitors, topoisomerase II inhibitors, anthracyclines, alkylating agents, particularly anemia secondary to inflammatory, aging and / or chronic diseases. PHD1 inhibition can also be used to treat anemia symptoms, including chronic fatigue, pallor and dizziness.

[0408] In other embodiments, the compounds disclosed herein (e.g., a compound of any one of formulas (A) and (I) to (XIII), such as any one of Compounds 1 to 33) or a pharmaceutically acceptable salt thereof can be used to treat or prevent metabolic disorders, including but not limited to diabetes and obesity.

[0409] In still other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat or prevent vascular diseases. These include but are not limited to hypoxia or wound healing-related diseases that require angiogenic mediators for angiogenesis, neovascularization, and arteriogenesis.

[0410] In yet other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat or prevent ischemia-reperfusion injury. These include but are not limited to stroke, myocardial infarction, and acute kidney injury.

[0411] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat inflammatory bowel disease. These include but are not limited to ulcerative colitis and Crohn's disease.

[0412] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat cancers such as colorectal cancer.

[0413] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat atherosclerosis.

[0414] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat cardiovascular diseases.

[0415] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat diseases or conditions of the eye. These include but are not limited to radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

[0416] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat diseases associated with hyperoxia.

[0417] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat bronchopulmonary dysplasia (BPD).

[0418] In yet other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat heart diseases. The conditions include but are not limited to myocardial ischemia after pancreatic surgery, myocardial injury after percutaneous coronary intervention (PCI), myocardial injury after non-cardiac surgery, perioperative myocardial ischemia during elective surgery for abdominal aortic aneurysm, myocardial injury after PCI, myocardial injury in patients undergoing coronary artery bypass graft (CABG) surgery, minimally invasive mitral valve (MIMV) repair or replacement, adult patients undergoing open heart surgery, chronic heart failure, NYHA II-IV class.

[0419] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat lung diseases. The conditions include but are not limited to lung injury during elective lobectomy, lung injury during CABG surgery, lung transplantation.

[0420] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat liver diseases. The conditions include but are not limited to non-alcoholic steatohepatitis (NASH).

[0421] In other embodiments, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used to treat kidney diseases. The conditions include but are not limited to contrast-induced acute kidney injury, stage III to IV chronic kidney disease in patients undergoing planned coronary angiography, acute kidney injury in patients undergoing cardiac valve surgery, non-dialysis-dependent chronic kidney disease, chronic kidney disease patients starting dialysis, non-dialysis-dependent chronic kidney disease.

[0422] In addition, the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof can be used in combination with additional active ingredients to treat the above-mentioned conditions. The additional compounds can be co-administered alone with the compounds disclosed herein (e.g., compounds of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or pharmaceutically acceptable salts thereof or can be included in a pharmaceutical composition according to the invention together with the additional active ingredients. In an exemplary embodiment, the additional active ingredients are those active ingredients known or found to be effective in treating conditions mediated by PHD enzymes, disorders, or having activity against another target associated with a particular condition, disorder, or disease, such as alternative PHD modulators. The combination can be used to increase efficacy (e.g., by including in the combination a compound that enhances the potency or effectiveness of the compounds according to the invention), reduce one or more side effects, or reduce the required dose of the compounds according to the invention.

[0423] The compounds of the present invention are used alone or in combination with one or more other active ingredients for formulating the pharmaceutical compositions of the present invention. The pharmaceutical compositions of the present invention comprise: (a) an effective amount of a compound disclosed herein (e.g., a compound of any one of formulas (A) and (I) to (XIII), such as any one of compounds 1 to 33) or a pharmaceutically acceptable salt, pharmaceutically acceptable prodrug, or pharmaceutically active metabolite thereof; and (b) a pharmaceutically acceptable excipient.

[0424] "Pharmaceutically acceptable excipient" refers to a non-toxic, biologically tolerable, and otherwise biologically suitable substance for administration to a subject, such as an inert substance, that is added to a pharmacological composition or otherwise used as an agent, carrier, or diluent to facilitate the administration of a medicament and is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars, and various types of starches, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Suitable excipients can also include antioxidants. Such antioxidants can be used in pharmaceutical compositions or storage media to extend the shelf life of the pharmaceutical product.

[0425] Pharmaceutical Formulations and Routes of Administration

[0426] As is well known in the art, the compounds and compositions of the present invention can be delivered directly or in a pharmaceutical composition or medicament together with a suitable carrier or excipient. The treatment methods of the present invention can include administering to a subject in need an effective amount of a compound of the present invention. In a preferred embodiment, the subject is a mammalian subject, and in the most preferred embodiment, the subject is a human subject.

[0427] The effective amount of such compounds, compositions or medicaments can be readily determined by routine experimentation, such as the most effective and convenient route of administration and the most appropriate formulation. A variety of formulations and drug delivery systems are available in the art. See, for example, Gennaro, A.R., ed. (1995) Remington's Pharmaceutical Sciences, supra.

[0428] Suitable routes of administration can include, for example, oral, rectal, topical, nasal, pulmonary, ocular, enteral and parenteral administration. The principal routes of parenteral administration include intravenous, intramuscular and subcutaneous administration. The secondary routes of administration include intraperitoneal, intraarterial, intraarticular, intracardiac, intracisternal, intradermal, intralesional, intraocular, intrathoracic, intrathecal, intrauterine and intraventricular administration. The type of formulation and route of administration to be used, as well as whether local or systemic delivery is preferred, are determined by the indication to be treated and the physical, chemical and biological properties of the medicament.

[0429] The pharmaceutical dosage forms of the compounds of the present invention can be provided in the form of immediate release, controlled release, sustained release or targeted drug delivery systems. Commonly used dosage forms include, for example, solutions and suspensions, (micro)emulsions, ointments, gels and patches, liposomes, tablets, pills, soft or hard shell capsules, suppositories, ovules, implants, amorphous or crystalline powders, aerosols and lyophilized formulations. Depending on the route of administration used, special devices may be required to apply or administer the medicament, such as syringes and needles, inhalers, pumps, injection pens, applicators or special flasks. Pharmaceutical dosage forms generally consist of a medicament, excipients and a container / closure system. One or more excipients, also known as inactive ingredients, can be added to the compounds of the present invention to improve or facilitate the manufacture, stability, administration and safety of the medicament and can provide a means of achieving the desired drug release profile. Thus, the type of excipient to be added to the medicament can depend on various factors, such as the physical and chemical properties of the medicament, the route of administration and the manufacturing procedure. Pharmaceutically acceptable excipients are available in the art and include those listed in various pharmacopoeias. See, for example, the United States Pharmacopeia (USP), the Japanese Pharmacopoeia (JP), the European Pharmacopoeia (EP) and the British Pharmacopoeia (BP); the U.S. Food and Drug Administration

[0430] (www.fda.gov) Publications of the Center for Drug Evaluation and Research (CEDR), such as the Inactive Ingredient Guide (1996); Ash and Ash, eds. (2002) Handbook of Pharmaceutical Additives, Synapse Information Resources, Inc., Endicott NY; etc.).

[0149] The pharmaceutical dosage forms of the compounds of the present invention can be manufactured by any method well known in the art, such as by conventional mixing, sieving, dissolving, melting, granulating, pelletizing, tableting, suspending, extruding, spray drying, milling, emulsifying, (nano / micro) encapsulating, entrapping or lyophilizing methods. As described above, the compositions of the present invention can contain one or more physiologically acceptable inactive ingredients, which facilitate the processing of the active molecules into formulations for pharmaceutical use.

[0431] The appropriate formulation depends on the desired route of administration. For example, for intravenous injection, the composition can be formulated in an aqueous solution, using, if necessary, a physiologically compatible buffer, containing, for example, phosphate, histidine or citrate to adjust the pH of the formulation, and a tonicity agent, such as sodium chloride or glucose. For transmucosal or nasal administration, semi-solid, liquid formulations or patches may be preferred and may contain permeation enhancers. Such permeation enhancers are known in the art. For oral administration, the compounds can be formulated into liquid or solid dosage forms, as well as immediate-release or controlled-release / sustained-release formulations. Suitable dosage forms for oral ingestion by a subject include tablets, pills, pellets, hard and soft shell capsules, liquids, gels, syrups, slurries, suspensions and emulsions. The compounds can also be formulated into rectal compositions (such as suppositories or retention enemas) containing, for example, conventional suppository bases such as cocoa butter or other glycerides.

[0432] Solid oral dosage forms can be obtained using excipients, which can include fillers, disintegrants, binders (dry and wet), dissolution retardants, lubricants, glidants, anti-adhesion agents, cation exchange resins, wetting agents, antioxidants, preservatives, colorants, and flavorants. These excipients can be of synthetic or natural origin. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatin, magnesium carbonate, magnesium lauryl sulfate / sodium, mannitol, polyethylene glycol, polyvinylpyrrolidone, silicates, silica, sodium benzoate, sorbitol, starch, stearic acid or its salts, sugars (i.e., glucose, sucrose, lactose, etc.), talc, tragacanth mucilage, vegetable oils (hydrogenated), and waxes. Ethanol and water can be used as granulation aids. In some cases, it is desirable to coat tablets with, for example, taste-masking films, anti-gastric acid films, or controlled-release films. Natural and synthetic polymers in combination with colorants, sugars, and organic solvents or water are commonly used to coat tablets, resulting in sugar-coated pills. When capsules are preferred over tablets, the drug powder, its suspension, or solution can be delivered in compatible hard or soft shell capsules.

[0433] In one embodiment, the compounds of the present invention can be administered topically, such as by a skin patch, semi-solid or liquid formulation, e.g., a gel, (micro)emulsion, ointment, solution, (nano / micro)suspension, or foam. The penetration of the drug through the skin and into the subcutaneous tissue can be modulated, for example, by using penetration enhancers; appropriately selecting and combining lipophilic, hydrophilic, and amphiphilic excipients, including water, organic solvents, waxes, oils, synthetic and natural polymers, surfactants, emulsifiers; adjusting the pH; and using complexing agents. Other techniques, such as iontophoresis, can be used to modulate the skin penetration of the compounds of the present invention. For example, in cases where local delivery with minimal systemic exposure is desired, transdermal or topical administration is preferred.

[0434] For inhalation or nasal administration, the compounds used according to the present invention are conveniently delivered from a pressurized package or nebulizer in the form of a solution, suspension, emulsion, or semi-solid aerosol, typically using a propellant, such as a halogenated carbon derived from methane and ethane, carbon dioxide, or any other suitable gas. Hydrocarbons, such as butane, isobutane, and pentane, are useful for topical aerosols. In the case of a pressurized aerosol, appropriate dosage units can be determined by providing a valve for delivering a metered amount. Capsules and cartridges, e.g., of gelatin, can be formulated for use in inhalers or insufflators. These typically contain a powder mixture of the compound and a suitable powder matrix, such as lactose or starch.

[0435] Compounds and compositions formulated for parenteral administration by injection are generally sterile and may be presented in unit dosage forms, for example in ampoules, syringes, pen injectors or multi-dose containers, the latter usually containing a preservative. The compositions may be in such forms as suspensions, solutions or emulsions in oily or aqueous vehicles and may contain formulating agents such as buffers, tonicity agents, viscosity enhancers, surfactants, suspending and dispersing agents, antioxidants, biocompatible polymers, chelating agents and preservatives. Depending on the site of injection, the vehicle may contain water, synthetic or vegetable oils and / or organic co-solvents. In some cases, such as with lyophilized products or concentrates, the parenteral formulation will be reconstituted or diluted prior to administration. Depot formulations providing controlled or sustained release of the compounds of the invention may comprise injectable suspensions of nano / microparticles or nano / microparticles or non-micronized crystals. Polymers such as poly(lactic acid), poly(glycolic acid) or copolymers thereof, among others well known in the art, may be used as controlled / sustained release matrices. Other depot delivery systems may be presented in the form of implants and pumps that require an incision.

[0436] Suitable carriers for intravenous injection of the compounds of the invention are well known in the art and include aqueous solutions containing a base such as sodium hydroxide to form the ionized compound; sucrose or sodium chloride as a tonicity agent; and buffers such as buffers containing phosphate or histidine. Co-solvents such as polyethylene glycol may be added. These aqueous systems are effective in dissolving the compounds of the invention and produce low toxicity upon systemic administration. The proportions of the components of the solution system may vary widely without destroying the solubility and toxicity characteristics. In addition, the characteristics of the components may vary. For example, low toxicity surfactants such as polysorbates or poloxamers may be used, polyethylene glycol or other co-solvents may be used, biocompatible polymers such as polyvinylpyrrolidone may be added, and other sugars and polyols may replace glucose.

[0437] The therapeutically effective dose can be initially estimated using a variety of techniques well known in the art. The initial dose used in animal studies may be based on the effective concentration established in cell culture assays. For example, data obtained from animal studies and cell culture assays can be used to determine the dose range suitable for human subjects. In some embodiments, the compounds of the present disclosure are formulated for oral administration. Exemplary doses of the compounds of the present disclosure in pharmaceutical formulations for oral administration are from about 0.5 to about 10 mg / kg of subject body weight. In some embodiments, the pharmaceutical formulation comprises from about 0.7 to about 5.0 mg / kg of subject body weight, or alternatively, from about 1.0 to about 2.5 mg / kg of subject body weight. Typical dosing regimens for oral administration are administration of the pharmaceutical formulation for oral administration three times a week, twice a week, once a week or daily.

[0438] An effective amount or a therapeutically effective amount or dose of an agent, such as a compound of the present invention, is the amount of the agent or compound that results in amelioration of symptoms or prolongation of survival in a subject. The toxicity and therapeutic efficacy of such molecules can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., to determine the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio LD50 / ED50. Agents that exhibit high therapeutic indices are preferred.

[0439] An effective amount or a therapeutically effective amount is the amount of a compound or pharmaceutical composition that a researcher, veterinarian, physician, or other clinician is seeking to elicit a biological or medical response in a tissue, system, animal, or human. The dose specifically falls within a circulating concentration range that has little or no toxicity and includes the ED50. The dose can vary within this range depending on the dosage form employed and / or the route of administration utilized. Given the particular circumstances of the subject's medical condition, the precise formulation, route of administration, dose, and dosing interval should be selected according to methods known in the art.

[0440] The dose and interval can be adjusted individually to provide a plasma level of the active moiety sufficient to achieve the desired effect; i.e., the minimum effective concentration (MEC). For each compound, the MEC will vary, but can be estimated from, e.g., in vitro data and animal experimentation. The dose required to achieve the MEC will depend on the individual characteristics and the route of administration. In the case of topical administration or selective uptake, the effective local concentration of the drug may be independent of the plasma concentration.

[0441] The amount of the compound or composition administered can depend on a variety of factors, including the sex, age, and weight of the subject to be treated, the severity of the affliction, the mode of administration, and the judgment of the prescribing physician.

[0442] If desired, the compounds and compositions of the present invention can be present in a package or dispenser device that contains one or more unit dosage forms of the active ingredient. Such a package or device can include, for example, a metal or plastic foil, such as a blister pack, or glass, and a rubber stopper, such as in a vial. The package or dispenser device can be accompanied by instructions for administration. Compositions can also be prepared that include the compounds of the present invention formulated in a compatible pharmaceutical carrier, placed in a suitable container, and labeled for the treatment of the designated medical condition.

[0443] In view of the disclosure herein, these and other embodiments of the present invention will be readily apparent to and specifically contemplated by those of ordinary skill in the art.

[0444] Illustrative

[0445] Purity Determination Using HPLC

[0446] The purity of the compound and its synthetic intermediates was determined by reverse-phase HPLC using any one of the methods described below:

[0447] Method A: Mobile phase: A: water (0.01% TFA); B: acetonitrile (0.01% TFA); Gradient phase: increase from 5% B to 95% B in 1.4 minutes, 95% B for 1.6 minutes (total running time: 3 minutes); Flow rate: 2.3 mL / min. Column: SunFire C18, 4.6 * 50 mm, 3.5 μm; Column temperature: 50 °C. Detector: ADC ELSD, DAD (214 nm and 254 nm), ES-API.

[0448] Method B: Mobile phase: A: water (10 mM NH4HCO3); B: acetonitrile; Gradient phase: increase from 5% B to 95% B in 1.5 minutes, 95% B for 1.5 minutes (total running time: 3 minutes); Flow rate: 2.0 mL / min; Column: XBridge C18, 4.6 * 50 mm, 3.5 um; Column temperature: 40 °C. Detector: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API).

[0449] Synthesis of Exemplary Compounds

[0450] Example 1: Preparation of Compound 1

[0451] tert-Butyl 6-chloronicotinate

[0452]

[0453] To a solution of 6-fluoronicotinic acid (5.0 g, 6.37 mmol) and 4-dimethylaminopyridine (0.39 g, 0.64 mmol) in tetrahydrofuran (50.0 mL) was added di-tert-butyl dicarbonate (10.41 g, 47.77 mmol). The reaction mixture was refluxed for 4 hours and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 10 / 1) to give tert-butyl 6-chloronicotinate as a yellow solid (5.5 g, 5.17 mmol, 81.12% yield). LCMS: m / z = 214.0 (M + H) + , retention time was 1.83 minutes (Method A).

[0454] tert-Butyl 6-hydrazinonicotinate

[0455]

[0456] To a solution of tert-butyl 6-chloronicotinate (5.5 g, 25.82 mmol) in ethanol (25.0 mL) was added hydrazine hydrate (6.46 g, 129.11 mmol, 85% in water). The mixture was stirred at 100 °C for 2 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give tert-butyl 6-hydrazinonicotinate (5.0 g, 23.9 mmol, 92.76% yield) as a yellow solid. LCMS: m / z = 210.0 (M+H) + , retention time 1.19 min (Method A).

[0457] tert-butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0458]

[0459] To a solution of (E)-ethyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (2.5 g, 10.86 mmol) and tert-butyl 6-hydrazinonicotinate (2.27 g, 10.86 mmol) in ethanol (25.0 mL) was added p-toluenesulfonic acid monohydrate (410 mg, 2.17 mmol). The mixture was stirred at 80 °C for 12 h and concentrated to dryness. The residue was purified by flash chromatography (methanol / dichloromethane = 1 / 8) to give tert-butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (3.0 g, 8.35 mmol, 76.92% yield) as a yellow solid. LCMS: m / z = 363.1 (M+H) + , retention time 1.98 min (Method A).

[0460] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0461]

[0462] To a solution of tert-butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (1.00 g 2.76 mmol) in dichloromethane (10.0 mL) was added trifluoroacetic acid (5.0 mL). The mixture was stirred at 40 °C for 2 h and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (900 mg, crude) as a yellow solid. LCMS: m / z = 307.0 (M+H) + , retention time 1.77 min (Method A).

[0463] 6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinoyl chloride

[0464]

[0465] To a solution of 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (900 mg, 2.94 mmol) in dichloromethane (10.0 mL) was added thionyl chloride (10.0 mL). The mixture was stirred at 40 °C for 3 h and concentrated to dryness. The crude product (900 mg) was obtained and used in the next step. LCMS: m / z = 325.1 (M+H) + , retention time was 1.96 min (Method A).

[0466] 6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-methoxy-N-methylnicotinamide

[0467]

[0468] To a solution of N,O-dimethylhydroxylamine hydrochloride (407.43 mg, 4.16 mmol) and N,N-diisopropylethylamine (1.07 g, 8.31 mmol) in dichloromethane (5.0 mL) at 0 °C was added 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinoyl chloride (900 mg, 2.77 mmol). The mixture was stirred at 0 °C for 3 h and concentrated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 10 / 1) to obtain 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-methoxy-N-methylnicotinamide as a yellow solid (900 mg, 2.58 mmol, 93.17% yield). LCMS: m / z = 350.1 [M+H] + , retention time was 1.63 min (Method A).

[0469] 4-(1-(5-Acetylpyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0470]

[0471] To a solution of methylmagnesium bromide (0.76 mL, 2.29 mmol, 3 M in diethyl ether) at -20 °C in anhydrous tetrahydrofuran (5.0 mL) was added 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-methoxy-N-methylnicotinamide (200 mg, 0.57 mmol). The mixture was warmed to 0 °C and stirred for an additional hour. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to afford 4-(1-(5-acetylpyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (35 mg, 0.11 mmol, 20.23% yield) as a white solid. LCMS: m / z = 305.0 (M+H) + , retention time 4.504 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 9.00 (s, 1H), 8.58 - 8.43 (m, 3H), 8.18 - 8.09 (m, 3H), 7.73 - 7.69 (m, 2H), 2.63 (s, 3H).

[0472] Example 2: Preparation of Compound 2

[0473] 4-(5-Hydroxy-1-(5-propionylpyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0474]

[0475] To a solution of ethylmagnesium bromide (0.76 mL, 2.29 mmol, 3 M in diethyl ether) at -20 °C in anhydrous tetrahydrofuran (5.0 mL) was added 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-methoxy-N-methylnicotinamide (intermediate from Example 1) (200 mg, 0.57 mmol). The mixture was warmed to 0 °C and stirred for an additional hour. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to afford 4-(5-hydroxy-1-(5-propionylpyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (25 mg, 0.08 mmol, 13.81% yield) as a white solid. LCMS: m / z = 319.0 (M+H) + , retention time 5.01 min (Method A). 1 H NMR (400 MHz, DMSO-d 6)δ13.56(s,1H),9.02(s,1H),8.67(s,1H),8.49(d,J=8.2Hz,2H),8.15 - 8.10(m,2H),7.79(d,J=4.2Hz,2H),3.14 - 3.02(m,2H),1.18 - 1.09(m,3H).

[0476] Example 3: Preparation of Compound 3

[0477] 4-(5-Hydroxy-1-(5-isobutyrylpyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0478]

[0479] To a solution of ethylmagnesium bromide isopropylmagnesium chloride (2.29 mL, 2.29 mmol, 1 M in tetrahydrofuran) in anhydrous tetrahydrofuran (5.0 mL) at -20 °C was added 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-methoxy-N-methylnicotinamide (intermediate from Example 1) (200 mg, 0.57 mmol). The mixture was warmed to 0 °C and stirred for an additional hour. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by reverse-phase preparative HPLC to give 4-(5-hydroxy-1-(5-isobutyrylpyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile as a white solid (16.5 mg, 0.05 mmol, 9.5% yield). LCMS: m / z = 333.1 (M+H) + , retention time 5.10 minutes (Method A). 1 H NMR (400 MHz, DMSO-d 6 )δ9.01(s,1H),8.54(s,1H),8.40(d,J=7.8Hz,1H),8.27(s,1H),8.14(s,1H),8.05(d,J=8.5Hz,2H),7.63(d,J=8.5Hz,2H),3.63(s,1H),1.12(d,J=6.7Hz,6H).

[0480] Example 4: Preparation of Compound 4

[0481] Methyl 2-(4-cyanophenyl)acetate

[0482]

[0483] At 0 °C, hydrochloric acid-containing methanol (20.0 mL, 3.0 N) was added to a mixture of 2-(4-cyanophenyl)acetic acid (5.0 g, 31.0 mmol) in methanol (10.0 mL). The mixture was stirred at 70 °C for 3 hours and cooled to precipitate a solid. The solid was filtered, washed with methanol and dried to give methyl 2-(4-cyanophenyl)acetate (5.0 g, 28.4 mmol, 92% yield) as a yellow solid. LCMS: m / z = 176.0 [M+H] + , retention time 1.54 minutes (Method A).

[0484] (E)-Methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate

[0485]

[0486] To a solution of methyl 2-(4-cyanophenyl)acetate (5.0 g, 28.5 mmol) in N,N-dimethylformamide (25.0 mL) was added N,N-dimethylformamide diethyl acetal (14.0 g, 114.16 mmol). The mixture was stirred at 100 °C for 16 hours and cooled. Ethyl acetate and water were added to the solution and the layers were separated. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (5.20 g, 25.4 mmol, 89% yield) as a yellow solid. LCMS: m / z = 231.0 [M+H] + , retention time 1.70 minutes (Method A). The product was pure enough and used directly in the next step.

[0487] 2-Bromo-5-(methylsulfonyl)pyridine

[0488]

[0489] Under nitrogen, isopropylmagnesium chloride (8.25 mL, 16.5 mmol, 2.0 N in hexanes) was added to a solution of 3,6-dibromopyridine (2.5 g, 12.7 mmol) in anhydrous tetrahydrofuran (10.0 mL) at 0 °C. The mixture was stirred at 0 °C for 45 minutes and then a solution of methanesulfonyl chloride (1.89 g, 16.5 mmol) in anhydrous tetrahydrofuran (5.0 mL) was added. The mixture was warmed to room temperature and stirred for an additional hour. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 3 / 1) to afford 2-bromo-5-(methylsulfonyl)pyridine as a yellow solid (1.4 g, 5.98 mmol, 47.1% yield). LCMS: m / z = 236.0 (M+H) + , retention time 1.54 minutes (Method A).

[0490] 2-Hydrazino-5-(methylsulfonyl)pyridine

[0491]

[0492] Hydrazine hydrate (1.0 g, 17.0 mmol, 85% in water) was added to a solution of 2-bromo-5-(methylsulfonyl)pyridine (1.0 g, 4.25 mmol) in ethanol (10.0 mL). The mixture was stirred at 80 °C for 4 hours. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 2-hydrazino-5-(methylsulfonyl)pyridine as a white solid (1.2 g, 6.4 mmol, 75% yield). LCMS: m / z = 188.0 (M+H)+, retention time 0.43 minutes (Method A).

[0493] 4-(5-Hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0494]

[0495] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (230 mg, 1.0 mmol) and 2-hydrazino-5-(methylsulfonyl)pyridine (187 mg, 1.0 mmol) in ethanol (3.0 mL) was added p-toluenesulfonic acid monohydrate (38 mg, 0.2 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (58 mg, 0.17 mmol, 17.0% yield) as a white solid. LCMS: m / z = 341.0 (M+H) + , retention time 3.93 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.68 (s, 1H), 8.93 (s, 1H), 8.68 - 8.76 (m, 2H), 8.49 - 8.51 (m, 2H), 8.15 - 8.17 (d, J = 6.5 Hz, 2H), 7.78 - 7.80 (d, J = 6.9 Hz, 2H), 3.35 (s, 3H).

[0496] Example 5: Preparation of Compound 5

[0497] Ethyl 2-(3-cyanophenyl)acetate

[0498]

[0499] To a mixture of ethyl 2-(3-bromophenyl)acetate (2.5 g, 10.3 mmol) and zinc cyanide (1.20 g, 10.3 mmol) in N,N-dimethylformamide (30.0 mL) was added tetrakis(triphenylphosphine)palladium (1.16 g, 1.0 mmol). The mixture was stirred under nitrogen at 90 °C for 18.0 h and cooled to room temperature. The mixture was diluted with water and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 9 / 1) to afford ethyl 2-(3-cyanophenyl)acetate (1.72 g, 9.1 mmol, 88.3% yield) as a yellow oil. LC-MS: m / z = 190.0 (M+H) + , retention time 1.65 min (Method A).

[0500] (E)-Ethyl 2-(3-cyanophenyl)-3-(dimethylamino)acrylate

[0501]

[0502] To a solution of ethyl 2-(3-cyanophenyl)acetate (1.2 g, 6.35 mmol) in N,N-dimethylformamide (8.0 mL) was added N,N-dimethylformamide diethyl acetal (4.7 g, 31.74 mmol). The mixture was stirred at 100 °C for 16.0 h and cooled. Ethyl acetate and water were added to the solution and the layers were separated. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give ethyl (E)-2-(3-cyanophenyl)-3-(dimethylamino)acrylate (1.34 g, 5.49 mmol, 86.5% yield). LC-MS: m / z = 245.0 [M+H] + , retention time 1.50 min (Method A).

[0503] 3-(5-Hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0504]

[0505] To a solution of ethyl (E)-2-(3-cyanophenyl)-3-(dimethylamino)acrylate (200 mg, 0.82 mmol) and 2-hydrazino-5-(methylsulfonyl)pyridine (intermediate from Example 4) (153 mg, 0.82 mmol) in ethanol (3.0 mL) was added p-toluenesulfonic acid monohydrate (38 mg, 0.2 mmol). The mixture was stirred at 90 °C in a sealed tube for 12.0 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 3-(5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile as a white solid (60 mg, 0.17 mmol, 21.5% yield). LC-MS: m / z = 341.1 (M+H) + , retention time 1.57 min (Method A). 1 HNMR (400 MHz, DMSO-d 6 ) δ 13.43 (s, 1H), 8.94 (s, 1H), 8.66 - 8.73 (m, 2H), 8.49 - 8.52 (d, J = 10.8 Hz, 2H), 8.41 (s, 1H), 8.30 - 8.31 (d, J = 6.5 Hz, 1H), 7.55 - 7.62 (m, 2H), 3.36 (s, 3H).

[0506] Example 6: Preparation of Compound 6

[0507] Ethyl 2-(4-cyano-2-methylphenyl)acetate

[0508]

[0509] A mixture of 4-bromo-3-methylbenzonitrile (5.0 g, 25.6 mmol), diethyl malonate (27 g, 168 mmol), tris(dibenzylideneacetone)dipalladium(0) (0.24 g, 0.26 mmol), tri-tert-butylphosphine tetrafluoroborate (0.08 g, 0.26 mmol), potassium carbonate (5.3 g, 38.4 mmol) and potassium bicarbonate (3.84 g, 38.4 mmol) was stirred at 160 ° C for 12.0 hours. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate=1 / 1) to give ethyl 2-(4-cyano-2-methylphenyl)acetate (2.0 g, 8.11 mmol, 31.7% yield) as a yellow oil. LC-MS: m / z=204.1 (M+H) + , retention time 1.87 min (Method A).

[0510] (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino) ethyl acrylate:

[0511]

[0512] N,N-dimethylformamide diethyl acetal (2.9 g, 25.0 mmol) was added to a solution of ethyl 2-(4-cyano-2-methylphenyl)acetate (1.0 g, 5.0 mmol) in N,N-dimethylformamide (10.0 mL). The mixture was stirred overnight at 100 ° C and cooled to room temperature. Ethyl acetate and water were added to the solution and the layers were separated. The organic layer was washed with brine, dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (dichloromethane / methanol=98 / 2) to give (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino) ethyl acrylate (600 mg, 2.36 mmol, 47.2% yield) as a yellow oil. LC-MS: m / z=259.0[M+H] + , retention time 1.68 min (Method B).

[0513] 4-(5-Hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)-3-methylbenzonitrile:

[0514]

[0515] To a solution of ethyl (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate (391 mg, 1.60 mmol) and 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (300 mg, 1.60 mmol) in ethanol (10.0 mL) was added 4-methylbenzenesulfonic acid (34.4 mg, 0.2 mmol). The mixture was stirred at 90 °C for 12.0 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to afford 4-(5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)-3-methylbenzonitrile (77 mg, 0.21 mmol, 13.6% yield) as a white solid. LC-MS: m / z = 355.1 (M+H) + , retention time 1.79 min (Method A). 1 HNMR (400 MHz, DMSO-d 6 ) δ 13.19 (s, 1H), 8.95 (s, 1H), 8.64 - 8.68 (m, 1H), 8.48 - 8.51 (d, J = 8.1 Hz, 1H), 8.28 (s, 1H), 7.60 - 7.80 (m, 3H), 3.35 (s, 3H), 2.36 (s, 3H).

[0516] Example 7: Preparation of Compound 7

[0517] Ethyl 5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate

[0518]

[0519] A mixture of 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (0.4 g, 2.1 mmol), diethyl 2-(ethoxymethylene)malonate (1.15 g, 5.3 mmol) and potassium carbonate (0.73 g, 5.3 mmol) in water / ethanol (30.0 mL / 10.0 mL) was stirred overnight at 60 °C. Aqueous hydrochloric acid (10.0 mL, 3 N) was added to the solution and a solid precipitated. The solid was filtered, washed with water and dried to afford ethyl 5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate (0.4 g, 1.3 mmol, 61.5% yield) as a yellow solid. LCMS: m / z = 312.0 (M+H) + , retention time 1.63 min (Method A).

[0520] Ethyl 5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate

[0521]

[0522] To a solution of ethyl 5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate (373 mg, 1.2 mmol) in dichloromethane / methanol (10.0 mL / 1.0 mL) was added (diazo methyl)trimethylsilane (1.61 mL, 3.23 mmol, 2N in hexanes). The mixture was stirred overnight at 25 °C and concentrated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 100 / 2) to afford ethyl 5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate as a yellow solid (0.3 g, 0.92 mmol, 76.9% yield). LCMS: m / z = 326.0 [M+H] + , retention time 1.75 min (Method A).

[0523] 5-Methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid

[0524]

[0525] To a solution of ethyl 5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylate (0.3 g, 0.92 mmol) in 1,4-dioxane (10.0 mL) at 0 °C was added aqueous lithium hydroxide (3.0 mL, 3.0 mmol, 1.0 N). The mixture was warmed to room temperature and stirred for 18 h. The solution was acidified with 1 N hydrochloric acid and extracted with dichloromethane. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated under reduced pressure to afford 5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid as a yellow solid (0.2 g, 0.61 mmol, 66.8% yield). LCMS: m / z = 326.0 (M+H) + , retention time 1.25 min (Method A).

[0526] 2-(4-Bromo-5-methoxy-1H-pyrazol-1-yl)-5-(methylsulfonyl)pyridine

[0527]

[0528] A mixture of 5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazole-4-carboxylic acid (0.2 g, 0.61 mmol), N-bromosuccinimide (0.18 g, 1.0 mmol) and sodium bicarbonate (0.17 g, 2.0 mmol) in N,N-dimethylformamide (10.0 mL) was stirred at 25 °C for 2 h. The reaction was diluted with water and extracted with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 10 / 1) to afford 2-(4-bromo-5-methoxy-1H-pyrazol-1-yl)-5-(methylsulfonyl)pyridine as a yellow solid (0.16 g, 0.47 mmol, 78% yield). LCMS: m / z = 332.0 (M+H) + , retention time 1.55 min (Method A).

[0529] 2-(4-(5-Methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)phenyl)acetonitrile

[0530]

[0531] A mixture of 2-(4-bromo-5-methoxy-1H-pyrazol-1-yl)-5-(methylsulfonyl)pyridine (0.1 g, 0.3 mmol), 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)acetonitrile (0.15 g, 0.6 mmol), sodium carbonate (64 mg, 0.60 mmol), tetrakis(triphenylphosphine)palladium (35 mg, 0.03 mmol) in 1,4-dioxane / water (10.0 mL / 1.0 mL) was stirred at 110 °C overnight. The mixture was cooled and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to give 2-(4-(5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)phenyl)acetonitrile as a yellow solid (0.1 g, 0.27 mmol, 90% yield). LCMS: m / z = 369.0 [M+H] + , retention time 1.55 min (Method B).

[0532] 2-(4-(5-Hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)phenyl)acetonitrile

[0533]

[0534] A mixture of 2-(4-(5-methoxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)phenyl)acetonitrile (0.1 g, 0.3 mmol) and lithium chloride (0.04 g, 0.90 mmol) in N,N-dimethylformamide (5.0 mL) was stirred overnight at 60 °C. The solution was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and evaporated to dryness. The residue was purified by reverse-phase preparative HPLC to give 2-(4-(5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)phenyl)acetonitrile as a white solid (55 mg, 0.15 mmol, 51.8% yield). LCMS: m / z = 355.0 (M+H) + , retention time 3.42 min (Method B). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.18 (s, 1H), 8.94 (s, 1H), 8.47 - 8.55 (m, 3H), 7.95 - 7.97 (d, J = 6.5 Hz, 2H), 7.32 - 7.34 (d, J = 6.5 Hz, 2H), 4.01 (s, 2H), 3.29 (s, 3H).

[0535] Example 8: Preparation of Compound 8

[0536] Ethyl 2-(2-methoxypyridin-4-yl)acetate

[0537]

[0538] To a solution of 2-methoxy-4-methylpyridine (2.0 g, 16.2 mmol) in anhydrous tetrahydrofuran (50.0 mL) at -78 °C under a nitrogen atmosphere was added lithium diisopropylamide (16.0 mL, 32.0 mmol, 2.0 N in heptane). The mixture was stirred at -78 °C for 10 min and diethyl carbonate (3.78 g, 32.0 mmol) was added. The mixture was warmed to room temperature and stirred for 2 h. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 10 / 1) to afford ethyl 2-(4-cyano-2-methylphenyl)-3-oxobutanoate as a yellow oil (2.0 g, 10.2 mmol, 63.4% yield). LCMS: m / z = 196.1 (M+H) + , retention time 1.97 min (Method A).

[0539] (E)-Ethyl 3-(dimethylamino)-2-(2-methoxypyridin-4-yl)acrylate

[0540]

[0541] To a solution of ethyl 2-(2-methoxypyridin-4-yl)acetate (1.95 g, 10 mmol) in N,N-dimethylformamide (3.0 mL) was added N,N-dimethylformamide diethyl acetal (5.95 g, 50 mmol). The mixture was stirred at 100 °C for 12 h and cooled. Ethyl acetate and water were added to the solution and the layers were separated. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (dichloromethane / methanol = 98 / 2) to give ethyl (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)acrylate (1.1 g, 4.4 mmol, 44% yield). LCMS: m / z = 251.0 [M+H] + , retention time 1.68 min (Method B).

[0542] 4-(2-Methoxypyridin-4-yl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol

[0543]

[0544] To a solution of ethyl (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)acrylate (0.25 g, 1.0 mmol) and 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (0.2 g, 1.0 mmol) in ethanol (3.0 mL) was added p-toluenesulfonic acid monohydrate (0.19 g, 1.0 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(2-methoxypyridin-4-yl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol as a white solid (90 mg, 0.26 mmol, 26% yield). LCMS: m / z = 347.0 (M+H) + , retention time 2.28 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.94 - 8.97 (d, J = 8.5 Hz, 2H), 8.67 - 8.70 (d, J = 8.5 Hz, 1H), 8.49 - 8.52 (d, J = 7.5 Hz, 1H), 8.18 - 8.19 (d, J = 7.6 Hz, 1H), 7.77 - 7.83 (m, 2H), 4.03 (s, 3H), 3.36 (s, 3H).

[0545] Example 9: Preparation of Compound 9

[0546] (E)-Ethyl 2-(4-bromophenyl)-3-(dimethylamino)acrylate

[0547]

[0548] To a solution of ethyl 2-(4-bromophenyl)acetate (1.5 g, 6.2 mmol) in N,N-dimethylformamide (3.0 mL) was added N,N-dimethylformamide diethyl acetal (3.7 g, 31 mmol). The mixture was stirred at 100 °C for 12 h and cooled. Ethyl acetate and water were added to the solution and the layers were separated. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give (E)-ethyl 2-(4-bromophenyl)-3-(dimethylamino)acrylate (1.1 g, 3.7 mmol, 59.5% yield) as a yellow oil. LCMS: m / z = 298.0 [M+H] + , retention time 2.08 min (Method A). The product was pure enough and used directly in the next step.

[0549] 4-(4-Bromophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol

[0550]

[0551] To a solution of (E)-ethyl 2-(4-bromophenyl)-3-(dimethylamino)acrylate (0.5 g, 1.7 mmol) and 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (0.31 g, 1.7 mmol) in ethanol (10.0 mL) was added p-toluenesulfonic acid monohydrate (64 mg, 0.34 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(4-bromophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol (0.3 g, 0.76 mmol, 44.9% yield) as a white solid. LCMS: m / z = 394.0 (M+H) + , retention time 1.88 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.28 (s, 1H), 8.93 (s, 1H), 8.50 - 8.53 (m, 3H), 7.91 - 7.93 (d, J = 8.5 Hz, 2H), 7.53 - 7.55 (d, J = 8.9 Hz, 2H), 3.35 (s, 3H).

[0552] Example 10: Preparation of Compound 10

[0553] 2-Bromo-5-(1H-pyrazol-1-yl)pyridine

[0554]

[0555] A mixture of 2-bromo-5-iodopyridine (1.00 g, 3.52 mmol), 1H-pyrazole (239.8 mg, 3.52 mmol), copper(I) iodide (67.09 mg, 0.35 mmol), potassium phosphate (1.87 g, 8.81 mmol), and (1R,2R)-cyclohexane-1,2-diamine (45.6 mg, 0.4 mmol) in 1,4-dioxane (10.0 mL) was stirred at room temperature for 12 h. The reaction solution was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 6 / 1) to give 2-bromo-5-(1H-pyrazol-1-yl)pyridine as a yellow oil (220 mg, 2.85 mmol, 81.12% yield). LCMS: m / z = 224.1 (M+H) + , retention time 1.55 min (Method A).

[0556] 2-Hydrazino-5-(1H-pyrazol-1-yl)pyridine

[0557]

[0558] Hydrazine hydrate (223.2 mg, 4.46 mmol, 85% in water) was added to a solution of 2-bromo-5-(1H-pyrazol-1-yl)pyridine (200 mg, 0.89 mmol) in ethanol (2.0 mL). The mixture was stirred in a sealed tube at 100 °C for 2 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 2-hydrazino-5-(1H-pyrazol-1-yl)pyridine as a yellow solid (140 mg, 0.80 mmol, 90.32% yield). LCMS: m / z = 176.1 (M+H) + , retention time 1.01 min (Method B).

[0559] 4-(1-(5-(1H-Pyrazol-1-yl)pyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0560]

[0561] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (210.29 mg, 0.91 mmol) and 2-hydrazino-5-(1H-pyrazol-1-yl)pyridine (160.00 mg, 0.91 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (34 mg, 0.18 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to afford 4-(1-(5-(1H-pyrazol-1-yl)pyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (74 mg, 0.22 mmol, 24.8% yield) as a white solid. LCMS: m / z = 329.1 (M+H) + , retention time 4.50 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.44 (s, 1H), 8.99 (t, J = 1.2 Hz, 1H), 8.62 (d, J = 6.6 Hz, 2H), 8.50 - 8.48 (m, 2H), 8.16 (d, J = 4.0 Hz, 2H), 7.85 - 7.79 (m, 3H), 6.64 (t, J = 2.0 Hz, 1H).

[0562] Example 11: Preparation of Compound 11

[0563] 2-(6-Chloropyridin-3-yl)oxazole

[0564]

[0565] Under a nitrogen atmosphere, n-butyllithium (4.26 mL, 10.65 mmol, 2.5 N in hexanes) was added to a solution of 2-chloro-5-iodopyridine (1.5 g, 6.26 mmol) in anhydrous tetrahydrofuran (5.0 mL) at -78 °C. The mixture was stirred at -78 °C for 30 min and then zinc chloride (18.79 mg, 18.79 mmol, 1.0 N in dichloromethane) was added. The mixture was warmed to room temperature and a solution of tetrakis(triphenylphosphine)palladium (361.78 mg, 0.31 mmol) and oxazole (605.65 mg, 8.77 mmol) in anhydrous tetrahydrofuran (10.0 mL) was added. The mixture was stirred at 60 °C for 4 h and concentrated. The residue was partitioned between ethyl acetate and water. The organic layer was washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 5 / 1) to afford 2-(6-chloropyridin-3-yl)oxazole (900 mg, 4.94 mmol, 79.93% yield) as a white solid. LCMS: m / z = 181.1 (M+H)+ , retention time is 1.51 minutes (Method A).

[0566] 2-(6-Hydrazinopyridin-3-yl)oxazole

[0567]

[0568] To a solution of 2-(6-chloropyridin-3-yl)oxazole (700 mg, 3.13 mmol) in ethanol (5.0 mL) was added hydrazine hydrate (781.25 mg, 15.63 mmol, 85% in water). The mixture was stirred in a sealed tube at 100 °C for 2 hours. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 2-(6-hydrazinopyridin-3-yl)oxazole (400 mg, 2.25 mmol, 72.72% yield) as a yellow solid. LCMS: m / z = 176.1 (M+H) + , retention time is 0.99 minutes (Method B).

[0569] 4-(5-Hydroxy-1-(5-(oxazol-2-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0570]

[0571] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (328.57 mg, 1.43 mmol) and 2-(6-hydrazinopyridin-3-yl)oxazole (250.00 mg, 1.43 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (55 mg, 0.29 mmol). The mixture was stirred at reflux for 12 hours and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(5-hydroxy-1-(5-(oxazol-2-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (150 mg, 0.46 mmol, 31.91% yield) as a white solid. LCMS: m / z = 330.0 (M+H) + , retention time is 1.74 minutes (Method B). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.54 (s, 1H), 9.04 (s, 1H), 8.64 (s, 1H), 8.61–8.46 (m, 2H), 8.32 (d, J = 0.7 Hz, 1H), 8.14 (d, J = 8.4 Hz, 2H), 7.78 (d, J = 8.6 Hz, 2H), 7.46 (d, J = 0.7 Hz, 1H).

[0572] Example 12: Preparation of Compound 12

[0573] 2-(6-Chloropyridin-3-yl)thiazole

[0574]

[0575] To a solution of 5-bromo-2-chloropyridine (500.0 mg, 2.60 mmol) and 2-(tributylstannyl)thiazole (1458.2 mg, 3.90 mmol) in N,N-dimethylformamide (10.0 mL) was added bis(triphenylphosphine)palladium(II) dichloride (182.37 mg, 0.26 mmol). The reaction was stirred in a sealed tube at 100 °C for 3 hours. The mixture was cooled to room temperature and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 20 / 1) to give 2-(6-chloropyridin-3-yl)thiazole (350 mg, 1.77 mmol, 68% yield). LCMS: m / z = 197.0 [M+H] + , retention time 1.719 min (Method A).

[0576] 2-(6-Hydrazinopyridin-3-yl)thiazole

[0577]

[0578] A mixture of 2-(6-chloropyridin-3-yl)thiazole (300.0 mg, 1.53 mmol) in ethanol (3.0 mL) and hydrazine hydrate (3.0 mL, 85% in water) was stirred in a sealed tube at 110 °C for 3 hours. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 2-(6-hydrazinopyridin-3-yl)thiazole (185 mg, 0.96 mmol, 63% yield). LCMS: m / z = 193.0 [M+H] + , retention time 1.120 min (Method B). The product was pure enough and used directly for the next step.

[0579] 4-(5-Hydroxy-1-(5-(thiazol-2-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0580]

[0581] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (200.0 mg, 0.87 mmol) in ethanol (8 mL) was added 2-(6-hydrazinopyridin-3-yl)thiazole (166.97 mg, 0.87 mmol) and p-toluenesulfonic acid monohydrate (17 mg, 0.09 mmol). The reaction was stirred at 90 °C for 16 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(5-hydroxy-1-(5-(thiazol-2-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (80 mg, 0.23 mmol, 27% yield) as a yellow solid. LCMS: m / z = 346.0 [M+H] + , retention time = 1.893 min (Method B). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.51 (br, 1H), 9.05 (s, 1H), 8.65 (s, 1H), 8.53 (s, 2H), 8.14 (d, J = 7.5 Hz, 2H), 8.01 (d, J = 2.5 Hz, 1H), 7.90 (d, J = 3.0 Hz, 1H), 7.79 (d, J = 8.0 Hz, 2H).

[0582] Example 13: Preparation of Compound 13

[0583] 6'-Chloro-2,3'-bipyridine

[0584]

[0585] A mixture of 2-bromopyridine (390 mg, 2.5 mmol), (6-chloropyridin-3-yl)boronic acid (470 mg, 3.0 mmol), potassium carbonate (828 mg, 6.0 mmol), palladium(II) acetate (56 mg, 0.6 mmol) in 1,2-dimethoxyethane / water (10.0 mL / 2.0 mL) was stirred overnight at 90 °C. The mixture was cooled and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 3 / 1) to give 6'-chloro-2,3'-bipyridine (380 mg, 2.0 mmol, 80% yield) as a yellow solid. LCMS: m / z = 191.0 [M+H] + , retention time = 2.20 min (Method A).

[0586] 6'-Hydrazino-2,3'-bipyridine

[0587]

[0588] To a solution of 6'-chloro-2,3'-bipyridine (380 mg, 2.0 mmol) in ethanol (10.0 mL) was added hydrazine hydrate (5.0 mL, 85% in water). The mixture was stirred overnight at 120 °C in a sealed tube. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give 6'-hydrazino-2,3'-bipyridine as a yellow oil (200 mg, 1.07 mmol, 54% yield). LCMS: m / z = 187.1 [M+H] + , retention time 0.54 min (Method A). The product was used directly in the next step.

[0589] 4-(1-([2,3'-Bipyridin]-6'-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0590]

[0591] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (240 mg, 1.07 mmol) and 6'-hydrazino-2,3'-bipyridine (200 mg, 1.07 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (38.2 mg, 0.20 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(1-([2,3'-bipyridin]-6'-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile as a white solid (35 mg, 0.10 mmol, 9.64% yield). LCMS: m / z = 340.2 (M+H) + , retention time 4.71 min (Method A). 1 H NMR (500 MHz, DMSO-d 6 ) δ 9.19 (s, 1H), 8.81–8.66 (m, 2H), 8.60 (s, 1H), 8.49 (s, 1H), 8.12 (dd, J = 17.7, 7.9 Hz, 3H), 7.96 (t, J = 7.6 Hz, 1H), 7.79 (d, J = 8.3 Hz, 2H), 7.49–7.38 (m, 1H).

[0592] Example 14: Preparation of Compound 14

[0593] 4-(5-Hydroxy-1-(5-(trifluoromethyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0594]

[0595] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (130 mg, 0.56 mmol) and 2-hydrazinyl-5-(trifluoromethyl)pyridine (100 mg, 0.56 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (21 mg, 0.11 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(5-hydroxy-1-(5-(trifluoromethyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (70.8 mg, 0.21 mmol, 38% yield) as a white solid. LCMS: m / z = 331.0 (M+H) + , retention time = 5.08 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.54 (s, 1H), 8.86 (s, 1H), 8.73 (s, 1H), 8.65 (d, J = 8.6 Hz, 1H), 8.41 (dd, J = 8.9, 2.1 Hz, 1H), 8.15 (d, J = 8.3 Hz, 2H), 7.78 (d, J = 8.4 Hz, 2H).

[0596] Example 15: Preparation of Compound 15

[0597] 1-(6-Bromopyridin-3-yl)pyrrolidin-2-one

[0598]

[0599] A mixture of 2-bromo-5-iodopyridine (2.0 g, 7.04 mmol), potassium phosphate (4.5 g, 21.13 mmol), copper(I) iodide (134 mg, 0.70 mmol), pyrrolidin-2-one (1.2 g, 14.09 mmol) and ethylene glycol (44 mg, 0.70 mmol) in isopropanol (20.0 mL) was stirred in a sealed tube at 110 °C for 12 h. The mixture was cooled to room temperature and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to give 1-(6-bromopyridin-3-yl)pyrrolidin-2-one (0.8 g, 9.93 mmol, 47% yield) as a white solid. LCMS: m / z = 241.1 [M+H] + , retention time = 1.67 min (Method A).

[0600] 1-(6-Hydrazinylpyridin-3-yl)pyrrolidin-2-one

[0601]

[0602] To a solution of 1-(6-bromopyridin-3-yl)pyrrolidin-2-one (400 mg, 1.66 mmol) in ethanol (4.0 mL) was added hydrazine hydrate (2.0 mL, 85% in water). The mixture was stirred in a sealed tube at 130 °C for 18 h. The mixture was cooled and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to afford 1-(6-hydrazinopyridin-3-yl)pyrrolidin-2-one as a yellow oil (160 mg, 0.83 mmol, 50% yield). LCMS: m / z = 193.2 [M+H] + , retention time = 0.69 min (Method B).

[0603] 4-(5-Hydroxy-1-(5-(2-oxopyrrolidin-1-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0604]

[0605] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (120 mg, 0.52 mmol) and 1-(6-hydrazinopyridin-3-yl)pyrrolidin-2-one (100 mg, 0.52 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (19 mg, 0.10 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was purified by reverse-phase preparative HPLC to afford 4-(5-hydroxy-1-(5-(2-oxopyrrolidin-1-yl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile as a white solid (9.0 mg, 0.03 mmol, 5.0% yield). LCMS: m / z = 346.4 (M+H) + , retention time was 4.08 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.38 (s, 1H), 8.99–8.70 (m, 1H), 8.70–8.41 (m, 1H), 8.31 (d, 2H), 8.11 (d, J = 7.2 Hz, 2H), 7.79 (d, J = 8.5 Hz, 2H), 3.90 (t, J = 7.0 Hz, 2H), 2.56–2.51 (m, 2H), 2.20–2.03 (m, 2H).

[0606] Example 16: Preparation of Compound 16

[0607] 5-Cyclopropyl-2-fluoropyridine

[0608]

[0609] A mixture of 5-bromo-2-fluoropyridine (435 mg, 2.5 mmol), cyclopropylboronic acid (260 mg, 3.0 mmol), potassium phosphate (1.27 g, 6.0 mmol), palladium(II) acetate (56 mg, 0.6 mmol) and tricyclohexylphosphine (340 mg, 1.2 mmol) in 1,2-dimethoxyethane / water (10.0 mL / 2.0 mL) was stirred overnight at 80 °C. The mixture was cooled and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 3 / 1) to give 5-cyclopropyl-2-fluoropyridine as a yellow solid (246 mg, 1.8 mmol, 72% yield). LCMS: m / z = 138.1 [M+H] + , retention time = 2.25 min (Method A).

[0610] 5-Cyclopropyl-2-hydrazinopyridine

[0611]

[0612] Hydrazine hydrate (5.0 mL, 85% in water) was added to a solution of 5-cyclopropyl-2-fluoropyridine (246 mg, 1.8 mmol) in ethanol (10.0 mL). The mixture was stirred overnight at 120 °C in a sealed tube. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give 5-cyclopropyl-2-hydrazinopyridine as a yellow oil (150 mg, 1.00 mmol, 55% yield). LCMS: m / z = 150.1 [M+H] + , retention time was 0.50 min (Method A). The product was used directly in the next step.

[0613] 4-(1-(5-Cyclopropylpyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0614]

[0615] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (230 mg, 1.00 mmol) and 5-cyclopropyl-2-hydrazinopyridine (150 mg, 1.00 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (38.0 mg, 0.20 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(1-(5-cyclopropylpyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (36 mg, 0.12 mmol, 12% yield) as a white solid. LCMS: m / z = 303.3 (M+H) + , retention time 5.189 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 8.30 (s, 1H), 8.15 (s, 1H), 8.14–8.07 (m, 1H), 8.02 (d, J = 7.5 Hz, 2H), 7.67–7.53 (m, 3H), 1.96–1.82 (m, 1H), 0.95 (d, J = 7.0 Hz, 2H), 0.67 (d, J = 7.0 Hz, 2H).

[0616] Example 17: Preparation of Compound 17

[0617] 5-Fluoro-2-hydrazinopyridine

[0618]

[0619] To a solution of 2,5-difluoropyridine (500 mg, 4.34 mmol) in ethanol (2.0 mL) was added hydrazine hydrate (434 mg, 8.69 mmol, 85% in water). The mixture was stirred overnight in a sealed tube at 120 °C. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 5-fluoro-2-hydrazinopyridine (270 mg, 2.13 mmol, 49% yield) as a white solid. LCMS: m / z = 128.1 (M+H) + , retention time 0.33 min (Method A).

[0620] 4-(1-(5-Fluoropyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0621]

[0622] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (181 mg, 0.79 mmol) and 5-fluoro-2-hydrazinopyridine (100 mg, 0.79 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (30.4 mg, 0.16 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(1-(5-fluoropyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (94.6 mg, 0.34 mmol, 43% yield) as a white solid. LCMS: m / z = 281.3 (M+H) + , retention time 4.38 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.30 (s, 1H), 8.61 (s, 1H), 8.52 (d, J = 2.7 Hz, 1H), 8.46 (m, 1H), 8.14 (d, J = 8.1 Hz, 2H), 8.00 (td, J = 8.8, 2.9 Hz, 1H), 7.78 (d, J = 8.4 Hz, 2H).

[0623] Example 18: Preparation of Compound 18

[0624] 4-(1-(5-Chloropyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0625]

[0626] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (150.0 mg, 0.65 mmol) in ethanol (3.0 mL) was added 5-chloro-2-hydrazinopyridine (93.52 mg, 0.65 mmol) and p-toluenesulfonic acid monohydrate (13.3 mg, 0.07 mmol). The reaction was stirred in a sealed tube at 90 °C for 16 h. The reaction was cooled and concentrated to dryness. The residue was purified by flash chromatography (dichloromethane / ethyl acetate = 100 / 1) to give 4-(1-(5-chloropyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (77 mg, 0.26 mmol, 40% yield) as a yellow solid. LCMS: m / z = 297.0 [M+H] + , retention time 5.095 min (Method A). 1 H NMR (500 MHz, DMSO-d 6)δ 13.35 (s, 1H), 8.65 (s, 1H), 8.55 (d, J = 2.5 Hz, 1H), 8.46 (s, 1H), 8.16 - 8.12 (m, 3H), 7.78 (d, J = 6.8 Hz, 2H).

[0627] Example 19: Preparation of Compound 19

[0628] 4-(4-Chlorophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol

[0629]

[0630] To a solution of ethyl (E)-3-(dimethylamino)-2-(4-chlorophenyl)acrylate (200 mg, 0.79 mmol) and 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (147.83 mg, 0.79 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (30.4 mg, 0.16 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(4-chlorophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol as a white solid (95.00 mg, 0.27 mmol, 34.54% yield). LCMS: m / z = 350.1 (M + H) + , retention time 1.93 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 )δ 8.94 (d, J = 2.8 Hz, 1H), 8.81–8.56 (m, 2H), 8.49 (dd, J = 8.9, 2.4 Hz, 1H), 7.98 (d, J = 8.1 Hz, 2H), 7.42 (d, J = 8.6 Hz, 2H), 3.38 (s, 3H).

[0631] Example 20: Preparation of Compound 20

[0632] 4-(4-Fluorophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol

[0633]

[0634] To a solution of ethyl (E)-3-(dimethylamino)-2-(4-fluorophenyl)acrylate (127 mg, 0.53 mmol) and 2-hydrazinyl-5-(methylsulfonyl)pyridine (intermediate from Example 4) (100 mg, 0.53 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (20.9 mg, 0.11 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(4-fluorophenyl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol (93.8 mg, 52% yield) as a white solid. LCMS: m / z = 334.3 (M+H) + , retention time 4.02 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 13.01 (s, 1H), 8.93 (d, J = 2.1 Hz, 1H), 8.67 (m, 1H), 8.58–8.37 (m, 2H), 7.98 (dd, J = 8.3, 5.7 Hz, 2H), 7.20 (t, J = 8.9 Hz, 2H), 3.35 (s, 3H)

[0635] Example 21: Preparation of Compound 21

[0636] 4-(1-(5-Bromopyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0637]

[0638] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (612 mg, 2.66 mmol) and 5-bromo-2-hydrazinylpyridine (500 mg, 2.66 mmol) in ethanol (10.0 mL) was added p-toluenesulfonic acid monohydrate (51 mg, 0.27 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(1-(5-bromopyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (680 mg, 1.99 mmol, 75% yield) as a white solid. LCMS: m / z = 340.9 (M+H) + , retention time 4.99 min (Method A). 1 HNMR (500 MHz, DMSO-d 6)δ 13.35 (s, 1H), 8.69–8.58 (m, 2H), 8.41 (s, 1H), 8.25 (dd, J = 8.9, 2.3 Hz, 1H), 8.14 (d, J = 8.2 Hz, 2H), 7.78 (d, J = 8.4 Hz, 2H).

[0639] Example 22: Preparation of Compound 22

[0640] N-(6-Fluoropyridin-3-yl)cyclopropanecarboxamide

[0641]

[0642] To a solution of 6-fluoropyridin-3-amine (500.0 mg, 4.46 mmol) in dichloromethane (20.0 mL) at 0 °C was added cyclopropanecarbonyl chloride (559.4 mg, 5.35 mmol, 0.48 mL) and triethylamine (902.6 mg, 8.92 mmol, 1.24 mL). The mixture was warmed to room temperature and stirred for 2 h. The reaction was diluted with water and extracted twice with dichloromethane. The organic layer was washed with brine, dried over sodium sulfate and concentrated to give N-(6-fluoropyridin-3-yl)cyclopropanecarboxamide (850 mg of crude product). LCMS: m / z = 181.0 [M+H] + , retention time 1.577 min (Method A). The product was pure enough and used directly for the next step.

[0643] N-(6-Hydrazinopyridin-3-yl)cyclopropanecarboxamide

[0644]

[0645] A mixture of N-(6-fluoropyridin-3-yl)cyclopropanecarboxamide (850.0 mg, 4.72 mmol) in ethanol (5.0 mL) and hydrazine hydrate (5.0 mL, 85% in water) was stirred in a sealed tube at 110 °C for 3 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give N-(6-hydrazinopyridin-3-yl)cyclopropanecarboxamide (312 mg of crude product). LCMS: m / z = 193.0 [M+H] + , retention time 0.867 min (Method A). The product was pure enough and used directly for the next step.

[0646] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)cyclopropanecarboxamide

[0647]

[0648] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (200.0 mg, 0.87 mmol) and N-(6-hydrazinopyridin-3-yl)cyclopropanecarboxamide (166.9 mg, 0.87 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (17.1 mg, 0.09 mmol). The mixture was stirred at 90 °C for 3 h, cooled to room temperature and evaporated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 20 / 1) to afford N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)cyclopropanecarboxamide (78 mg, 0.22 mmol, 26% yield) as a yellow solid. LCMS: m / z = 346.0 [M+H] + , retention time 4.330 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.56 (s, 1H), 8.77 (d, J = 2.4 Hz, 1H), 8.51 - 8.44 (m, 1H), 8.27 - 8.24 (m, 1H), 8.17 - 8.13 (m, 1H), 8.09 (d, J = 8.0 Hz, 2H), 7.78 (d, J = 8.4 Hz, 2H), 1.82 - 1.78 (m, 1H), 0.85 (d, J = 6.0 Hz, 4H).

[0649] Example 23: Preparation of Compound 23

[0650] N-(6-Fluoropyridin-3-yl)propanamide

[0651]

[0652] To a solution of 6-fluoropyridin-3-amine (500.0 mg, 4.46 mmol) in dichloromethane (20 mL) at 0 °C was added propanoyl chloride (495.16 mg, 5.35 mmol, 0.47 mL) and triethylamine (902.60 mg, 8.92 mmol, 1.24 mL). The mixture was warmed to room temperature and stirred for 2 h. The reaction was diluted with water and extracted with dichloromethane. The organic layer was washed with brine, dried over sodium sulfate and concentrated to afford N-(6-fluoropyridin-3-yl)propanamide (808 mg crude product). LCMS: m / z = 169.0 [M+H] + , retention time 1.498 min (Method A). The product was pure enough and used directly in the next step.

[0653] N-(6-Hydrazinopyridin-3-yl)propanamide

[0654]

[0655] A mixture of N-(6-fluoropyridin-3-yl)propanamide (800.00 mg, 4.76 mmol) in ethanol (5 mL) and hydrazine hydrate (5.0 mL, 85% in water) was stirred in a sealed tube at 110 °C for 3 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give N-(6-hydrazinopyridin-3-yl)propanamide (294 mg of crude product). LCMS: m / z = 181.0 [M+H] + , retention time 0.317 min (Method A). The product was pure enough and used directly in the next step.

[0656] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)propanamide

[0657]

[0658] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (200.0 mg, 0.87 mmol) and N-(6-hydrazinopyridin-3-yl)propanamide (156.5 mg, 0.87 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (14.9 mg, 0.09 mmol). The mixture was stirred at 90 °C for 3 h, cooled to room temperature and evaporated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 20 / 1) to give N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)propanamide as a yellow solid (23 mg, 0.07 mmol, 8.0% yield). LCMS: m / z = 334.0 [M+H] + , retention time 4.322 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.23 (s, 1H), 8.78 (s, 1H), 8.51 - 8.44 (m, 1H), 8.26 - 8.21 (m, 1H), 8.18 - 8.15 (m, 1H), 8.09 (d, J = 8.0 Hz, 2H), 7.78 (d, J = 8.4 Hz, 2H), 2.37 (q, J = 7.6 Hz, 2H), 1.58 (t, J = 7.6 Hz, 3H).

[0659] Example 24: Preparation of Compound 24

[0660] N-(6-Fluoropyridin-3-yl)methanesulfonamide

[0661]

[0662] To a solution of 6-fluoropyridin-3-amine (500 mg, 4.46 mmol) in pyridine (0.5 mL) at 0 °C was added methanesulfonyl chloride (2.5 mL). The mixture was warmed to room temperature and stirred for an additional hour. The reaction was diluted with water and extracted twice with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and concentrated to afford N-(6-fluoropyridin-3-yl)methanesulfonamide as a yellow oil (600 mg, 3.12 mmol, 70% yield). LCMS: m / z = 190.9 [M+H] + , retention time 1.38 min (Method A).

[0663] N-(6-Hydrazinopyridin-3-yl)methanesulfonamide

[0664]

[0665] To a solution of N-(6-fluoropyridin-3-yl)methanesulfonamide (600 mg, 3.15 mmol) in ethanol (4.0 mL) was added hydrazine hydrate (2.0 mL, 85% in water). The mixture was stirred overnight at 110 °C in a sealed tube. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and concentrated to afford N-(6-hydrazinopyridin-3-yl)methanesulfonamide as a yellow oil (300 mg, 1.48 mmol, 47% yield). LCMS: m / z = 203.2 [M+H] + , retention time 0.34 min (Method A). The product was pure enough and used directly in the next step.

[0666] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)methanesulfonamide

[0667]

[0668] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (341 mg, 1.48 mmol) and N-(6-hydrazinylpyridin-3-yl)methanesulfonamide (300 mg, 1.48 mmol) in ethanol (4.0 mL) was added p-toluenesulfonic acid monohydrate (28.5 mg, 0.15 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to afford N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)methanesulfonamide (250 mg, 0.72 mmol, 49% yield) as a white solid. LCMS: m / z = 356.4 (M+H) + , retention time 3.74 min (Method A). 1 H NMR (500 MHz, DMSO-d 6 ) δ 10.03 (s, 1H), 8.51 (s, 1H), 8.32 (d, J = 13.4 Hz, 2H), 8.11 (d, J = 7.8 Hz, 2H), 7.86 (d, J = 8.1 Hz, 1H), 7.77 (d, J = 7.9 Hz, 2H), 3.08 (s, 3H).

[0669] Example 25: Preparation of Compound 25

[0670] N-(6-Fluoropyridin-3-yl)ethanesulfonamide

[0671]

[0672] To a solution of 6-fluoropyridin-3-amine (500 mg, 4.46 mmol) in pyridine (5.0 mL) at 0 °C was added ethanesulfonyl chloride (689.71 mg, 5.36 mmol). The mixture was warmed to room temperature and stirred for an additional hour. The reaction was diluted with water and extracted twice with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and concentrated to afford N-(6-fluoropyridin-3-yl)ethanesulfonamide (900 mg, 4.38 mmol, 98.26% yield) as a white solid. LCMS: m / z = 205.1 [M+H] + , retention time 1.32 min (Method A).

[0673] N-(6-Hydrazinylpyridin-3-yl)ethanesulfonamide

[0674]

[0675] To a solution of N-(6-fluoropyridin-3-yl)ethanesulfonamide (910 mg, 4.46 mmol) in ethanol (5.0 mL) was added hydrazine hydrate (1.05 g, 17.84 mmol, 85% in water). The mixture was stirred in a sealed tube at 100 °C for 4 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give N-(6-hydrazinopyridin-3-yl)ethanesulfonamide as a white solid (810.0 mg, 3.75 mmol, 84.11%). LCMS: m / z = 217.0 (M+H) + , retention time 0.36 min (Method A).

[0676] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)ethanesulfonamide

[0677]

[0678] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (212.96 mg, 0.93 mmol) and N-(6-hydrazinopyridin-3-yl)ethanesulfonamide (200.00 mg, 0.93 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (36.1 mg, 0.19 mmol). The mixture was stirred at reflux for 12 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)ethanesulfonamide as a white solid (75 mg, 0.20 mmol, 21.86%). LCMS: m / z = 370.0 (M+H) + , retention time 4.01 min (Method A). 1 H NMR (400 MHz, DMSO-d 6 ) δ 10.13 (s, 1H), 8.55 (s, 1H), 8.34 (s, 1H), 8.32–8.18 (m, 1H), 8.11 (d, J = 7.5 Hz, 2H), 7.86 (dd, J = 9.0, 2.5 Hz, 1H), 7.79 (d, J = 8.4 Hz, 2H), 3.18 (dd, J = 14.6, 7.3 Hz, 2H), 1.24 (t, J = 7.3 Hz, 3H).

[0679] Example 26: Preparation of Compound 26

[0680] 2-Chloro-5-(methylthio)pyridine

[0681] Under nitrogen, n-butyllithium (7.5 mL, 12.0 mmol, 1.6 M in hexanes) was added to a solution of 5-bromo-2-chloropyridine (1.92 g, 10.0 mmol) and N,N,N',N'-tetramethylethylenediamine (1.51 g, 13.0 mmol) in anhydrous tetrahydrofuran (15.0 mL) at -78 °C. The mixture was stirred at -78 °C for 50 minutes and dimethyldisulfide (1.13 g, 12.0 mmol) was added. The mixture was warmed to 20 °C and stirred for an additional hour. The reaction was quenched with saturated ammonium chloride solution and extracted twice with ethyl acetate. The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 50 / 1) to afford 2-chloro-5-(methylthio)pyridine as a yellow oil (1.0 g, 6.29 mmol, 62.9% yield). LC-MS: m / z = 160 (M+H) + , retention time 0.85 min (Method A).

[0682] 2-Chloro-5-(methylsulfinyl)pyridine

[0683]

[0684] 3-Chloroperoxybenzoic acid (1.26 g, 6.22 mmol, 85%) was added to a solution of 2-chloro-5-(methylthio)pyridine (900 mg, 5.66 mmol) in dichloromethane (10.0 mL) at 0 °C. The mixture was stirred at room temperature for 1 hour. The reaction was basified with 10% sodium hydroxide solution and extracted twice with dichloromethane. The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate = 2 / 1) to afford 2-chloro-5-(methylsulfinyl)pyridine as a white solid (700 mg, 4.0 mmol, 70.6% yield). LC-MS: m / z = 176.1 (M+H) + , retention time 0.55 min (Method A).

[0685] 2-Hydrazino-5-(methylsulfinyl)pyridine

[0686]

[0687] To a solution of 2-chloro-5-(methylsulfinyl)pyridine (700 mg, 4.0 mmol) in ethanol (10.0 mL) was added hydrazine hydrate (1.23 g, 20.0 mmol, 85% in water). The mixture was stirred at 80 °C for 4.0 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 2-hydrazino-5-(methylsulfinyl)pyridine as a yellow solid (400 mg, 2.34 mmol, 58.5% yield). LC-MS: m / z = 172.0 (M+H)+, retention time 0.38 min (Method A).

[0688] (6-Chloropyridin-3-yl)(imino)(methyl)-λ 6 -sulfone

[0689]

[0690] To a mixture of 2-chloro-5-(methylsulfinyl)pyridine (200 mg, 1.14 mmol) (intermediate of Example 12) and sodium azide (223 mg, 3.43 mmol) in chloroform (5.0 mL) at 0 °C was added concentrated sulfuric acid (1.0 mL). The mixture was stirred at 55 °C for 16.0 h and cooled. The reaction was diluted with ice water and the organic layer was removed. The aqueous phase was made basic by addition of ammonium hydroxide solution, whereby an oil was separated, which was extracted with dichloromethane. The organic layer was separated, washed with brine, dried over sodium sulfate and concentrated to give (6-chloropyridin-3-yl)(imino)(methyl)-λ 6 -sulfone as a yellow solid (120 mg, 0.63 mmol, 55.4% yield). LC-MS: m / z = 191.0 (M+H) + , retention time 1.3 min (Method A).

[0691] (6-Hydrazinopyridin-3-yl)(imino)(methyl)-λ 6 -sulfone

[0692]

[0693] To (6-chloropyridin-3-yl)(imino)(methyl)-λ 6A solution of -sulfone (120 mg, 0.63 mmol) in ethanol (10.0 mL) was added with hydrazine hydrate (200 mg, 3.15 mmol, 85% in water). The mixture was stirred at 80 °C for 4.0 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give (6-hydrazinopyridin-3-yl)(imino)(methyl)-λ 6 -sulfone (100 mg, 0.54 mmol, 85.3% yield). LC-MS: m / z = 187.0 (M+H)+, retention time 0.36 min (Method A).

[0694] 4-(5-Hydroxy-1-(5-(S-methylsulfinyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile

[0695]

[0696] To a solution of (E)-2-(4-cyanophenyl)-3-(dimethylamino)methyl acrylate (150 mg, 0.65 mmol) and (6-hydrazinopyridin-3-yl)(imino)(methyl)-λ 6 -sulfone (121 mg, 0.65 mmol) in ethanol (6.0 mL) was added p-toluenesulfonic acid monohydrate (24.7 mg, 0.13 mmol). The mixture was stirred overnight at 90 °C in a sealed tube and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 4-(5-hydroxy-1-(5-(S-methylsulfinyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (formate) (14 mg, 0.04 mmol, 5.5% yield) as a white solid. LC-MS: m / z = 340.0 (M+H) + , retention time 3.50 min (Method A). 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.90 (s, 1H), 8.61 - 8.63 (m, 1H), 8.30 - 8.33 (m, 2H), 8.14 (s, 1H), 8.03 - 8.05 (d, J = 7.4 Hz, 2H), 7.61 - 7.63 (d, J = 8.9 Hz, 2H), 3.13 (s, 3H).

[0697] Example 27: Preparation of Compound 27

[0698] 6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-(methylsulfonyl)nicotinamide

[0699]

[0700] To a solution of methanesulfonamide (35.3 mg, 0.37 mmol) and triethylamine (74.9 mg, 0.74 mmol) in dichloromethane (5.0 mL) at 0 °C was added 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinoyl chloride (intermediate from Example 1) (100 mg, 0.31 mmol). The mixture was stirred overnight at room temperature and concentrated to dryness. The residue was purified by reverse-phase preparative HPLC to afford 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-N-(methylsulfonyl)nicotinamide as a white solid (17.6 mg, 0.05 mmol, 6.2% yield). LC-MS: m / z = 384.1 (M+H) + , retention time 4.24 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 ) δ 8.28 - 8.20 (m, 3H), 8.01 - 7.99 (m, 4H), 7.55 - 7.50 (m, 2H), 2.89 (s, 3H).

[0701] Example 28: Preparation of Compound 28

[0702] tert-Butyl 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinate

[0703]

[0704] To a solution of tert-butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (intermediate from Example 1) (1.0 g, 2.76 mmol) in dichloromethane / methanol (29.0 mL / 5.0 mL) was added (diazo methyl)trimethylsilane (2.07 mL, 4.14 mmol, 2 M in hexanes). The mixture was stirred overnight at 25 °C and concentrated to dryness. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 3 / 1) to afford tert-butyl 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinate as a yellow solid (200 mg, 0.53 mmol, 19.2% yield). LC-MS: m / z = 377.0 [M+H] + , retention time 2.40 minutes (Method A).

[0705] 6-(4-(4-Cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinic acid

[0706]

[0707] To a solution of tert-butyl 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinate (200 mg, 0.53 mmol) in dichloromethane (10.0 mL) was added trifluoroacetic acid (5.0 mL). The mixture was stirred at 40 °C for 2.0 h and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinic acid as a yellow solid (150 mg, 0.47 mmol, 88.4% yield). LC-MS: m / z = 321.0 (M+H) + , retention time 1.98 min (Method A).

[0708] 4-(1-(5-Isocyanatopyridin-2-yl)-5-methoxy-1H-pyrazol-4-yl)benzonitrile

[0709]

[0710] A mixture of 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)nicotinic acid (150 mg, 0.47 mmol), diphenylphosphoryl azide (194 mg, 0.71 mmol) and triethylamine (95 mg, 0.94 mmol) in toluene (5.0 mL) was stirred at 110 °C for 3 h. The reaction was diluted with water and extracted with ethyl acetate. The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated under reduced pressure. The crude product 4-(1-(5-isocyanatopyridin-2-yl)-5-methoxy-1H-pyrazol-4-yl)benzonitrile was obtained as a yellow syrup (150 mg, crude). LC-MS: m / z = 345.9 (M+H) + , retention time 2.01 min (Method A). The crude product was used in the next step.

[0711] N-(6-(4-(4-Cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide

[0712]

[0713] A mixture of 4-(1-(5-isocyanatopyridin-2-yl)-5-methoxy-1H-pyrazol-4-yl)benzonitrile (150 mg, crude product) and morpholine (174 mg, 2.0 mmol) in dichloromethane (5.0 mL) was stirred overnight at room temperature. The reaction was diluted with water and extracted with dichloromethane. The organic layer was separated, washed with brine solution, dried over sodium sulfate and concentrated to dryness. The residue was purified by flash chromatography (methanol / dichloromethane = 1 / 10) to give N-(6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide (50 mg, 0.12 mmol, 26.3% yield) as a yellow solid. LC-MS: m / z = 405.1 [M+H] + , retention time 1.96 min (Method A). The product was used in the next step.

[0714] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide

[0715]

[0716] Lithium chloride (50.4 mg, 1.2 mmol) was added to a solution of N-(6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide (50 mg, 0.12 mmol) in N,N-dimethylformamide (6.0 mL). The mixture was stirred overnight at 60 °C. The solution was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and evaporated to dryness. The residue was purified by reverse-phase preparative HPLC to give N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide (formate) (9.7 mg, 0.02 mmol, 18.6% yield) as a white solid. LC-MS: m / z = 391.0 (M+H) + , retention time 4.27 min (Method A). 1 1HNMR (400 MHz, DMSO-d 6 ) δ 8.87 (s, 1H), 8.64 (s, 1H), 8.37 (s, 1H), 8.17 (s, 1H), 8.08 - 8.03 (m, 3H), 7.74 - 7.72 (m, 2H), 3.63 - 3.62 (m, 4H), 3.47 - 3.45 (m, 4H).

[0717] Example 29: Preparation of Compound 29

[0718] 6-Hydrazinylpyridine-3-sulfonamide

[0719]

[0720] Hydrazine hydrate (5.0 mL, 85% in water) was added to a solution of 6-chloropyridine-3-sulfonamide (1.63 g, 8.5 mmol) in ethanol (5.0 mL). The mixture was stirred in a sealed tube at 100 °C for 4 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give 6-hydrazinopyridine-3-sulfonamide (600 mg, 3.20 mmol, 37.7% yield) as a yellow solid. LCMS: m / z = 189.0 (M+H) + , retention time 0.32 min (Method A).

[0721] 6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide

[0722]

[0723] p-Toluenesulfonic acid monohydrate (76.4 mg, 0.4 mmol) was added to a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (452 mg, 1.97 mmol) and 6-hydrazinopyridine-3-sulfonamide (370 mg, 1.97 mmol) in ethanol (8.0 mL). The mixture was stirred at 90 °C for 16.0 h and cooled to precipitate a solid. The solid was filtered, washed with ethanol and dried to give 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide (340 mg, 1.0 mmol, 50.8% yield) as a white solid. LC-MS: m / z = 342.0 (M+H) + , retention time 1.76 min (Method A).

[0724] 6-(4-(4-Cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide and 6-(4-(4-cyano-phenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridine-3-sulfonamide

[0725]

[0726] To a solution of 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide (340 mg, 1.0 mmol) in dichloromethane / methanol (15.0 mL / 3.0 mL) was added (diazomethyl)trimethylsilane (0.75 mL, 1.5 mmol, 2 M in hexanes). The mixture was stirred overnight at 25 °C and concentrated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 100 / 2) to afford the two isomers 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide and 6-(4-(4-cyanophenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridine-3-sulfonamide as a yellow solid (245 mg, 0.69 mmol, 69% yield). LC-MS: m / z = 356.0 [M+H] + , retention time 1.66 min (Method A). The two isomers were used in the next step without separation.

[0727] N-((6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide and N-((6-(4-(4-cyanophenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide

[0728]

[0729] To a solution of 6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridine-3-sulfonamide and 6-(4-(4-cyanophenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridine-3-sulfonamide (245 mg, 0.69 mmol) in anhydrous tetrahydrofuran (10.0 mL) at 0 °C was added triethylamine (140 mg, 1.38 mmol) and acetyl chloride (69 mg, 0.90 mmol). The mixture was stirred overnight at room temperature and concentrated to dryness. The residue was purified by flash chromatography (dichloromethane / methanol = 20 / 1) to afford the two isomers N-((6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide and N-((6-(4-(4-cyanophenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide as a yellow solid (200 mg, 0.40 mmol, 58.3% yield). LC-MS: m / z = 398.0 [M+H] +, the retention time is 1.73 minutes (Method A). The two isomers are used in the next step without separation.

[0730] N-((6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide

[0731]

[0732] To a solution of N-((6-(4-(4-cyanophenyl)-5-methoxy-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide and N-((6-(4-(4-cyanophenyl)-2-methyl-5-oxo-2,5-dihydro-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide (200 mg, 0.40 mmol) in N,N-dimethylformamide (10.0 mL) was added lithium chloride (168 mg, 4.0 mmol). The mixture was stirred overnight at 60 °C. The solution was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate and evaporated to dryness. The residue was purified by reverse-phase preparative HPLC to give N-((6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)sulfonyl)acetamide (formate) as a white solid (15.3 mg, 0.04 mmol, 8.92% yield). LC-MS: m / z = 384.0 (M+H) + , the retention time is 3.42 minutes (Method A). 1 HNMR(500MHz,DMSO-d 6 ) δ 12.77 (br, 1H), 8.89 (s, 1H), 8.66 (d, J = 8.0 Hz, 1H), 8.48 (s, 1H), 8.34 (dd, J = 8.5 Hz, J = 2.0 Hz, 1H), 8.09 (d, J = 8.0 Hz, 2H), 7.69 (d, J = 8.5 Hz, 2H), 1.93 (s, 3H).

[0733] Example 30: Preparation of Compound 30

[0734] (6-Bromopyridin-3-yl)dimethylphosphine oxide

[0735]

[0736] A mixture of 2-bromo-5-iodopyridine (500 mg, 1.76 mmol), dimethylphosphine oxide (275 mg, 3.53 mmol), potassium phosphate (1.12 g, 5.28 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (203 mg, 0.35 mmol) and palladium(II) acetate (156 mg, 0.7 mmol) in 1,4-dioxane (15.0 mL) was stirred overnight at 100 °C under nitrogen. The reaction mixture was filtered through Celite, and the filtrate was concentrated under reduced pressure. The residue obtained was purified by flash chromatography (petroleum ether / ethyl acetate = 1 / 1) to give (6-bromopyridin-3-yl)dimethylphosphine oxide as a yellow oil (50 mg, 0.21 mmol, 12.1% yield). LC-MS: m / z = 234 [M+H] + , retention time = 1.36 min (Method A).

[0737] (6-Hydrazinopyridin-3-yl)dimethylphosphine oxide

[0738]

[0739] Hydrazine hydrate (160 mg, 2.55 mmol, 85% in water) was added to a solution of (6-bromopyridin-3-yl)dimethylphosphine oxide (120 mg, 0.51 mmol) in ethanol (5.0 mL). The mixture was stirred at 80 °C for 4.0 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate and concentrated. The residue was triturated with petroleum ether and filtered to give (6-hydrazinopyridin-3-yl)dimethylphosphine oxide as a yellow solid (80 mg, 0.43 mmol, 80% yield). LC-MS: m / z = 186.0 (M+H )+ , retention time was 0.36 min (Method A).

[0740] 4-(1-(5-(Dimethylphosphoryl)pyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile

[0741]

[0742] To a solution of methyl (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (100 mg, 0.43 mmol) and (6-hydrazinopyridin-3-yl)dimethylphosphine oxide (80.4 mg, 0.43 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (19 mg, 0.1 mmol). The mixture was stirred overnight at 100 °C in a sealed tube and cooled to precipitate a solid. The solid was purified by reverse-phase preparative HPLC to give 4-(1-(5-(dimethylphosphoryl)pyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)benzonitrile (formate) as a white solid (15.0 mg, 0.04 mmol, 9.3% yield). LC-MS: m / z = 339.0 (M+H) + , retention time 3.48 min (Method A). 1 HNMR (400 MHz, DMSO-d 6 ) δ 8.79 - 8.77 (m, 1H), 8.51 - 8.45 (m, 2H), 8.31 - 8.26 (m, 1H), 8.10 - 8.08 (m, 2H), 7.72 - 7.70 (m, 2H), 1.74 - 1.70 (m, 6H).

[0743] Example 31: Preparation of Compound 31

[0744] 4-(5-Chloropyridin-2-yl)-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-5-ol

[0745]

[0746] The compound was synthesized using 5-chloropyridine-2-acetic acid according to the procedure for the preparation of 4-(5-hydroxy-1-(5-(methylsulfonyl)pyridin-2-yl)-1H-pyrazol-4-yl)benzonitrile (Example 4). LCMS (ESI+): m / z 365 (M+H) + ; 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.91 (s, 1H), 8.70 (d, J = 5.7 Hz, 1H), 8.55 (d, J = 1.5 Hz, 1H), 8.46 (d, J = 8.7 Hz, 1H), 8.27 (d, J = 8.7 Hz, 1H), 7.51 (dd, J = 8.7 Hz, J = 2.4 Hz, 1H), 3.34 (s, 3H), 2.73 (s, 3H).

[0747] Example 32: Preparation of Compound 32

[0748] 3-Methyl-4-vinylbenzonitrile

[0749]

[0750] Under a nitrogen atmosphere, 4-bromo-3-methylbenzonitrile (6.14 g, 31.34 mmol), Cs 2 CO 3 (40.87 g, 125.38 mmol), potassium trifluoro(vinyl)borate (8.40 g, 62.69 mmol), and Pd(dppf)Cl 2 (2.29 g, 3.13 mmol) in a mixture of THF (300 mL) and water (30 mL) were stirred at 75 °C for 6 h. After completion of the reaction as indicated by TLC analysis, the resulting mixture was diluted with brine (80 mL) and extracted with ethyl acetate (200 mL × 2). The combined organic phases were dried over Na 2 SO 4 (30 g), filtered, and concentrated. The residue was purified by silica gel column chromatography (EtOAc:Hex = 1:50) to afford 6.74 g of the desired product as an oil. GC-MS: m / z 143 (M) + .

[0751] 2-(4-Cyano-2-methylphenyl)acetic acid

[0752]

[0753] 3-Methyl-4-vinylbenzonitrile (6.74 g, 47.13 mmol) and I 2 (1.20 g, 4.71 mmol) in a mixture of DME (400 mL) and water (96 mL) were stirred at room temperature for 5 min. Potassium bisulfate (57.97 g, 94.26 mmol) was added in one portion. The reaction was stirred at room temperature overnight. After completion of the reaction as indicated by TLC analysis, the resulting mixture was diluted with an aqueous Na 2 S 2 O 3 solution (100 mL) and extracted with ethyl acetate (50 mL × 5). The combined organic phases were dried over Na 2 SO 4 (30 g), filtered, and concentrated. The residue was purified by slurry to afford 3.4 g of the crude product as a solid, which was used directly in the next step without further purification.

[0754] Methyl 2-(4-cyano-2-methylphenyl)acetate

[0755]

[0756] Over 5 minutes, SOCl 2 (7 mL) was added dropwise to a mixture of 2-(4-cyano-2-methylphenyl)acetic acid (3.40 g, 21.66 mmol) in MeOH (60 mL). The mixture was stirred at room temperature for 40 minutes. After completion of the reaction as indicated by TLC analysis, the resulting mixture was concentrated directly. The residue was purified by silica gel column chromatography (EtOAc:n-Hex = 1:50 to 1:10) to give 1.66 g of the desired product as an oil. 1 1H-NMR (300 MHz, CDCl 3 ) δ 7.46 (m, 2H), 7.30 (d, J = 8.1 Hz, 1H), 3.71 (s, 3H), 3.69 (s, 2H), 2.34 (s, 3H).

[0757] (Z)-Methyl 2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate

[0758]

[0759] A mixture of methyl 2-(4-cyano-2-methylphenyl)acrylate (1.66 g, 8.78 mmol) in DMF-DMA (6 mL) was stirred at 100 °C for 5 hours. After completion of the reaction as indicated by TLC analysis, the resulting mixture was concentrated directly. The residue was purified by silica gel column chromatography (EtOAc:n-Hex = 1:50 to 1:5) to give 2.0 g of the desired product as an oil. LCMS (ESI+): m / z 245 (M+H) + ; 1 1H-NMR (300 MHz, CDCl 3 ) δ 7.61 (s, 1H), 7.46 (s, 1H), 7.42 (d, J = 8.1 Hz, 1H), 7.22 (d, J = 8.1 Hz, 1H), 3.61 (s, 3H), 2.66 (s, 6H), 2.22 (s, 3H).

[0760] 4-(5-Hydroxy-1H-pyrazol-4-yl)-3-methylbenzonitrile

[0761]

[0762] To a solution of (Z)-methyl 2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate (0.34 g, 1.39 mmol) in EtOH (10 mL) was added N 2 H 4 .H 2O (0.64 g, 10.20 mmol). The reaction was stirred overnight at 70 °C. After completion of the reaction as indicated by TLC, the resulting mixture was directly concentrated. The residue was purified by silica gel column chromatography (EtOAc:n-Hex = 1:50 to 1:10) to afford 287 mg of the desired product as an oil. 1 H-NMR (300 MHz, DMSO-d 6 ) δ 11.96 (brs, 1H), 10.21 (brs, 1H), 7.74 (s, 1H), 7.67 (s, 1H), 7.62 (d, J = 8.1 Hz, 2H), 2.36 (s, 3H).

[0763] 2,3-Dichloro-5-(methylthio)pyridine

[0764]

[0765] To a mixture of 5,6-dichloropyridin-3-amine (0.50 g, 3.07 mmol) in concentrated HCl (2.5 mL) at 0 °C was added dropwise an aqueous solution of NaNO 2 (0.32 g, 4.60 mmol, 1 mL of water) over 5 minutes. The reaction was stirred at 0 °C for 1 hour. The reaction was filtered to remove inorganic salts. The solution was added dropwise over 10 minutes to a suspension of MeSNa (1.29 g, 3.68 mmol, 20%) and NaBF 4 (3.4 mg, 0.031 mmol) in acetonitrile (2.5 ml) at 0 °C. The resulting orange suspension was stirred at room temperature for 2 hours. After completion of the reaction as indicated by TLC, the reaction was quenched with aqueous NaOH solution (1 N, 5 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic phases were dried over Na 2 SO 4 (20 g), filtered and concentrated to afford 456 mg of the desired product as a black solid. 1 H-NMR (300 MHz, CDCl 3 ) δ 8.15 (d, J = 2.1 Hz, 1H), 7.62 (d, J = 2.4 Hz, 1H), 2.50 (d, J = 6.0 Hz, 3H).

[0766] 2,3-Dichloro-5-(methylsulfonyl)pyridine

[0767]

[0768] To a solution of 2,3-dichloro-5-(methylthio)pyridine (0.46 g, 2.35 mmol) in DCM (30 mL) was added m-CPBA (0.85 g, 4.94 mmol) in one portion. The reaction was then stirred at room temperature for 3.5 h. After completion of the reaction as indicated by TLC, the mixture was concentrated directly. The residue-containing EtOAc (50 mL) was washed with aqueous Na 2 S 2 O 3 aqueous solution (10 mL). The aqueous phase was extracted with EtOAc (2 × 50 mL). The combined organic phases were dried over Na 2 SO 4 (30 g), filtered and concentrated. The residue was purified by silica gel column chromatography (EtOAc:n-Hex = 1:50 to 1:20) to afford 460 mg of the desired product as a solid. 1 H-NMR (300 MHz, CDCl 3 ) δ 8.82 (d, J = 2.1 Hz, 1H), 8.29 (d, J = 2.4 Hz, 1H), 3.15 (s, 3H).

[0769] 4-(1-(3-chloro-5-(methylsulfonyl)pyridin-2-yl)-5-hydroxy-1H-pyrazol-4-yl)-3-methylbenzonitrile

[0770]

[0771] NaH (60 mg, 1.50 mmol) was added portionwise over 5 min to a mixture of 4-(5-hydroxy-1H-pyrazol-4-yl)-3-methylbenzonitrile (0.15 g, 0.75 mmol) in DMF (3 mL). The reaction was stirred at room temperature for 40 min. 2,3-Dichloro-5-(methylsulfonyl)pyridine (0.25 g, 1.13 mmol) was added and the reaction was stirred at room temperature overnight. After completion of the reaction as indicated by TLC analysis, the resulting mixture was concentrated directly to dryness. The residue was purified by preparative HPLC to afford 11 mg of the desired product as a solid. LCMS (ESI+): m / z 389 (M+H) + ; 1 H-NMR (300 MHz, CDCl 3 ) δ 10.23 (brs, 1H), 8.44 (d, J = 2.1 Hz, 1H), 8.18 (d, J = 2.1 Hz, 1H), 7.69 (s, 1H), 7.50 (s, 1H), 7.38 - 7.45 (m, 2H), 3.06 (s, 3H), 2.41 (s, 3H).

[0772] Example 33: Preparation of Compound 33

[0773] N-(6-(4-(4-Cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)piperazine-1-carboxamide

[0774]

[0775] The compound was synthesized using tert-butyl piperazine-1-carboxylate according to the procedure for the preparation of N-(6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)pyridin-3-yl)morpholine-4-carboxamide (Example 28).

[0776] In Vitro Assays Demonstrate PHD Inhibition

[0777] The enzymatic half-maximal inhibitory concentration (IC 50 ) values were determined for selected compounds of the present invention.

[0778] Time-resolved fluorescence resonance energy transfer (TR-FRET) assays were used to determine the enzymatic half-maximal inhibitory concentration (IC 50 ) values of PHD inhibitors against the full-length human prolyl-4-hydroxylase domain (PHD) enzymes, PHD1, PHD2, and PHD3. The TR-FRET assay was developed based on the specific binding of the hydroxylated HIF-1α peptide to the complex formed by VHL, EloB, and EloC (VBC), resulting in a fluorescent signal. The terbium (Tb)-donor (monoclonal antibody anti-6His-Tb-cryptate gold) and D2-acceptor (streptavidin [SA]-D2) of TR-FRET were linked to the VBC complex and the HIF-1α peptide, respectively. The VBC complex binds specifically to the HIF-1α peptide upon hydroxylation, enabling energy transfer from the TR-FRET donor to the acceptor ( Figure 1 ).

[0779] Materials and Methods

[0780] Unless otherwise stated, all chemicals and materials were of standard laboratory grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).

[0781] Reagents

[0782] TR-FRET Reagents

[0783] The monoclonal antibodies anti-6His-Tb-cryptand gold (Catalog No. 61HI2TLA) and streptavidin (SA)-D2 (Catalog No. 610SADLA) were purchased from CisBio International (Bedford, MA, USA).

[0784] The N-terminal biotinylated HIF-1α C35 synthetic peptide representing amino acids 547 to 581 and containing the proline 564 PHD2 hydroxylation site was purchased from California Peptide Research (Salt Lake City, UT, USA).

[0785] Recombinant protein

[0786] VBC complex

[0787] The His-tagged recombinant VHL protein, EloB, EloC complex (His-VBC) was provided by Axxam (Milan, Italy). Recombinant human VHL (National Center for Biotechnology Information [NCBI] accession number NP_00542.1) contains a His-tag at the C-terminus of amino acids 55 to 213 and is designated VHL-His. VHL-His was co-expressed with full-length human EloB (NCBI accession number Q15370.1) and full-length human EloC (NCBI accession number Q15369.1) in Escherichia coli and purified as the His-VBC complex by affinity chromatography on a nickel-nitrilotriacetic acid (Ni-NTA) column. The purity was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) (approx. 80%).

[0788] PHD1

[0789] The recombinant human PHD1 protein (Catalog No. 81064, Lot No. 24717001) was purchased from ActiveMotif (Carlsbad, CA, USA). PHD1 was expressed as a full-length protein (NCBI accession number NP_542770.2) with an N-terminal FLAG-tag (molecular weight 44.9 kDa) in the baculovirus expression system. The purity was evaluated by SDS-PAGE (>90%).

[0790] PHD2

[0791] The full-length human PHD2 enzyme was produced using a baculovirus-infected insect cell (BIIC) expression system from Beryllium (Bedford, MA, USA). The PHD2 construct contained amino acids 1 to 426 of PHD2 (UniProt Knowledgebase [UniProtKB] / Swiss-Prot accession number Q9GZT9.1) along with a His-tag at the N-terminus and a tobacco etch virus (TEV) protease cleavage site. The construct was expressed in Sf9 insect cells, purified by Ni-NTA column and digested with TEV protease to remove the His-tag. The purity of the final cleaved protein was assessed by SDS-PAGE and found to be >94%.

[0792] PHD3

[0793] Recombinant human PHD3 protein (molecular weight 31.1 kDa) was purchased from Active Motif (Carlsbad, CA, USA). The recombinant human PHD3 protein was expressed in Escherichia coli as a full-length protein (NCBI accession number NP_071356.1) with an N-terminal 6-His tag (catalog number 81033, lot number 24417001). The purity was assessed by SDS-PAGE and found to be >75%.

[0794] PHD inhibitor.

[0795] Synthetic small molecule PHD inhibitors were made and their identity confirmed as described herein.

[0796] TR-FRET assay procedure

[0797] In a white 384-well Optiplate microplate (catalog number 6007290, Perkin Elmer, Waltham, MA, USA), PHD inhibitor compounds were pre-incubated with PHD enzymes in a 10 μL reaction volume. For this, 5 μL of PHD inhibitor compound was serially diluted with dilution buffer (50 mM HEPES [4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid] pH 7.5, 50 mM sodium chloride [NaCl], 0.01% Tween-20, 0.01% purified bovine serum albumin [BSA]) and mixed with 5 μL of PHD enzyme mixture prepared as a 4X concentrate in dilution buffer containing PHD enzymes (60 nM PHD1, 20 nM PHD2, 140 nM PHD3), 40 μM ammonium ferrous sulfate (FAS), 4 mM sodium ascorbate (Na). The plate was incubated at room temperature for 30 minutes without shaking.

[0798] Then add 5 μL of the VBC / anti-6His-Tb-cavitand gold mixture, which is prepared as a 4X concentrate in a dilution buffer containing 20 nM His-VBC and 1.32 nM monoclonal antibody anti-6His-Tb-cavitand gold. Immediately after this step, add 5 μL of the HIF-1αC35 substrate mixture to achieve a final reaction volume of 20 μL, which is prepared as a 4X concentrate in a dilution buffer containing 120 nM biotinylated HIF-1αC35, 132 nM SA-D2, and 4 μM 2-oxoglutarate (2-OG).

[0799] The final assay reaction contains 50 mM HEPES, pH 7.5; 50 mM NaCl; 1 μM 2-OG; 10 μM FAS; 1 mM sodium ascorbate; 0.01% Tween-20; 0.01% purified BSA; 30 nM biotinylated HIF-1αC35; 5 nM His-VBC; 0.33 nM monoclonal antibody anti-6His-Tb-cavitand gold; 33 nM SA-D2 and PHD enzyme (15 nM PHD1, 5 nM PHD2, or 35 nM PHD3) and the diluted compound.

[0800] To measure the IC 50 of the PHD inhibitor compound, the reaction was incubated at room temperature for 10 minutes and then read on a PerkinElmer EnVision (Waltham, MA, USA) at an excitation wavelength of 340 nm and emission wavelengths of 615 nm and 665 nm. The data represent the quotient of the signal intensities at 665 nm and 615 nm, which is automatically calculated by the Envision Manager software (PerkinElmer, Inc., Waltham, MA, USA). The IC 50 values (mean, standard deviation, standard error of the mean, geometric mean, and 95% confidence interval) were determined using GraphPad Prism 7.0 (GraphPad, La Jolla, CA, USA) using four-parameter curve fitting and represent the compound concentration plotted against the calculated ratio for 665 nm and 615 nm. The TR-FRET assay was performed in triplicate at each compound concentration, and the assay was independently repeated three times.

[0801] Ki was calculated from the IC 50 based on the Cheng Prussoff equation as follows:

[0802] Ki = IC50 / (1 + [2-OG] / Km)

[0803] The final concentration of 2-OG in both PHD1 and PHD2 assays was 1 μM. The Km determination of 2-OG for PHD1 was 12.7 nM, while the Km determination of 2-OG for PHD2 was 22.6 nM.

[0804] Exemplary compounds

[0805]

[0806]

[0807]

[0808]

[0809]

[0810] Based on the ongoing description, those skilled in the art can easily determine the necessary characteristics of the present invention, and various changes and modifications can be made to the present invention without departing from the spirit and scope of the present invention to adapt it to various usage and conditions.

[0811] All references, patents or applications cited in this application, whether US or foreign, are incorporated herein by reference as if written in their entirety herein. In case of any inconsistency, the materials literally disclosed herein shall prevail.

Claims

1. A compound having the structure according to Formula III: or a pharmaceutically acceptable salt thereof, wherein: Each occurrence, R 1 is independently selected from the group consisting of hydrogen and C 1-3 alkyl; R 1a is CN, a halogen or a C 1-3 alkyl group optionally substituted by CN; Each occurrence, R 2 is independently selected from the group consisting of hydrogen and C 1-3 alkyl; R 3 is SO 2 R 6 、SOR 7 R 8 、SOR 9 、COR 10 、NHR 11 、POR 12 R 13 、halogen, cyclopropyl, optionally substituted by C 1-3 alkyl thiazole, oxazole, pyridine, triazole, tetrazole or pyrazole, or C 1-3 alkyl optionally substituted by one or more halogens; R 6 is C 1-3 alkyl or NHCOR 15 ; R 7 is C 1-3 alkyl; R 8 is NH or NCH 3 ; R 9 is C 1-3 alkyl; R 10 is C 1-3 alkyl or NHSO 2 R 20 ; R 11 is COR 21 ; R 12 and R 13 each independently is C 1-3 alkyl; R 15 is C 1-3 alkyl; R 20 is C 1-3 alkyl; R 21 is cyclopropyl or C 1-3 alkyl; m is 1 or 2; and n is 0 or 1.

2. The compound according to claim 1, having the structure of Formula IVa: or a pharmaceutically acceptable salt thereof, wherein: R 2 selected from the group consisting of hydrogen or C 1-3 alkyl groups; R 7 is C 1-3 alkyl; and R 8 is NH or NCH 3 .

3. The compound according to claim 2, wherein R 1a is CN.

4. The compound according to claim 2, wherein R 7 is C 1-3 alkyl.

5. The compound according to claim 4, wherein R 7 is CH 3 .

6. The compound according to claim 1, which has the structure of formula VIa: or a pharmaceutically acceptable salt thereof, wherein: R 2 is hydrogen or C 1-3 alkyl; and R 3 is cyclopropyl or 7. The compound according to claim 6, wherein R 2 is H.

8. The compound according to claim 6, wherein R 3 is 9. The compound according to claim 1, having the structure of Formula VIIa: or a pharmaceutically acceptable salt thereof, wherein: R 2 is hydrogen or C 1-3 alkyl; R 11 is COR 21 ; R 21 is cyclopropyl or C 1-3 alkyl.

10. The compound according to claim 9, wherein R 2 is H.

11. The compound according to claim 10, wherein R 11 is COR 21 and wherein R 21 is cyclopropyl.

12. The compound according to claim 10, wherein R 21 is C 1-3 alkyl.

13. The compound according to claim 12, wherein R 21 is CH 2 CH 3 .

14. The compound according to claim 1, having the structure of Formula VIIIa: or a pharmaceutically acceptable salt thereof, wherein: R 3 is an optionally C 1-3 alkyl-substituted thiazole, oxazole, pyridine, triazole, tetrazole or pyrazole.

15. The compound according to claim 14, wherein R 3 is thiazole, oxazole, pyridine, triazole, tetrazole or pyrazole.

16. The compound according to claim 15, wherein R 3 is 17. The compound according to claim 15, wherein R 3 is 18. The compound according to claim 15, wherein R 3 is 19. The compound according to claim 15, wherein R 3 is 20. The compound according to claim 1, having the structure of Formula IXa: or a pharmaceutically acceptable salt thereof, wherein: R 10 is C 1-3 alkyl or NHSO 2 R 20 ; and R 20 is C 1-3 alkyl group.

21. The compound according to claim 20, wherein R 1a is CN.

22. The compound according to claim 21, wherein R 10 is C 1-3 alkyl.

23. The compound according to claim 22, wherein R 10 is CH 3 .

24. The compound according to claim 22, wherein R 10 is CH(CH 3 ) 2 .

25. The compound according to claim 22, wherein R 10 is CH 2 CH 3 .

26. The compound according to claim 21, wherein R 10 is NHSO 2 R 20 , and wherein R 20 is C 1-3 alkyl.

27. The compound according to claim 26, wherein R 20 is CH 3 .

28. The compound according to claim 1, wherein: R 3 is a halogen.

29. The compound according to claim 28, wherein R 1a is CN.

30. The compound according to claim 29, wherein R 1 is H.

31. The compound according to claim 30, wherein R 2 is H.

32. The compound according to claim 31, wherein R 3 is Cl.

33. The compound according to claim 31, wherein R 3 is Br.

34. The compound according to claim 31, wherein R 3 is F.

35. The compound according to claim 1, having the structure according to Formula XIII: or a pharmaceutically acceptable salt thereof, wherein: R 12 is C 1-3 alkyl; and R 13 is C 1-3 alkyl group.

36. The compound according to claim 35, wherein R 1a is CN.

37. The compound according to claim 36, wherein R 1 is H.

38. The compound according to claim 37, wherein R 12 is C 1-3 alkyl.

39. The compound according to claim 38, wherein R 12 is CH 3 .

40. The compound according to claim 39, wherein R 13 is C 1-3 alkyl.

41. The compound according to claim 40, wherein R 13 is CH 3 .

42. A compound selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

43. A pharmaceutical composition comprising the compound according to any one of claim 42 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

44. Use of the compound according to any one of claim 42 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease mediated by PHD activity, wherein the disease mediated by PHD activity is selected from the group consisting of ischemia-reperfusion injury, inflammatory bowel disease, cancer, liver disease, cardiovascular disease, diseases or conditions of the eye, anemia, chronic kidney disease, and bronchopulmonary dysplasia (BPD).

45. The use according to claim 44, wherein the disease mediated by PHD activity is ischemia-reperfusion injury.

46. The use according to claim 45, wherein the ischemia-reperfusion injury is selected from the group consisting of stroke, myocardial infarction, and acute kidney injury.

47. The use according to claim 44, wherein the disease mediated by PHD activity is inflammatory bowel disease.

48. The use according to claim 47, wherein the inflammatory bowel disease is ulcerative colitis.

49. The use according to claim 47, wherein the inflammatory bowel disease is Crohn's disease.

50. The use according to claim 44, wherein the disease mediated by PHD activity is cancer.

51. The use according to claim 50, wherein the cancer is colorectal cancer.

52. The use according to claim 44, wherein the disease mediated by PHD activity is liver disease.

53. The use according to claim 44, wherein the disease mediated by PHD activity is cardiovascular disease.

54. The use according to claim 53, wherein the cardiovascular disease is atherosclerosis.

55. The use according to claim 44, wherein the disease mediated by PHD activity is diseases or conditions of the eye.

56. Use according to claim 55, wherein the eye disease or condition is selected from radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

57. Use according to claim 44, wherein the disease is anemia.

58. Use according to claim 57, wherein the anemia is anemia associated with chronic kidney disease.

59. Use according to claim 44, wherein the disease is chronic kidney disease.

60. Use according to claim 44, wherein the disease mediated by PHD activity is bronchopulmonary dysplasia (BPD).

61. Use of a compound according to any one of claims 1 to 42 or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a disease mediated by PHD activity, wherein the disease mediated by PHD activity is selected from ischemic heart disease, valvular heart disease, congestive heart failure, acute lung injury, pulmonary fibrosis, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), acute liver failure, liver fibrosis, and cirrhosis.

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