PHD inhibitor compounds, compositions and uses

Abstract

The present invention provides, in part, a novel small molecule PHD inhibitor, or a pharmaceutically acceptable salt thereof, having a structure according to Formula (A) and its sub-formula. Compounds provided herein are useful for the treatment of diseases, the diseases include heart diseases (e.g., ischemic heart disease, congestive heart failure and valvular heart disease), lung diseases (e.g., acute lung injury, pulmonary arterial hypertension, pulmonary fibrosis and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure and liver fibrosis and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease). # imgabs0 #

Description

[0001] This application is a divisional application of Chinese invention patent application 202180034124.X, filed on March 9, 2021, and entitled “PHD Inhibitor Compounds, Compositions and Uses”.

[0002] Cross-reference to related applications

[0003] This application claims priority to U.S. Provisional Patent Application No. 62 / 992,585, filed on March 20, 2020, which is hereby incorporated by reference in its entirety. Background Art

[0004] Hypoxia is a condition or state in which the oxygen supply is insufficient to maintain normal life functions, for example, insufficient arterial oxygen supply. Hypoxia may lead to impaired cell function and structural tissue damage. The activation of cellular defense mechanisms during hypoxia is mediated by HIF (hypoxia-inducible factor) proteins. Under hypoxic conditions, HIFα levels in most cells increase due to reduced HIFα prolyl hydroxylation. The prolyl hydroxylation of HIFα is completed by a series of proteins, which are called prolyl hydroxylase domain-containing proteins (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 isomers play different roles in their regulation of HIF and may have other non-HIF-related regulatory effects.

[0005] Indeed, many studies have shown that stabilization of HIF can suppress tissue inflammation and promote its repair. Therefore, compounds that can inhibit the activity of PHD proteins may be particularly beneficial in new therapies (Lee et al. (2019) Experimental and Molecular Medicine (Exp. Mol. Med. 51:68)

[0006] Described herein are novel small molecule PHD inhibitors that are useful for treating diseases including cardiac diseases (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), pulmonary diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure and liver fibrosis and cirrhosis), and kidney diseases (e.g., acute kidney injury and chronic kidney disease). Summary of the invention

[0007] The present invention provides novel small molecule PHD inhibitors, and can be used to treat diseases including but not limited to cardiac diseases (e.g., ischemic heart disease, congestive heart failure and valvular heart disease), pulmonary diseases (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis and chronic obstructive pulmonary disease), liver diseases (e.g., acute liver failure and liver fibrosis and cirrhosis) and kidney diseases (e.g., acute kidney injury and chronic kidney disease).

[0008] In one aspect, provided herein are compounds having a structure according to Formula (A),

[0009]

[0010] or a pharmaceutically acceptable salt thereof, wherein:

[0011] Ar 1 is phenyl or a six-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl or C 1-3 Alkoxy substitution;

[0012] R 2 H or C 1-3 alkyl;

[0013] Ar 2 is a six-membered nitrogen-containing heteroaryl group, which is optionally substituted by halogen, OH, amine or C 1-3 Alkyl substitution;

[0014] R 4 is hydrogen or C 1-4 Alkyl; and

[0015] The formula (A) does not include the following compounds:

[0016]

[0017] In an embodiment, R 2 For H.

[0018] In an embodiment, R 2 C 1-3 alkyl;

[0019] In the embodiment, Ar 1 for

[0020] in

[0021] X, Y and Z are independently CH or N, wherein N is optionally oxidized;

[0022] Each R 1independently 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; and

[0023] m is 1, 2, 3 or 4.

[0024] In the embodiment, Ar 1 is a substituted phenyl group. 1 By at least one R 1 Substitution, where R 1 It is CN halogen.

[0025] In the embodiment, Ar 1 By one or two R 1 A group is substituted, the group being independently selected from C optionally substituted by one or more halogens 1-3 Alkyl, halogen, CN or OH.

[0026] In the embodiment, Ar 1 is a pyridyl N-oxide or is optionally substituted by at least one R 1 Substituted pyridyl, wherein R1 is C 1-3 Alkoxy or halogen.

[0027] In an embodiment, each R 1 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;

[0028] In the embodiment, Ar 2 for

[0029] in

[0030] A and B are independently CH or N, wherein N is optionally oxidized;

[0031] Each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; and

[0032] n is 0, 1 or 2.

[0033] In the embodiment, Ar 2 is a group which is pyridyl or pyrazinyl, and wherein the group is unsubstituted or comprises a substituent which is halogen, C 1-3 Alkyl or OH.

[0034] In an embodiment, each R 3independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl.

[0035] In an embodiment, R 4 is H. In the embodiment, R 4 C 1-4 alkyl.

[0036] In embodiments, the compound of formula (A) is not

[0037]

[0038] In the embodiment, the compound of formula (A) does not include the following compounds

[0039]

[0040]

[0041] In an embodiment, the compound of formula (A) has the following structure,

[0042] or a pharmaceutically acceptable salt thereof.

[0043] In an embodiment, X, Y, Z, A and B are independently CH or N, wherein N is optionally oxidized; m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0044] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0045] or a pharmaceutically acceptable salt thereof.

[0046] In an embodiment, X, Y and Z are independently CH or N, wherein N is optionally oxidized; m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0047] In embodiments, the compound of formula (I) is not

[0048]

[0049]

[0050] In the embodiments, the compound of formula (I) does not include the following compounds

[0051]

[0052] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0053] or a pharmaceutically acceptable salt thereof.

[0054] In an embodiment, A and B are independently CH or N, wherein N is optionally oxidized; m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0055] In embodiments, the compound of formula (Ib) is not

[0056]

[0057] In the embodiment, the compound of formula (Ib) does not include the following compounds

[0058]

[0059]

[0060] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0061] or a pharmaceutically acceptable salt thereof.

[0062] In an embodiment, X, Y, Z, A and B are independently CH or N, wherein N is optionally oxidized; m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0063] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0064] or a pharmaceutically acceptable salt thereof.

[0065] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0066] In embodiments, the compound of formula (II) is not

[0067]

[0068] In the embodiment, the compound of formula (II) does not include the following compounds

[0069]

[0070]

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

[0072] or a pharmaceutically acceptable salt thereof.

[0073] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2 is hydrogen or C 1-3 alkyl; and each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl.

[0074] In embodiments, the compound of formula (IIa) is not .

[0075] In the embodiment, the compound of formula (IIa) does not include the following compounds

[0076]

[0077] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0078] or a pharmaceutically acceptable salt thereof.

[0079] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; R 2 is hydrogen or C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0080] In embodiments, the compound of formula (IIb) is not

[0081]

[0082]

[0083] In the embodiment, the compound of formula (IIb) does not include the following compounds

[0084]

[0085] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0086] or a pharmaceutically acceptable salt thereof.

[0087] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; and R 2 is hydrogen or C 1-3 alkyl.

[0088] In embodiments, the compound of formula (IIc) is not

[0089]

[0090] In the embodiment, the compound of formula (IIc) does not include the following compounds

[0091]

[0092] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0093] or a pharmaceutically acceptable salt thereof.

[0094] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; and each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl.

[0095] In embodiments, the compound of formula (IId) is not

[0096]

[0097] In the embodiment, the compound of formula (IId) does not include the following compounds

[0098]

[0099]

[0100] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0101] or a pharmaceutically acceptable salt thereof.

[0102] In an embodiment, m is 1, 2, 3 or 4; each R1 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0103] In embodiments, the compound of formula (IIe) is not

[0104]

[0105] In an embodiment, the compound of formula (IIe) does not include the following compounds

[0106]

[0107] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0108] or a pharmaceutically acceptable salt thereof.

[0109] In an embodiment, m is 1, 2, 3 or 4; and each R 1 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.

[0110] In embodiments, the compound of formula (IIf) is not

[0111]

[0112]

[0113] In an embodiment, the compound of formula (IIf) does not include the following compounds

[0114]

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

[0116] or a pharmaceutically acceptable salt thereof.

[0117] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R1 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; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 4 is hydrogen or C 1-4 alkyl.

[0118] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0119] or a pharmaceutically acceptable salt thereof.

[0120] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; R 4 is hydrogen or C 1-4 alkyl; and R 5 is CN or halogen.

[0121] In embodiments, the compound of formula (III) is not

[0122]

[0123] In the embodiment, the compound of formula (III) does not include the following compounds

[0124]

[0125]

[0126] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0127] or a pharmaceutically acceptable salt thereof.

[0128] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1independently 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; R 2 is hydrogen or C 1-3 Alkyl; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 5 is CN or halogen.

[0129] In embodiments, the compound of formula (IIIa) is not

[0130]

[0131]

[0132] In the embodiment, the compound of formula (IIIa) does not include the following compounds

[0133]

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

[0135] or a pharmaceutically acceptable salt thereof.

[0136] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; R 2 is hydrogen or C 1-3 Alkyl; R 4 is hydrogen or C 1-4 alkyl; and R 5 is CN or halogen.

[0137] In embodiments, the compound of formula (IIIb) is not

[0138]

[0139] In the embodiment, the compound of formula (IIIb) does not include the following compounds

[0140]

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

[0142] or a pharmaceutically acceptable salt thereof.

[0143] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; R 2 is hydrogen or C 1-3 alkyl; and R 5 is CN or halogen.

[0144] In embodiments, the compound of formula (IIIc) is not

[0145]

[0146] In the embodiment, the compound of formula (IIIc) does not include the following compounds

[0147]

[0148]

[0149] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0150] or a pharmaceutically acceptable salt thereof.

[0151] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; and R 5 is CN or halogen.

[0152] In embodiments, the compound of formula (IIId) is not

[0153]

[0154] In the embodiment, the compound of formula (IIId) does not include the following compounds

[0155]

[0156] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0157] or a pharmaceutically acceptable salt thereof.

[0158] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; R 4 is hydrogen or C 1-4 alkyl; and R 5 is CN or halogen.

[0159] In embodiments, the compound of formula (IIIe) is not

[0160]

[0161]

[0162] In the embodiment, the compound of formula (IIIe) does not include the following compounds

[0163]

[0164] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0165] or a pharmaceutically acceptable salt thereof.

[0166] In an embodiment, m is 1, 2, 3 or 4; each R 1 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; and R 5 is CN or halogen.

[0167] In embodiments, the compound of formula (IIIf) is not

[0168]

[0169] In the embodiment, the compound of formula (IIIf) does not include the following compounds

[0170]

[0171] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0172] or a pharmaceutically acceptable salt thereof.

[0173] In an embodiment, m is 1, 2, 3 or 4; n is 0, 1 or 2; each R 1 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; each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; R 4 is hydrogen or C 1-4 alkyl; and R 5 is CN or halogen.

[0174] In embodiments, X is CH. In embodiments, X is N, wherein N is optionally oxidized.

[0175] In an embodiment, Y is CH. In an embodiment, Y is N.

[0176] In an embodiment, Z is CH. In an embodiment, Z is N.

[0177] In an embodiment, A is CH. In an embodiment, A is N.

[0178] In an embodiment, B is CH. In an embodiment, B is N.

[0179] In an embodiment, m is 1. In an embodiment, m is 2. In an embodiment, m is 3. In an embodiment, m is 4.

[0180] In an embodiment, n is 0. In an embodiment, n is 1. In an embodiment, n is 2.

[0181] In an embodiment, R 1 is hydrogen. In an embodiment, R 1 is CN. In the embodiment, R 1 For OH.

[0182] In an embodiment, R 1 In the embodiment, R 1 is F. In the embodiment, R 1 In the embodiment, R 1 For Br.

[0183] In an embodiment, R 1 is C optionally substituted by one or more halogens1-3 In the embodiment, R 1 C 1-3 In the embodiment, R 1 In the embodiment, R 1 In the embodiment, R 1 For CF 3 .

[0184] In an embodiment, R 1 C 1-3 In the embodiment, R 1 It is a methoxy group.

[0185] In an embodiment, R 2 For hydrogen.

[0186] In an embodiment, R 2 C 1-3 In the embodiment, R 2 It is methyl.

[0187] In an embodiment, R 3 For hydrogen.

[0188] In an embodiment, R 3 In the embodiment, R 3 For F.

[0189] In an embodiment, R 3 For OH.

[0190] In an embodiment, R 3 In the embodiment, R 3 For NH 2 .

[0191] In an embodiment, R 3 C 1-3 In the embodiment, R 3 It is methyl.

[0192] In an embodiment, R 4 For hydrogen.

[0193] In an embodiment, R 4 C 1-4 In the embodiment, R 4 In the embodiment, R 4 In the embodiment, R 4 is isopropyl. 4 It is tert-butyl.

[0194] In an embodiment, R 5 is F. In the embodiment, R 5 In the embodiment, R5 For Br.

[0195] In an embodiment, R 5 For CN.

[0196] In some embodiments, the compound is any one of Compounds 1-44 or a pharmaceutically acceptable salt thereof:

[0197]

[0198]

[0199]

[0200] In an embodiment, in the compound of formula (A) and (I)-(III), such as any one of compounds 1-44, or a pharmaceutically acceptable salt thereof, at least one hydrogen atom is replaced by a deuterium atom.

[0201] In another aspect, the invention features a pharmaceutical composition comprising any compound described herein (e.g., a compound of Formula (A) and (I)-(III), such as any of Compounds 1-44), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0202] In another aspect, the invention features a method for treating a disease mediated by PHD activity, the method comprising administering to a subject any compound described herein (e.g., a compound of Formula (A) and (I)-(III), such as any of Compounds 1-44), or a pharmaceutically acceptable salt thereof.

[0203] In embodiments, the disease mediated by PHD activity is ischemia-reperfusion injury (eg, stroke, myocardial infarction, or acute kidney injury).

[0204] In embodiments, the disease mediated by PHD activity is inflammatory bowel disease (eg, ulcerative colitis or Crohn's disease).

[0205] In embodiments, the disease mediated by PHD activity is cancer (eg, colorectal cancer).

[0206] In embodiments, the disease mediated by PHD activity is a liver disease.

[0207] In embodiments, the disease mediated by PHD activity is atherosclerosis.

[0208] In embodiments, the disease mediated by PHD activity is a cardiovascular disease.

[0209] In embodiments, the disease mediated by PHD activity is an ocular disease or condition (eg, radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia).

[0210] In embodiments, the disease mediated by PHD activity is anemia (eg, anemia associated with chronic kidney disease).

[0211] In embodiments, the disease mediated by PHD activity is associated with hyperoxia

[0212] In embodiments, the disease mediated by PHD activity is retinopathy of prematurity.

[0213] In embodiments, the disease mediated by PHD activity is bronchopulmonary dysplasia (BPD).

[0214] In embodiments, the disease mediated by PHD activity is 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

[0215] Figure 1 FIG. 5 is an exemplary schematic diagram showing the principle of TR-FRET assay for PHD enzymes (PHD1, PHD2, and PHD3). 2 In the case of hydroxylation, PHD enzyme hydroxylates proline 564 of biotinylated HIF-1α peptide, thereby generating 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 SA-D2 complex bound to HIF-1α-hydroxyproline generates a detectable and quantifiable fluorescence resonance energy transfer signal. DETAILED DESCRIPTION

[0216] definition

[0217] In order to make the present invention more easily understood, some terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification. Publications and other reference materials cited herein that are used to describe the background technology of the present invention and to provide additional details about its implementation are hereby incorporated by reference.

[0218] 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., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, a cow, a primate, and / or a pig). In some embodiments, the animal includes, but is not limited to, a mammal, a bird, a reptile, an amphibian, a fish, an insect, and / or a worm. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and / or a clone.

[0219] Approximately or approximately: As used herein, when applied to one or more values ​​of interest, the term "approximately" or "approximately" refers to a value similar to the reference value. In certain embodiments, the term "approximately" 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 value, unless otherwise specified or otherwise apparent from the context (unless the number exceeds 100% of the possible value).

[0220] As used in the specification and the appended claims, the singular forms "a", "an", and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a composition" includes a mixture of two or more such compositions.

[0221] Throughout the description and claims of this specification, the word "comprise" and other forms of the word, 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.

[0222] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0223] Improvement, increase or decrease: As used herein, the terms "improvement," "increase," or "decrease," or grammatical equivalents, refer to values ​​relative to a baseline measurement, such as a measurement in the same individual prior to starting a treatment described herein, or a measurement in a control subject (or multiple control subjects) in the absence of a treatment described herein. A "control subject" is a subject having the same form of disease as the subject being treated, who is approximately the same age as the subject being treated.

[0224] 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., rather than in vivo in a multicellular organism.

[0225] In vivo: As used herein, the term "in vivo" refers to events that occur within a multicellular organism (such as a human or non-human animal). In the context of a cell-based system, the term can be used to refer to events that occur within living cells (as opposed to, for example, in vitro systems).

[0226] Patient: As used herein, the term "patient" or "subject" refers to any organism to which the provided compositions can be administered, e.g., for experimental, diagnostic, preventive, 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.

[0227] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" refers to substances that are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio and within the scope of sound medical judgment.

[0228] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66: 1–19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts with an amino group 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 adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, gluconate, heptadecanoate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. 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. Where appropriate, 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 are carried out using appropriate electrophilic reagents (e.g., alkyl halides) to form quaternized alkylated amino salts.

[0229] 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. The subject may be a patient, which refers to a person presented to a medical provider for diagnosis or treatment of a disease. The term "subject" is used interchangeably herein with "individual" or "patient". A subject may suffer from or be susceptible to a disease or condition, but may or may not show symptoms of the disease or condition.

[0230] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting the full or nearly full extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, reach completion and / or proceed to completion or achieve or avoid an absolute result. Thus, the term "substantially" is used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0231] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" of a therapeutic agent means an amount that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, diagnose, prevent, and / or delay the onset of symptoms of the disease, disorder, and / or condition. One 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.

[0232] Treatment: As used herein, the term "treat", "treatment" or "treating" refers to any method used to partially or completely alleviate, improve, alleviate, 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. In order to reduce the risk of developing pathology associated with a disease, treatment can be administered to subjects who do not show signs of the disease and / or only show early signs of the disease.

[0233] Aliphatic: As used herein, the term aliphatic refers to C 1 –C 40 Hydrocarbons include saturated hydrocarbons and unsaturated hydrocarbons. Aliphatic hydrocarbons 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 or branched chain C 1 –C 20 Saturated alkyl), C 2 –C 20 Alkenyl (e.g., straight or branched C 4 –C 20 Diene, straight or branched chain C 6 –C 20 triene, etc.) and C 2 –C 20 Alkynyl (e.g., straight or branched C 2 –C 20 C 1 –C 20 Aliphatic can contain C 3 –C 20Cyclic aliphatic (e.g., C 3 –C 20 Cycloalkyl, C 4 –C 20 Cycloalkenyl or C 8 –C 20 In certain embodiments, an aliphatic group may include one or more cycloaliphatic groups and / or one or more heteroatoms, such as oxygen, nitrogen, or sulfur, and may be optionally substituted with one or more substituents, such as alkyl, halo, alkoxy, hydroxy, amino, aryl, ether, ester, or amide. An aliphatic group is unsubstituted or substituted with one or more substituents as described herein. For example, an aliphatic group may be substituted with 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' is substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein 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 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 In some embodiments, R' is independently unsubstituted C 1 –C 3 In some embodiments, the aliphatic group is unsubstituted. In some embodiments, the aliphatic group does not contain any heteroatoms.

[0234] Alkyl: As used herein, the term "alkyl" means non-cyclic straight chain and branched chain hydrocarbon groups, such as "C 1 –C 20"Alkyl" refers to an alkyl group having 1-20 carbons. The alkyl group can be straight or branched. 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 an alkyl straight or branched chain alkyl group having 1 to 6 carbon atoms. Other alkyl groups will be apparent to those skilled in the art in view of the benefits of this disclosure. 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 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' is substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein 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 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 In some embodiments, R' is independently unsubstituted C 1 –C 3 Alkyl. In some embodiments, the alkyl group is substituted (e.g., substituted with 1, 2, 3, 4, 5, or 6 substituents as described herein). In some embodiments, the alkyl group is substituted with an -OH group and may also be referred to herein as a "hydroxyalkyl" group, wherein the prefix represents an -OH group and "alkyl" is as described herein. In some embodiments, the alkyl group is substituted with an -OR' group and may also be referred to herein as an "alkoxy" group.

[0235] Appending the suffix "-ene" to a group indicates that the group is a divalent moiety, for example, arylene is a divalent moiety of aryl, and heteroarylene is a divalent moiety of heteroaryl.

[0236] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent straight or branched hydrocarbon group, and is exemplified by methylene, ethylene, isopropylene, and the like. Similarly, as used herein, the term "alkenylene" refers to an unsaturated divalent straight or branched 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" refers herein to an unsaturated divalent straight or branched 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, an alkylene, alkenylene, or alkynylene group may include one or more cyclic aliphatic groups and / or one or more heteroatoms, such as oxygen, nitrogen, or sulfur, and may be optionally substituted with one or more substituents, such as alkyl, halide, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide. For example, an alkylene, alkenylene, or alkynylene group may be substituted with halogen, -COR', -CO', or -COR'. 2 H, -CO 2 R', -CN, -OH, -OR', -OCOR', -OCO 2 R', -NH 2 、-NHR'、-N(R') 2 , -SR' or -SO 2 R' is substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein 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 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 In some embodiments, R' is independently unsubstituted C 1 –C 3 In some embodiments, the alkylene, alkenylene or alkynylene group is unsubstituted. In some embodiments, the alkylene, alkenylene or alkynylene group does not contain any heteroatoms.

[0237] Alkenyl: As used herein, "alkenyl" means any straight or branched hydrocarbon chain having one or more unsaturated carbon-carbon double bonds which may be present at any stable point along the chain, e.g., "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, the alkenyl group includes 1, 2, or 3 carbon-carbon double bonds. In some embodiments, the alkenyl group includes a single carbon-carbon double bond. In some embodiments, multiple double bonds (e.g., 2 or 3) are conjugated. The alkenyl group can be unsubstituted or substituted with one or more substituents as described herein. For example, the alkenyl group can be substituted with 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' is substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein 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 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 In some embodiments, R' is independently unsubstituted C 1 -C 3In some embodiments, the alkenyl group is an alkyl group. In some embodiments, the alkenyl group is unsubstituted. In some embodiments, the alkenyl group is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituents as described herein). In some embodiments, the alkenyl group is substituted with an -OH group and may also be referred to herein as a "hydroxyalkenyl" group, wherein the prefix represents an -OH group and the "alkenyl" group is as described herein.

[0238] Alkynyl: As used herein, "alkynyl" means any hydrocarbon chain in a straight or branched configuration having one or more carbon-carbon triple bonds at any stable point along the chain, e.g., "C 2 -C 20 "Alkynyl" refers to an alkynyl group having 2 to 20 carbons. Examples of alkynyl groups include prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl, hex-5-ynyl, and the like. In some embodiments, the alkynyl group includes a carbon-carbon triple bond. The alkynyl group may be unsubstituted or substituted with one or more substituents as described herein. For example, the alkynyl group may be substituted with 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' is substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein 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 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 In some embodiments, R' is independently unsubstituted C 1 –C 3In some embodiments, the alkynyl group is unsubstituted. In some embodiments, the alkynyl group is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituents as described herein).

[0239] Aryl: The term "aryl" used alone or as part of a larger moiety as in "aralkyl" refers to a monocyclic, bicyclic, or tricyclic carbon ring system having a total of six to fourteen ring members, wherein the ring system has a single point of attachment to the rest of the molecule, at least one ring in the system is aromatic, and wherein each ring in the system contains 4 to 7 ring members. In some embodiments, an aryl group has 6 ring carbon atoms ("C 6 In some embodiments, an aryl group has 10 ring carbon atoms (“C 10 In some embodiments, an aryl group has 14 ring carbon atoms (“C 14 "Aryl"; e.g., anthracenyl). "Aryl" also includes ring systems in which an aromatic ring, as defined above, is fused to one or more carbocyclic or heterocyclic groups, wherein the radical or point of attachment is on the aromatic ring, and in such a case, the number of carbon atoms continues to refer to the number of carbon atoms in the aromatic ring system. Exemplary aryl groups include phenyl, naphthyl, and anthracene.

[0240] Arylene: As used herein, the term "arylene" refers to a divalent aromatic group (ie, having two points of attachment to the molecule). Exemplary arylene groups include phenylene (eg, unsubstituted phenylene or substituted phenylene).

[0241] Halogen or halo: As used herein, the term "halogen" or "halo" means fluoro, chloro, bromo, or iodo.

[0242] Amide: Unless otherwise specified in the specification, the term "amide" or "amide group" refers to a amide 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, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl or heterocycloalkyl (bonded through a ring carbon), as described herein, each of which itself can be optionally substituted, or two R' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring.

[0243] Amino: Unless otherwise specified in the specification, 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, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), sulfonyl or carbonyl, each of which itself may be optionally substituted as described herein, or two R' may be combined with the nitrogen atom to form a 3-, 4-, 5-, 6- or 7-membered ring. In an embodiment, amino is -NHR', wherein R' is aryl ("arylamino"), heteroaryl ("heteroarylamino") or alkyl ("alkylamino").

[0244] Sulfonyl: Unless otherwise specified in the specification, the term "sulfonyl" refers to -S(=O) 2 R' or -S(=O) 2 -group, wherein R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), each of which moieties can itself be optionally substituted as described herein.

[0245] Sulfinyl: Unless stated otherwise in the specification, the term "sulfinyl" refers to a chemical moiety having the formula -S(=O)R', -S(=O)-, or -S(=O)(=NR')-, wherein R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), each of which moiety itself can be optionally substituted as described herein.

[0246] Carbonyl: Unless stated otherwise in the specification, the term "carbonyl" refers to a -C(=O)R' or a -C(=O)- group, where R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon), cycloalkyl, aryl, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, heterocycloalkyl (bonded through a ring carbon), each of which moiety can itself be optionally substituted as described herein.

[0247] Phosphoryl: Unless otherwise specified in the specification, the term "phosphoryl" refers to -P(=O)(R') 2 or a -P(=O)(R')- group, wherein each R' is selected from hydrogen, alkyl, alkenyl, alkynyl, heteroalkyl (bonded through a chain carbon or through a heteroatom), cycloalkyl, aryl, arylalkyl, heteroaryl (bonded through a ring carbon), heteroarylalkyl, or heterocycloalkyl (bonded through a ring carbon), as described herein, each of which itself can be optionally substituted, or two R's can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring.

[0248] Assorted alkyl: The term "assorted alkyl" means a branched or unbranched alkyl, alkenyl or alkynyl group having 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. Assorted alkyl includes tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiester, phosphoramidates, sulfonamides and disulfides. Assorted alkyl can optionally include monocyclic, bicyclic or tricyclic rings, wherein each ring ideally has three to six members. Examples of assorted alkyl include polyethers, such as methoxymethyl and ethoxyethyl.

[0249] Heteroalkylene: As used herein, the term "heteroalkylene" refers to a divalent form of a heteroalkyl group as described herein.

[0250] Heteroaryl: As used herein, the term "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 rest 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.

[0251] Heterocycloalkyl: As used herein, the term "heterocycloalkyl" is a non-aromatic ring in which 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 can be substituted or unsubstituted.

[0252] Deuterium: The term "deuterium" ("D" or " 2 Deuterium is an isotope of hydrogen whose nucleus consists of one proton and one neutron, and which is twice as massive as the nucleus of ordinary hydrogen (one proton).

[0253] Isotope: The term "isotope" refers to a variant of a particular chemical element in which the number of protons and, therefore, nucleons differs. All isotopes of a given element have the same number of protons but different numbers of neutrons in each atom.

[0254] The term "substituted" means that the specified group or moiety bears one or more substituents. The term "unsubstituted" means that the specified group bears no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted with one or more substituents. Where the term "substituted" is used to describe a structural system, substitution is intended to occur at any valence-permitted position on the system, e.g., substitution produces a stable compound (e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction). Where a specified moiety or group is not explicitly noted as being optionally substituted or substituted with any specified substituent, it is understood that such moiety or group is intended to be unsubstituted.

[0255] When a ring system (e.g., cycloalkyl, heterocyclyl, aryl, or heteroaryl) is substituted with multiple substituents that vary within well-defined limits, it is understood that the total number of substituents does not exceed the normal available valences under 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 encompass only those combinations of substituents and variables that produce stable or chemically feasible compounds. A stable compound or chemically feasible compound is one that has stability sufficient to allow its preparation and testing, among other factors.

[0256] A variety of substituents are well known, as are methods of forming and introducing a variety of substituents into a variety of parent groups. Representative substituents include, but are not limited to, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, aralkyl, alkaryl, aryl, arylalkoxy, arylamino, heteroarylamino, heteroaryl, heteroarylalkoxy, heterocycloalkyl, hydroxyalkyl, aminoalkyl, haloalkyl, thioalkyl, alkylthioalkyl, carboxyalkyl, imidazolidinyl, indolylalkyl, mono-, di- and tri-haloalkyl, mono-, di- and tri-haloalkoxy, 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 (halogen) 2 ,═S,═O,—CON(R 50 R 51 ),—OCOR 50 、—OCON(R 50 R 51 ),—N(R 52 )CO(R 50 ),—N(R 52 )COOR 50 and—N(R 52 )CON(R 50 (R 51 ), where R 50 , R 51 and R 52 can be independently selected from the following: hydrogen atoms and branched or straight chain, C 1–6-alkyl, C 3–6 -Cycloalkyl, C 4–6 - heterocycloalkyl, heteroaryl and aryl, with or without substituents. Where permitted, R 50 and R 51 They may be linked together to form carbocyclic or heterocyclic ring systems.

[0257] 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 In certain embodiments thereof, R' is independently unsubstituted alkyl (eg, unsubstituted C 1 –C 20 Alkyl, C 1 –C 15 Alkyl, C 1 –C 10 Alkyl or C 1 –C 3 Preferably, R' is independently an unsubstituted C 1 –C 3 alkyl.

[0258] Any formula given herein is intended to represent a compound having a structure depicted by a structural formula and some variations or forms. Specifically, the compound of any formula given herein may have an asymmetric center and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of the general formula compound and their mixtures are considered to be within the scope of the formula. Therefore, any formula given herein is intended to represent a racemate, one or more enantiomeric forms, one or more diastereomeric forms, one or more atropisomer forms, and mixtures thereof. In addition, some structures may exist as geometric isomers (i.e., cis and trans isomers), as tautomers or as atropisomers. In addition, any formula given herein is intended to include hydrates, solvates and polymorphs of such compounds, and mixtures thereof.

[0259] Compounds of the present invention

[0260] Disclosed herein are compounds that are effective inhibitors of PHD. In some embodiments, the compounds of the invention have an enzymatic half maximal inhibitory concentration (IC) of less than 100 μM for any of PHD1, PHD2, and PHD3. 50 ) value. In some embodiments, the compounds of the present invention have an IC of less than 50 μM for any one of PHD1, PHD2 and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 25 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 20 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 15 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 10 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 5 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of less than 1 μM for any of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 3 nM to about 5 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 5 nM to about 10 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 10 nM to about 20 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 20 nM to about 50 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 50 nM to about 100 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 100 nM to about 200 nM for any one of PHD1, PHD2, and PHD3. 50In some embodiments, the compounds of the present invention have an IC value of about 200 nM to about 500 nM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, the compounds of the present invention have an IC value of about 500 nM to about 1000 nM for any one of PHD1, PHD2, and PHD3. 50 value.

[0261] Representative examples from this class show inhibitory activity against PHD1, PHD2, and PHD3 in vitro.

[0262] Exemplary compounds are described herein. Specifically, these selective inhibitors can be characterized by a pyrazole moiety (eg, a 5-hydroxy substituted pyrazole) linking two aromatic moieties.

[0263] Compounds of formula (A) and (I)-(III)

[0264] In one aspect, provided herein are compounds having a structure according to Formula (A):

[0265]

[0266] or a pharmaceutically acceptable salt thereof, wherein:

[0267] Ar 1 is phenyl or a six-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl or C 1-3 Alkoxy substitution;

[0268] R 2 H or C 1-3 alkyl;

[0269] Ar 2 is a six-membered nitrogen-containing heteroaryl group, which is optionally substituted by halogen, OH, amine or C 1-3 Alkyl substituted; and

[0270] R 4 is hydrogen or C 1-4 Alkyl, and

[0271] In embodiments of the formulae described herein (eg, formula (A)), the compound (eg, compound of formula (A)) is not

[0272]

[0273] In embodiments of the formulae described herein (eg, formula (A)), compounds (eg, compounds of formula (A)) do not include compounds

[0274]

[0275]

[0276] In an embodiment, R 2 For H.

[0277] In an embodiment, R 2 C 1-3 In the embodiment, R 2 CH 3 In an embodiment, R 2 CH 2 CH 3 In an embodiment, R 2 CH 2 CH 2 CH 3 In an embodiment, R 2 for CH(CH 3 ) 2 .

[0278] In an embodiment, R 4 For H.

[0279] In an embodiment, R 4 C 1-4 In the embodiment, R 4 CH 3 In an embodiment, R 4 CH 2 CH 3 In an embodiment, R 4 CH 2 CH 2 CH 3 In an embodiment, R 4 for CH(CH 3 ) 2 In an embodiment, R 4 CH 2 CH 2 CH 2 CH 3 In an embodiment, R 4 CH(CH 3 )(CH 2 CH 3 ). In an embodiment, R 4 C(CH 3 ) 3 .

[0280] In the embodiment, Ar 1 is an unsubstituted aryl group. 1 is a substituted aryl group.1 is an unsubstituted phenyl group. 1 is a substituted phenyl group.

[0281] In the embodiment, Ar 1 is an unsubstituted 6-membered nitrogen-containing heteroaryl group. 1 It is a substituted 6-membered nitrogen-containing heteroaryl group.

[0282] In the embodiment, Ar 1 is selected from hydrogen, halogen, CN, OH, C optionally substituted by one or more halogens 1-3 Alkyl and C 1-3 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 In some embodiments, Ar 1 Substituted with 4 substituents.

[0283] In the embodiment, Ar 1 Include one or more R 1 A group in which each R 1 are independently selected from hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl and C 1-3 In the embodiment, Ar 1 Including R represented by m 1 The number of groups, where m is 1, 2, 3 or 4. 1 When present, R 1 Can replace the hydrogen in the parent molecular structure. In the embodiment, when R 1 When present and is a non-hydrogen moiety, R 1 In the embodiment, R 1 independently selected from halogen, CN, OH, C optionally substituted by CN or one or more halogens 1-3 Alkyl and C 1-3 Alkoxy.

[0284] Thus, it is also understood that for any value of m described herein, hydrogen is present where appropriate to form a 1 The valence requirements are fulfilled on the constituent atoms of the molecule such that the molecule is a stable compound (e.g., a molecule that does not spontaneously transform, such as by rearrangement, cyclization, elimination, or other reactions). 1 , R 1 and exemplary embodiments of m.

[0285] In the embodiment, Ar 1 for

[0286] in

[0287] X, Y and Z are independently CH or N, wherein N is optionally oxidized;

[0288] Each R 1 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; and

[0289] m is 1, 2, 3 or 4.

[0290] In an embodiment, R 1 In an embodiment, when R 1 When present and is a non-hydrogen moiety, R 1 represents a substituent.

[0291] In embodiments, the value of m is based on the number of nitrogen atoms present in the ring. In embodiments, when two and only two of X, Y, and Z are N, m is 1, 2, or 3. In embodiments, when each of X, Y, and Z is N, m is 1 or 2.

[0292] In an embodiment, m is 1. In an embodiment, m is 2. In an embodiment, m is 3. In an embodiment, m is 4.

[0293] In embodiments, X, Y, and Z are all N, and m is 1 or 2. In embodiments, m is 1, and any remaining unsubstituted carbon ring atoms are assumed to be bonded with hydrogen to fill the valence. In embodiments, m is 2.

[0294] In an embodiment, one of X, Y, and Z is CH, and the others are N, wherein N is optionally oxidized, and m is 1, 2, or 3. In an embodiment, m is 1, and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 2, and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3.

[0295] In an embodiment, two of X, Y, and Z are CH, and the other is N, wherein N is optionally oxidized, 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 valences. In an embodiment, m is 2, and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 3, and any remaining unsubstituted carbon ring atoms are assumed to be bonded to hydrogen to fill the valences. In an embodiment, m is 4.

[0296] In the embodiment, Ar 1 The N atoms in the ions are not oxidized.

[0297] In the embodiment, Ar 1 The N atoms in the ions are oxidized.

[0298] In the embodiment, Ar 1 is a substituted phenyl group. 1 By at least one R 1 Substitution, where R 1 It is CN halogen.

[0299] In the embodiment, Ar 1 By one or two R 1 A group is substituted, the group being independently selected from C optionally substituted by one or more halogens 1-3 Alkyl, halogen, CN or OH.

[0300] In the embodiment, Ar 1 is a pyridyl N-oxide or is optionally substituted by at least one R 1 Substituted pyridyl, wherein R1 is C 1-3 Alkoxy or halogen.

[0301] In the embodiment, each R 1 For hydrogen.

[0302] In the embodiment, each R 1 For CN.

[0303] In the embodiment, each R 1 For OH.

[0304] In the embodiment, each R 1 In an embodiment, the halogen is Cl. In an embodiment, the halogen is Br. In an embodiment, the halogen is I.

[0305] In the embodiment, each R 1 C 1-3 alkyl.

[0306] In the embodiment, each R 1 For unsubstituted C 1-3 In the embodiments, each R 1 CH 3 In the embodiment, each R 1 CH 2 CH 3 .

[0307] In the embodiment, each R 1 is substituted C 1-3 In the embodiments, each R 1 is C substituted by one or more halogens 1-3 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.

[0308] In the embodiment, each R 1 For CF 3 .

[0309] In the embodiment, each R 1 C 1-3 In the embodiment, each R 1 It is OMe.

[0310] In some embodiments, Ar 1 Selected from:

[0311]

[0312]

[0313] In some embodiments, Ar 1 Selected from:

[0314] In some embodiments, Ar 1 Selected from:

[0315]

[0316] In some embodiments, Ar 1 Selected from:

[0317] In some embodiments, Ar 1 Selected from:

[0318]

[0319] In some embodiments, Ar 1 Selected from:

[0320]

[0321] In the embodiment, Ar 2 It is an unsubstituted six-membered nitrogen-containing heteroaryl group.

[0322] In the embodiment, Ar 2 For halogen, OH, amine or C 1-3 alkyl-substituted six-membered nitrogen-containing heteroaryl.2 is substituted with 1 substituent as described herein. 2 Substituted with 2 substituents as described herein.

[0323] In the embodiment, Ar 2 for

[0324] in

[0325] A and B are independently CH or N, wherein N is optionally oxidized;

[0326] Each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; and

[0327] n is 0, 1 or 2.

[0328] In embodiments, the value of n is based on the number of nitrogen atoms present in the ring. In embodiments, when one and only one of A and B is N, n is 0, 1 or 2. In embodiments, when both A and B are N, n is 0 or 1.

[0329] In an embodiment, n is 0. In an embodiment, n is 1. In an embodiment, n is 2.

[0330] In an embodiment, A and B are both N, wherein N is optionally oxidized, and n is 0 or 1. In an embodiment, n is 0. In an embodiment, n is 1.

[0331] In an embodiment, one of A and B is CH and the other is N, wherein N is optionally oxidized, and n is 0, 1 or 2. In an embodiment, n is 0. In an embodiment, n is 1. In an embodiment, n is 2.

[0332] In an embodiment, A and B are both CH, and n is 0, 1 or 2. In an embodiment, n is 0. In an embodiment, n is 1. In an embodiment, n is 2.

[0333] In the embodiment, Ar 2 The N atoms in the ions are not oxidized.

[0334] In the embodiment, Ar 2 The N atoms in the ions are oxidized.

[0335] In the embodiment, each R 3 For hydrogen.

[0336] In the embodiment, each R 3 For OH.

[0337] In the embodiment, each R 3In 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.

[0338] In the embodiment, each R 3 In the embodiment, each R 3 For NH 2 .

[0339] In the embodiment, each R 3 C 1-3 alkyl.

[0340] In the embodiment, each R 3 For unsubstituted C 1-3 In the embodiments, each R 3 CH 3 .

[0341] In the embodiment, Ar 2 Selected from the group consisting of:

[0342]

[0343] In an embodiment, the compound of formula (A) has the following structure,

[0344] or a pharmaceutically acceptable salt thereof, wherein A, B, X, Y, Z, R 1 , R 2 , R 3 and R 4 As defined anywhere herein.

[0345] In embodiments of the formulae described herein (eg, formula (I)), the compound (eg, compound of formula (I)) is not

[0346]

[0347]

[0348] In the embodiments of the formulae described herein (eg, formula (I)), the compounds (eg, compounds of formula (I)) do not include the following compounds

[0349]

[0350] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0351] or a pharmaceutically acceptable salt thereof, wherein X, Y, Z, R 1 , R 2 , R3 and R 4 As defined anywhere herein.

[0352] In embodiments of the formulae described herein (eg, formula (Ia)), the compound (eg, compound of formula (Ia)) is not

[0353]

[0354] In the embodiments of the formulae described herein (eg, formula (Ia)), the compounds (eg, compounds of formula (Ia)) do not include the following compounds

[0355]

[0356]

[0357] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0358] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 and R 4 As defined anywhere herein.

[0359] In embodiments of the formulae described herein (eg, formula (Ib)), the compound (eg, compound of formula (Ib)) is not

[0360]

[0361] In embodiments of the formulae described herein (eg, formula (Ib)), compounds (eg, compounds of formula (Ib)) do not include compounds

[0362]

[0363] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0364] or a pharmaceutically acceptable salt thereof, wherein A, B, X, Y, Z, R 1 , R 3 and R 4 As defined anywhere herein.

[0365] In an embodiment, the compound of formula (A) or the compound of formula (I) has the following structure,

[0366] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R3 and R 4 As defined anywhere herein.

[0367] In embodiments of the formulae described herein (eg, formula (II)), the compound (eg, compound of formula (II)) is not

[0368]

[0369] In the embodiments of the formula (eg, formula (II)) described herein, the compound (eg, compound of formula (II)) does not include the following compounds

[0370]

[0371]

[0372] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0373] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 3 As defined anywhere herein.

[0374] In embodiments of the formulae described herein (eg, formula (IIa)), the compound (eg, compound of formula (IIa)) is not

[0375]

[0376] In the embodiments of the formula (eg, formula (IIa)) described herein, the compound (eg, compound of formula (IIa)) does not include the following compounds

[0377]

[0378] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0379] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 and R 4 As defined anywhere herein.

[0380] In embodiments of the formulae described herein (eg, formula (IIb)), the compound (eg, compound of formula (IIb)) is not

[0381]

[0382]

[0383] In the embodiments of the formula (eg, formula (IIb)) described herein, the compound (eg, compound of formula (IIb)) does not include the following compounds

[0384]

[0385] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0386] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 As defined anywhere herein.

[0387] In embodiments of the formulae described herein (eg, formula (IIc)), the compound (eg, compound of formula (IIc)) is not

[0388]

[0389] In embodiments of the formulae described herein (eg, formula (IIc)), compounds (eg, compounds of formula (IIc)) do not include compounds

[0390]

[0391] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0392] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 3 As defined anywhere herein.

[0393] In embodiments of the formulae described herein (eg, formula (IId)), the compound (eg, compound of formula (IId)) is not

[0394]

[0395] In embodiments of the formulae described herein (eg, formula (IId)), compounds (eg, compounds of formula (IId)) do not include compounds

[0396]

[0397]

[0398] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0399] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 4 As defined anywhere herein.

[0400] In embodiments of the formulae described herein (eg, formula (IIe)), the compound (eg, compound of formula (IIe)) is not

[0401]

[0402] In embodiments of the formulae described herein (eg, formula (IIe)), compounds (eg, compounds of formula (IIe)) do not include compounds

[0403]

[0404] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0405] or a pharmaceutically acceptable salt thereof, wherein R 1 As defined anywhere herein.

[0406] In embodiments of the formulae described herein (eg, Formula (IIf)), the compound (eg, compound of Formula (IIf)) is not

[0407]

[0408]

[0409] In embodiments of the formulae described herein (eg, formula (IIf)), compounds (eg, compounds of formula (IIf)) do not include compounds

[0410]

[0411] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0412] or a pharmaceutically acceptable salt thereof wherein R 1 , R 3 and R 4 As defined anywhere herein.

[0413] In an embodiment, the compound of formula (A), the compound of formula (I) or the compound of formula (II) has the following structure,

[0414] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2, R 3 and R 4 as defined anywhere herein, and wherein

[0415] R 5 is CN or halogen.

[0416] In an embodiment, R 5 For CN.

[0417] In an embodiment, R 5 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.

[0418] In embodiments of the formulae described herein (eg, formula (III)), the compound (eg, compound of formula (III)) is not

[0419]

[0420] In the embodiments of the formula (eg, formula (III)) described herein, the compound (eg, compound of formula (III)) does not include the following compounds

[0421]

[0422]

[0423] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0424] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 and R 5 As defined anywhere herein.

[0425] In embodiments of the formulae described herein (eg, formula (IIIa)), the compound (eg, compound of formula (IIIa)) is not

[0426]

[0427]

[0428] In the embodiments of the formula (eg, formula (IIIa)) described herein, the compound (eg, compound of formula (IIIa)) does not include the following compounds

[0429]

[0430] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0431] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 4 and R 5 As defined anywhere herein.

[0432] In embodiments of the formulae described herein (eg, formula (IIIb)), the compound (eg, compound of formula (IIIb)) is not

[0433]

[0434] In the embodiments of the formula (eg, formula (IIIb)) described herein, the compound (eg, compound of formula (IIIb)) does not include the following compounds

[0435]

[0436] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0437] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , and R 5 As defined anywhere herein.

[0438] In embodiments of the formulae described herein (eg, formula (IIIc)), the compound (eg, compound of formula (IIIc)) is not

[0439]

[0440] In embodiments of the formula (eg, formula (IIIc)) described herein, the compound (eg, compound of formula (IIIc)) does not include the following compounds

[0441]

[0442]

[0443] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0444] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 3 , and R5 As defined anywhere herein.

[0445] In embodiments of the formulae described herein (eg, formula (IIId)), the compound (eg, compound of formula (IIId)) is not

[0446]

[0447] In embodiments of the formulae described herein (eg, formula (IIId)), compounds (eg, compounds of formula (IIId)) do not include compounds

[0448]

[0449] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0450] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 4 , and R 5 As defined anywhere herein.

[0451] In embodiments of the formulae described herein (eg, formula (IIIe)), the compound (eg, compound of formula (IIIe)) is not

[0452]

[0453]

[0454] In embodiments of the formulae described herein (eg, formula (IIIe)), compounds (eg, compounds of formula (IIIe)) do not include compounds

[0455]

[0456] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0457] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 5 As defined anywhere herein.

[0458] In embodiments of the formulae described herein (eg, formula (IIIf)), the compound (eg, compound of formula (IIIf)) is not

[0459]

[0460] In embodiments of the formulae described herein (eg, formula (IIIf)), compounds (eg, compounds of formula (IIIf)) do not include compounds

[0461]

[0462] In an embodiment, the compound of formula (A), the compound of formula (I), the compound of formula (II) or the compound of formula (III) has the following structure,

[0463] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 3 , R 4 and R 5 As defined anywhere herein.

[0464] Exemplary compounds

[0465] In some embodiments, the PHD inhibitor compound is any one of Compounds 1-44 or a pharmaceutically acceptable salt thereof.

[0466]

[0467]

[0468]

[0469] isotope

[0470] It should be understood that in the compounds described herein (e.g., compounds of any of Formulas (A) and (I)-(III), such as any of Compounds 1-44), atoms may be present in their natural isotopic abundance, or one or more atoms may be artificially enriched with a particular isotope having the same atomic number but an atomic mass or mass ordinal number different from the atomic mass or mass ordinal number predominantly found in nature. The present invention is intended to include all suitable isotopic variations of the compounds described herein (e.g., compounds of any of Formulas (A) and (I)-(III), such as any of Compounds 1-44). For example, different isotopic forms of hydrogen (H) include protium ( 1 H), deuterium ( 2 H) and tritium ( 3 H). Protium is the major isotope of hydrogen found in nature.

[0471] In some embodiments, one or more hydrogens of a compound described herein (e.g., a compound of any one of formula (A) and (I)-(III), such as any one of compounds 1-44) are replaced by deuterium. Enrichment with deuterium can provide certain therapeutic advantages, such as increasing half-life in vivo or reducing dosage requirements, or can provide a compound that can be used as a standard for characterizing a biological sample. In some embodiments, one or more hydrogens of a compound described herein (e.g., a compound of any one of formula (A) and (I)-(III), such as any one of compounds 1-44) are replaced by tritium. Tritium is radioactive and can therefore provide a radiolabeled compound that can be used as a tracer in metabolic or kinetic studies.

[0472] Isotopic enrichment of the compounds disclosed herein (e.g., compounds of any of Formulas (A) and (I)-(III), such as any of Compounds 1-44) can be achieved without undue experimentation by conventional techniques well known to those skilled in the art or by methods analogous to those described in the Schemes and Examples herein using appropriate isotopically enriched reagents and / or intermediates.

[0473] The term "isotopologue" refers to a substance having the same chemical structure and chemical formula as a specific compound provided herein, for example, hydrogen and deuterium, except for the isotope substitution position and / or isotope enrichment level at one or more positions. Therefore, as used herein, the term "compound" encompasses a collection of molecules having the same chemical structure but also having isotopic changes between the constituent atoms of the molecule. Therefore, it will be clear to those skilled in the art that a compound represented by a specific chemical structure containing an indicated deuterium atom will also contain a smaller amount of isotopologues having hydrogen atoms at one or more designated deuterium positions in the structure. The relative amount of such isotopologues in the provided compound depends on many factors, including but not limited to the isotopic purity of the deuterated reagent used to prepare the compound and the efficiency of deuterium incorporation in each synthesis step used to prepare the compound.

[0474] When a position is designated as "H" or "hydrogen", the position is understood to have hydrogen according to its natural abundance isotopic composition. When a position is designated as "D" or "deuterium", the position is understood to have deuterium at an abundance at least 3340 times greater than the natural abundance of deuterium, which is 0.015% (i.e., the term "D" or "deuterium" means at least 50.1% deuterium incorporation).

[0475] In embodiments, the compounds provided herein can have an isotopic enrichment factor for each deuterium present at a site designated as a potential deuteration site on the compound of 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 incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0476] Synthesis of the compounds of the present invention

[0477] Compounds described herein (e.g., compounds of any of Formulas A and I-III, such as any of Compounds 1-44) can be prepared according to methods known in the art, including the exemplary syntheses of the examples provided herein, such as the synthesis shown in Scheme A.

[0478] The purity of the compounds and their synthetic intermediates was determined by reverse phase HPLC using any of the following methods:

[0479] Method A: Mobile phase: A: water (0.01% TFA) B: acetonitrile (0.01% TFA); Gradient step: 5% B increased to 95% B in 1.4 minutes, 95% B for 1.6 minutes (total run time: 3 minutes); Flow rate: 2.3 ml / min. Column: SunFire C18, 4.6*50 mm, 3.5 μm; Column temperature: 50°C. Detectors: ADC ELSD, DAD (214 nm and 254 nm), ES-API.

[0480] Method B: Mobile phase: A: water (10 mM NH 4 HCO 3 ) B: acetonitrile; gradient stage: from 5% to 95% B in 1.5 minutes, 95% B for 1.5 minutes (total run time: 3 minutes); flow rate: 2.0 ml / min; column: XBridge C18, 4.6*50 mm, 3.5 um; column temperature: 40°C. Detectors: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API).

[0481] Abbreviations and acronyms used in this article include the following:

[0482]

[0483]

[0484] Plan A

[0485]

[0486] Compounds of formula (IId) are prepared according to Scheme A using commercially available materials. Compounds of formula (IId) are prepared according to Scheme A using commercially available materials. Reaction of esters of formula (B) with N,N-dimethylformamide dimethyl acetal produces enamine compounds of formula (C). Cyclization of (C) and hydrazide (D) provides pyrazole compounds having formula (IId).

[0487] Compositions and methods

[0488] The present invention provides a compound of any one of formula (A) and (I)-(III) or a pharmaceutically acceptable salt thereof for use in the preparation of a medicament for treating various conditions or disorders as described herein. In one embodiment, a pharmaceutical composition is provided, comprising at least one compound of any one of formula (A) and (I)-(III) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient or carrier. In various embodiments, the medicament or pharmaceutical composition may further comprise at least one additional therapeutic agent or be used in combination with at least one additional therapeutic agent.

[0489] The compounds of the present invention or the drugs or compositions comprising the compounds can be used to inhibit the activity of PHD. The inhibition of PHD may be particularly beneficial in the treatment of 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). 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)-(III), or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising one or more compounds of any one of formulas (A) and (I)-(III).

[0490] The present invention also relates to a method for inhibiting PHD activity. In one embodiment, the method comprises contacting PHD with an effective amount of one or more compounds or pharmaceutically acceptable salts thereof, wherein the one or more compounds are selected from the group consisting of compounds of any one of formula (A) and (I)-(III).

[0491] In still other embodiments, the compounds disclosed herein (e.g., compounds of any one of Formulas (A) and (I)-(III), such as any one of Compounds 1-44) or pharmaceutically acceptable salts 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, bone marrow transplant 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, nocturnal paroxysmal hemoglobinuria, myelofibrosis, pancytopenia, pure red cell aplasia, purpura Schoenlein-Henoch, refractory anemia with excess blasts, rheumatoid arthritis, Shwachman syndrome. syndrome), sickle cell disease, thalassemia major, thalassemia minor, thrombocytopenic purpura, anemic or non-anemic patients undergoing surgery, trauma-related or secondary anemia, sideroblastic anemia, anemia secondary to other treatments, including reverse transcriptase inhibitors used to treat HIV, corticosteroids, chemotherapy drugs with or without cisplatin, vinca alkaloids, mitotic inhibitors, topoisomerase II inhibitors, anthracyclines, alkylating agents, specifically, anemia secondary to inflammation, aging, and / or chronic disease. PHD1 inhibition can also be used to treat anemic symptoms, including chronic fatigue, pallor, and dizziness.

[0492] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat or prevent metabolic disorders, including but not limited to diabetes and obesity.

[0493] In yet other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat or prevent vascular disorders. These include, but are not limited to, hypoxia or wound healing related diseases that require pro-angiogenic mediators for angiogenesis, vasculogenesis, and arteriogenesis.

[0494] In still other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) 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.

[0495] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat inflammatory bowel diseases. These include, but are not limited to, ulcerative colitis and Crohn's disease.

[0496] In other embodiments, the compounds disclosed herein (eg, compounds of Formula (A) and (I)-(III), such as any of Compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat cancer, such as colorectal cancer.

[0497] In other embodiments, the compounds disclosed herein (eg, compounds of Formula (A) and (I)-(III), such as any of Compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat atherosclerosis.

[0498] In other embodiments, the compounds disclosed herein (eg, compounds of Formula (A) and (I)-(III), such as any one of Compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat cardiovascular disease.

[0499] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat ocular diseases or conditions. These include, but are not limited to, radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

[0500] In other embodiments, the compounds disclosed herein (eg, compounds of Formula (A) and (I)-(III), such as any of Compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat diseases associated with hyperoxia.

[0501] In other embodiments, the compounds disclosed herein (eg, compounds of Formula (A) and (I)-(III), such as any of Compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat bronchopulmonary dysplasia (BPD).

[0502] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat heart disease. These 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 abdominal aortic aneurysm surgery, myocardial injury after PCI, myocardial injury in patients undergoing coronary artery bypass grafting (CABG), minimally invasive mitral valve (MIMV) repair or replacement, adult patients undergoing open heart surgery, chronic heart failure, NYHA class II-IV.

[0503] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat pulmonary diseases. These conditions include, but are not limited to, lung injury during elective lobectomy, lung injury during CABG surgery, and lung transplantation.

[0504] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat liver diseases. These conditions include but are not limited to non-alcoholic steatohepatitis (NASH).

[0505] In other embodiments, the compounds disclosed herein (e.g., compounds of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used to treat kidney disease. These conditions include, but are not limited to, acute kidney injury caused by contrast agents, stage III-IV chronic kidney disease planned for coronary angiography, acute kidney injury in patients undergoing heart valve surgery, non-dialysis dependent chronic kidney disease, chronic kidney disease patients starting dialysis, non-dialysis dependent chronic kidney disease.

[0506] In addition, the compounds disclosed herein (e.g., compounds of any one of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof can be used in combination with additional active ingredients to treat the above-mentioned conditions. The additional compound can be co-administered separately with the compounds disclosed herein (e.g., compounds of any one of formula (A) and (I)-(III), such as any one of compounds 1-44) or pharmaceutically acceptable salts thereof, or included in a pharmaceutical composition according to the present invention together with the additional active ingredient. In an exemplary embodiment, the additional active ingredient is one that is known or found to be effective in treating a condition, disorder or disease mediated by a PHD enzyme or is active against another target associated with a particular condition, disorder or disease, such as an alternative PHD modulator. The combination can be used to increase efficacy (e.g., by including a compound in the combination that enhances the potency or effectiveness of a compound according to the present invention), reduce one or more side effects, or reduce the required dose of a compound according to the present invention.

[0507] The compounds of the present invention are used alone or in combination with one or more other active ingredients to prepare the pharmaceutical compositions of the present invention. The pharmaceutical compositions of the present invention include: (a) an effective amount of a compound disclosed herein (e.g., a compound of any one of formulas (A) and (I)-(III), such as any one of compounds 1-44) or a pharmaceutically acceptable salt, a pharmaceutically acceptable prodrug or a pharmaceutically active metabolite thereof; and (b) a pharmaceutically acceptable excipient.

[0508] "Pharmaceutically acceptable excipient" refers to a non-toxic, biologically tolerable, and otherwise biologically suitable substance for administration to a patient, such as an inert substance, that is added to a pharmacological composition or otherwise used as a vehicle, carrier, or diluent to facilitate administration of a medicament and is compatible therewith. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Suitable excipients may also include antioxidants. Such antioxidants may be used in pharmaceutical compositions or storage media to extend the shelf life of the drug product.

[0509] Drug formulations and routes of administration

[0510] The compounds and compositions of the present invention can be delivered directly or in a pharmaceutical composition or medicine together with a suitable carrier or excipient, as is well known in the art. The therapeutic methods of the present invention may include administering an effective amount of the compounds of the present invention to a subject in need thereof. In a preferred embodiment, the subject is a mammalian subject, and in a most preferred embodiment, the subject is a human subject.

[0511] The effective amount of such compounds, compositions or drugs can be easily determined by routine experiments, and the most effective and convenient route of administration and the most appropriate formulation can also be determined. Various formulations and drug delivery systems are available in the art. See, for example, editor: Gennaro, AR, (1995) "Remington's Pharmaceutical Sciences", supra.

[0512] Suitable routes of administration may for example include oral, rectal, topical, nasal, pulmonary, ocular, intestinal and parenteral administration. The primary routes of parenteral administration include intravenous, intramuscular and subcutaneous administration. Secondary routes of administration include intraperitoneal, intraarterial, intraarticular, intracardiac, intracisternal, intradermal, intralesional, intraocular, intrapleural, intrathecal, intrauterine and intraventricular administration. The physical, chemical and biological properties of the indication to be treated and the drug determine the type of formulation and route of administration to be used, and whether local or systemic administration is preferred.

[0513] The pharmaceutical dosage form of the compounds of the present invention can be provided with quick release, controlled release, sustained release or targeted drug delivery system. Common dosage forms include, for example, solutions and suspensions, (micro) emulsions, ointments, gels and patches, liposomes, tablets, dragees, soft or hard shell capsules, suppositories, ovules, implants, amorphous or crystalline powders, aerosols and freeze-dried formulations. Depending on the route of administration used, special devices may be required to apply or administer the drug, such as, for example, syringes and needles, inhalers, pumps, injection pens, applicators or special flasks. Pharmaceutical dosage forms are generally composed of drugs, excipients and containers / closed systems. One or more excipients (also referred to as inactive ingredients) can be added to the compounds of the present invention to improve or promote the manufacture, stability, administration and safety of the drug, and the means for achieving the desired drug release profile can be provided. Therefore, the type of excipient to be added to the drug may depend on various factors, such as the physical and chemical properties of the drug, the route of administration and the manufacturing procedure. Pharmaceutically acceptable excipients are available in the art and are included in those excipients listed in various pharmacopoeias. See, e.g., the United States Pharmacopoeia (USP), the Japanese Pharmacopoeia (JP), the European Pharmacopoeia (EP), and the British Pharmacopoeia (BP); U.S. Food and Drug Administration (www.fda.gov) Center for Drug Evaluation and Research (CEDR) publications, e.g., Inactive Ingredient Guide (1996); Editors: Ash and Ash, (2002) Handbook of Pharmaceutical Additives, Synapse Information Resources, Inc., Endicott NY; etc.) Pharmaceutical dosage forms of the compounds of the invention may be prepared by any method well known in the art, such as by conventional mixing, sieving, dissolving, melting, granulating, preparing dragees, tableting, suspending, extruding, spray drying, grinding, emulsifying, (nano / micro) encapsulation, embedding, or lyophilizing processes. As described above, the compositions of the invention may include one or more physiologically acceptable inactive ingredients that aid in processing the active molecule into a formulation for pharmaceutical use.

[0515] Suitable formulations depend on the desired route of administration. For example, for intravenous injection, the composition can be formulated in an aqueous solution, if necessary, using a physiologically compatible buffer, including, for example, phosphates, histidine or citrates for adjusting the pH of the formulation, and tonicity agents, such as, for example, sodium chloride or glucose. For transmucosal or nasal administration, semisolid, liquid formulations or patches may be preferred, and may contain a penetration enhancer. Such permeabilizing agents are generally known in the art. For oral administration, the compound can be formulated into liquid or solid dosage forms, and quick-release or controlled-release / slow-release formulations. Suitable dosage forms for oral ingestion by a subject include tablets, pills, dragees, hard and soft shell capsules, liquids, gels, syrups, slurries, suspensions and emulsions. The compound can also be formulated into, for example, rectal compositions (such as suppositories or retention enemas) containing conventional suppository bases (such as cocoa butter or other glycerides).

[0516] Solid oral dosage forms can be obtained using excipients, which may include fillers, disintegrants, binders (dry and wet), dissolution retardants, lubricants, glidants, antiadherents, cation exchange resins, wetting agents, antioxidants, preservatives, colorants and flavorings. These excipients may be synthetic or natural sources. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatin, magnesium carbonate, magnesium lauryl sulfate / sodium, mannitol, polyethylene glycol, polyvinyl pyrrolidone, silicates, silicon dioxide, sodium benzoate, sorbitol, starch, stearic acid or salts, including sugars (i.e. glucose, sucrose, lactose, etc.), talcum powder, tragacanth mucus, vegetable oils (hydrogenated) and waxes. Ethanol and water may be used as granulation aids. In some cases, it is desirable to coat tablets with, for example, a taste masking film, a gastric acid resistant film or a sustained release film. Natural and synthetic polymers, combined with colorants, sugars, and organic solvents or water, are often used to coat tablets to make dragees. When capsules are preferred over tablets, drug powders, suspensions, or solutions can be delivered in compatible hard or soft shell capsules.

[0517] In one embodiment, the compounds of the present invention can be topically applied, such as by skin patches, semisolid or liquid formulations, such as gels, (micro) emulsions, ointments, solutions, (nano / micro) suspensions or foams. The penetration of drugs to the skin and underlying tissues can be regulated, for example, using penetration enhancers; appropriate selection and combination of lipophilic, hydrophilic and amphiphilic excipients, including water, organic solvents, waxes, oils, synthetic and natural polymers, surfactants, emulsifiers; by pH adjustment; and the use of complexing agents. Other techniques (such as iontophoresis) can be used to regulate the skin penetration of the compounds of the present invention. For example, in the case of local delivery with minimal systemic exposure, transdermal or topical administration is preferred.

[0518] For administration by inhalation or nasal administration, the compound used according to the invention is conveniently delivered in the form of a solution, suspension, emulsion or semisolid aerosol from a pressurized package or sprayer, usually using a propellant, for example, a halocarbon derived from methane and ethane, carbon dioxide or any other suitable gas. For topical aerosols, hydrocarbons such as butane, isobutylene and pentane are useful. In the case of pressurized aerosols, a suitable dosage unit can be determined by providing a valve for delivering a metered amount. Capsules and cartridges such as gelatin can be formulated for use in inhalers or insufflators. These usually comprise a powder mixture of the compound and a suitable powder base (such as lactose or starch).

[0519] Compounds and compositions formulated for parenteral administration by injection are usually sterile and can exist in unit dosage form, for example, in ampoule, syringe, injection pen or multi-dose container, the latter usually containing preservative. Composition can adopt the form of suspension, solution or emulsion in oily or aqueous medium, and can contain formulating agent, such as buffer, tonicity agent, viscosity enhancer, surfactant, suspending agent and dispersant, antioxidant, biocompatible polymer, chelating agent and preservative. According to injection site, medium may contain water, synthetic oil or vegetable oil and / or organic cosolvent. In some cases, such as using lyophilized product or concentrate, parenteral formulation will be reconstructed or diluted before application. The long-acting formulation of controlled release or sustained release of the compounds of the present invention can include injectable suspension of nanometer / micron particles or nanometer / micron or non-micronized crystals. In addition to other substrates known in the art, polymers such as poly (lactic acid), poly (glycolic acid) or its copolymer can be used as controlled release / slow release substrate. Other long-acting delivery systems may come in the form of implants and pumps that require an incision.

[0520] 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, for example, sodium hydroxide, for forming ionized compounds; sucrose or sodium chloride, as a tonicity agent; and buffers, such as those containing phosphate or histidine. A cosolvent such as, for example, polyethylene glycol may be added. These aqueous systems are effective in dissolving the compounds of the invention and produce low toxicity when administered systemically. The proportions of the components of the solution system may vary widely without destroying the solubility and toxicity characteristics. In addition, the identity of the components may be altered. For example, low toxicity surfactants such as polysorbates or poloxamers may be used, polyethylene glycol or other cosolvents may also be used, biocompatible polymers such as polyvinyl pyrrolidone may be added, and other sugars and polyols may be used in place of glucose.

[0521] 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 can be based on the effective concentration established in the cell culture assay. For example, the data obtained from animal studies and cell culture assays can be used to determine a dosage range suitable for human subjects. In certain embodiments, the compounds of the present disclosure are formulated for oral administration. An exemplary dose of the compounds of the present disclosure in a pharmaceutical formulation for oral administration is about 0.5 to about 10 mg / kg subject body weight. In certain embodiments, the pharmaceutical formulation includes about 0.7 to about 5.0 mg / kg subject body weight, or alternatively, about 1.0 to about 2.5 mg / kg subject body weight. A typical dosing regimen for oral administration is three times a week, twice a week, once a week, or daily administration of a pharmaceutical formulation for oral administration.

[0522] The effective amount or therapeutically effective amount or dosage of an agent (e.g., a compound of the present invention) refers to the amount of an agent or compound that results in improved symptoms or prolonged 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, for example, by determining LD50 (a dose lethal to 50% of the population) and ED50 (a dose effective for treatment in 50% of the population). The dose ratio of toxic effects to therapeutic effects is the therapeutic index, which can be expressed as the ratio LD50 / ED50. Agents that exhibit a high therapeutic index are preferred.

[0523] An effective amount or therapeutically effective amount is the amount of a compound or pharmaceutical composition that will elicit a biological response or medical response in a tissue, system, animal or human that a researcher, veterinarian, doctor or other clinician is seeking. The dosage specifically falls within a circulating concentration range that includes an ED50 with little toxicity or no toxicity. The dosage can vary within this range depending on the dosage form used and / or the route of administration utilized. In view of the specific circumstances of the subject's condition, the precise formulation, route of administration, dosage and dosage interval should be selected according to methods known in the art.

[0524] Dosage and interval can be adjusted individually to provide a plasma level of the active moiety sufficient to achieve the desired effect; that is, the minimum effective concentration (MEC). The MEC will be different for each compound, but can be estimated from, for example, in vitro data and animal experiments. The dosage required to achieve the MEC will depend on individual characteristics and route of administration. In the case of local administration or selective uptake, the effective local concentration of the drug may be independent of the plasma concentration.

[0525] The amount of compound or composition administered may depend on a variety of factors, including the sex, age, and weight of the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.

[0526] If desired, the compounds and compositions of the present invention may be present in a package or dispenser device containing one or more unit dosage forms containing the active ingredient. Such a package or device may, for example, include 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 may be accompanied by instructions for administration. Compositions comprising the compounds of the present invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of a specified condition.

[0527] These and other embodiments of the present invention will readily occur to those of ordinary skill in the art in view of the disclosure herein and are specifically contemplated.

[0528] Example

[0529] Synthesis of Exemplary Compounds

[0530] Example 1: Preparation of Compound 1

[0531] tert-Butyl 6-chloronicotinate

[0532]

[0533] To a solution of 6-chloronicotinic 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 stirred at reflux for 4 hours and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate=10 / 1) to give tert-butyl 6-chloronicotinate (5.5 g, 81.12% yield) as a yellow solid. LC-MS: m / z=214.0 (M+H) + , retention time 1.83 min (Method A).

[0534] tert-Butyl 6-hydrazinonicotinate

[0535]

[0536] 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 hours 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, 92.76% yield) as a yellow solid. LC-MS: m / z = 210.0 (M + H) +, retention time 1.19 min (Method A).

[0537] (E)-3-(Dimethylamino)-2-(p-tolyl) ethyl acrylate

[0538]

[0539] To a solution of ethyl 2-(p-tolyl)acetate (1.00 g, 5.61 mmol) in N,N-dimethylformamide (10.0 mL) was added N,N-dimethylformamide diethyl acetal (3.34 g, 28.05 mmol, 3.73 mL). The reaction was stirred at 100 ° C for 3 hours and cooled to 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 to give (E)-3-(dimethylamino)-2-(p-tolyl)ethyl acrylate (1.1 g, 4.71 mmol, 84% yield). LC-MS: m / z=234.0[M+H] + , retention time 2.064 min (Method B). The product was pure enough to be used directly in the next step.

[0540] 6-(5-Hydroxy-4-(p-tolyl)-1H-pyrazol-1-yl)nicotinate tert-butyl ester

[0541]

[0542] To a solution of (E)-3-(dimethylamino)-2-(p-tolyl) ethyl acrylate (50.00 mg, 2.14 mmol) and tert-butyl 6-hydrazinonicotinate (448.4 mg, 2.14 mmol) in ethanol (10.0 mL) was added p-toluenesulfonic acid monohydrate (40 mg, 0.21 mmol). The reaction was stirred at 90 ° C for 16 hours and cooled to precipitate a solid. The crude solid was purified by flash chromatography (dichloromethane / ethyl acetate=10 / 1) to give tert-butyl 6-(5-hydroxy-4-(p-tolyl)-1H-pyrazol-1-yl)nicotinate (393 mg, 1.11 mmol, 52% yield) as a yellow solid. LC-MS: m / z=352.0[M+H] + , retention time 6.78 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ12.86(s,1H),8.92(s,1H),8.80-8.15(m,3H),7.78(s,2H),7.17(d,J=8.0Hz,2H),2.29(s,3H),1.58(s,9H).

[0543] Example 2: Preparation of Compound 2

[0544] (E)-2-(4-bromophenyl)-3-(dimethylamino) ethyl acrylate

[0545]

[0546] To a solution of ethyl 2-(4-bromophenyl)acetate (1.01 g, 4.15 mmol) in N,N-dimethylformamide (20.0 mL) was added N,N-dimethylformamide diethyl acetal (2.47 g, 20.7 mmol). The mixture was stirred at 100 ° C for 16 hours and cooled to 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 to give (E)-2-(4-bromophenyl)-3-(dimethylamino) ethyl acrylate (980 mg, 3.32 mmol, 80% yield) as a yellow oil. LC-MS: m / z=298.0[M+H] + , retention time 1.710 min (Method B). The product was pure enough to be used directly in the next step.

[0547] tert-Butyl 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0548]

[0549] To a solution of (E)-2-(4-bromophenyl)-3-(dimethylamino)ethyl acrylate (300.0 mg, 1.01 mmol) and tert-butyl 6-hydrazinonicotinate (210.5 mg, 1.01 mmol) in ethanol (10.0 mL) was added p-toluenesulfonic acid monohydrate (38 mg, 0.2 mmol). The mixture was stirred at 80 ° C for 16 hours and cooled to precipitate a solid. The crude product was purified by flash chromatography (dichloromethane / methanol=100 / 3) to give tert-butyl 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (286 mg, 0.69 mmol, 68% yield) as a yellow solid. LC-MS: m / z=416.0 (M+H) + , retention time 6.76 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.13(br,1H),8.91(s,1H),8.80-8.25(m,3H),7.89(br,2H),7.53(d,J=8.0Hz,2H),1.58(s,9H).

[0550] Example 3: Preparation of Compound 3

[0551] 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0552]

[0553] To a solution of tert-butyl 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (80.00 mg, 0.19 mmol) in dichloromethane (10.0 mL) was added trifluoroacetic acid (0.5 mL). The mixture was stirred at room temperature overnight and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (67 mg, 0.18 mmol, 97% yield) as a yellow solid. LC-MS: m / z=360.0 (M+H) + , retention time 4.995 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.29(br,2H),8.96(s,1H),8.51-8.44(m,3H),7.89(d,J=7.5Hz,2H),7.53(d,J=8.5Hz,2H).

[0554] Example 4: Preparation of Compound 4

[0555] (E)-3-(Dimethylamino)-2-(4-chlorophenyl) ethyl acrylate

[0556]

[0557] To a solution of ethyl 2-(4-chlorophenyl)acetate (5.0 g, 25.25 mmol) in N,N-dimethylformamide (25.0 mL) was added N,N-dimethylformamide diethyl acetal (12.0 g, 101.01 mmol). The mixture was stirred at 100 ° C overnight 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 to give (E)-ethyl 3-(dimethylamino)-2-(4-chlorophenyl)acrylate (3.5 g, 14.04 mmol, 55.60% yield) as a colorless oil. LC-MS: m / z=254.1 [M+H] + , retention time 2.030 min (Method A). The product was pure enough to be used directly in the next step.

[0558] tert-Butyl 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0559]

[0560] To a solution of (E)-3-(dimethylamino)-2-(4-chlorophenyl)ethyl acrylate (0.83 g, 3.95 mmol) and tert-butyl 6-hydrazinonicotinate (1.00 g, 3.95 mmol) in ethanol (10.0 mL) was added p-toluenesulfonic acid monohydrate (150 mg, 0.79 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 tert-butyl 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (800.0 mg, 2.16 mmol, 54.79% yield) as a white solid. LC-MS: m / z=372.1 (M+H) + , retention time 6.645 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ9.04–8.86(m,1H),8.62–8.33(m,3H),7.95(d,J=7.2Hz,2H),7.41(d,J=8.6Hz,2H),1.58(s,9H).

[0561] Example 5: Preparation of Compound 5

[0562] 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0563]

[0564] To a solution of tert-butyl 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (200.00 mg, 0.54 mmol) in dichloromethane (10.0 mL) was added trifluoroacetic acid (5.0 mL). The mixture was stirred at 40 °C for 2 hours and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (74.0 mg, 0.24 mmol, 43.52% yield) as a white solid. LC-MS: m / z=316.0 (M+H) + , retention time 4.718 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.23(s,1H),8.97(s,1H),8.67-8.47(m,3H),8.04-7.97(m,2H),7.41(d,J=3.8Hz,2H).

[0565] Example 6: Preparation of Compound 6

[0566] (E)-3-(Dimethylamino)-2-(4-fluorophenyl) ethyl acrylate

[0567]

[0568] To a solution of ethyl 2-(4-fluorophenyl)acetate (2.0 g, 10.98 mmol) in N,N-dimethylformamide (5.0 mL) was added N,N-dimethylformamide diethyl acetal (8.07 g, 54.8 mmol). The mixture was stirred at 100 ° C overnight 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 to give (E)-3-(dimethylamino)-2-(4-fluorophenyl) ethyl acrylate (2.0 g, 8.45 mmol, 77% yield) as a colorless oil. LC-MS: m / z=238.0 [M+H] + , retention time 1.89 minutes (method B). The product was pure enough to be used directly in the next step

[0569] tert-Butyl 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0570]

[0571] To a solution of (E)-3-(dimethylamino)-2-(4-fluorophenyl)ethyl acrylate (567.0 mg, 2.39 mmol) and tert-butyl 6-hydrazinonicotinate (500.0 mg, 2.39 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (91.2 mg, 0.48 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 tert-butyl 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (350.0 mg, 0.98 mmol, 41% yield) as a white solid. LC-MS: m / z=356.0 (M+H) + , retention time 6.23 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ8.91(s,1H),8.41(s,3H),7.93(s,2H),7.19(s,2H),1.57(s,9H).

[0572] Example 7: Preparation of Compound 7

[0573] 6-(4-(4-Fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0574]

[0575] To a solution of tert-butyl 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (150.0 mg, 0.42 mmol) in dichloromethane (5.0 mL) was added trifluoroacetic acid (2.0 mL). The mixture was stirred at 40 °C for 2 hours and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (103.2 mg, 0.34 mmol, 81% yield) as a white solid. LC-MS: m / z=300.0 (M+H) + , retention time 4.23 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ8.95(s,1H),8.45(s,3H),7.94(s,2H),7.20-7.17(m,2H).

[0576] Example 8: Preparation of Compound 8

[0577] Ethyl 2-(4-cyano-2-methylphenyl)acetate

[0578]

[0579] 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 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, 31.7% yield) as a yellow oil. LC-MS: m / z 204.1 (M+H) + .

[0580] (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino) ethyl acrylate

[0581]

[0582] To a solution of ethyl 2-(4-cyano-2-methylphenyl)acetate (1.0 g, 5.0 mmol) in N,N-dimethylformamide (10.0 mL) was added N,N-dimethylformamide diethyl acetal (2.9 g, 25.0 mmol). The mixture was stirred at 100 ° C overnight 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)-ethyl 2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate (600 mg, 47.2% yield) as a yellow oil. LC-MS: m / z 259.0 (M+H) + .

[0583] tert-Butyl 6-chloronicotinate

[0584]

[0585] To a solution of 6-chloronicotinic acid (5.0 g, 6.37 mmol) and 4-dimethylaminopyridine (0.39 g, 0.64 mmol) in THF (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 (5.5 g, 81.12% yield) as a yellow solid. LC-MS: m / z 214.0 (M+H) + .

[0586] tert-Butyl 6-hydrazinonicotinate

[0587]

[0588] 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 hours and concentrated. 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, 92.76% yield) as a yellow solid. LC-MS: m / z 210.0 (M+H) + .

[0589] tert-Butyl 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0590]

[0591] To a solution of (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)ethyl acrylate (260 mg, 1.0 mmol) and tert-butyl 6-hydrazinonicotinate (200 mg, 1.0 mmol) in ethanol (10.0 mL) was added 4-methylbenzenesulfonic acid (34.4 mg, 0.2 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 tert-butyl 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (190 mg, 50% yield) as a white solid. LC-MS: m / z 377.0 (M+H) + .

[0592] Example 9: Preparation of Compound 9

[0593] 3-Acetoxy-2-(4-cyanophenyl)but-2-enoic acid ethyl ester

[0594]

[0595] In N 2 Atmosphere, LHMDS (28.00mL, 27.75mmol) was added dropwise to a mixture of ethyl 2-(4-cyanophenyl)acetate (3.50g, 18.50mmol) and anhydrous THF (100ml) at -30°C in 20 minutes. The reaction was allowed to warm to 0°C and stirred at 0°C for 30 minutes. The reaction was then cooled to -30°C again and a solution of acetyl chloride (2.18g, 27.75mmol) in THF (8mL) was added dropwise to the solution in 15 minutes. The reaction was slowly warmed to room temperature and stirred for 4 hours. After the reaction was completed by TLC, the mixture was washed with cold NH 4 The solution was quenched with aqueous Cl solution (50 mL) and extracted with EtOAc (3 x 80 ml). The combined organic phases were washed with anhydrous Na 2 SO 4 (30 g) was dried, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EA:Hex=1:50 to 1:5) to give the desired product (1.9 g) as a yellow oil. 1 H NMR δppm (300MHz, CDCl 3 )7.68(dd,J=6.6,1.8Hz,2H),7.41(dd,J=6.6,1.8Hz,2H),4.11-4.18(q,J=6.9Hz,2H),2.25(s,3H),1.88(s,3H),1.20(t,J=6.9Hz,3H).

[0596] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0597]

[0598] 4-Hydrazinobenzoic acid (1.06 g, 6.91 mmol) was added to a mixture of ethyl 2-(4-cyanophenyl)-3-oxobutanoate (0.30 g, 1.38 mmol) in HOAc (24 ml). The mixture was stirred overnight at 100 ° C. After the reaction was completed by TLC, the reaction was quenched with water (20 mL), and a large amount of solids were precipitated. After filtration, the solid was dissolved in TEA (5 equivalents), water (20 mL) and MeOH (20 mL). The solution was extracted with EtOAc (3 x 10 mL), and the aqueous phase was adjusted to pH 3 with a diluted HCl solution. A large amount of solids were precipitated. The solid was collected by filtration and dried to obtain the desired product (128 mg) as a yellow solid. LC-MS (ESI+): m / z 321 (M+H) + ; HPLC purity: 97.3%; 1 H NMR (300 MHz, DMSO-d 6 )δppm13.25(brs,2H),8.95(d,J=1.5Hz,1H),8.56(d,J=8.7Hz,1H),8.41(dd,J=8.7Hz,1H),7.89(d,J=8.1Hz,2H),7.81(d,J=8.1Hz,2H),2.51(s,3H).

[0599] Example 10: Preparation of Compound 10

[0600] tert-Butyl 6-hydrazinonicotinate

[0601]

[0602] To a solution of tert-butyl 6-chloronicotinate (0.20 g, 0.94 mmol) in 1,4-dioxane (10 mL) was added a portion of hydrazine (0.35 g, 5.62 mmol). The mixture was stirred overnight at 80 ° C. After the reaction was completed by TLC analysis, the mixture was concentrated in vacuo. The residue was purified by preparative HPLC to obtain 180 mg of the title compound (180 mg) as an oil. 1 HNMR (300MHz, CDCl 3)δppm 8.71(d,J=1.8Hz,1H),8.01(dd,J=8.1,1.8Hz,1H),6.70(d,J=8.7Hz,1H),6.24(brs,1H),3.90(brs,2H),1.60(s,9H).

[0603] tert-Butyl 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinate

[0604]

[0605] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using tert-butyl 6-hydrazinonicotinate. LC-MS (ESI-): m / z 375 (MH) - HPLC purity: 96.3%; 1 H NMR (300 MHz, CDCl 3 )δppm 13.10(brs,1H),8.88(d,J=2.1Hz,1H),8.42(dd,J=8.7,2.1Hz,1H),7.96(d,J=8.7Hz,1H),7.69(s,4H),2.46(s,3H),1.63(s,9H).

[0606] Example 11: Preparation of Compound 11

[0607] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinate

[0608]

[0609] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using 6-hydrazinonicotinic acid methyl ester. LC-MS: m / z=335 (M+H) + ;1H NMR(300MHz,DMSO-d6)δppm 13.11(s,1H),8.95(d,J=1.6Hz,1H),8.58(d,J=8.8Hz,1H),8.43(dd,J=7.0,2.3Hz,1H) ,7.90(dd,J=17.6,8.3Hz,2H),7.79(dd,J=14.5,8.4Hz,2H),3.90(s,3H),2.48(s,3H).

[0610] Example 12: Preparation of Compound 12

[0611] Di-tert-butyl 2-(4-cyano-2,5-difluorophenyl)malonate

[0612]

[0613] Under nitrogen atmosphere, di-tert-butyl malonate (8.26 g, 31.19 mmol) was added dropwise to a solution of sodium hydride (60%, dispersed in mineral oil) (5.09 g, 127.31 mmol) in anhydrous DMF (100 mL) at 0 ° C over 15 minutes. After the reaction was stirred at 0 ° C for 15 minutes, 2,4,5-trifluorobenzonitrile (5.00 g, 31.83 mmol) was added to the reaction in one portion. The mixture was stirred at 0 ° C for 15 minutes and then stirred at 60 ° C overnight. After the reaction was completed by TLC analysis, the mixture was washed with saturated NH 4 The mixture was quenched with aqueous Cl solution (800 mL) and extracted with EtOAc (200 mL x 3). The combined organic phases were washed with water (100 mL x 2) and washed with anhydrous Na 2 SO 4 (50 g) was dried, filtered and concentrated in vacuo. 12.60 g of the crude title compound was obtained as a yellow oil which was used in the next step without further purification. LC-MS (ESI+): m / z 239 (M-(t-Bu) 2 ) + .

[0614] 2-(4-Cyano-2,5-difluorophenyl)acetic acid

[0615]

[0616] To a solution of di-tert-butyl 2-(4-cyano-2,5-difluorophenyl)malonate (12.30 g, 34.80 mmol) in DCM (35 mL) was added a portion of TFA (35 mL). The reaction was stirred at room temperature overnight. After the reaction was completed by TLC analysis, the mixture was concentrated in vacuo. Toluene (50 mL) was added to the residue, stirred and concentrated to dryness again. 6.07 g of crude title compound as a white solid was obtained, which was used in the next step without further purification. LC-MS (ESI+): m / z 216 (M+H+H 2 O) + .

[0617] Methyl 2-(4-cyano-2,5-difluorophenyl)acetate

[0618]

[0619] To a solution of 2-(4-cyano-2,5-difluorophenyl)acetic acid (4.00 g, 20.29 mmol) in MeOH (160 mL) was added SOCl over 5 min at room temperature. 2 (4.0 mL). The reaction was stirred at 70 ° C for 3 hours. After the reaction was completed by TLC analysis, 80% of the reaction solvent was removed by evaporation. The residue was quenched with ice water (180 mL) and extracted with EtOAc (50 mL x 3). The combined organic phase was washed with anhydrous Na 2 SO 4 (50 g) was dried, filtered and concentrated in vacuo. 4.30 g of the crude title compound was obtained as a yellow oil which was used in the next step without further purification. LC-MS (ESI+): m / z 230 (M+H+H 2 O) + .

[0620] (E)-2-(4-cyano-2,5-difluorophenyl)-3-(dimethylamino)acrylate

[0621]

[0622] The compound was synthesized according to the procedure for preparing ethyl (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-cyano-2,5-difluorophenyl)acetate. LC-MS (ESI+): m / z 267 (M+H) + .

[0623] 6-(4-(4-cyano-2,5-difluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0624]

[0625] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (E)-2-(4-cyano-2,5-difluorophenyl)-3-(dimethylamino)acrylate. LC-MS (ESI-): m / z 341 (MH) - , HPLC purity: 95.4%, 1 H NMR (300 MHz, DMSO-d 6 )δppm 13.41(brs,1H),8.96(d,J=2.1Hz,1H),8.58–8.42(m,3H),8.31(d,J=3.0Hz,1H),7.93(dd,J=10.8,5.4Hz,1H).

[0626] Example 13: Preparation of Compound 13

[0627] Methyl 2-(3-fluoro-4-hydroxyphenyl)acetate

[0628]

[0629] To a mixture of 2-(3-fluoro-4-hydroxyphenyl)acetic acid (2.0 g, 11.76 mmol) in MeOH (20 ml) was added concentrated H 2 SO 4 The reaction mixture was stirred at 65 ° C for 2 hours. After the reaction was completed by TLC, the mixture was quenched with water (40 mL) and extracted with EtOAc (3 x 50 mL). The combined organic phase was dried, filtered and directly concentrated to obtain the title product (2.49 g) as an oil. 1 H NMR (300 MHz, CDCl 3 )δppm7.03(d,J=1.5Hz,1H),6.91(dd,J=3.9,1.5Hz,2H),3.70(s,3H),3.54(s,2H).

[0630] Methyl 2-(3-fluoro-4-(((trifluoromethyl)sulfonyl)oxy)phenyl)acetate

[0631]

[0632] In N 2 To a mixture of methyl 2-(3-fluoro-4-hydroxyphenyl)acetate (2.49 g, 13.50 mmol) in DCM (52 mL) was added trifluoromethanesulfonic anhydride (5.71 g, 20.25 mmol) dropwise at 0 ° C. under atmosphere over 15 minutes. TEA (4.10 g, 40.50 mmol) was added dropwise to the reaction over 10 minutes. The reaction was stirred at 0 ° C. for 4 hours. After the reaction was completed by TLC, the mixture was washed with NaHCO 3 The mixture was quenched with aqueous solution (30 mL) and extracted with DCM (3 x 50 mL). The combined organic phases were washed with anhydrous Na 2 SO 4 (50 g) was dried, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EA:n-Hex=1:20) to give the desired product (2.18 g) as a yellow oil. 1 H NMR (300 MHz, CDCl 3 )δppm 7.30(d,J=8.1Hz,1H),7.24(dd,J=3.3,1.11Hz,1H),7.13(d,J=8.1Hz,1H),3.73(s,3H),3.65(s,2H).

[0633] Methyl 2-(4-cyano-3-fluorophenyl)acetate

[0634]

[0635] In N 2 To a mixture of methyl 2-(3-fluoro-4-(((trifluoromethyl)sulfonyl)oxy)phenyl)acetate (2.18 g, 6.90 mmol) in anhydrous DMF (40 mL) was added Zn(CN) 2 (0.49 g, 4.14 mmol) and Pd(PPh 3 ) 4 (0.80 g, 0.69 mmol). The reaction was stirred at 80 ° C for 4 hours. After the reaction was completed by TLC, the mixture was quenched with water (50 mL) and extracted with EtOAc (3 x 50 mL). The combined organic phase was washed with water (2 x 20 mL) and washed with anhydrous Na 2 SO 4 (50 g) was dried, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EA:Hex=1:40 to 1:8) to give the desired product (1.27 g) as a yellow oil. 1 H NMR (300 MHz, CDCl 3 )δppm7.59(t,J=7.5Hz,1H),7.20(d,J=8.7Hz,2H),3.73(s,3H),3.70(s,2H).

[0636] (E)-2-(4-cyano-3-fluorophenyl)-3-(dimethylamino)acrylate

[0637]

[0638] The compound was synthesized according to the procedure for preparing ethyl (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-cyano-3-fluorophenyl)acetate. LC-MS (ESI+): m / z 249 (M+H) + .

[0639] 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0640]

[0641] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (E)-2-(4-cyano-3-fluorophenyl)-3-(dimethylamino)acrylate. LC-MS (ESI-): m / z 323 (MH) - ; 1 H-NMR (300 MHz, DMSO-d 6 )δppm 13.32(brs,1H),8.77(d,J=1.2Hz,1H),8.55(s,1H),8.34(d,J=8.4Hz,1H),8.26(dd,J =8.4, 2.1Hz, 1H), 7.88 (d, J = 12.3Hz, 1H), 7.78 (d, J = 8.1Hz, 1H), 7.65 (t, J = 7.5Hz, 1H).

[0642] Example 14: Preparation of Compound 14

[0643] 5-Fluoro-6-hydrazinonicotinic acid

[0644]

[0645] To a solution of 6-chloro-5-fluoronicotinic acid (0.46 g, 2.59 mmol) in THF (22 mL) was added N 2 H 4 .H 2 O (0.81 g, 12.95 mmol). After the mixture was stirred at 65 ° C overnight, a large amount of solid precipitated. The suspension was filtered and the solid was slurried in MeOH (3 mL) for 1 hour. After filtration, 440 mg of the desired product was obtained as a white solid. LC-MS (ESI+): m / z 172 (M+H) + .

[0646] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-5-fluoronicotinic acid

[0647]

[0648] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using 5-fluoro-6-hydrazinonicotinic acid and (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate methyl ester. LC-MS (ESI+): m / z 325 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6)δppm8.78(s,1H),8.08(d,J=9.9Hz,1H),7.90(d,J=8.4Hz,2H),7.84(s,1H),7.46(d,J=8.4Hz,2H),7.12(brs,2H).

[0649] Example 15: Preparation of Compound 15

[0650] Methyl 2-(4-chlorophenyl)-3-oxobutanoate

[0651]

[0652] Under nitrogen atmosphere, LHMDS (8.1 mL, 8.11 mmol) was added dropwise to a solution of 2-(4-chlorophenyl) methyl acetate (1.00 g, 5.41 mmol) in anhydrous THF (50 mL) at -40 ° C in 10 minutes. After the resulting mixture was stirred at -40 ° C for 1 hour, 1-(1H-imidazole-1-yl) ethane-1-one (0.89 g, 8.11 mmol) was added in batches within 10 minutes. After addition, the mixture was warmed to room temperature and stirred for 1 hour. After TLC analysis showed that the reaction was complete, the residue was diluted with water (50 mL) and extracted with ethyl acetate (50 mL x 3). The combined organic phase was washed with Na 2 O 4 % 4-nitropropene and 4-nitropropene. The resulting mixture was stirred for 1 hour at -40 ° C for 1 hour. After 3 hours, the mixture was stirred for 2 hours ... 2 SO 4 (30 g) was dried, filtered and concentrated to dryness to give the desired product (1.6 g) as an oil. LC-MS (ESI+): m / z 249 (M+Na) + .

[0653] 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0654]

[0655] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-chlorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 330 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δppm 12.77-13.27(m,2H),8.95(d,J=1.5Hz,1H),8.55(brs,1H),8.41(dd,J=9.0 Hz, J=2.1Hz, 1H), 7.66 (d, J=8.4Hz, 2H), 7.44 (d, J=8.4Hz, 2H), 2.42 (s, 3H).

[0656] Example 16: Preparation of Compound 16

[0657] Methyl 2-(4-(Benzyloxy)-3-methylphenyl)acetate

[0658]

[0659] To a solution of methyl 2-(4-hydroxy-3-methylphenyl)acetate (300 mg, 1.70 mmol) in DMF (10 ml) was added benzyl bromide (292 mg, 1.70 mmol) and Cs 2 CO 3 (1.66 g, 5.10 mmol). The mixture was stirred at room temperature for 1 hour. After the reaction was completed by TLC analysis, the mixture was quenched with water (50 mL) and extracted with EtOAc (50 mL x 3). The combined organic phases were washed with anhydrous Na 2 SO 4 (30 g) was dried, filtered and concentrated in vacuo. The residue was purified by silica gel column chromatography (PE:EtOAc=100:1-50:1) to give 498 mg of the title compound. 1 H NMR (300 MHz, CDCl 3 )δppm 7.29-7.45(m,5H),7.03-7.08(m,2H),6.82(d,J=8.1Hz,1H),5.06(s,2H),3.68(s,3H),3.54(s,2H),2.27(s,3H).

[0660] (Z)-2-(4-(Benzyloxy)-3-methylphenyl)-3-(dimethylamino)acrylate

[0661]

[0662] The compound was synthesized according to the procedure for preparing ethyl (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-(benzyloxy)-3-methylphenyl)acetate. LC-MS (ESI+): m / z 326 (M+H + ).

[0663] 6-(4-(4-(Benzyloxy)-3-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0664]

[0665] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (Z)-2-(4-(benzyloxy)-3-methylphenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 402 (M+H + ).

[0666] 6-(5-Hydroxy-4-(4-hydroxy-3-methylphenyl)-1H-pyrazol-1-yl)nicotinic acid

[0667]

[0668] A solution of 6-(4-(4-(benzyloxy)-3-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (60 mg, 0.12 mmol) and Pd / C (10 mg) in MeOH (6.5 mL) was stirred from a balloon under a hydrogen atmosphere for 7 hours. After TLC analysis showed that the reaction was complete, the suspension was filtered through a diatomaceous earth packing, and the filter cake was washed with MeOH (5 mL). The combined filtrate was concentrated and purified by preparative HPLC to give 9 mg of the title compound as a solid. LC-MS (ESI-): m / z 310 (MH - ), 1 HNMR (300MHz, CD 3 OD)δppm 8.94(s,1H),8.45(d,J=8.7Hz,1H),8.11(d,J=8.7Hz,1H),7.90(s,1H),7.48( s,1H),7.41(d,J=8.7Hz,1H),6.75(s,1H),6.65(d,J=6.9Hz,1H),2.22(s,3H).

[0669] Example 17: Preparation of Compound 17

[0670] (Z)-2-(6-chloropyridin-3-yl)-3-(dimethylamino) ethyl acrylate

[0671]

[0672] The compound was synthesized according to the procedure for preparing ethyl (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate using ethyl 2-(6-chloropyridin-3-yl)acetate. LC-MS (ESI+): m / z 241 (M+H + ), 1 H NMR (300 MHz, CDCl 3)δppm 8.18(d,J=2.4Hz,1H),7.66(s,1H),7.51(dd,J=2.4,8.1Hz,1H),

[0673] 7.26(d,J=8.4Hz,1H),3.64(s,3H),2.75(s,6H).

[0674] 6-(4-(6-chloropyridin-3-yl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0675]

[0676] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using ethyl (Z)-2-(6-chloropyridin-3-yl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 317 (M+H + ), 1 H NMR(300MHz,DMSO-d6)δ13.44(brs,2H),8.94(d,J=16.8Hz,2H),8.67(s,1H), 8.55(s,1H),8.44-8.48(m,1H),8.38(d,J=7.8Hz,1H),7.51(d,J=8.4Hz,1H).

[0677] Example 18: Preparation of Compound 18

[0678] 6-Hydrazino-4-methylnicotinic acid

[0679]

[0680] To a solution of 6-fluoro-4-methylnicotinic acid (0.50 g, 3.23 mmol) in THF (50 ml) was added hydrazine (2.00 g, 32.26 mmol). The reaction was stirred at 66 ° C for 2 hours. After LC-MS analysis showed that the reaction was complete, the mixture was diluted with ethanol (6 mL) and a large amount of solid was precipitated. After filtration, 725 mg of the desired product was obtained. LC-MS (ESI+): m / z 168 (M+H + ), 1 H NMR (300 MHz, DMSO-d 6 )δ8.44(s,1H),7.56(brs,1H),6.43(s,1H),5.90(brs,2H),2.44(s,3H)

[0681] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid

[0682]

[0683] To a solution of 6-hydrazino-4-methylnicotinic acid (0.30 g, 1.80 mmol) in acetic acid (20 ml) was added 2-(4-cyanophenyl)-3-oxobutanoic acid ethyl ester (0.43 g, 1.98 mmol). The reaction was stirred overnight at 100 ° C. After LC-MS analysis showed that the reaction was complete, the mixture was cooled to room temperature and a large amount of solid was precipitated. After filtration, the filter cake was further purified by preparative HPLC to obtain 36 mg of the desired product. LC-MS (ESI+): m / z 335 (M+H + ), 1 H-NMR (300 MHz, DMSO-d 6 )δ12.97-13.21(brs,2H),8.87(s,1H),8.45(brs,1H),8.01(d,J=8.1Hz,2H),7.82(d,J=8.1Hz,2H),2.78(s,3H),2.52(s,3H).

[0684] Example 19: Preparation of Compound 19

[0685] 6-Hydrazino-2-methylnicotinic acid

[0686]

[0687] The compound was synthesized according to the procedure for the preparation of 6-hydrazino-4-methylnicotinic acid using 6-fluoro-2-methylnicotinic acid. 1 H-NMR (300MHz, D 2 O) δ7.63(d,J=8.7Hz,1H),6.55(d,J=8.7Hz,1H),2.37(s,3H).

[0688] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-2-methylnicotinic acid

[0689]

[0690] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid using 6-hydrazino-2-methylnicotinic acid. LC-MS (ESI+): m / z 335 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6)δ12.95(brs,1H),8.28-8.35(m,2H),7.89(d,J=8.7Hz,2H),7.79(d,J=8.7Hz,2H),2.80(s,3H),2.58(s,3H).

[0691] Example 20: Preparation of Compound 20

[0692] Ethyl 2-(5-chloropyridin-2-yl)-3-oxobutanoate

[0693]

[0694] In N 2 Atmosphere, LHMDS (5.0 mL, 5.0 mmol) was added dropwise to a mixture of ethyl 2-(5-chloropyridin-2-yl)acetate (500 mg, 2.5 mmol) in anhydrous THF (15 mL) at -55 ° C in 20 minutes. After the reaction was stirred at 0 ° C for 1 hour, a solution of 1-(1H-imidazole-1-yl)ethane-1-one (412 mg, 3.75 mmol) in THF (15 mL) was added dropwise at -55 ° C in 15 minutes. The reaction was slowly warmed to room temperature and stirred at room temperature for 1 hour. After TLC analysis showed that the reaction was complete, the mixture was washed with NH 4 Cl aqueous solution (50 mL) was quenched and extracted with EtOAc (3 x 20 mL). The combined organic phases were washed with anhydrous Na 2 SO 4 (20 g) was dried, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc:Hex=1:100) to give the title compound (152 mg) as a solid. LC-MS (ESI+): m / z 242 (M+H) + .

[0695] 6-(4-(5-chloropyridin-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0696]

[0697] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using ethyl 2-(5-chloropyridin-2-yl)-3-oxobutanoate. LC-MS (ESI+): m / z 331 (M+H) + ; 1 H NMR (300 MHz, DMSO-d 6)δ13.20(brs,2H),8.95(d,J=1.8Hz,1H),8.53-8.61(m,1H),8.52(s,1H),

[0698] 8.41(dd,J=8.7Hz,J=2.1Hz,1H),8.32(d,J=5.1

[0699] Hz, 1H), 7.87 (dd, J=8.7Hz, J=2.7Hz, 1H), 2.62 (s, 3H).

[0700] Example 21: Preparation of Compound 21

[0701] Methyl 2-(4-bromo-2-methoxyphenyl)acetate

[0702]

[0703] To a solution of 2-(4-bromo-2-methoxyphenyl)acetic acid (2.00 g, 8.16 mmol) in MeOH (35 ml) was added SOCl dropwise over 5 min at room temperature. 2 (6 mL, 82.71 mmol). After addition, the mixture was stirred at 55 °C overnight. After TLC analysis showed that the reaction was complete, the reaction mixture was directly concentrated to dryness. The residue was washed with NaHCO 3 The solution was diluted with aqueous solution (50 mL) and extracted with EtOAc (100 mL x 3). The combined organic phases were washed with Na 2 SO 4 Dry, filter and concentrate to give the desired product as a yellow oil (2.05 g). LC-MS (ESI+): m / z 281 (M+Na) + ; 1 H-NMR (300MHz, CDCl 3 )δ7.02-7.08(m,2H),7.00(s,1H),3.93(s,3H),3.81(s,3H),3.57(s,2H).

[0704] Methyl 2-(4-cyano-2-methoxyphenyl)acetate

[0705]

[0706] Under nitrogen protection, Zn(CN) was added to a solution of methyl 2-(4-bromo-2-methoxyphenyl)acetate (3.02 g, 11.66 mmol) in DMF (20 ml). 2 (2.74 g, 23.32 mmol), Pd 2 (dba) 3(0.11 g, 0.12 mmol) and S-Phos (0.48 g, 1.17 mmol). The reaction was stirred at 110 ° C for 4 hours. After TLC analysis showed that the reaction was complete, the reaction was filtered to remove undissolved solids. The filtrate was directly concentrated to dryness. The residue was purified by flash silica gel chromatography (EtOAc / PE=1 / 50 to 1 / 10) to give the desired product (2.04 g) as a white solid. LC-MS (ESI+): m / z206 (M+H) + ; 1 H-NMR (300MHz, CDCl 3 )δ7.23-7.30(m,3H),3.86(s,3H),3.71(s,3H),3.67(s,2H).

[0707] (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate

[0708]

[0709] A solution of methyl 2-(4-cyano-2-methoxyphenyl)acetate (0.94 g, 4.57 mmol) in DMF-DMA (27.4 g) was stirred at 100° C. overnight. After TLC analysis indicated that the reaction was complete, the mixture was diluted with ice water (80 mL) and extracted with EtOAc (20 mL×3). The combined organic phases were washed with Na 2 SO 4 Dry, filter and concentrate to give the desired product as a brown oil (1.23 g). LC-MS (ESI+): m / z 261 (M+H) + ;

[0710] 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0711]

[0712] To a suspension of (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate (0.32 g, 1.24 mmol) in i-PrOH (6 mL) was added 6-hydrazinonicotinic acid (0.19 g, 1.24 mmol) and HCl (1.25 mL, 1 M, 1.24 mmol). After the reaction was stirred at room temperature for 6 hours, a large amount of solid was precipitated. The suspension was filtered. The filter cake was dissolved in i-PrOH (6 mL), water (1 mL) and DIEA (320 mg). The resulting mixture was stirred at 50 ° C overnight. Diluted HCl solution (1 M, 5 mL) was added to the reaction, and the reaction was continued to stir for another 20 minutes at room temperature. A large amount of solid was precipitated. After filtration, the filter cake was slurried in methanol (5 mL) overnight. After filtration and drying, 185 mg of the title compound was obtained. LC-MS (ESI+): m / z 335 (MH) - ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.5(brs,1H),8.97(s,1H),8.56-8.68(brs,2H),8.45-8.47(m,2H),7.44-7.49(m,2H),3.96(s,3H).

[0713] 6-(4-(4-cyano-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0714]

[0715] Under nitrogen protection, a portion of EtSNa (0.11 g, 1.34 mmol) was added to a solution of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid (0.15 g, 0.45 mmol) in DMF (8 mL). The reaction was stirred at 150 ° C for 48 hours. After TLC analysis showed that the reaction was complete, the mixture was diluted with water (80 mL) and the pH was adjusted to 2 with a dilute HCl solution. A large amount of solid was precipitated. After filtration, the solid was purified by preparative HPLC to give 20 mg of the title compound as a yellow solid. LC-MS (ESI+): m / z 321 (MH) - ; 1 H-NMR (300MHz, CD 3 OD)δ9.10(s,1H),8.30-8.50(m,2H),8.20(m,1H),7.82(m,1H),7.12(m,1H),7.02(s,1H)

[0716] Example 22: Preparation of Compound 22

[0717] 1-Chloro-2-methoxy-4-methylbenzene

[0718]

[0719] To a solution of 2-chloro-5-methylphenol (1 g, 6.99 mmol) in DMF (4 mL) was added K 2 CO 3 The product was stirred for about 6 hours at room temperature. After TLC analysis showed that the reaction was completed, the reaction was diluted with water (20mL) and extracted with EtOAc (20mL x3). The organic phase merged was washed with water (10mL), and Na 2 SO 4 Dry and concentrate to dryness to obtain 1.05 g of the desired crude product. GC-MS (EI+): 156; 1 H-NMR (300MHz, CDCl 3 )δ7.22(d,J=8.1Hz,1H),6.69-6.74(m,2H),3.88(s,3H),2.33(s,3H).

[0720] 4-(Bromomethyl)-1-chloro-2-methoxybenzene

[0721]

[0722] 1-Chloro-2-methoxy-4-methylbenzene (20.98 g, 0.13 mol) was added to CCl 4 NBS (26.24 g, 0.15 mol) and BPO (1.62 g, 6.7 mmol) were added to the solution in (200 mL). The resulting mixture was stirred overnight at 80 ° C. After the reaction was completed by TLC analysis, the mixture was filtered and diluted with diluted HCl solution (20 mL, 1 N), and extracted with DCM (200 mL x3). The combined organic phase was saturated with NaHCO 3 Solution (40 mL) was washed with Na 2 SO 4 (60 g) was dried, filtered and concentrated to give 41.44 g of a crude product. The crude product was further purified by silica gel column chromatography (PE / EtOAc=100 / :1 to 60 / 1) to give 32.6 g of the title compound. 1 H-NMR (300MHz, CDCl 3)δ7.32(d,J=8.1Hz,1H),6.90-6.96(m,2H),4.54(s,2H),3.93(s,3H).

[0723] 2-(4-Chloro-3-methoxyphenyl)acetonitrile

[0724]

[0725] To 4-(bromomethyl)-1-chloro-2-methoxybenzene (10 g, 42.7 mmol) in EtOH (50 mL) and H 2 O (10mL) was added with a portion of NaCN (3.14 g, 64.05 mmol). The resulting mixture was stirred at 80 ° C overnight. After the reaction was completed by TLC analysis, the reaction was diluted with water (100 mL) and extracted with DCM (200 mL x 2). The combined organic phase was washed with saturated NaHCO 3 The solution (50 mL) and brine (50 mL), dried and concentrated to give 7.58 g of a crude product. The crude product was purified by column chromatography (PE:EtOAc=60 / 1 to 15 / 1) to give 5.58 g of the title compound. 1 H-NMR (300MHz, CDCl 3 )δ7.34(d,J=8.1Hz,1H),6.83-6.88(m,2H),3.91(s,3H),3.73(s,2H).

[0726] 2-(4-Chloro-3-methoxyphenyl)acetic acid

[0727]

[0728] To 2-(4-chloro-3-methoxyphenyl)acetonitrile (5.58g, 31mmol) in ethanol (120mL) and water (40mL) solution, add a portion of KOH (8.68g, 155mol). The reaction was stirred for about 3 hours under reflux. After TLC analysis showed that the reaction was completed, the concentrated reaction was to remove most of ethanol. The pH of the residue was adjusted to 3 with diluted HCl solution, then extracted with EtOAc (200mL x 3). The organic phase merged was washed with water (30mL), dried and concentrated to dryness, thereby obtaining 7.29g crude product. 1 H-NMR (300 MHz, DMSO-d 6 )δ12.40(brs,1H),7.34(d,J=7.8Hz,1H),7.05(d,J=1.5Hz,1H),6.84(d,J=7.8,1.5Hz,1H),3.92(s,3H),3.72(s,2H).

[0729] Methyl 2-(4-chloro-3-methoxyphenyl)acetate

[0730]

[0731] To a solution of 2-(4-chloro-3-methoxyphenyl)acetic acid (7.29 g, 36.45 mmol) in methanol (100 mL) was added concentrated H 2 SO 4 4.68g title compound was obtained by purifying the residue by column chromatography (PE / EtOAc=60 / 1 to 20 / 1). 1 H-NMR (300MHz, CDCl 3 )δ7.29(d,J=8.1Hz,1H),6.86(d,J=1.5Hz,1H),6.80(d,J=7.8,1.5Hz,1H),3.90(s,3H),3.72(s,3H),3.60(s,2H).

[0732] (Z)-2-(4-chloro-3-methoxyphenyl)-3-(dimethylamino)acrylate

[0733]

[0734] The compound was synthesized according to the procedure for preparing methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-chloro-3-methoxyphenyl)acetate. LC-MS (ESI+): m / z 270 (M+H) + ;

[0735] 6-(4-(4-chloro-3-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0736]

[0737] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (Z)-2-(4-chloro-3-methoxyphenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 346 (M+H) + ;

[0738] 6-(4-(4-chloro-3-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0739]

[0740] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using 6-(4-(4-chloro-3-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid. LC-MS (ESI+): m / z 332 (M+H) + ; 1 H-NMR (300MHz, CDCl 3 )δδ13.30(brs,1H),10.07(s,1H),8.95(s,1H),8.42-8.52(m,2H),8.33(s,1H),7.70(s,1H),7.25(s,2H).

[0741] Example 23: Preparation of Compound 23

[0742] Methyl 2-(4-bromo-3-fluorophenyl)acetate

[0743]

[0744] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-bromo-2-methoxyphenyl)acetate using 2-(4-bromo-3-fluoro)acetic acid. 1 H-NMR (300MHz, CDCl 3 )δ7.49(t,J=8.1Hz,1H),7.08(d,J=9.0Hz,1H),6.95(d,J=8.1Hz,1H),3.71(s,3H),3.59(s,2H).

[0745] Methyl 2-(4-cyano-3-fluorophenyl)acetate

[0746]

[0747] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-methoxyphenyl)acetate using methyl 2-(4-bromo-3-fluorophenyl)acetate. 1 H-NMR (300MHz, CDCl 3 )δ7.59(t,J=8.1Hz,1H),7.19(d,J=8.7Hz,2H),3.73(s,3H),3.70(s,2H).

[0748] Methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate

[0749]

[0750] Under nitrogen protection, LiHMDS (2.9mmol, 2.9mL) was added dropwise to a solution of 2-(4-cyano-3-fluorophenyl)acetic acid methyl ester (363mg, 1.88mmol) in THF (15mL) at -78°C within 10 minutes. After the resulting mixture was stirred at -78°C for 30 minutes, a solution of 1-acetylimidazole (249mg, 2.25mmol) in THF (5mL) was added dropwise to the reaction within 10 minutes. The resulting mixture was continued to be stirred at -78°C for 1 hour. After TLC analysis showed that the reaction was complete, the reaction was heated with saturated NH 4 The reaction mixture was quenched with Cl solution (20 mL) and extracted with EtOAc (30 mL x 2). The combined organic phases were dried and concentrated to dryness to give 410 mg of the crude title compound, which was used in the next step without further purification. LC-MS (ESI-): m / z 234 (MH) – ;

[0751] 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0752]

[0753] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid using methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 337 (M+H + ), 1 H NMR (300 MHz, DMSO-d 6 )δ8.99(d,J=1.5Hz,1H),8.34(m,1H),8.26(d,J=8.1Hz,2H),7.86(d,J=11.7Hz,1H),7.68(m,1H),7.57(m,1H),2.44(s,3H).

[0754] Example 24: Preparation of Compound 24

[0755] 3-Methyl-4-vinylbenzonitrile

[0756]

[0757] Under nitrogen protection, to a solution of 4-bromo-3-methylbenzonitrile (5 g, 25.51 mmol) and potassium vinyl trifluoroborate (6.84 g, 51.02 mmol) in THF / water (250 mL / 25 mL) was added Cs 2 CO 3 (33.3 g, 102.04 mmol) and Pd(dppf)Cl 2 (1.8g, 2.55mmol). The resulting mixture was stirred at 70 ° C overnight. After TLC analysis showed that the reaction was complete, the reaction was quenched with water (100mL) and extracted with EtOAc (50mL x 2). The combined organic phase was dried and concentrated. The residue was purified by column chromatography (PE: EtOAc = 20 / 1 to 10 / 1) to obtain 3.2g of the title compound as a light yellow oil. GC-MS (EI +): m / z 143 (M + ).

[0758] 2-(4-Cyano-2-methylphenyl)acetic acid

[0759]

[0760] To a solution of 3-methyl-4-vinylbenzonitrile (3.2 g, 22.38 mmol) in DME (200 mL) and water (48 mL) was added 1 portion of 2 (0.57 g, 2.24 mmol). Oxone (27.52 g, 44.76 mmol) was added to the resulting mixture in portions at room temperature over 10 minutes. After addition, the reaction was stirred at room temperature overnight. After TLC analysis indicated that the reaction was complete, the suspension was filtered to remove undissolved solids, and the filtrate was concentrated in vacuo to remove most of the organic solvent. The residue was washed with saturated Na 2 S 2 O 3 In 4% paraformaldehyde (2-nitropropene) 4-nitropropene (4 ...

[0761] Methyl 2-(4-cyano-2-methylphenyl)acetate

[0762]

[0763] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-bromo-2-methoxyphenyl)acetate using 2-(4-bromo-2-methyl)acetic acid. 1 H NMR (300 MHz, CDCl 3)δ7.45-7.48(m,2H),7.30(d,J=8.1Hz,1H),3.71(s,3H),3.69(s,2H),2.34(s,3H).

[0764] Methyl 2-(4-cyano-2-methylphenyl)-3-oxobutanoate

[0765]

[0766] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-cyano-2-methylphenyl)acetate. LC-MS (ESI+): m / z 232 (M+H) + ;

[0767] 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0768]

[0769] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-cyano-2-methylphenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 332 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ12.91(brs,2H),8.95(s,1H),8.52(d,J=8.7Hz,1H),8.39(d,J=8.7Hz,1H),8.23(s,1H),7.80(s,1H),6.78(s,1H).

[0770] Example 25: Preparation of Compound 25

[0771] Methyl 2-(4-bromo-2-fluorophenyl)acetate

[0772]

[0773] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-chloro-3-methoxyphenyl)acetate using 2-(4-bromo-2-fluorophenyl)acetic acid. 1 H NMR (300 MHz, CDCl 3 )δ7.24-7.27(m,2H),7.14(t,J=8.4Hz,1H),3.71(s,3H),3.63(s,2H).

[0774] Methyl 2-(4-cyano-2-fluorophenyl)acetate

[0775]

[0776] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)acetate using methyl 2-(4-bromo-2-fluorophenyl)acetate. 1 H NMR (300 MHz, CDCl 3 )δ7.86(dd,J=8.7,1.2Hz,1H),7.70(d,J=8.1Hz,1H),7.60(t,J=7.5Hz,1H),3.87(s,2H),3.64(s,3H).

[0777] Methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate

[0778]

[0779] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-cyano-2-fluorophenyl)acetate. LC-MS (ESI+): m / z 258 (M+Na) + .

[0780] 6-(4-(4-cyano-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0781]

[0782] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 339 (M+H + ), 1 HNMR (300 MHz, DMSO-d 6 )δ13.22(brs,1H),8.94(d,J=1.8Hz,1H),8.59(d,J=9.3Hz,1H),8.38(d,J=9.3Hz,1H),7.69-7.94(m,3H),2.23(s,3H).

[0783] Example 26: Preparation of Compound 26

[0784] Methyl 2-(4-chloro-3-fluorophenyl)acetate

[0785]

[0786] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-chloro-3-methoxyphenyl)acetate using 2-(4-chloro-3-fluorophenyl)acetic acid. 1 H NMR (300 MHz, CDCl 3 )δ7.34(t,J=7.8Hz,1H),7.10(dd,J=8.1,1.8Hz,1H),7.01(d,J=8.1Hz,1H),3.71(s,3H),3.6(s,2H).

[0787] Methyl 2-(4-chloro-3-fluorophenyl)-3-oxobutanoate

[0788]

[0789] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate using methyl 2-(4-chloro-3-fluorophenyl)acetate. LC-MS (ESI+): m / z 267 (M+Na) + ;

[0790] 6-(4-(4-chloro-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0791]

[0792] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-chloro-3-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 348 (M+Na) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.30(brs,1H),13.01(brs,1H),8.95(s,1H),8.56(brs,1H),8.41(dd, J=9.0,2.1Hz,1H),7.74(d,J=11.7Hz,1H),7.55-7.58(m,2H),2.47(s,3H).

[0793] Example 27: Preparation of Compound 27

[0794] 4-Amino-6-hydrazinonicotinic acid

[0795]

[0796] The compound was synthesized according to the procedure for preparing 6-hydrazino-4-methylnicotinic acid using 4-amino-6-fluoronicotinic acid. LC-MS (ESI+): m / z 169 (M+H) + ;

[0797] 4-Amino-6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0798]

[0799] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using 4-amino-6-hydrazinonicotinic acid. LC-MS (ESI+): m / z 336 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ8.63(s,1H),8.08(s,2H),7.88(d,J=8.4Hz,2H),7.76(d,J=8.4Hz,2H),7.37(brs,1H),2.39(s,3H).

[0800] Example 28: Preparation of Compound 28

[0801] 6-(4-(4-chloro-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0802]

[0803] The compound was synthesized according to the procedure used to prepare 6-(4-(4-chloro-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid. LC-MS (ESI+): m / z 348 (M+Na) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.36(brs,1H),12.82(brs,1H),8.95(d,J=1.5Hz,1H),8.58(brs,1H),8.40(m,1H),7.48-7.56(m,2H),7.33(m,1H),2.21(s,3H).

[0804] Example 29: Preparation of Compound 29

[0805] 2-Methyl-4-vinylbenzonitrile

[0806]

[0807] The compound was synthesized according to the procedure for the preparation of 3-methyl-4-vinylbenzonitrile using 4-bromo-2-methylbenzonitrile. 1 H NMR (300 MHz, CDCl 3 )δ7.55(d,J=7.8Hz,1H),7.26-7.32(m,2H),6.70(m,1H),5.85(d,J=17.4Hz,1H),5.42(d,J=10.8Hz,1),2.54(s,3H).

[0808] Methyl 2-(4-cyano-3-methylphenyl)acetate

[0809]

[0810] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-methylphenyl)acetate using 2-methyl-4-vinylbenzonitrile. 1 H NMR (300 MHz, CDCl 3 )δ7.56(d,J=7.8Hz,1H),7.18-7.25(m,2H),3.71(s,3H),3.64(s,2H),2.54(s,3H).

[0811] Methyl 2-(4-cyano-3-methylphenyl)-3-oxobutanoate

[0812]

[0813] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-cyano-3-methylphenyl)acetate. LC-MS (ESI-): m / z 230 (MH) - ;

[0814] 6-(4-(4-cyano-3-methylphenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0815]

[0816] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid using methyl 2-(4-cyano-3-methylphenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 333 (MH) – ; 1 H-NMR (300MHz, CD 3OD)δ8.96(s,1H),8.40(d,J=8.7,2.1Hz,1H),8.39(d,J=8.7Hz,1H),7.71(s,1H),7.58-7.66(m,2H),2.55(s,3H),2.45(s,3H).

[0817] Example 30: Preparation of Compound 30

[0818] Methyl 2-(4-bromo-2-chlorophenyl)acetate

[0819]

[0820] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-bromo-2-fluorophenyl)acetate using 2-(4-bromo-2-fluorophenyl)acetic acid. 1 H-NMR (300MHz, CDCl 3 )δ7.55(d,J=1.8Hz,1H),7.37(dd,J=8.1,1.8Hz,1H),7.16(d,J=1.8Hz,1H),3.71(s,3H),3.67(s,2H).

[0821] Methyl 2-(2-chloro-4-cyanophenyl)acetate

[0822]

[0823] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-fluorophenyl)acetate using methyl 2-(4-bromo-2-chlorophenyl)acetate. GC-MS (EI+): m / z 209.

[0824] (Z)-2-(2-chloro-4-cyanophenyl)-3-(dimethylamino)acrylate methyl ester

[0825]

[0826] The compound was synthesized according to the procedure for preparing methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using methyl 2-(2-chloro-4-cyanophenyl)acetate. LC-MS (ESI+): m / z 265 (M+H) +

[0827] 6-(4-(2-chloro-4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0828]

[0829] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (Z)-2-(2-chloro-4-cyanophenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 341 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.42(brs,2H),8.97(s,1H),8.44-8.60(m,3H),8.28(d,J=8.1Hz,1H),8.06(s,1H),7.81(d,J=8.1Hz,1H).

[0830] Example 31: Preparation of Compound 31

[0831] 3,5-Dimethyl-4-vinylbenzonitrile

[0832]

[0833] The compound was synthesized according to the procedure for the preparation of 3-methyl-4-vinylbenzonitrile using 4-bromo-3,5-dimethylbenzonitrile. 1 H-NMR (300MHz, CDCl 3 )δ7.32(s,2H),6.64(m,1H),5.54(dd,J=11.7,1.5Hz,1H),5.32(dd,J=17.7,1.5Hz,1H),2.32(s,6H).

[0834] Methyl 2-(4-cyano-2,6-dimethylphenyl)acetate

[0835]

[0836] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-methylphenyl)acetate using 3,5-dimethyl-4-vinylbenzonitrile. 1 H-NMR (300MHz, CDCl 3 )δ7.33(s,2H),3.72(s,2H),3.70(s,3H),2.35(s,6H).

[0837] (Z)-2-(4-cyano-2,6-dimethylphenyl)-3-(dimethylamino)acrylate

[0838]

[0839] The compound was synthesized according to the procedure for preparing methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-cyano-2,6-dimethylphenyl)acetate. 1 H-NMR (300MHz, CDCl 3 )δ7.62(s,1H),7.28(s,2H),3.59(s,3H),2.64(s,6H),2.20(s,6H).

[0840] 6-(4-(4-cyano-2,6-dimethylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0841]

[0842] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (Z)-2-(4-cyano-2,6-dimethylphenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 335 (M+H) + ; 1 H-NMR (300MHz, CD 3 OD)δ8.03(s,1H),8.52(m,2H),7.75(s,1H),7.40-7.47(m,2H),2.30(s,6H).

[0843] Example 32: Preparation of Compound 32

[0844] Methyl 2-(4-methoxy-3-methylphenyl)acetate

[0845]

[0846] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-bromo-2-methoxyphenyl)acetate using 2-(4-methoxy-3-methylphenyl)acetic acid. 1 H-NMR (300MHz, CD 3 OD)δ7.05-7.07(m,2H),6.77(d,J=7.8Hz,1H),3.81(1,3H),3.76(s,3H),3.54(s,2H),2.20(s,3H).

[0847] Methyl 2-(4-methoxy-3-methylphenyl)-3-oxobutanoate

[0848]

[0849] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-methoxy-3-methylphenyl)acetate. LC-MS (ESI+): m / z 259 (M+Na) + ;

[0850] 6-(5-Hydroxy-4-(4-methoxy-3-methylphenyl)-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0851]

[0852] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-methoxy-3-methylphenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 340 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.30(brs,1H),12.35(brs,1H),8.93(s,1H),8.58(s,1H),8.39(d,J=6.9Hz,1H ),7.34-7.36(m,2H),6.96(d,J=9.0Hz,1H),3.80(s,3H),2.35(s,3H),2.18(s,3H).

[0853] Example 33: Preparation of Compound 33

[0854] Diethyl 2-(2-bromo-4-cyanophenyl)malonate

[0855]

[0856] Under nitrogen protection, NaH (600mg, 15mmol) was added to a solution of diethyl malonate (2g, 12.5mmol) in DMF in batches at 0°C within 5 minutes. After the reaction was stirred at 0°C for 30 minutes, 3-bromo-4-fluorobenzonitrile (2.08g, 10mmol) was added to the reaction in one portion, and the reaction was stirred at 80°C for 1.5 hours. After TLC analysis showed that the reaction was complete, the reaction was quenched with water (20mL), and the pH was adjusted to 5 with a diluted HCl solution. The resulting mixture was then extracted with EtOAc (50mL x 2). The combined organic phases were washed with water (20mL) and brine (20mL), dried and concentrated to dryness. The residue was purified by column chromatography (PE / EtOAc=20 / 1 to 10 / 1) to obtain 1.8g of the title compound. LC-MS (ESI+): m / z 340,342 (M+H)+ . 1 H-NMR (300MHz, CDCl 3 )δ7.90(s,2H),7.64(s,2H),5.24(s,1H),4.21-4.32(m,4H),1.29(t,J=7.2Hz,6H).

[0857] Ethyl 2-(2-bromo-4-cyanophenyl)acetate

[0858]

[0859] To 2-(2-bromo-4-cyanophenyl) diethyl malonate (1.8g, 5.3mmol) in DMSO (20mL) and water (96mg, 5.3mmol) solution, add a portion of LiCl (339mg, 8mmol). The resulting mixture is stirred overnight at 120°C. After TLC analysis shows that the reaction is completed, the reaction is quenched by water (20mL), pH is adjusted to 6 with diluted HCl solution, and extracted with EtOAc (20mL x 2). The combined organic phase is washed with water (20mL) and brine (20mL), dried and concentrated. The residue is purified by column chromatography (PE / EtOAc=20 / 1 to 10 / 1) to obtain 780mg title compound. 1 H-NMR (300MHz, CDCl 3 )δ7.87(s,1H),7.59(d,J=7.8Hz,1H),7.42(d,J=7.8Hz,1H),4.20(q,J=6.9Hz,2H),3.84(s,2H),1.27(t,J=6.9Hz,3H)

[0860] Ethyl 2-(4-cyano-2-vinylphenyl)acetate

[0861]

[0862] The compound was synthesized according to the procedure for the preparation of 3-methyl-4-vinylbenzonitrile using ethyl 2-(2-bromo-4-cyanophenyl)acetate. 1 H NMR (300 MHz, CDCl 3 )δ7.7(s,1H),7.52(d,J=7.8Hz,1H),7.33(d,J=7.8Hz,1H),6.89(m,1H),5.70(d,J=17.4 Hz, 1H), 5.42 (d, J = 10.8Hz, 1), 4.15 (q, J = 7.2Hz, 1H), 3.73 (s, 3H), 1.24 (q, J = 7.2Hz, 1H).

[0863] Ethyl 2-(4-cyano-2-ethylphenyl)acetate

[0864]

[0865] Pd / C (56 mg) is added to a solution of 2-(4-cyano-2-vinylphenyl) methyl acetate (560 mg, 2.6 mmol) in methanol (10 mL). The suspension is stirred for about 1 hour from a balloon under a hydrogen atmosphere. After TLC analysis shows that the reaction has been completed, the suspension is filtered through diatomite packaging, and the filter cake is washed with methanol (10 mL). The filtrate is concentrated to dryness, thereby obtaining 540 mg crude title compound. 1 H-NMR (300MHz, CDCl 3 )δ7.61(s,1H),7.49(d,J=8.1Hz,1H),7.31(d,J=8.1Hz,1H),4.16(q,J=7.2Hz,2H),3.69(s,2H),2.68(q,J=7.5Hz,2H),1.21-1.28(m,6H).

[0866] (Z)-2-(4-cyano-2-ethylphenyl)-3-(dimethylamino) ethyl acrylate

[0867]

[0868] The compound was synthesized according to the procedure for the preparation of methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using ethyl 2-(4-cyano-2-ethylphenyl)acetate. 1 H-NMR (300MHz, CDCl 3 )δ7.60(s,1H),7.49(s,1H),7.41(d,J=8.1Hz,1H),7.23(m,1H),4.12(q,J=6.9Hz,2H),3.60(s,3H),2.67(s,6H),1.14(t,J=6.9Hz,3H)

[0869] 6-(4-(4-cyano-2-ethylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0870]

[0871] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using ethyl (Z)-2-(4-cyano-2-ethylphenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 335 (M+H) + ;1 H-NMR (300 MHz, DMSO-d 6 )δ8.97(s,1H),8.52(m,2H),8.12(s,1H),7.75(s,1H),7.60-7.69(m,2H),2.78(q,J=7.5Hz,2H),1.17(q,J=7.5Hz,3H)

[0872] Example 34: Preparation of Compound 34

[0873] 5-Hydrazinopyrazine-2-carboxylic acid

[0874]

[0875] The compound was synthesized according to the procedure for preparing 6-hydrazino-4-methylnicotinic acid using 5-fluoropyrazine-2-carboxylic acid. LC-MS (ESI+): m / z 155 (M+H) + ;

[0876] 5-(4-(4-Cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)pyrazine-2-carboxylic acid

[0877]

[0878] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using 5-hydrazinopyrazine-2-carboxylic acid. LC-MS (ESI+): m / z 322 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ9.80(s,1H),9.04(s,1H),7.91(d,J=8.4Hz,2H),7.83(d,J=8.4Hz,2H),2.51(s,3H).

[0879] Example 35: Preparation of Compound 35

[0880] 2-(2-methoxypyridin-4-yl)acetic acid ethyl ester

[0881]

[0882] Under nitrogen, lithium diisopropylamide (16.0 mL, 32.0 mmol, 2.0 M in n-heptane) was added to a solution of 2-methoxy-4-methylpyridine (2.0 g, 16.2 mmol) in anhydrous tetrahydrofuran (50.0 mL) at -78 ° C. The mixture was stirred at -78 ° C for 10 minutes and diethyl carbonate (3.78 g, 32.0 mmol) was added. The mixture was allowed to warm to room temperature and stirred for 2.0 hours. The reaction was quenched with water and extracted twice with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash chromatography (petroleum ether / ethyl acetate=10 / 1) to obtain ethyl 2-(2-methoxypyridin-4-yl)acetate (2.0 g, 10.2 mmol, 63.4% yield) as a yellow oil. LC-MS: m / z=196.1 (M+H) + , retention time 1.97 min (Method A).

[0883] (E)-3-(Dimethylamino)-2-(2-methoxypyridin-4-yl)ethyl acrylate

[0884]

[0885] 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 12.0 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 under reduced pressure. The crude product was purified by flash chromatography (dichloromethane / methanol=98 / 2) to obtain (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)ethyl acrylate (1.1 g, 4.4 mmol, 44% yield) as a colorless oil. LC-MS: m / z=251.0[M+H] + , retention time 1.68 min (Method B).

[0886] tert-Butyl 6-(5-hydroxy-4-(2-methoxypyridin-4-yl)-1H-pyrazol-1-yl)nicotinate

[0887]

[0888] To a solution of (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)ethyl acrylate (0.25 g, 1.0 mmol) and tert-butyl 6-hydrazinonicotinate (0.21 g, 1.0 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (19 mg, 0.1 mmol). The mixture was stirred at reflux temperature for 12.0 hours and cooled to precipitate a solid. The solid was filtered, washed with ethanol, and dried to give tert-butyl 6-(5-hydroxy-4-(2-methoxypyridin-4-yl)-1H-pyrazol-1-yl)nicotinate (210 mg, 0.57 mmol, 57% yield) as a white solid. LC-MS: m / z=369.0 (M+H) + , retention time 4.38 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.53(m,1H),8.91(s,1H),8.39-8.67(m,2H),8.04-8.06(d,J=5.0Hz,1H),7.39-7.50(m,2H),3.86(m,3H),1.58(s,9H).

[0889] Example 36: Preparation of Compound 36

[0890] 6-Hydrazinonicotinate Ethyl Ester

[0891]

[0892] To a solution of ethyl 6-chloronicotinate (1.0 g, 5.40 mmol) in tetrahydrofuran (6.0 mL) was added hydrazine hydrate (300 mg, 5.94 mmol, 85% in water). The mixture was stirred at reflux temperature overnight. 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 crude product (600 mg, 3.31 mmol, 61.4% yield) was obtained as a yellow oil. LC-MS: m / z=182.1 (M+H) + , retention time 0.37 min (Method A). The product was pure enough to be used directly in the next step.

[0893] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0894]

[0895] To a solution of (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (300 mg, 1.30 mmol) and ethyl 6-hydrazinonicotinate (235 mg, 1.30 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (25 mg, 0.13 mmol). The reaction was stirred overnight at 90 ° C and cooled to precipitate a solid. The solid was filtered, washed with ethanol, and dried to obtain ethyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (51 mg, 0.15 mmol, 11.7% yield) as a yellow solid. LC-MS: m / z=335.1 [M+H] + , retention time 5.05 min (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.58(s,1H),8.97(s,1H),8.70-8.47(m,3H),8.15(s,2H),7.79-7.77(m,2H),4.39-4.34(m,2H),1.37-1.22(m,3H).

[0896] Example 37: Preparation of Compound 37

[0897] Isopropyl 6-chloronicotinate

[0898]

[0899] To a solution of 6-chloronicotinic acid (3.0 g, 19.2 mmol) in dichloromethane (50.0 mL) was added carbonyl diimidazole (3.42 g, 21.1 mmol) at room temperature. The mixture was stirred for 1.0 hours, and isopropanol (3.78 g, 32.0 mmol) was added. Dichloromethane was removed under vacuum. A catalytic amount of sodium isopropoxide (164 mg, 2.0 mmol) was added, and the mixture was heated at 90 ° C for 1.0 hours. The solution was concentrated under vacuum. The resulting residue was slurried with water and extracted with ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, and concentrated to give 6-chloronicotinic acid isopropyl ester (3.4 g, 17.1 mmol, 89% yield) as a yellow solid. LC-MS: m / z=200.0[M+H] + , retention time 2.04 minutes (method A). The product was pure enough to be used directly in the next step

[0900] 6-Hydrazinonicotinate Isopropyl

[0901]

[0902] To a solution of isopropyl 6-chloronicotinate (1.0 g, 5.03 mmol) in ethanol (10.0 mL) was added hydrazine hydrate (1.0 g, 20.1 mmol, 85% in water). The mixture was stirred at 80 ° C overnight. 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 isopropyl 6-hydrazinonicotinate (800 mg, 4.1 mmol, 81.6% yield) as a yellow solid. LC-MS: m / z=196.0 (M+H) + , retention time 0.39 min (Method A).

[0903] Isopropyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0904]

[0905] To a solution of (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (300 mg, 1.30 mmol) and 6-hydrazinonicotinic acid isopropyl ester (254 mg, 1.30 mmol) in ethanol (5.0 mL) was added p-toluenesulfonic acid monohydrate (25 mg, 0.13 mmol). The reaction was stirred overnight at 90 ° C and cooled to precipitate a solid. The solid was filtered, washed with ethanol, and dried to obtain 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate isopropyl ester (53 mg, 0.15 mmol, 11.7% yield) as a yellow solid. LC-MS: m / z=349.0[M+H] + , retention time 5.44 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.55(s,1H),8.95(s,1H),8.69(s,1H),8.53-8.44(m,2H),8.14-8.12(m,2H),8.79-8.77(m,2H),5.21-5.15(m,1H),1.36-1.34(m,6H).

[0906] Example 38: Preparation of Compound 38

[0907] tert-Butyl 6-chloronicotinate

[0908]

[0909] To a solution of 6-chloronicotinic 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 4.0 hours and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate=10 / 1) to give tert-butyl 6-chloronicotinate (5.5 g, 81.12% yield) as a yellow solid. LC-MS: m / z=214.0 (M+H) + , retention time 1.83 min (Method A).

[0910] tert-Butyl 6-hydrazinonicotinate

[0911]

[0912] 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 2.0 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 tert-butyl 6-hydrazinonicotinate (5.0 g, 92.76% yield) as a yellow solid. LC-MS: m / z=210.0 (M+H) + , retention time 1.19 min (Method A).

[0913] Methyl 2-(4-cyanophenyl)acetate

[0914]

[0915] To a mixture of 2-(4-cyanophenyl)acetic acid (5.0 g, 31.0 mmol) in methanol (10.0 mL) was added hydrochloric acid (20.0 mL, 3.0 M) in methanol at 0 ° C. The mixture was stirred at 70 ° C for 3.0 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. LC-MS: m / z=176.0 [M+H] + , retention time 1.54 min (Method A).

[0916] (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate

[0917]

[0918] 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 16.0 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)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (5.20 g, 25.4 mmol, 89% yield) as a yellow solid. LC-MS: m / z=231.0 [M+H] + , retention time 1.70 minutes (method A). The product was pure enough to be used directly in the next step

[0919] tert-Butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate

[0920]

[0921] To a solution of (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate (2.0 g, 8.70 mmol) and tert-butyl 6-hydrazinonicotinate (1.82 g, 8.70 mmol) in ethanol (20.0 mL) was added p-toluenesulfonic acid monohydrate (171 mg, 0.9 mmol). The reaction was stirred at 90 ° C for 16.0 hours and cooled to precipitate a solid. The solid was filtered, washed with ethanol, and dried to obtain tert-butyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (2.3 g, 6.35 mmol, 73% yield) as a yellow solid. LC-MS: m / z=363.1 (M+H) + , retention time 5.82 minutes (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.58(s,1H),8.92(s,1H),8.69(s,1H),8.51(s,1H),8.44-8.41(m,1H),8.15-8.13(m,2H),7.81-7.78(m,2H),1.58(s,9H).

[0922] Example 39: Preparation of Compound 39

[0923] Methyl 2-(4-chloro-2-methoxyphenyl)acetate

[0924]

[0925] The compound was synthesized according to the procedure for preparing methyl 2-(4-bromo-2-methoxyphenyl)acetate using 2-(4-chloro-2-methoxyphenyl)acetic acid. LC-MS (ESI+): m / z 237 (M+Na) + ; 1 H-NMR (300MHz, CDCl 3 )δ7.10(d,J=8.1Hz,1H),6.85-6.92(m,2H),3.86(s,3H),3.71(s,3H),3.67(s,2H).

[0926] (Z)-2-(4-chloro-2-methoxyphenyl)-3-(dimethylamino)acrylate

[0927]

[0928] The compound was synthesized according to the procedure for preparing methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-chloro-2-methoxyphenyl)acetate. LC-MS (ESI+): m / z 270 (M+H) + .

[0929] 6-(4-(4-chloro-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0930]

[0931] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using methyl (Z)-2-(4-chloro-2-methoxyphenyl)-3-(dimethylamino)acrylate. LC-MS (ESI+): m / z 346 (M+H) + ;

[0932] 6-(4-(4-chloro-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0933]

[0934] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using 6-(4-(4-chloro-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid. LC-MS (ESI+): m / z 332 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6)δ12.91(brs,2H),8.95(s,1H),8.52(d,J=8.7Hz,1H),8.39(d,J=8.7Hz,1H),8.23(s,1H),7.80(s,1H),6.78(s,2H).

[0935] Example 40: Preparation of Compound 40

[0936] Diethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)malonate

[0937]

[0938] Under nitrogen protection, a solution of ethyl 3-(ethylperoxy)-3-oxopropanoate (10.20 g, 63.50 mmol) in DMF (100 ml) was added to Cs 2 CO 3 (45.50g, 139.70mmol). After the reaction was stirred at 70°C for 10 minutes, 4-fluoro-3-(trifluoromethyl)benzonitrile (12.00g, 63.50mmol) was added to the reaction. The resulting mixture was stirred at 70°C for 2 hours. After TLC showed that the reaction was complete, the reaction was quenched with water (300mL) and extracted with EtOAc (500mL×2). The combined organic phase was washed with water (50mL) and washed with Na 2 SO 4 Drying, filtration and concentration gave the desired product as an oil (20.8 g). 1 H-NMR (300MHz, CDCl 3 )δ7.99(s,1H),7.86-7.92(m,2H),5.11(s,1H),4.20-4.33(m,4H),1.28(t,J=7.2Hz,6H).

[0939] Ethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)acetate

[0940]

[0941] A solution of diethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)malonate (15.00 g, 45.60 mmol) in DMSO (150 ml) was added to LiCl (2.90 g, 68.40 mmol) and water (0.80 mL, 45.6 mmol). The mixture was stirred overnight at 120 ° C. After TLC showed that the reaction was complete, the reaction was quenched with water (300 mL) and extracted with EtOAc (150 mL × 3). The combined organic phase was washed with water (100 mL) and purified by Na 2 SO4 Dried, filtered and concentrated.The residue was purified by silica gel chromatography (EA / PE=1 / 25) to give the desired product (8.28 g) as a white solid. 1 H-NMR (300MHz, CDCl 3 )δ7.97(d,J=1.8Hz,1H),7.83(d,J=7.8Hz,1H),7.57(d,J=7.8Hz,1H),4.18(q,J=7.2Hz,2H),3.88(s,2H),1.26(t,J=7.2Hz,3H).

[0942] (Z)-2-(4-cyano-2-(trifluoromethyl)phenyl)-3-(dimethylamino) ethyl acrylate

[0943]

[0944] A solution of ethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)acetate (4.00 g, 15.56 mmol) and DME-DMA (15 mL) in a sealed tube was stirred overnight at 150 °C. After TLC indicated that the reaction was complete, the mixture was diluted with water (100 mL) and extracted with EtOAc (150 mL x 3). The combined organic phases were washed with NaOH and 4% paraffin. 2 SO 4 Dried, filtered and concentrated.The residue was purified by silica gel chromatography (EA / PE=1 / 9) to give the desired product (3.2 g) as a brown oil. 1 H-NMR (300MHz, CDCl 3 )δ7.94(s,1H),7.75(d,J=7.8Hz,1H),7.59(s,1H),7.46(d,J=7.8Hz,1H),3.95-4.08(m,2H),2.66(brs,6H),1.10(t,J=7.2Hz,3H).

[0945] 6-(4-(4-cyano-2-(trifluoromethyl)phenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid

[0946]

[0947] To a solution of (Z)-2-(4-cyano-2-(trifluoromethyl)phenyl)-3-(dimethylamino)ethyl acrylate (1.00 g, 3.20 mmol) in i-PrOH (38 mL) was added 6-hydrazinonicotinic acid (0.59 g, 0.38 mmol) and diluted HCl solution (38 mL, 1 M). The reaction was stirred at room temperature for 24 hours. After LCMS analysis showed that the reaction was complete, DIEA (15 equivalents) was added, and the mixture was stirred at room temperature for 2 days. After the pH of the mixture was adjusted to 3, a large amount of solids was precipitated. After filtration and slurry purification, 90 mg of the desired product was obtained as a solid. HPLC purity was 93.1%. LC-MS (ESI+): m / z 375 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ8.98(s,1H),8.47-8.57(m,2H),8.34(s,1H),8.17(d,J=8.4Hz,1H),7.97-8.02(m,2H).

[0948] Example 41: Preparation of Compound 41

[0949] Ethyl 3-oxo-2-(pyridin-4-yl)butanoate

[0950]

[0951] The compound was synthesized according to the procedure for preparing methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate using ethyl 3-oxo-2-(pyridin-4-yl)butanoate. LC-MS (ESI+): m / z 208 (M+H) +

[0952] 6-(5-Hydroxy-3-methyl-4-(pyridin-4-yl)-1H-pyrazol-1-yl)nicotinic acid

[0953]

[0954] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using ethyl 3-oxo-2-(pyridin-4-yl)butanoate. LC-MS (ESI+): m / z 297 (M+H) + ; 1 H-NMR (300MHz, CD 3 OD)δ9.00(s,1H),8.24-8.33(m,4H),7.84(d,J=6.0Hz,2H),2.44(s,3H)

[0955] 4-(1-(5-carboxypyridin-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-4-yl)pyridine 1-oxide

[0956]

[0957] To a solution of 6-(5-hydroxy-3-methyl-4-(pyridin-4-yl)-1H-pyrazol-1-yl)nicotinic acid (190 mg, 0.64 mmol) in DMF (10 mL) was added a portion of m-CPBA (166 mg, 0.96 mmol) at room temperature. The resulting mixture was stirred at room temperature for about 2 hours. After HPLC analysis indicated that the reaction was complete, the reaction mixture was directly purified by preparative HPLC to give 50 mg of the title compound. LC-MS (ESI+): m / z 313 (M+H) + ; 1 H-NMR (300MHz, CD 3 OD) δ9.03(d,J=2.1Hz,1H),8.61(d,J=6.3Hz,1H),8.45(m,1H),8.07(d,J=8.7Hz,1H),7.92(d,J=6.3Hz,1H),2.07(s,3H).

[0958] Example 42: Preparation of Compound 42

[0959] Methyl 2-(3-bromo-4-chlorophenyl)acetate

[0960]

[0961] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-bromo-2-methoxyphenyl)acetate using 2-(3-bromo-4-chlorophenyl)acetic acid. 1 H-NMR (300MHz, CDCl 3 )δ7.55(d,J=1.8Hz,1H),7.40(d,J=8.1Hz,1H),7.17(dd,J=8.1,1.8Hz,1H),3.71(s,3H),3.58(s,2H).

[0962] Methyl 2-(4-chloro-3-cyanophenyl)acetate

[0963]

[0964] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-fluorophenyl)acetate using methyl 2-(3-bromo-4-chlorophenyl)acetate. 1 H-NMR (300MHz, CDCl 3)δ7.61(s,1H),7.48(s,2H),3.73(s,3H),3.64(s,2H).

[0965] Methyl 2-(4-chloro-3-cyanophenyl)-3-oxobutanoate

[0966]

[0967] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-chloro-3-cyanophenyl)acetate. The crude product was used directly in the next step without further treatment.

[0968] 6-(4-(4-chloro-3-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[0969]

[0970] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using methyl 2-(4-chloro-3-cyanophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 355 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ8.95(s,1H),8.57(d,J=6.9Hz,1H),8.43(m,1H),8.22(s,1H),8.03(d,J=9.0Hz,1H),7.75(d,J=9.0Hz,1H),1.91(s,3H).

[0971] Example 43: Preparation of Compound 43

[0972] 2-(6-Oxo-1,6-dihydropyridin-3-yl)acetic acid:

[0973]

[0974] A mixture of 2-(6-chloropyridin-3-yl)acetic acid (1.0 g, 6.4 mmol) in glacial acetic acid (13.0 mL) and water (3.0 mL) was stirred in a sealed tube at 150 ° C for 3.0 days. The solution was cooled and evaporated to give a residue. The residue was re-evaporated from toluene to obtain 2-(6-oxo-1,6-dihydropyridin-3-yl)acetic acid (1.1 g, crude) as a brown solid. LC-MS: m / z=154.1 [M+H] + , retention time 1.63 min (Method A). The crude product was used in the next step.

[0975] Methyl 2-(6-oxo-1,6-dihydropyridin-3-yl)acetate:

[0976]

[0977] To a solution of 2-(6-oxo-1,6-dihydropyridin-3-yl)acetic acid (1.1 g, crude) in methanol (20.0 mL) was added concentrated sulfuric acid (0.5 mL). The mixture was stirred at room temperature for 15 hours and concentrated. The residue was distributed between ethyl acetate and saturated sodium bicarbonate solution. The organic layer was washed with brine, dried over sodium sulfate, and concentrated under reduced pressure to give methyl 2-(6-oxo-1,6-dihydropyridin-3-yl)acetate (800 mg, 4.79 mmol, 74.8% yield) as a white solid. LC-MS: m / z=168.1[M+H] + , retention time 1.35 min (Method A).

[0978] Methyl 2-(6-methoxypyridin-3-yl)acetate

[0979]

[0980] To a solution of 2-(6-oxo-1,6-dihydropyridin-3-yl)methyl acetate (450 mg, 2.69 mmol) and cesium carbonate (1.05 g, 3.23 mmol) in anhydrous tetrahydrofuran (20.0 mL) was added iodomethane (580 mg, 4.20 mmol). The mixture was stirred at room temperature for 18 hours. The 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=3 / 1) to give a mixture of 2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)methyl acetate and 2-(6-methoxypyridin-3-yl)methyl acetate as a yellow oil (400 mg, 2.21 mmol, 82.1% yield). LC-MS: m / z=182.1 (M+H) + , retention time 1.33 min (Method A).

[0981] (E)-3-(Dimethylamino)-2-(6-methoxypyridin-3-yl) methyl acrylate

[0982]

[0983] To a solution of 2-(1-methyl-6-oxo-1,6-dihydropyridine-3-yl)methyl acetate and 2-(6-methoxypyridine-3-yl)methyl acetate (400mg, 2.21mmol) in N,N-dimethylformamide (5.0mL) was added N,N-dimethylformamide diethyl acetal (1.63g, 11.1mmol). The mixture was stirred at 100 ° C overnight 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 obtain a mixture of (E)-3-(dimethylamino)-2-(1-methyl-6-oxo-1,6-dihydropyridine-3-yl)methyl acrylate and (E)-3-(dimethylamino)-2-(6-methoxypyridine-3-yl)methyl acrylate (400mg, 1.69mmol, 76.7% yield) as a yellow oil. LC-MS: m / z = 237.1 [M+H] + , retention time 1.08 min, 1.51 min (Method B). The mixture was used for the next step.

[0984] 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinate tert-butyl

[0985]

[0986] A mixture of (E)-3-(dimethylamino)-2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl) methyl acrylate and (E)-3-(dimethylamino)-2-(6-methoxypyridin-3-yl) methyl acrylate in ethanol (5.0 mL) (350 mg, 1.48 mmol) was stirred at 90 ° C for 16.0 hours in a sealed tube and cooled. The insoluble solid was filtered and the filtrate was concentrated to dryness. The two isomers were separated as white solids by reverse preparative HPLC. 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl) tert-butyl nicotinate (76 mg, 0.21 mmol, 27.9% yield). LC-MS: m / z=369.0[M+H] + , retention time 2.29 minutes (Method A). 1 HNMR (500 MHz, DMSO-d 6 )δ12.97(br,1H),8.91(d,J=1.5Hz,1H),8.70(s,1H),8.47-8.39(m,2H),8.19-8.14(m,1H),6.85-6.83(m,1H),3.85(s,3H),1.58(s,9H).

[0987] 6-(5-Hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinic acid

[0988]

[0989] To a solution of tert-butyl 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinate (76 mg, 0.21 mmol) in dichloromethane (6.0 mL) was added trifluoroacetic acid (3.0 mL). The mixture was stirred at room temperature overnight and concentrated. The residue was triturated with ethyl acetate and filtered to give 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinic acid (5.4 mg, 0.017 mmol, 8.2% yield) as a white solid. LC-MS: m / z=313.0 [M+H] + , retention time 3.73 min (Method A). 1 HNMR (400 MHz, DMSO-d 6 )δ13.47(br,1H),13.06(br,1H),8.96(s,1H),8.73-8.44(m,4H),8.21-8.18(m,1H),6.84(d,J=8.8Hz,1H),3.85(s,3H).

[0990] Example 44: Preparation of Compound 44

[0991] Diethyl 2-(3-chloro-4-cyanophenyl)malonate

[0992]

[0993] The compound was synthesized according to the procedure for the preparation of diethyl 2-(2-bromo-4-cyanophenyl)malonate using 2-chloro-4-fluorobenzonitrile. 1 H-NMR (300MHz, CDCl 3 )δ7.68(d,J=8.1Hz,1H),7.61(d,J=1.2Hz,1H),7.45(dd,J=8.1,1.2Hz,1H),4.63(s,1H),4.30(q,J=6.9Hz,4H),1.26(t,J=6.9Hz,6H).

[0994] Ethyl 2-(3-chloro-4-cyanophenyl)acetate

[0995]

[0996] The compound was synthesized according to the procedure for the preparation of ethyl 2-(2-bromo-4-cyanophenyl)acetate using diethyl 2-(3-chloro-4-cyanophenyl)malonate.1 H-NMR (300MHz, CDCl 3 )δ7.63(d,J=8.1Hz,1H),7.47(d,J=1.2Hz,1H),7.27(dd,J=8.1,1.2Hz,1H),4.22(q,J=6.9Hz,2H),3.66(s,2H),1.26(t,J=6.9Hz,3H).

[0997] (E)-3-Acetoxy-2-(3-chloro-4-cyanophenyl)but-2-enoic acid ethyl ester

[0998]

[0999] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using ethyl 2-(3-chloro-4-cyanophenyl)acetate. LC-MS (ESI+): m / z 330 (M+Na) + .

[1000] 6-(4-(3-chloro-4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid

[1001]

[1002] The compound was synthesized according to the procedure for preparing 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using (E)-3-acetoxy-2-(3-chloro-4-cyanophenyl)but-2-enoic acid ethyl ester. LC-MS (ESI+): m / z 355 (M+H) + ; 1 H-NMR (300 MHz, DMSO-d 6 )δ13.35(brs,2H),8.95(s,1H),8.57(d,J=9.0Hz,1H),8.42(d,J=9.0,1.8Hz ,1H),8.14(s,1H),7.92(d,J=8.4Hz,1H),7.82(d,J=8.4Hz,1H),2.53(s,3H).

[1003] Example 45: In vitro assay demonstrating PHD inhibition

[1004] The half-maximal inhibitory concentration (IC) of the selected compounds of the present invention was determined. 50 )value.

[1005] The half-maximal inhibitory concentration (IC) of PHD inhibitors against full-length human prolyl-4-hydroxylase domain (PHD) enzymes (PHD1, PHD2, and PHD3) was determined using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay. 50 ) value. The TR-FRET assay was developed based on the following: the hydroxylated HIF-1α peptide binds specifically to a complex formed by VHL, EloB, and EloC (VBC) to generate a fluorescent signal. The terbium (Tb)-donor (monoclonal antibody anti-6His-Tb-cryptate gold) and D2 receptor (streptavidin [SA]-D2) of TR-FRET are linked to the VBC complex and HIF-1α peptide, respectively. The VBC complex binds specifically to the HIF-1α peptide when hydroxylated, resulting in energy transfer from the TR-FRET donor to the acceptor ( Figure 1 ).

[1006] Materials and methods

[1007] Unless otherwise stated, all chemicals and materials were of standard laboratory grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).

[1008] Reagents

[1009] TR-FRET Reagents

[1010] Monoclonal antibodies anti-6His-Tb-cryptate gold (Catalog No. 61HI2TLA) and streptavidin (SA)-D2 (Catalog No. 610SADLA) were purchased from CisBio International (Bedford, MA, USA).

[1011] An N-terminally biotinylated HIF-1α C35 synthetic peptide representing amino acids 547 to 581 and including the PHD2 hydroxylation site at proline 564 was purchased from California Peptide Research (Salt Lake City, UT, USA).

[1012] Recombinant protein

[1013] VBC Complex

[1014] 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 referred to as 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 a His-VBC complex by affinity chromatography on a nickel-nitrilotriacetic acid (Ni-NTA) column. Purity (~80%) was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

[1015] PhD1

[1016] Recombinant human PHD1 protein (Catalog No. 81064, Lot No. 24717001) was purchased from Active Motif (Carlsbad, CA, USA). PHD1 was expressed in a baculovirus expression system as a full-length protein with an N-terminal FLAG tag (molecular weight 44.9 kDa) (NCBI accession No. NP_542770.2). Purity was assessed by SDS-PAGE (>90%).

[1017] PHD2

[1018] The full-length human PHD2 enzyme was produced using the baculovirus-infected insect cell (BIIC) expression system from Beryllium (Bedford, MA, USA). The PHD2 construct contains amino acids 1 to 426 of PHD2 (UniProt Knowledge Base [UniProtKB] / Swiss-Prot Accession No. Q9GZT9.1), as well as a His tag and a tobacco etch virus (TEV) protease cleavage site at the N-terminus. 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% pure.

[1019] PHD3

[1020] Recombinant human PHD3 protein (molecular weight 31.1 kDa) was purchased from Active Motif (Carlsbad, CA, USA). It was expressed in E. coli as a full-length protein with an N-terminal 6-His tag (NCBI accession number NP_071356.1) (Catalog number 81033, batch number 24417001). Purity was assessed by SDS-PAGE and found to be >75%.

[1021] PHD inhibitors.

[1022] Small molecule PHD inhibitors were synthesized and their identities confirmed as described herein.

[1023] TR-FRET Assay Procedure

[1024] PHD inhibitor compounds were pre-incubated with PHD enzyme in a 10 μL reaction volume in a white 384-well Optiplate plate (Cat. No. 6007290, Perkin Elmer, Waltham, MA, USA). To this end, 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, which was prepared as a 4X concentrate in dilution buffer containing PHD enzyme (60 nM PHD1, 20 nM PHD2, 140 nM PHD3), 40 μM ferrous ammonium sulfate (FAS), 4 mM sodium ascorbate (Na). The plate was incubated at room temperature for 30 minutes without rotation.

[1025] Five microliters of VBC / anti-6His-Tb-cryptate gold mixture, prepared as a 4X concentrate in dilution buffer containing 20 nM His-VBC, 1.32 nM monoclonal antibody anti-6His-Tb-cryptate gold, were then added. This step was immediately followed by the addition of 5 μL of HIF-1αC35 substrate mixture, prepared as a 4X concentrate in dilution buffer containing 120 nM biotinylated HIF-1αC35, 132 nM SA-D2, 4 μM 2-oxoglutarate (2-OG), to a final reaction volume of 20 μL.

[1026] The final assay reaction contained 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-1α C35, 5 nM His-VBC, 0.33 nM monoclonal antibody anti-6His-Tb-cryptate gold, 33 nM SA-D2, and PHD enzyme (15 nM PHD1, 5 nM PHD2, or 35 nM PHD3) and diluted compounds.

[1027] To measure the IC of PHD inhibitor compounds 50 The reaction was incubated at room temperature for 10 minutes and then read on a PerkinElmer EnVision (Waltham, MA, USA) with an excitation wavelength of 340 nm and an emission wavelength of 615 nm and 665 nm. The data represent the quotient of the signal intensity at 665 nm and 615 nm, automatically calculated by Envision Manager software (PerkinElmer, Waltham, MA, USA). IC was determined using a four-parameter curve fit using GraphPad Prism 7.0 (GraphPad, La Jolla, CA, USA). 50 Values ​​(mean, standard deviation, standard error of the mean, geometric mean and 95% confidence interval) representing compound concentrations plotted against the calculated ratio at 665 nm and 615 nm. TR-FRET assays were performed in triplicate at each compound concentration and the assays were repeated three times independently.

[1028] Based on Cheng Prussoff equation by IC 50 Calculate Kis:

[1029] Ki=IC50 / (1+[2-OG] / Km)

[1030] The final concentration of 2-OG in the PHD1 and PHD2 assays was 1 uM. The Km for 2-OG for PHD1 was determined to be 12.7 nM, while the Km for 2-OG for PHD2 was determined to be 22.6 nM.

[1031] Exemplary compounds

[1032]

[1033]

[1034]

[1035]

[1036]

[1037]

[1038] From the above description, one skilled in the art can easily ascertain the essential characteristics of this invention, and without departing from the spirit and scope of the invention, can make various changes and modifications of the invention to adapt it to various usages and conditions.

[1039] All references, patents, or applications cited in this application, whether U.S. or foreign, are incorporated herein by reference as if written in their entirety. In the event of any inconsistency, the actual disclosure herein controls.

Claims

1. A compound of formula A: or a pharmaceutically acceptable salt thereof, wherein: Ar 1 is phenyl or a six-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl is optionally substituted with halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl or C 1-3 Alkoxy substitution; R 2 H or C 1-3 alkyl; Ar 2 is a six-membered nitrogen-containing heteroaryl group, which is optionally substituted by halogen, OH, amine or C 1-3 Alkyl substitution; R 4 is hydrogen or C 1-4 Alkyl; and The formula (A) does not include the following compounds:

2. The compound according to claim 1, wherein Ar 1 for in X, Y and Z are independently CH or N, wherein N is optionally oxidized; Each R 1 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; and m is 1, 2, 3 or 4.

3. The compound according to claim 1 or 2, wherein Ar 1 For at least one R 1 Substituted phenyl, wherein R1 is CN or halogen.

4. The compound according to claim 3, wherein Ar 1 By one or two R 1 A group is substituted, the group being independently selected from C optionally substituted by one or more halogens 1-3 Alkyl, halogen, CN or OH.

5. The compound according to claim 1 or 2, wherein Ar 1 is a pyridyl N-oxide or is optionally substituted by at least one R 1 Substituted pyridyl, wherein R1 is C 1-3 Alkoxy or halogen.

6. A compound according to any one of claims 1 to 5, wherein Ar 2 for in A and B are independently CH or N, wherein N is optionally oxidized; Each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; and n is 0, 1 or 2.

7. A compound according to any one of claims 1 to 6, wherein Ar 2 is a group which is pyridyl or pyrazinyl, and wherein the group is unsubstituted or comprises a substituent which is halogen, C 1-3 Alkyl or OH.

8. A compound according to any one of claims 1 to 7, wherein R 2 It is H or CH3.

9. A compound according to any one of claims 1 to 8, wherein R 4 For H.

10. A compound according to any one of claims 1 to 8, wherein R 4 C 1-4 alkyl.

11. The compound according to claim 1, having a structure according to formula (I): or a pharmaceutically acceptable salt thereof, wherein: X, Y, Z, A and B are independently CH or N, wherein N is optionally oxidized; m is 1, 2, 3 or 4; n is 0, 1 or 2; Each R 1 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; R 2 is hydrogen or C 1-3 alkyl; Each R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 Alkyl; and R 4 is hydrogen or C 1-4 alkyl.

12. The compound according to claim 11, which has the structure of formula (Ia): or a pharmaceutically acceptable salt thereof.

13. The compound according to claim 11, which has the structure of formula (Ib): or a pharmaceutically acceptable salt thereof.

14. The compound according to claim 11, which has the structure of formula (Ic): or a pharmaceutically acceptable salt thereof.

15. The compound according to claim 1, having a structure according to formula (II): or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3 or 4; n is 0, 1 or 2; Each time R 1 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; R 2 is hydrogen or C 1-3 alkyl; Each time R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine or C 1-3 Alkyl; and R 4 is hydrogen or C 1-4 alkyl.

16. The compound according to claim 15, which has the structure of formula (IIa): or a pharmaceutically acceptable salt thereof.

17. The compound according to claim 15, which has the structure of formula (IIb): or a pharmaceutically acceptable salt thereof.

18. The compound according to claim 15, which has the structure of formula (IIc): or a pharmaceutically acceptable salt thereof.

19. The compound according to claim 15, which has the structure of formula (IId): or a pharmaceutically acceptable salt thereof.

20. The compound according to claim 15, which has the structure of formula (IIe): or a pharmaceutically acceptable salt thereof.

21. The compound according to claim 15, which has the structure of formula (IIf): or a pharmaceutically acceptable salt thereof.

22. The compound according to claim 15, which has the structure of formula (IIg): or a pharmaceutically acceptable salt thereof.

23. The compound according to claim 1, having a structure according to Formula III: or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3 or 4; n is 0, 1 or 2; Each time R 1 independently selected from the group consisting of hydrogen, halogen, OH, and C optionally substituted with one or more halogens. 1-3 alkyl; R 2 is hydrogen or C 1-3 alkyl; Each time R 3 independently selected from the group consisting of hydrogen, halogen, OH, amine and C 1-3 alkyl; R 4 is hydrogen or C 1-4 alkyl; R 5 is CN or halogen.

24. The compound according to claim 23, which has the structure of formula (IIIa): or a pharmaceutically acceptable salt thereof.

25. The compound according to claim 23, which has the structure of formula (IIIb): or a pharmaceutically acceptable salt thereof.

26. The compound according to claim 23, which has the structure of formula (IIIc): or a pharmaceutically acceptable salt thereof.

27. The compound according to claim 23, which has the structure of formula (IIId): or a pharmaceutically acceptable salt thereof.

28. The compound according to claim 23, which has the structure of formula (IIIe): or a pharmaceutically acceptable salt thereof.

29. The compound according to claim 23, which has the structure of formula (IIIf): or a pharmaceutically acceptable salt thereof.

30. The compound according to claim 23, which has the structure of Formula III(g): or a pharmaceutically acceptable salt thereof.

31. The compound of claim 11, 12 or 14, wherein X is CH.

32. The compound of claim 11, 12 or 14, wherein X is N.

33. The compound of claim 32, wherein N is optionally oxidized.

34. The compound of claim 11, 12 or 14, wherein Y is CH.

35. The compound of claim 11, 12 or 14, wherein Y is N.

36. The compound of claim 11, 12 or 14, wherein Z is CH.

37. The compound of claim 11, 12 or 14, wherein Z is N.

38. The compound of claim 11, 12 or 14, wherein A is CH.

39. The compound of claim 11, 12 or 14, wherein A is N.

40. The compound of claim 11, 12 or 14, wherein B is CH.

41. The compound of claim 11, 12 or 14, wherein B is N.

42. A compound according to any one of claims 11 to 30, wherein m is 1.

43. A compound according to any one of claims 11 to 30, wherein m is 2.

44. A compound according to any one of claims 11 to 30, wherein m is 3.

45. A compound according to any one of claims 11 to 22, wherein m is 4.

46. ​​A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein n is 0.

47. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein n is 1.

48. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein n is 2.

49. A compound according to any one of claims 11 to 30, wherein R 1 For hydrogen.

50. A compound according to any one of claims 11 to 30, wherein R 1 It is a halogen.

51. The compound according to claim 50, wherein R 1 For F.

52. The compound according to claim 50, wherein R 1 For Cl.

53. The compound according to claim 50, wherein R 1 For Br.

54. A compound according to any one of claims 11 to 30, wherein R 1 For CN.

55. A compound according to any one of claims 11 to 30, wherein R 1 For OH.

56. A compound according to any one of claims 11 to 30, wherein R 1 is C optionally substituted by one or more halogens 1-3 alkyl.

57. The compound according to claim 56, wherein R 1 C 1-3 alkyl.

58. The compound according to claim 56, wherein R 1 It is methyl.

59. The compound according to claim 56, wherein R 1 It is ethyl.

60. The compound according to claim 56, wherein R 1 It is CF3.

61. A compound according to any one of claims 11 to 30, wherein R 1 C 1-3 Alkoxy.

62. The compound according to claim 61, wherein R 1 It is a methoxy group.

63. according to the compound described in any one of claim 11 to 18 and 23 to 26, wherein R 2 For hydrogen.

64. according to the compound described in any one of claim 11 to 18 and 23 to 26, wherein R 2 C 1-3 alkyl.

65. The compound according to claim 64, wherein R 2 It is methyl.

66. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein R 3 For hydrogen.

67. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein R 3 It is a halogen.

68. The compound according to claim 56, wherein R 3 For F.

69. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein R 3 For OH.

70. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein R 3 For amine.

71. The compound according to claim 70, wherein R 3 is NH2.

72. A compound according to any one of claims 11 to 16, 19, 22 to 24, 27 and 30, wherein R 3 C 1-3 alkyl.

73. The compound according to claim 72, wherein R 3 It is methyl.

74. according to any one of claims 11 to 15, 17, 20, 22, 25, 28 and 30, wherein R 4 For hydrogen.

75. A compound according to any one of claims 11 to 15, 17, 20, 22, 25 and 30, wherein R 4 C 1-4 alkyl.

76. The compound according to claim 75, wherein R 4 It is methyl.

77. The compound according to claim 75, wherein R 4 It is ethyl.

78. The compound according to claim 75, wherein R 4 It is isopropyl.

79. The compound according to claim 75, wherein R 4 It is tert-butyl.

80. A compound according to any one of claims 23 to 30, wherein R 5 For F.

81. according to the compound described in any one of claims 23 to 30, wherein R 5 For Cl.

82. according to the compound described in any one of claims 23 to 30, wherein R 5 For Br.

83. according to the compound described in any one of claims 23 to 30, wherein R 5 For CN.

84. The compound according to claim 1, selected from the group consisting of: or a pharmaceutically acceptable salt thereof.

85. A compound according to any one of claims 1 to 84, or a pharmaceutically acceptable salt thereof, wherein at least one hydrogen atom is replaced by a deuterium atom.

86. A pharmaceutical composition comprising a compound according to any one of claims 1 to 85 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

87. A method for treating a disease mediated by PHD activity, the method comprising administering to a subject a compound according to any one of claims 1 to 85.

88. The method of claim 87, wherein the disease mediated by PHD activity is ischemia-reperfusion injury.

89. The method of claim 88, wherein the ischemic reperfusion injury is selected from the group consisting of stroke, myocardial infarction, and acute kidney injury.

90. The method of claim 87, wherein the disease mediated by PHD activity is inflammatory bowel disease.

91. The method of claim 90, wherein the inflammatory bowel disease is ulcerative colitis.

92. The method of claim 90, wherein the inflammatory bowel disease is Crohn's disease.

93. The method of claim 87, wherein the disease mediated by PHD activity is cancer.

94. The method of claim 93, wherein the cancer is colorectal cancer.

95. The method of claim 87, wherein the disease mediated by PHD activity is a liver disease.

96. The method of claim 87, wherein the disease mediated by PHD activity is atherosclerosis.

97. The method of claim 87, wherein the disease mediated by PHD activity is a cardiovascular disease.

98. The method of claim 87, wherein the disease mediated by PHD activity is an ocular disease or condition.

99. The method of claim 98, wherein the ocular disease or condition is selected from radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

100. The method of claim 87, wherein the disease is anemia.

101. The method of claim 100, wherein the anemia is anemia associated with chronic kidney disease.

102. The method of claim 87, wherein the disease is chronic kidney disease.

103. The method of claim 87, wherein the disease is associated with hyperoxia.

104. The method of claim 103, wherein the disease is retinopathy of prematurity.

105. The method of claim 103, wherein the disease is bronchopulmonary dysplasia (BPD).

106. The method of claim 87, wherein the disease 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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