PHD inhibitor compounds, compositions, and uses

Abstract

The present invention relates, in part, to novel small molecule inhibitors of PHDs having a structure according to formula (A), and subformulas thereof: [Formula 1] TIFF2023518953000310.tif29170 or a pharmaceutically acceptable salt thereof. The compounds provided herein may be useful in treating diseases, including diseases of the heart (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver (e.g., acute liver failure, liver fibrosis, and liver cirrhosis), and kidney (e.g., acute kidney injury and chronic kidney disease).

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 62 / 992,585, filed March 20, 2020, which is incorporated herein by reference in its entirety.

[0002] Hypoxia is a condition or state in which the supply of oxygen is insufficient for normal vital functions, e.g., low arterial oxygen supply. Hypoxia can lead to cellular dysfunction and structural tissue damage. The activation of cellular defense mechanisms during hypoxia is mediated by HIF (hypoxia-inducible factor) proteins. In response to hypoxia, HIFα levels increase in most cells due to a decrease in HIFα prolyl hydroxylation. Prolyl hydroxylation of HIFα is achieved by a family of proteins variously called prolyl hydroxylase domain-containing proteins (PHD1, 2, and 3) and also known as HIF prolyl hydroxylases (HPH-3, 2, and 1) or EGLN-2, 1, and 3. PHD proteins are oxygen sensors and regulate HIF stability in an oxygen-dependent manner. The three PHD isoforms function differently in HIF regulation and may have other non-HIF-related regulatory roles.

[0003] In fact, numerous studies have demonstrated that stabilizing HIF can attenuate tissue inflammation and promote tissue repair. Therefore, compounds that can inhibit the activity of PHD proteins may be particularly beneficial in novel therapeutic strategies (Lee et al. (2019) Exp. Mol. Med. 51:68).

[0004] Described herein are novel small molecule PHD inhibitors that have utility in the treatment of diseases, including diseases of the heart (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney (e.g., acute kidney injury and chronic kidney disease). Summary of the Invention

[0005] The present invention provides, inter alia, novel small molecule inhibitors of PHDs, which have utility in the treatment of diseases including, but not limited to, diseases of the heart (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney (e.g., acute kidney injury and chronic kidney disease).

[0006] In one aspect, a compound having a structure according to formula (A): [ka] or a pharmaceutically acceptable salt thereof, wherein: Ar 1 is phenyl or a 6-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl is selected from the group consisting of halogen, CN, OH, C optionally substituted with one or more halogens, 1-3 Alkyl, or C 1-3 optionally substituted with alkoxy; R 2 is H or C alkyl, Ar 2 is a halogen, OH, amine, or C 1-3 a 6-membered nitrogen-containing heteroaryl optionally substituted with alkyl; R 4 is hydrogen or C 1-4 is alkyl, Formula (A) excludes the following compounds: [ka]

[0007] In some embodiments, R 2 is H.

[0008] In some embodiments, R 2 is C 1-3It is alkyl.

[0009] In some embodiments, Ar 1 teeth, [ka] where: X, Y, and Z are independently CH or N, wherein N is optionally oxidized; Each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy; m is 1, 2, 3, or 4.

[0010] In some embodiments, Ar 1 is substituted phenyl. In some embodiments, Ar 1 At least one R 1 wherein R 1 is CN or a halogen.

[0011] In some embodiments, Ar 1 is C optionally substituted with one or more halogens 1-3 one or two R independently selected from alkyl, halogen, CN, or OH; 1 The group is substituted.

[0012] In some embodiments, Ar 1 is a pyridyl N-oxide, or C 1-3 At least one R is alkoxy or halogen 1 is pyridyl optionally substituted by

[0013] In some embodiments, each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy is selected from the group consisting of:

[0014] In some embodiments, Ar 2 teeth, [ka] where: A and B are independently CH or N, wherein N is optionally oxidized; Each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 is selected from the group consisting of alkyl, n is 0, 1, or 2.

[0015] In some embodiments, Ar 2 is a group that is pyridyl or pyrazinyl, the group being unsubstituted or containing halogen, C 1-3 It includes substituents that are alkyl, or OH.

[0016] In some embodiments, each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl.

[0017] In some embodiments, R 4 is H. In some embodiments, R 4 is C 1-4 It is alkyl.

[0018] In embodiments, the compound of formula (A) is [ka] isn't it.

[0019] In embodiments, compounds of Formula (A) exclude the following compounds: [ka]

[0020] In embodiments, the compound of Formula (A) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0021] In embodiments, X, Y, Z, A, and B are independently CH or N, where N is optionally oxidized, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0022] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0023] In embodiments, X, Y, and Z are independently CH or N, where N is optionally oxidized, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0024] In embodiments, the compound of formula (I) is [ka] isn't it.

[0025] In embodiments, compounds of formula (I) exclude the following compounds: [ka]

[0026] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0027] In embodiments, A and B are independently CH or N, where N is optionally oxidized, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0028] In embodiments, the compound of formula (Ib) is [ka] isn't it.

[0029] In embodiments, compounds of Formula (Ib) exclude the following compounds: [ka]

[0030] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0031] In embodiments, X, Y, Z, A, and B are independently CH or N, where N is optionally oxidized, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0032] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0033] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R4 is halogen or C 1-4 It is alkyl.

[0034] In embodiments, the compound of formula (II) is [ka] isn't it.

[0035] In embodiments, compounds of formula (II) exclude the following compounds: [ka]

[0036] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0037] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl.

[0038] In embodiments, the compound of formula (IIa) is [ka] isn't it.

[0039] In embodiments, compounds of Formula (IIa) exclude the following compounds: [ka]

[0040] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0041] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently 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 alkyl, and R 4 is hydrogen or C 1-4 It is alkyl.

[0042] In embodiments, the compound of formula (IIb) is [ka] isn't it.

[0043] In embodiments, compounds of Formula (IIb) exclude the following compounds: [ka]

[0044] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0045] In some embodiments, m is 1, 2, 3, or 4, and each R 1are independently 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 C1-3 alkyl.

[0046] In embodiments, the compound of formula (IIc) is [ka] isn't it.

[0047] In embodiments, compounds of Formula (IIc) exclude the following compounds: [ka]

[0048] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0049] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl.

[0050] In embodiments, the compound of formula (IId) is [ka] isn't it.

[0051] In embodiments, compounds of Formula (IId) exclude the following compounds: [ka]

[0052] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0053] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0054] In embodiments, the compound of formula (IIe) is [ka] isn't it.

[0055] In embodiments, compounds of Formula (IIe) exclude the following compounds: [ka]

[0056] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0057] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy is selected from the group consisting of:

[0058] In embodiments, the compound of formula (IIf) is [ka] isn't it.

[0059] In embodiments, compounds of Formula (IIf) exclude the following compounds: [ka]

[0060] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0061] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 It is alkyl.

[0062] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0063] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is halogen or C 1-4 alkyl, and R 5 is CN or a halogen.

[0064] In embodiments, the compound of formula (III) is [ka] isn't it.

[0065] In embodiments, compounds of formula (III) exclude the following compounds: [ka]

[0066] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0067] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently 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 alkyl, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 5 is CN or a halogen.

[0068] In embodiments, the compound of formula (IIIa) is [ka] isn't it.

[0069] In embodiments, compounds of Formula (IIIa) exclude the following compounds: [ka]

[0070] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0071] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently 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 alkyl, and R 4 is hydrogen or C 1-4 alkyl, and R 5is CN or a halogen.

[0072] In embodiments, the compound of formula (IIIb) is [ka] isn't it.

[0073] In embodiments, compounds of Formula (IIIb) exclude the following compounds: [ka]

[0074] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0075] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently 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 alkyl, and R 5 is CN or a halogen.

[0076] In embodiments, the compound of formula (IIIc) is [ka] isn't it.

[0077] In embodiments, compounds of Formula (IIIc) exclude the following compounds: [ka]

[0078] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0079] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 5 is CN or a halogen.

[0080] In embodiments, the compound of formula (IIId) is [ka] isn't it.

[0081] In embodiments, compounds of Formula (IIId) exclude the following compounds: [ka]

[0082] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the following structure: [ka] or a pharmaceutically acceptable salt thereof.

[0083] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens1-3 Alkyl, and C 1-3 alkoxy; R 4 is hydrogen or C 1-4 alkyl, and R 5 is CN or a halogen.

[0084] In embodiments, the compound of formula (IIIe) is [ka] isn't it.

[0085] In embodiments, compounds of Formula (IIIe) exclude the following compounds: [ka]

[0086] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0087] In some embodiments, m is 1, 2, 3, or 4, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy; R 5 is CN or a halogen.

[0088] In embodiments, the compound of formula (IIIf) is [ka] isn't it.

[0089] In embodiments, compounds of Formula (IIIf) exclude the following compounds: [ka]

[0090] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof.

[0091] In embodiments, m is 1, 2, 3, or 4, n is 0, 1, or 2, and each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy, and each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 alkyl; R 4 is hydrogen or C 1-4 alkyl, and R 5 is CN or a halogen.

[0092] In embodiments, X is CH. In embodiments, X is N, which is optionally oxidized.

[0093] In embodiments, Y is CH. In embodiments, Y is N.

[0094] In embodiments, Z is CH. In embodiments, Z is N.

[0095] In embodiments, A is CH. In embodiments, A is N.

[0096] In embodiments, B is CH. In embodiments, B is N.

[0097] In embodiments, m is 1. In embodiments, m is 2. In embodiments, m is 3. In embodiments, m is 4.

[0098] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0099] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is CN. In some embodiments, R 1 is OH.

[0100] In some embodiments, R 1 is halogen. In some embodiments, R 1 is F. In some embodiments, R 1 is Cl. In some embodiments, R 1 is Br.

[0101] In some embodiments, R 1 is C optionally substituted with one or more halogens 1-3 In some embodiments, R 1 is C 1-3 In some embodiments, R 1 is methyl. In some embodiments, R 1 is ethyl. In some embodiments, R 1 is CF3.

[0102] In some embodiments, R 1 is C 1-3 In some embodiments, R 1 is methoxy.

[0103] In some embodiments, R 2 is hydrogen.

[0104] In some embodiments, R2 is C 1-3 In some embodiments, R2 is methyl.

[0105] In some embodiments, R 3 is hydrogen.

[0106] In some embodiments, R 3 is halogen. In some embodiments, R 3 is F.

[0107] In some embodiments, R 3 is OH.

[0108] In some embodiments, R 3 is an amine. In some embodiments, R 3 is NH2.

[0109] In some embodiments, R 3 is C 1-3 In some embodiments, R 3 is methyl.

[0110] In some embodiments, R 4 is hydrogen.

[0111] In some embodiments, R 4 is C 1-4 In some embodiments, R 4 is methyl. In some embodiments, R 4 is ethyl. In some embodiments, R 4 is isopropyl. In some embodiments, R 4 is tert-butyl.

[0112] In some embodiments, R 5 is F. In some embodiments, R 5 is Cl. In some embodiments, R 5 is Br.

[0113] In some embodiments, R 5 is CN.

[0114] In embodiments, the compound is any one of compounds 1-44, or a pharmaceutically acceptable salt thereof. [Table 1] TIFF0007768890000060.tif219170TIFF0007768890000061.tif170170

[0115] In embodiments, at least one hydrogen atom in a compound of Formulas (A) and (I)-(III), such as any one of Compounds 1-44, or a pharmaceutically acceptable salt thereof, is replaced with a deuterium atom.

[0116] In another aspect, the invention features a pharmaceutical composition including any compound described herein (e.g., a compound of Formulas (A) and (I)-(III), such as any one of Compounds 1-44), or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0117] In another aspect, the invention features a method for treating a disease mediated by PHD activity, the method including administering to a subject any compound described herein (e.g., a compound of Formulas (A) and (I)-(III), such as any one of Compounds 1-44), or a pharmaceutically acceptable salt thereof.

[0118] In embodiments, the disease mediated by PHD activity is ischemia-reperfusion injury (eg, stroke, myocardial infarction, or acute kidney injury).

[0119] In embodiments, the disease mediated by PHD activity is inflammatory bowel disease (eg, ulcerative colitis or Crohn's disease).

[0120] In embodiments, the disease mediated by PHD activity is cancer (eg, colorectal cancer).

[0121] In embodiments, the disease mediated by PHD activity is a liver disease.

[0122] In embodiments, the disease mediated by PHD activity is atherosclerosis.

[0123] In embodiments, the disease mediated by PHD activity is a cardiovascular disease.

[0124] 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).

[0125] In embodiments, the disease mediated by PHD activity is anemia (eg, anemia associated with chronic kidney disease).

[0126] In embodiments, the disease mediated by PHD activity is associated with hyperoxia.

[0127] In embodiments, the disease mediated by PHD activity is retinopathy of prematurity.

[0128] In embodiments, the disease mediated by PHD activity is bronchopulmonary dysplasia (BPD).

[0129] 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 liver cirrhosis. [Brief explanation of the drawings]

[0130] [Figure 1]Figure 1 is an exemplary schematic diagram illustrating the principle of the TR-FRET assay for PHD enzymes (PHD1, PHD2, and PHD3). In the presence of 2-oxoglutarate and O, PHD enzymes hydroxylate proline 564 of the biotin-tagged HIF-1α peptide, resulting in the production of biotin-tagged HIF-1α-hydroxyproline, succinate, and CO. The donor fluorophore conjugate, monoclonal antibody anti-6His-terbium (Tb)-cryptate gold, bound to His-tagged VHL protein, EloB, and EloC complexes (His-VBC), and the acceptor fluorophore, SA-D2 conjugate, bound to HIF-1α-hydroxyproline, result in a fluorescence resonance energy transfer (FER) signal that can be detected and quantified. DETAILED DESCRIPTION OF THE INVENTION

[0131] definition In order that the present invention may be more readily understood, certain terms are first defined below. Additional definitions of these terms and other terms are set forth throughout the specification. Publications and other reference materials referred to herein to describe the background of the invention and to provide further details regarding its practice are incorporated herein by reference.

[0132] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, mouse, rat, rabbit, monkey, dog, cat, sheep, cow, primate, and / or pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or parasites. In some embodiments, the animal may be a transgenic animal, a genetically engineered animal, and / or a clone.

[0133] Approximately or About: As used herein, the term "approximately" or "about" as applied to one or more values ​​of interest refers to a value similar to the stated reference value. In certain embodiments, the term "approximately" or "about" refers to a range of values ​​that falls 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 of (greater or less than) the stated reference value, unless otherwise stated or clear from the context (except when such number exceeds 100% of possible values).

[0134] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, the term "a composition" includes a mixture of two or more such compositions.

[0135] 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 are not limited to, and are not intended to exclude, for example, other additives, components, integers, or steps.

[0136] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and that the description includes cases where the event or circumstance occurs and cases where it does not occur.

[0137] Improve, increase, or reduce: As used herein, "improve," "increase," or "reduce," or grammatical equivalents, refer to a value compared to a baseline measurement, e.g., a measurement in the same individual before initiation of a treatment described herein, or a measurement in a control subject (or control subjects) in the absence of a treatment described herein. A "control subject" is a subject suffering from the same form of disease as the subject being treated and who is approximately the same age as the subject being treated.

[0138] In vitro: As used herein, the term "in vitro" refers to events that take place not within a multicellular organism but in an artificial environment, for example, in a test tube or reaction vessel, in cell culture, etc.

[0139] In vivo: As used herein, the term "in vivo" refers to events that occur within multicellular organisms, such as humans and non-human animals. In the context of cell-based systems, the term can be used to refer to events that occur within living cells (as opposed to, for example, in vitro systems).

[0140] Patient: As used herein, the term "patient" or "subject" refers to any organism to which provided compositions can be administered, for example, for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is a human. Humans include prenatal and postnatal forms.

[0141] Pharmaceutically acceptable: The term "pharmaceutically acceptable," as used herein, refers to materials that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0142] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art.For example, S.M. Berge et al. describe pharmaceutically acceptable salts in J. Pharmaceutical Sciences (1977) 66:1-19.The 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 the salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N(C1-4 alkyl) salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc.Further pharmaceutically acceptable salts include non-toxic ammonium cations, quaternary ammonium cations, and amine cations, formed where appropriate using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates, and arylsulfonates. Further pharmaceutically acceptable salts include salts formed from the quaternization of amines using suitable electrophiles, for example, alkyl halides to form quaternized alkylated amino salts.

[0143] Subject: As used herein, the term "subject" refers to a human or any non-human animal (e.g., a mouse, rat, rabbit, dog, cat, cow, pig, sheep, horse, or primate). Human includes prenatal and postnatal forms. In many embodiments, a subject is a human. A subject may be a patient, and refers to a person who sees a healthcare provider for diagnosis or treatment of a disease. The term "subject" is used interchangeably herein with "individual" or "patient." A subject may be afflicted with or susceptible to a disease or disorder, but may or may not exhibit symptoms of the disease or disorder.

[0144] Substantially: As used herein, the term "substantially" refers to the qualitative state of exhibiting the full or nearly full extent or degree of a desired characteristic or property. Those skilled in the art of biology will understand that biological and chemical phenomena rarely, if ever, go to completion and / or reach completion, or achieve or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of completion inherent in many biological and chemical phenomena.

[0145] 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 symptoms of, and / or delay the onset of, the disease, disorder, and / or condition. Those skilled in the art will appreciate that a therapeutically effective amount is typically administered in a dosing regimen comprising at least one unit dose.

[0146] Treating: As used herein, the terms "treat," "treatment," or "treating" refer to any method used to partially or completely alleviate, ameliorate, relieve, 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. Treatment may be administered to subjects who do not exhibit signs of disease and / or who exhibit only early signs of disease, for the purpose of reducing the risk of developing conditions associated with the disease.

[0147] Aliphatic: As used herein, the term aliphatic refers to C1-C 40 Aliphatic refers to hydrocarbons, including both saturated and unsaturated hydrocarbons. Aliphatic groups can be linear, branched, or cyclic. For example, C1-C 20 Aliphatic includes C1-C 20 Alkyl (e.g., linear or branched C1-C 20 saturated alkyl), C2-C 20 Alkenyl (e.g., linear or branched C4-C 20 Dienyl, linear or branched C6-C 20 trienyl, etc.), and C2-C 20 Alkynyl (e.g., linear or branched C-C 20 C-C alkynyl) may be included. 20 Aliphatic includes C3-C 20 Cycloaliphatic (e.g., C3-C 20 Cycloalkyl, C4-C 20 Cycloalkenyl, or C8-C20 In certain embodiments, an aliphatic group can include one or more cycloaliphatic groups and / or one or more heteroatoms, such as oxygen, nitrogen, or sulfur, and can be optionally substituted with one or more substituents, such as alkyl, halo, alkoxyl, 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 can be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected) halogen, —COR′, —COH, —COR′, —CN, —OH, —OR′, —OCOR′, —OCOR′, —NH, —NHR′, —N(R′)′, —SR′, or —SOR′, where each instance of R′ is independently C1-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently unsubstituted C1-C3 alkyl. In some embodiments, the aliphatic is unsubstituted. In some embodiments, the aliphatic does not contain any heteroatoms.

[0148] Alkyl: As used herein, the term "alkyl" refers to acyclic straight and branched chain hydrocarbon groups, e.g., C1-C 20"Alkyl" refers to an alkyl group having 1 to 20 carbon atoms. The alkyl group can be straight-chained 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-pentylhexyl, isohexyl, and the like. The term "lower alkyl" refers to a straight-chain or branched alkyl of an alkyl group having 1 to 6 carbon atoms. Other alkyl groups will be readily apparent to those of skill in the art given the benefit of this disclosure. The alkyl group can be unsubstituted or substituted with one or more substituents described herein. For example, an alkyl group can be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, —COR′, —COH, —COR′, —CN, —OH, —OR′, —OCOR′, —OCOR′, —NH, —NHR′, —N(R′)2, —SR′, or —SOR′, where each instance of R′ is independently C1-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted C1-C3 alkyl. In some embodiments, the alkyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituents 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, where the prefix indicates the -OH group and "alkyl" is as described herein. In some embodiments, the alkyl is substituted with an -OR' group, and may also be referred to herein as an "alkoxy" group.

[0149] When the suffix "-ene" is added to a group, it becomes a divalent moiety. For example, an arylene is a divalent moiety of an aryl, and a heteroarylene is a divalent moiety of a heteroaryl.

[0150] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent straight- or branched-chain 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-chain hydrocarbon group having one or more unsaturated carbon-carbon double bonds that may occur at any stable point along the chain, and the term "alkynylene" refers to an unsaturated divalent straight- or branched-chain hydrocarbon group having one or more unsaturated carbon-carbon triple bonds that may occur at any stable point along the chain. In certain embodiments, alkylene, alkenylene, or alkynylene groups can contain one or more cyclic aliphatic and / or one or more heteroatoms such as oxygen, nitrogen, or sulfur, and can be optionally substituted with one or more substituents such as alkyl, halo, alkoxyl, hydroxy, amino, aryl, ether, ester, or amide. For example, the alkylene, alkenylene, or alkynylene can be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, —COR′, —COH, —COR′, —CN, —OH, —OR′, —OCOR′, —OCOR′, —NH, —NHR′, —N(R′)2, —SR′, or —SOR′, where each instance of R′ is independently C-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10In some embodiments, R' is independently unsubstituted C1-C3 alkyl. In certain embodiments, the alkylene, alkenylene, or alkynylene is unsubstituted. In certain embodiments, the alkylene, alkenylene, or alkynylene does not contain any heteroatoms.

[0151] Alkenyl: As used herein, "alkenyl" means any straight or branched hydrocarbon chain with one or more unsaturated carbon-carbon double bonds that may occur at any stable point along the chain, e.g., "C2-C 20 "Alkenyl" refers to an alkenyl group having 2 to 20 carbons. For example, alkenyl groups include prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, hex-5-enyl, 2,3-dimethylbut-2-enyl, and the like. In some embodiments, an alkenyl contains one, two, or three carbon-carbon double bonds. In some embodiments, an alkenyl contains a single carbon-carbon double bond. In some embodiments, multiple double bonds (e.g., two or three) are conjugated. An alkenyl group can be unsubstituted or substituted with one or more substituents described herein. For example, an alkenyl group can be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, —COR′, —COH, —COR′, —CN, —OH, —OR′, —OCOR′, —OCOR′, —NH, —NHR′, —N(R′)2, —SR′, or —SOR′, where each instance of R′ is independently C1-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10In some embodiments, R' is independently an unsubstituted C1-C3 alkyl. In some embodiments, the alkenyl is unsubstituted. In some embodiments, the alkenyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituents 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, where the prefix indicates the -OH group and "alkenyl" is as described herein.

[0152] Alkynyl: As used herein, "alkynyl" means any hydrocarbon chain in either a straight or branched configuration with one or more carbon-carbon triple bonds occurring at any stable point along the chain, e.g., "C2-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, an alkynyl group contains one carbon-carbon triple bond. An alkynyl group can be unsubstituted or substituted with one or more substituents described herein. For example, an alkynyl group can be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR', -COH, -COR', -CN, -OH, -OR', -OCOR', -OCOR', -NH, -NHR', -N(R')2, -SR', or -SOR', where each instance of R' is independently C1-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In some embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10In some embodiments, R' is independently unsubstituted C1-C3 alkyl. In some embodiments, alkynyl is unsubstituted. In some embodiments, alkynyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituents described herein).

[0153] 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 6 to 14 ring members, wherein said 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 ("C6 aryl," e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C6 aryl," e.g., phenyl). 10 aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("C 14 "Aryl"; e.g., anthracyl). "Aryl" also includes ring systems in which the aryl ring is fused with one or more carbocyclic or heterocyclyl groups, as defined above, in which the bonding radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms subsequently designates the number of carbon atoms in the aryl ring system. Examples of aryl include phenyl, naphthyl, and anthracene.

[0154] Arylene: As used herein, the term "arylene" refers to a divalent aryl group (i.e., having two points of attachment to the molecule). Examples of arylene include phenylene (e.g., unsubstituted phenylene or substituted phenylene).

[0155] Halogen or Halo: As used herein, the terms "halogen" or "halo" mean fluorine, chlorine, bromine, or iodine.

[0156] Amide: The term “amide” or “amido” refers to a group of amides of the formula —C(O)N(R ’ )2, -C(O)N(R ’ )-, -NR ’ C(O)R ’ , -NR ’ C(O)N(R ’ )2-, or -NR ’ refers to a chemical moiety having the formula C(O)—, where each R ’ are 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), unless stated otherwise in the specification, each of which can itself be optionally substituted as described herein, or two R' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring.

[0157] Amino: The term “amino” or “amine” refers to the group —N(R′) 2 , where each R ’ are 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 groups, unless stated otherwise in the specification, each of which can itself be optionally substituted as described herein, or two R' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. In embodiments, the amino group is -NHR', where R' is aryl (arylamino), heteroaryl (heteroarylarylamino), or alkyl (alkylamino).

[0158] Sulfonyl: The term “sulfonyl” refers to the group —S(═O)R′, or —S(═O)—, where R ’ is, unless otherwise stated in the specification, 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 may itself be optionally substituted as described herein.

[0159] Sulfinyl: The term "sulfinyl" refers to a chemical moiety having the formula -S(=O)R', -S(=O)-, or -S(=O)(=NR')-, where R ’ is, unless otherwise stated in the specification, 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 may itself be optionally substituted as described herein.

[0160] Carbonyl: The term “carbonyl” refers to the group —C(═O)R′, or —C(═O)—, where R ’ is, unless otherwise stated in the specification, 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 may itself be optionally substituted as described herein.

[0161] Phosphoryl: The term “phosphoryl” refers to the group —P(═O)(R′) or —P(═O)(R′)—, where R ’is, unless stated otherwise in the specification, 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) group, each of which can itself be optionally substituted as described herein, or two R' can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring.

[0162] Heteroalkyl: The term "heteroalkyl" refers to 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. Heteroalkyls include tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, phosphoramidates, sulfonamides, and disulfides. Heteroalkyl groups may optionally contain monocyclic, bicyclic, or tricyclic rings, each ring desirably having 3 to 6 members. Examples of heteroalkyls include polyethers such as methoxymethyl and ethoxyethyl.

[0163] Heteroalkylene: As used herein, the term "heteroalkylene" refers to a divalent form of the heteroalkyl groups described herein.

[0164] Heteroaryl: As used herein, the term "heteroaryl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system having a total of six to fourteen ring members, said ring system having a single point of attachment to the rest of the molecule, at least one ring in the system being aromatic, each ring in the system containing four to seven ring members, and at least one ring atom being a heteroatom, such as, but not limited to, nitrogen and oxygen.

[0165] Heterocycloalkyl: As used herein, the term "heterocycloalkyl" refers to 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 groups can be substituted or unsubstituted.

[0166] Deuterium: "Deuterium" ("D" or " 2 The term "H" is also known as heavy hydrogen. Deuterium is an isotope of hydrogen with a nucleus consisting of one proton and one neutron, which is twice the mass of a nucleus of ordinary hydrogen (one proton).

[0167] Isotope: The term "isotope" refers to variants of a particular chemical element that differ in the number of neutrons, and therefore the number of nucleons. All isotopes of a given component have the same number of protons but different numbers of neutrons in each atom.

[0168] The term "substituted" means that the specified group or moiety has one or more substituents. The term "unsubstituted" means that the specified group has no substituents. The term "optionally substituted" means that the specified group is unsubstituted or substituted with one or more substituents. When the term "substituted" is used to describe a structural system, substitution is meant to occur at any valence-allowed position on the system; for example, substitution results in a stable compound (e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction). When a specified moiety or group is not explicitly described as being optionally substituted or substituted with any specified substituents, it is understood that such moiety or group is intended to be unsubstituted.

[0169] When a ring system (e.g., cycloalkyl, heterocyclyl, aryl, or heteroaryl) is substituted with a different number of substituents within the explicitly defined range, it is understood that the total number of substituents does not exceed the normally available valences under the existing conditions. It is also understood that hydrogen atoms are presumed to be present to satisfy the remaining valences of the ring system. Substituents include only those combinations of substituents and variables that result in stable or chemically feasible compounds. A stable or chemically feasible compound is one that has sufficient stability to enable its preparation and detection, among other factors.

[0170] A wide variety of substituents are well known, as are methods for their formation and introduction into various parent groups. Representative substituents include alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, arylalkyl, alkylaryl, aryl, arylalkoxy, arylamino, heteroarylamino, heteroaryl, heteroarylalkoxy, heterocycloalkyl, hydroxyalkyl, aminoalkyl, haloalkyl, thioalkyl, alkylthioalkyl, carboxyalkyl, imidazolylalkyl, indolylalkyl, mono-, di-, and trihaloalkyl, mono-, di-, and trihaloalkoxy, amino, alkylamino, dialkylamino, amido, cyano, alkoxy, hydroxy, sulfonamido, halo (e.g., -Cl and -Br), nitro, oximino, -COOR, and the like. 50 , -COR 50 , -SO 0-2 R 50 , -SO2NR 50 R 51 , N.R. 52 SO2R 50 , =C(R 50 R 51 ), =N-OR 50 , =N-CN, =C(halo)2, =S, =O, -CON(R 50 R 51 ), -OCOR 50 , -OCON(R 50 R 51 ), -N(R 52 )CO(R50 ), -N(R 52 )COOR 50 , and -N(R 52 )CON(R 50 (R 51 ), but are not limited to, wherein R 50 , R 51 , and R 52 may be independently selected from: a hydrogen atom, and branched or straight chain C 1-6 -Alkyl, C 3-6 -cycloalkyl, C 4-6 -heterocycloalkyl, heteroaryl, and aryl groups (substituted or unsubstituted). Where permitted, R 50 and R 51 can be joined together to form a carbocyclic or heterocyclic ring system.

[0171] In preferred embodiments, the substituents are selected from halogen, —COR′, —COH, —COR′, —CN, —OH, —OR′, —OCOR′, —OCOR′, —NH, —NHR′, —N(R′)2, —SR′, and —SOR′, wherein each instance of R′ is independently C1-C 20 Aliphatic (e.g., C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 In certain embodiments, R' is independently an unsubstituted alkyl (e.g., an unsubstituted C1-C3 alkyl, or a C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 alkyl, or C1-C3 alkyl). Preferably, R' is independently unsubstituted C1-C3 alkyl.

[0172] Any formula provided herein is intended to represent a compound having the structure shown by the structural formula, as well as certain variations or forms. In particular, compounds of any formula provided herein may have asymmetric centers and therefore exist in different enantiomeric forms. All optical isomers and stereoisomers of compounds of the general formula, as well as mixtures thereof, are considered within the scope of the formula. Thus, any formula provided herein is intended to represent a racemate, one or more enantiomers, one or more diastereomeric forms, one or more atropisomeric forms, and mixtures thereof. Furthermore, a particular structure may exist as a geometric isomer (i.e., cis and trans isomers), as a tautomer, or as an atropisomer. Furthermore, any formula provided herein is intended to encompass hydrates, solvates, and polymorphs of such compounds, as well as mixtures thereof.

[0173] Compounds of the Invention Disclosed herein are compounds that are potent inhibitors of PHDs. In some embodiments, the compounds of the present invention exhibit an enzymatic half-maximal inhibitory concentration (IC) of less than 100 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 50 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 25 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 20 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 15 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 10 μM for any one of PHD1, PHD2, and PHD3. 50In some embodiments, compounds of the present invention have an IC value of less than 5 μM for any one of PHD1, PHD2, and PHD3. 50 In some embodiments, compounds of the present invention have an IC value of less than 1 μM for any one 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. 50 In 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 It has a value.

[0174] Representative examples from this class exhibit inhibitory activity against PHD1, PHD2, and PHD3 in vitro.

[0175] Exemplary compounds are described herein. In particular, these selective inhibitors can feature a pyrazole moiety (e.g., a 5-hydroxy-substituted pyrazole) linking two aromatic moieties.

[0176] Compounds of formula (A) and (I) to (III) In one aspect, a compound having a structure according to formula (A): [ka] or a pharmaceutically acceptable salt thereof, wherein: Ar 1 is phenyl or a 6-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl is selected from the group consisting of halogen, CN, OH, C optionally substituted with one or more halogens, 1-3 Alkyl, or C 1-3 optionally substituted with alkoxy; R 2 is H or C1-3 alkyl, Ar 2 is a halogen, OH, amine, or C 1-3 a 6-membered nitrogen-containing heteroaryl optionally substituted with alkyl; R 4 is hydrogen or C 1-4 It is alkyl.

[0177] In embodiments of the formulas described herein (e.g., Formula (A)), the compound (e.g., a compound of Formula (A)) may be: [ka] isn't it.

[0178] In embodiments of the formulas described herein (eg, Formula (A)), the compounds (eg, compounds of Formula (A)) exclude the following compounds: [ka]

[0179] In some embodiments, R 2 is H.

[0180] In some embodiments, R 2 is C alkyl. In some embodiments, R 2 is CH3. In some embodiments, R 2 is CH2CH3. In some embodiments, R 2 is CH2CH2CH3. In some embodiments, R 2 is CH(CH3)2.

[0181] In some embodiments, R 4 is H.

[0182] In some embodiments, R 4 is C alkyl. In some embodiments, R 4 is CH3. In some embodiments, R 4 is CH2CH3. In some embodiments, R 4 is CH2CH2CH3. In some embodiments, R 4 is CH(CH). In some embodiments, R 4 is CH2CH2CH2CH3. In some embodiments, R 4 is CH(CH3)(CH2CH3). In some embodiments, R 4 is C(CH3)3.

[0183] In some embodiments, Ar 1 is unsubstituted aryl. In some embodiments, Ar 1 is substituted aryl. In some embodiments, Ar 1 is unsubstituted phenyl. In some embodiments, Ar 1 is a substituted phenyl.

[0184] In some embodiments, Ar 1 is an unsubstituted 6-membered nitrogen-containing heteroaryl. In some embodiments, Ar 1 is a substituted 6-membered nitrogen-containing heteroaryl.

[0185] In some embodiments, Ar 1 is halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 In some embodiments, Ar is substituted with one or more groups selected from alkoxy. 1 is substituted with one substituent. In some embodiments, Ar 1 is substituted with two substituents. In some embodiments, Ar 1 is substituted with three substituents. In some embodiments, Ar 1 is substituted with four substituents.

[0186] In some embodiments, Ar 1 is one or more R 1 groups, and each R 1 is hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 In some embodiments, Ar is independently selected from alkoxy. 1 is represented by m 1 R includes the amount of groups, and m is 1, 2, 3, or 4. 1 If there is R 1 can replace a hydrogen in the parent molecular structure. 1 is present and is a non-hydrogen moiety, R 1 represents a substituent. In some embodiments, R 1 is halogen, CN, OH, CN or C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy.

[0187] Thus, for any value of m described herein, the hydrogens should be such that the molecule is a stable compound (e.g., a compound that does not spontaneously undergo transformation, such as by rearrangement, cyclization, elimination, or other reaction). 1It is also understood that Ar is present as needed to satisfy valence requirements at a constituent atom of Ar. 1 , R 1 , and m are described herein.

[0188] In some embodiments, Ar 1 teeth, [ka] where: X, Y, and Z are independently CH or N, wherein N is optionally oxidized; Each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy; m is 1, 2, 3, or 4.

[0189] In some embodiments, R 1 is not hydrogen. In some embodiments, R 1 is present and is a non-hydrogen moiety, R 1 represents a substituent.

[0190] 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.

[0191] In embodiments, m is 1. In embodiments, m is 2. In embodiments, m is 3. In embodiments, m is 4.

[0192] 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 considered to be bonded to hydrogen to satisfy valence. In embodiments, m is 2.

[0193] In embodiments, one of X, Y, and Z is CH and the other is N, where N is optionally oxidized, and m is 1, 2, or 3. In embodiments, m is 1, and any remaining unsubstituted carbon ring atoms are considered to be bonded to hydrogen to satisfy valences. In embodiments, m is 2, and any remaining unsubstituted carbon ring atoms are considered to be bonded to hydrogen to satisfy valences. In embodiments, m is 3.

[0194] In embodiments, two of X, Y, and Z are CH and the other is N, where N is optionally oxidized, and m is 1, 2, 3, or 4. In embodiments, m is 1, and any remaining unsubstituted carbon ring atoms are considered to be bonded to hydrogen to satisfy valences. In embodiments, m is 2, and any remaining unsubstituted carbon ring atoms are considered to be bonded to hydrogen to satisfy valences. In embodiments, m is 3, and any remaining unsubstituted carbon ring atoms are considered to be bonded to hydrogen to satisfy valences. In embodiments, m is 4.

[0195] In some embodiments, Ar 1 The N atom in the

[0196] In some embodiments, Ar 1 The N atom in the

[0197] In some embodiments, Ar 1 is substituted phenyl. In some embodiments, Ar 1 At least one R 1 wherein R 1 is CN or a halogen.

[0198] In some embodiments, Ar 1 is C optionally substituted with one or more halogens 1-3one or two R independently selected from alkyl, halogen, CN, or OH; 1 The group is substituted.

[0199] In some embodiments, Ar 1 is a pyridyl N-oxide, or C 1-3 At least one R is alkoxy or halogen 1 is pyridyl optionally substituted by

[0200] In some embodiments, R 1 is always hydrogen.

[0201] In some embodiments, R 1 is always CN.

[0202] In some embodiments, R 1 is OH every time.

[0203] In some embodiments, R 1 is a halogen. In embodiments, the halogen is Cl. In embodiments, the halogen is Br. In embodiments, the halogen is I.

[0204] In some embodiments, R 1 Every time, C 1-3 It is alkyl.

[0205] In some embodiments, R 1 is always a non-substituted C 1-3 In some embodiments, R 1 is CH3 each time. In some embodiments, R 1 is always CH2CH3.

[0206] In some embodiments, R 1 is always replaced by C 1-3 In some embodiments, R 1 Each time, C is substituted with one or more halogens. 1-3In embodiments, the halogen is alkyl. In embodiments, the halogen is F. In embodiments, the halogen is Cl. In embodiments, the halogen is Br. In embodiments, the halogen is I.

[0207] In some embodiments, R 1 is always CF3.

[0208] In some embodiments, R 1 Every time, C 1-3 In some embodiments, R 1 is OMe every time.

[0209] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0210] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0211] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0212] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0213] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0214] In some embodiments, Ar 1 teeth, [ka] is selected from.

[0215] In some embodiments, Ar 2 is an unsubstituted 6-membered nitrogen-containing heteroaryl.

[0216] In some embodiments, Ar 2 is a halogen, OH, amine, or C 1-3 In some embodiments, Ar is a 6-membered nitrogen-containing heteroaryl substituted with alkyl. 2 is substituted with one of the substituents described herein. In some embodiments, Ar 2 is substituted with two substituents as described herein.

[0217] In some embodiments, Ar 2 teeth, [ka] where: A and B are independently CH or N, wherein N is optionally oxidized; Each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 is selected from the group consisting of alkyl, n is 0, 1, or 2.

[0218] 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, A and B are both N, and n is 0 or 1.

[0219] In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0220] In embodiments, A and B are both N, where N is optionally oxidized and n is 0 or 1. In embodiments, n is 0. In embodiments, n is 1.

[0221] In some embodiments, one of A and B is CH and the other is N, where N is optionally oxidized and n is 0, 1, or 2. In some embodiments, n is 0. In some embodiments, n is 1. In some embodiments, n is 2.

[0222] In embodiments, A and B are both CH and n is 0, 1, or 2. In embodiments, n is 0. In embodiments, n is 1. In embodiments, n is 2.

[0223] In some embodiments, Ar 2 The N atom in the

[0224] In some embodiments, Ar 2 The N atom in the

[0225] In some embodiments, R 3 is always hydrogen.

[0226] In some embodiments, R 3 is OH every time.

[0227] In some embodiments, R 3 is a halogen each time. In embodiments, the halogen is F. In embodiments, the halogen is Cl. In embodiments, the halogen is Br. In embodiments, the halogen is I.

[0228] In some embodiments, R 3 is an amine each time.3 is always NH2.

[0229] In some embodiments, R 3 Every time, C 1-3 It is alkyl.

[0230] In some embodiments, R 3 is always a non-substituted C 1-3 In some embodiments, R 3 is CH3 every time.

[0231] In some embodiments, Ar 2 is selected from the group consisting of: [ka]

[0232] In embodiments, the compound of Formula (A) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein A, B, X, Y, Z, R 1 , R 2 , R 3 , and R 4 is as defined elsewhere herein.

[0233] In embodiments of the formulas described herein (e.g., Formula (I)), the compound (e.g., a compound of Formula (I)) is [ka] isn't it.

[0234] In embodiments of the formulas described herein (eg, Formula (I)), the compounds (eg, compounds of Formula (I)) exclude the following compounds: [ka]

[0235] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein X, Y, Z, R 1 , R 2 , R 3 , and R 4 is as defined elsewhere herein.

[0236] In embodiments of the formulas described herein (e.g., Formula (Ia)), the compound (e.g., a compound of Formula (Ia)) is [ka] isn't it.

[0237] In embodiments of the formulas described herein (eg, Formula (Ia)), the compounds (eg, compounds of Formula (Ia)) exclude the following compounds: [ka]

[0238] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , and R 4 is as defined elsewhere herein.

[0239] In embodiments of the formulas described herein (e.g., Formula (Ib)), the compound (e.g., a compound of Formula (Ib)) is [ka] isn't it.

[0240] In embodiments of the formulas described herein (eg, Formula (Ib)), the compounds (eg, compounds of Formula (Ib)) exclude the following compounds: [ka]

[0241] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein A, B, X, Y, Z, R 1 , R 3 , and R 4 is as defined elsewhere herein.

[0242] In embodiments, the compound of Formula (A) or Formula (I) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , and R 4 is as defined elsewhere herein.

[0243] In embodiments of the formulas described herein (e.g., Formula (II)), the compound (e.g., a compound of Formula (II)) may be: [ka] isn't it.

[0244] In embodiments of the formulas described herein (eg, Formula (II)), the compounds (eg, compounds of Formula (II)) exclude the following compounds: [ka]

[0245] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , and R 3 is as defined elsewhere herein.

[0246] In embodiments of the formulas described herein (e.g., Formula (IIa)), the compound (e.g., a compound of Formula (IIa)) is [ka] isn't it.

[0247] In embodiments of the formulas described herein (eg, Formula (IIa)), the compounds (eg, compounds of Formula (IIa)) exclude the following compounds: [ka]

[0248] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , and R 4 is as defined elsewhere herein.

[0249] In embodiments of the formulas described herein (e.g., Formula (IIb)), the compound (e.g., a compound of Formula (IIb)) is [ka] isn't it.

[0250] In embodiments of the formulas described herein (eg, Formula (IIb)), the compounds (eg, compounds of Formula (IIb)) exclude the following compounds: [ka]

[0251] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 2 is as defined elsewhere herein.

[0252] In embodiments of the formulas described herein (e.g., Formula (IIc)), the compound (e.g., a compound of Formula (IIc)) is [ka] isn't it.

[0253] In embodiments of the formulas described herein (eg, Formula (IIc)), the compounds (eg, compounds of Formula (IIc)) exclude the following compounds: [ka]

[0254] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 3 is as defined elsewhere herein.

[0255] In embodiments of the formulas described herein (e.g., Formula (IId)), the compound (e.g., a compound of Formula (IId)) may be: [ka] isn't it.

[0256] In embodiments of the formulas described herein (eg, Formula (IId)), the compounds (eg, compounds of Formula (IId)) exclude the following compounds: [ka]

[0257] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 4 is as defined elsewhere herein.

[0258] In embodiments of the formulas described herein (e.g., Formula (IIe)), the compound (e.g., a compound of Formula (IIe)) is [ka] isn't it.

[0259] In embodiments of the formulas described herein (eg, Formula (IIe)), the compounds (eg, compounds of Formula (IIe)) exclude the following compounds: [ka]

[0260] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 is as defined elsewhere herein.

[0261] In embodiments of the formulas described herein (e.g., Formula (IIf)), the compound (e.g., a compound of Formula (IIf)) may be: [ka] isn't it.

[0262] In embodiments of the formulas described herein (eg, Formula (IIf)), the compounds (eg, compounds of Formula (IIf)) exclude the following compounds: [ka]

[0263] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 3 , and R 4 is as defined elsewhere herein.

[0264] In embodiments, the compound of Formula (A), Formula (I), or Formula (II) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , and R 4 is as defined elsewhere herein. R 5 is CN or a halogen,

[0265] In some embodiments, R 5 is CN.

[0266] In some embodiments, R 5 is a halogen. In embodiments, the halogen is F. In embodiments, the halogen is Cl. In embodiments, the halogen is Br. In embodiments, the halogen is I.

[0267] In embodiments of the formulas described herein (e.g., Formula (III)), the compound (e.g., a compound of Formula (III)) may be: [ka] isn't it.

[0268] In embodiments of the formulas described herein (eg, Formula (III)), the compounds (eg, compounds of Formula (III)) exclude the following compounds: [ka]

[0269] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 and R 5 is as defined elsewhere herein.

[0270] In embodiments of the formulas described herein (e.g., Formula (IIIa)), the compound (e.g., a compound of Formula (IIIa)) may be: [ka] isn't it.

[0271] In embodiments of the formulas described herein (eg, Formula (IIIa)), the compounds (eg, compounds of Formula (IIIa)) exclude the following compounds: [ka]

[0272] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 4 , and R 5 is as defined elsewhere herein.

[0273] In embodiments of the formulas described herein (e.g., Formula (IIIb)), the compound (e.g., a compound of Formula (IIIb)) is [ka] isn't it.

[0274] In embodiments of the formulas described herein (eg, Formula (IIIb)), the compounds (eg, compounds of Formula (IIIb)) exclude the following compounds: [ka]

[0275] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , and R 5 is as defined elsewhere herein.

[0276] In embodiments of the formulas described herein (e.g., Formula (IIIc)), the compound (e.g., a compound of Formula (IIIc)) is [ka] isn't it.

[0277] In embodiments of the formulas described herein (eg, Formula (IIIc)), the compounds (eg, compounds of Formula (IIIc)) exclude the following compounds: [ka]

[0278] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 3 , and R 5 is as defined elsewhere herein.

[0279] In embodiments of the formulas described herein (e.g., Formula (IIId)), the compound (e.g., a compound of Formula (IIId)) may be: [ka] isn't it.

[0280] In embodiments of the formulas described herein (eg, Formula (IIId)), the compounds (eg, compounds of Formula (IIId)) exclude the following compounds: [ka]

[0281] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 4 , and R 5 is as defined elsewhere herein.

[0282] In embodiments of the formulas described herein (e.g., Formula (IIIe)), the compound (e.g., a compound of Formula (IIIe)) is [ka] isn't it.

[0283] In embodiments of the formulas described herein (eg, Formula (IIIe)), the compounds (eg, compounds of Formula (IIIe)) exclude the following compounds: [ka]

[0284] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 and R 5 is as defined elsewhere herein.

[0285] In embodiments of the formulas described herein (e.g., Formula (IIIf)), the compound (e.g., a compound of Formula (IIIf)) is [ka] isn't it.

[0286] In embodiments of the formulas described herein (eg, Formula (IIIf)), the compounds (eg, compounds of Formula (IIIf)) exclude the following compounds: [ka]

[0287] In embodiments, the compound of Formula (A), Formula (I), Formula (II), or Formula (III) has the structure: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 3 , R 4 , and R 5 is as defined elsewhere herein.

[0288] Exemplary Compounds In some embodiments, the PHD inhibitor compound is any one of compounds 1-44, or a pharmaceutically acceptable salt thereof. [Table 2] TIFF0007768890000129.tif221170TIFF0007768890000130.tif223170TIFF0007768890000131.tif47170

[0289] Isotopic substitution It should be understood that in the compounds described herein (e.g., compounds of any one of Formulas (A) and (I)-(III), such as any one of Compounds 1-44), atoms may exhibit their natural isotopic abundance, or one or more of the atoms may be artificially enriched in a particular isotope having the same atomic number but an atomic mass or mass number different from that found predominantly in nature. The present invention is intended to include all suitable isotopic variations of the compounds described herein (e.g., compounds of any one of Formulas (A) and (I)-(III), such as any one of Compounds 1-44). For example, different isotopic forms of hydrogen (H) are available, such as protium ( 1H), deuterium ( 2 H), and tritium ( 3 Protium is the predominant hydrogen isotope found in nature.

[0290] In some embodiments, one or more hydrogens of a compound described herein (e.g., a compound of any one of Formulas (A) and (I)-(III), such as any one of Compounds 1-44) are replaced with deuterium. Deuterium enrichment may provide certain therapeutic advantages, such as increased in vivo half-life or reduced dosage requirements, or may provide a compound useful as a standard for characterization of biological samples. In some embodiments, one or more hydrogens of a compound described herein (e.g., a compound of any one of Formulas (A) and (I)-(III), such as any one of Compounds 1-44) are replaced with tritium. Tritium is radioactive and may provide a radiolabeled compound useful as a tracer in metabolic or kinetic studies.

[0291] Isotopic enrichment of the compounds disclosed herein (e.g., compounds of any one of Formulae (A) and (I)-(III), such as any one of Compounds 1-44) can be achieved without undue experimentation by conventional techniques well known to those of skill in the art or by processes analogous to those described in the schemes and examples herein using appropriate isotopically enriched reagents and / or intermediates.

[0292] The term "isotopic substitution" refers to a species having the same chemical structure and formula as a specific compound provided herein, except for the position of isotopic substitution and / or the level of isotopic enrichment at one or more positions, e.g., hydrogen versus deuterium. Thus, as used herein, the term "compound" encompasses a collection of molecules having the same chemical structure but also isotopic variation among the constituent atoms of the molecule. Thus, it will be apparent to those skilled in the art that a compound represented by a specific chemical structure containing a deuterium atom as shown will also contain lesser amounts of isotopic substitutions having hydrogen atoms at one or more of the designated deuterium positions in that structure. The relative amounts of such isotopic substitutions in a provided compound depend on many factors, including, but not limited to, the isotopic purity of the deuteration reagent used to make the compound and the efficiency of deuterium incorporation in the various synthetic steps used to prepare the compound.

[0293] When a position is designated as "H" or "hydrogen," the position is understood to have hydrogen at its natural abundance isotopic composition. When a position is designated as "D" or "deuterium," the position is understood to have deuterium at an abundance that is at least 3340 times the natural abundance of deuterium, which is 0.015% (i.e., the term "D" or "deuterium" indicates at least 50.1% deuterium incorporation).

[0294] In embodiments, the compounds provided herein may have an isotopic enrichment factor for each deuterium present at a site designated as a potential site for deuteration 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), 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).

[0295] Synthesis of Compounds of the Invention The compounds described herein (e.g., compounds of any one of Formulas A and I-III, such as any one of Compounds 1-44) can be prepared according to methods known in the art, including exemplary syntheses of the Examples provided herein, such as the synthesis shown in Scheme A.

[0296] The purity of compounds and their synthetic intermediates was determined by reverse phase HPLC using one of the methods described below:

[0297] Method A: Mobile phase: A: Water (0.01% TFA), B: Acetonitrile (0.01% TFA); Gradient phase: 5% B to 95% B in 1.4 min, increasing to 95% B in 1.6 min (total run time: 3 min); 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.

[0298] Method B: Mobile phase: A: Water (10 mM NH₄HCO₃), B: Acetonitrile; 5% to 95% B in 1.5 min, 95% B in 1.5 min (total run time: 3 min); Flow rate: 2.0 mL / min; Column: XBridge C18, 4.6*50 mm, 3.5 μm; Column temperature: 40°C. Detectors: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API). Abbreviations and acronyms used herein include the following: [Table 3] Scheme A [ka]

[0299] 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 an ester of formula (B) with N,N-dimethylformamide dimethyl acetal provides an enamine compound of formula (C). Cyclization of (C) and hydrazide (D) provides a pyrazole compound of formula (IId).

[0300] Compositions and Methods The present invention provides the use of a compound of any one of Formulas (A) and (I)-(III), or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for use in treating various conditions or disorders described herein. Pharmaceutical compositions are provided comprising at least one compound of any one of Formulas (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, or be used in combination with, at least one additional therapeutic agent.

[0301] The compounds of the present invention, or pharmaceutical or compositional agents containing the compounds, can be used to inhibit the activity of PHD. Inhibition of PHD can be particularly useful in treating diseases, including those of the heart (e.g., ischemic heart disease, congestive heart failure, and valvular heart disease), lung (e.g., acute lung injury, pulmonary hypertension, pulmonary fibrosis, and chronic obstructive pulmonary disease), liver (e.g., acute liver failure, liver fibrosis, and cirrhosis), and kidney (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 containing one or more compounds of any one of Formulas (A) and (I)-(III).

[0302] The present invention is also directed to a method for inhibiting the activity of PHD. In one embodiment, the method comprises contacting PHD with an effective amount of one or more compounds selected from the group consisting of compounds of any one of Formulas (A) and (I)-(III), or a pharmaceutically acceptable salt thereof.

[0303] In yet other embodiments, a compound disclosed herein (e.g., a compound of any one of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or a pharmaceutically acceptable salt thereof, is useful in treating anemic 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 anemia, Felty syndrome, graft-versus-host disease, hematopoietic stem cell transplantation, hemolytic uremic syndrome, myelodysplastic syndrome, paroxysmal nocturnal hemoglobinuria, myelofibrosis, pancytopenia, pure red cell aplasia, Schönlein-Henoch purpura, hyperblastemia, and hyperblastemia. PHD1 inhibition is useful for the treatment or prevention of anemia, including refractory anemia with rheumatoid arthritis, Shwachman syndrome, sickle cell disease, thalassemia major, thalassemia minor, thrombocytopenic purpura, anemic or non-anemic patients undergoing surgery, anemia associated with or secondary to trauma, sideroblastic anemia, and anemia secondary to other treatments, including reverse transcriptase inhibitors for treating HIV, corticosteroid hormones, cyclic cisplatin or non-cisplatin-containing chemotherapeutics, vinca alkaloids, mitotic inhibitors, topoisomerase II inhibitors, anthracyclines, alkylating agents, particularly for the treatment of anemia secondary to inflammatory, age-related, and / or chronic disease. PHD1 inhibition may also be used to treat symptoms of anemia, including chronic fatigue, pallor, and dizziness.

[0304] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating or preventing metabolic disorders, including, but not limited to, diabetes and obesity.

[0305] In still other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating or preventing vascular disorders, including, but not limited to, hypoxia- or wound healing-related disorders that require pro-angiogenic mediators for vasculogenesis, angiogenesis, and arteriogenesis.

[0306] In still other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating or preventing ischemia-reperfusion injury, including, but not limited to, stroke, myocardial infarction, and acute kidney injury.

[0307] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating inflammatory bowel diseases, including, but not limited to, ulcerative colitis and Crohn's disease.

[0308] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in the treatment of cancer, such as colorectal cancer.

[0309] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating atherosclerosis.

[0310] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating cardiovascular disease.

[0311] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in treating ophthalmic diseases or conditions, including, but not limited to, radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

[0312] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful for treating disorders associated with hyperoxia.

[0313] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or a pharmaceutically acceptable salt thereof, are useful for treating bronchopulmonary dysplasia (BPD).

[0314] In still other embodiments, the compounds disclosed herein (e.g., compounds of Formulas (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in treating cardiac conditions, including, but not limited to, postoperative myocardial ischemia in pancreatic surgery, myocardial injury after percutaneous coronary intervention (PCI), myocardial injury after non-cardiac surgery, perioperative myocardial ischemia in elective abdominal aortic aneurysm surgery, myocardial injury after PCI, myocardial injury in patients undergoing coronary artery bypass graft (CABG) surgery, minimally invasive mitral valve (MIMV) repair or replacement, adult patients undergoing open-heart surgery, chronic heart failure, and NYHA class II-IV.

[0315] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in treating pulmonary conditions, including, but not limited to, lung injury during elective lobectomy, lung injury during CABG surgery, and lung transplantation.

[0316] In other embodiments, compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in treating liver disease conditions, including, but not limited to, nonalcoholic steatohepatitis (NASH).

[0317] In other embodiments, the compounds disclosed herein (e.g., compounds of Formulae (A) and (I)-(III), such as any one of Compounds 1-44), or pharmaceutically acceptable salts thereof, are useful in treating renal disease conditions, including, but not limited to, contrast-induced acute kidney injury, stage III-IV chronic kidney disease undergoing planned coronary angiography, acute kidney injury in patients undergoing valvular heart surgery, non-dialysis-dependent chronic kidney disease, chronic kidney disease patients initiating dialysis, and non-dialysis-dependent chronic kidney disease.

[0318] In addition, 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 in combination with an additional active ingredient in the treatment of the above-described conditions. The additional compound can be separately co-administered with 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 a pharmaceutically acceptable salt thereof, or can be included with the additional active ingredient in a pharmaceutical composition according to the invention. In exemplary embodiments, the additional active ingredient is an active ingredient that is known or discovered to be effective in treating a condition, disorder, or disease mediated by a PHD enzyme, or that is active against another target associated with the particular condition, disorder, or disease, such as an alternative PHD modulator. The combination can serve to increase efficacy (e.g., by including in the combination a compound that enhances the potency or efficacy of a compound according to the invention), reduce one or more side effects, or reduce the required dose of a compound according to the invention.

[0319] The compounds of the present invention are used alone or in combination with one or more other active ingredients to formulate pharmaceutical compositions of the present invention, which comprise (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, pharmaceutically acceptable prodrug, or pharmaceutically active metabolite thereof, and (b) a pharmaceutically acceptable excipient.

[0320] "Pharmaceutically acceptable excipient" refers to a substance that is non-toxic, biologically tolerable, and otherwise biologically suitable for administration to a subject, such as an inert substance added to a pharmacological composition or used as a vehicle, carrier, or diluent to facilitate and be compatible with the administration of a drug. Examples of excipients include calcium carbonate, calcium phosphate, various sugars and starch types, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols. Suitable excipients may also contain antioxidants. Such antioxidants can be used in pharmaceutical compositions or storage media to extend the shelf life of pharmaceutical products.

[0321] Pharmaceutical Formulations and Routes of Administration The compounds and compositions of the present invention can be delivered directly or in pharmaceutical compositions or medicaments together with suitable carriers or excipients, as is well known in the art.The therapeutic method of the present invention can 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 the most preferred embodiment, the subject is a human subject.

[0322] The effective amount of such a compound, composition, or agent can be easily determined by routine experimentation, as can the most effective and convenient route of administration and the most suitable formulation. A variety of formulations and drug delivery systems are available in the art. See, for example, Gennaro, AR, ed. (1995) Remington's Pharmaceutical Sciences (see above).

[0323] Suitable administration routes may include, for example, oral, rectal, topical, nasal, pulmonary, ocular, intestinal, and parenteral administration. Primary routes of parenteral administration include intravenous, intramuscular, and subcutaneous administration. Secondary administration routes include intraperitoneal, intraarterial, intraarticular, intracardiac, intravesical, intradermal, intralesional, intraocular, intrapleural, intrathecal, intrauterine, and intracerebroventricular administration. The indication to be treated, along with the physical, chemical, and biological properties of the drug, determine the type of formulation and administration route used, and whether local or systemic delivery is preferred.

[0324] Pharmaceutical dosage forms of the compounds of the present invention can be provided in immediate-release, controlled-release, sustained-release, or targeted drug delivery systems. Commonly used dosage forms include, for example, solutions and suspensions, (micro)emulsions, ointments, gels and patches, liposomes, tablets, dragees, soft or hard-shell capsules, suppositories, vaginal suppositories, implants, amorphous or crystalline powders, aerosols, and lyophilized formulations. Depending on the route of administration used, special devices may be required for drug application or administration, such as syringes and needles, inhalers, pumps, injection pens, applicators, or special flasks. Pharmaceutical dosage forms often consist of a drug, excipients, and a container / closure system. One or more excipients, also known as inactive ingredients, may be added to the compounds of the present invention to improve or facilitate drug manufacturing, stability, administration, and safety, and to provide a means for achieving a desired drug release profile. Therefore, the type of excipient added to a 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 include those listed in various pharmacopeias, such as the United States Pharmacopoeia (USP), the Japanese Pharmacopoeia (JP), the European Pharmacopoeia (EP), and the British Pharmacopoeia (BP); the US Food and Drug Administration.

[0325] See the publications of the Center for Drug Evaluation and Research (CEDR) of the Food and Drug Administration (www.fda.gov), e.g., Inactive Ingredient Guide (1996); Ash and Ash, Eds. (2002) Handbook of Pharmaceutical Additives, Synapse Information Resources, Inc., Endicott, NY. Pharmaceutical dosage forms of the compounds of the present invention can be prepared by any method well known in the art, such as, for example, conventional mixing, sieving, dissolving, melting, granulating, dragee-making, tabletting, suspending, extruding, spray-drying, micronizing, emulsifying, (nano / micro)encapsulating, entrapping, or lyophilizing processes. As mentioned above, the compositions of the present invention may contain one or more physiologically acceptable inactive ingredients that facilitate processing of the active molecule into preparations for pharmaceutical use.

[0326] The appropriate formulation depends on the desired route of administration.For intravenous injection, for example, the composition can be formulated in an aqueous solution, using physiologically compatible buffers, including, for example, phosphate, histidine, or citrate to adjust the formulation pH, and isotonicity agents, such as sodium chloride or dextrose, as needed.For transmucosal or nasal administration, semi-solid, liquid formulations, or patches may be preferred, and may optionally contain penetration enhancers.Such penetrants are generally known in the art.For oral administration, the compound can be formulated in liquid or solid dosage forms and as immediate or controlled / sustained release formulations.Dosage forms suitable for oral ingestion by subjects include tablets, pills, dragees, hard and soft shell capsules, liquids, gels, syrups, slurries, suspensions, and emulsions.The compound can also be formulated into rectal compositions, such as suppositories or retention enemas, containing conventional suppository bases, such as cocoa butter or other glycerides.

[0327] Solid oral dosage forms can be obtained using excipients, which can include fillers, disintegrants, binders (dry and wet), dissolution retarders, lubricants, glidants, anti-adherents, cationic exchange resins, wetting agents, antioxidants, preservatives, colorants, and flavoring agents. These excipients can be of synthetic or natural origin. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatin, magnesium carbonate, magnesium / sodium lauryl sulfate, mannitol, polyethylene glycol, polyvinylpyrrolidone, silicates, silicon dioxide, sodium benzoate, sorbitol, starch, stearic acid or its salts, sugars (i.e., dextrose, sucrose, lactose, etc.), talc, tragacanth mucilage, vegetable oils (hydrogenated), and waxes. Ethanol and water can act as granulation aids. In certain cases, for example, it is desirable to use taste masking film, gastric acid resistant film or release retarding film to coat tablet.Coloring agent, sugar and natural and synthetic polymers combined with organic solvent or water are often used to coat tablet, resulting in sugar-coated tablet.When capsule is more preferable than tablet, its drug powder, suspension or solution can be delivered in compatible hard or soft shell capsule.

[0328] In one embodiment, the compounds of the present invention can be administered topically, such as through a skin patch, semi-solid, or liquid formulation, such as a gel, (micro)emulsion, ointment, solution, (nano / micro)suspension, or foam. The penetration of the drug into the skin and underlying tissues can be controlled, for example, by the use of penetration enhancers, the appropriate selection and combination of lipophilic, hydrophilic, and amphiphilic excipients, including water, organic solvents, waxes, oils, synthetic and natural polymers, surfactants, and emulsifiers, pH adjustment, and the use of complexing agents. Other techniques, such as iontophoresis, can be used to control the skin penetration of the compounds of the present invention. Transdermal or topical administration is preferred, for example, in situations where local delivery with minimal systemic exposure is desired.

[0329] For inhalation or nasal administration, the compounds for use according to the present invention are usually conveniently delivered in the form of solution, suspension, emulsion, or semi-solid aerosol from a pressurized pack or nebulizer, using a propellant such as, for example, halogenated carbons derived from methane and ethane, carbon dioxide, or any other suitable gas.For local aerosols, hydrocarbons such as butane, isobutene, and pentane are useful.For pressurized aerosols, the appropriate dosage unit can be determined by providing a valve to deliver a metered amount.For example, gelatin capsules and cartridges can be formulated for use in inhalers or insufflators.These typically contain a powder mix of the compound and a suitable powder base such as lactose or starch.

[0330] Compounds and compositions formulated for parenteral administration by injection are usually sterile and may be presented in unit dosage forms, such as ampoules, syringes, injection pens, or multi-dose containers, the latter usually containing preservatives. The compositions may take the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulating agents such as buffers, isotonicity agents, viscosity enhancers, surfactants, suspending and dispersing agents, antioxidants, biocompatible polymers, chelating agents, and preservatives. Depending on the injection site, the vehicle may contain water, synthetic or vegetable oils, and / or organic cosolvents. In certain cases, such as with lyophilized products or concentrates, parenteral formulations are reconstituted or diluted prior to administration. Depot formulations providing controlled or sustained release of the compounds of the present invention may include injectable suspensions of nano / microparticles or nano / micro or non-micronized crystals. Polymers such as poly(lactic acid), poly(glycolic acid), or copolymers thereof, can act as controlled / sustained release matrices, in addition to others known in the art. Other depot delivery systems can be presented in the form of implants and pumps that require incision.

[0331] Suitable carriers for intravenous injection of the compounds of the present invention are well known in the art and include, for example, aqueous solutions containing a base such as sodium hydroxide to form ionizable compounds, sucrose or sodium chloride as an isotonic agent, and buffers such as phosphate or histidine-containing buffers. Cosolvents such as polyethylene glycol may be added. These aqueous systems are effective in dissolving the compounds of the present invention and result in low toxicity upon systemic administration. The proportions of the components of the solution system may vary considerably without destroying the solubility and toxicity characteristics. Furthermore, the identity of the components may be varied. For example, low-toxicity surfactants such as polysorbates or poloxamers may be used, as may polyethylene glycol or other cosolvents, biocompatible polymers such as polyvinylpyrrolidone, and other sugars and polyols may be substituted for dextrose.

[0332] A therapeutically effective dose can be initially estimated using various techniques known in the art. Initial doses used in animal studies may be based on effective concentrations established in cell culture assays. A suitable dosage range for human subjects can be determined, for example, using data obtained from animal studies and cell culture assays. In certain embodiments, the compounds of the present disclosure are formulated for oral administration. An exemplary dose of a compound of the present disclosure in a pharmaceutical formulation for oral administration is about 0.5 to about 10 mg / kg of subject body weight. In some embodiments, the pharmaceutical formulation contains about 0.7 to about 5.0 mg / kg of subject body weight, or alternatively, about 1.0 to about 2.5 mg / kg of subject body weight. Typical dosing regimens for oral administration are three times a week, twice a week, once a week, or daily administration of the pharmaceutical formulation for oral administration.

[0333] The effective amount or therapeutically effective amount or dose of a drug, for example, a compound of the present invention, refers to the amount of drug or compound that can improve symptoms or prolong survival in a subject.The toxicity and therapeutic efficacy of such molecules can be determined by standard pharmaceutical procedures in cell culture or experimental animals, for example, by determining LD50 (the dose that is lethal to 50% of the population) and ED50 (the dose that is therapeutically effective in 50% of the population).The dose ratio of toxicity to therapeutic effect is therapeutic index, and this can be expressed as the ratio of LD50 / ED50.Preferably, a drug exhibits a high therapeutic index.

[0334] An effective or therapeutically effective amount is the amount of a compound or pharmaceutical composition that elicits the biological or medical response in a tissue, system, animal, or human that is desired by a researcher, veterinarian, physician, or other clinician. The dosage is particularly within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending on the dosage form used and / or the route of administration utilized. The exact formulation, route of administration, dosage, and administration interval must be selected according to methods known in the art, taking into account the specifics of the subject's condition.

[0335] The dosage and interval can be individually adjusted to provide a plasma level of the active moiety that is sufficient to achieve the desired effect, i.e., the minimum effective concentration (MEC). The MEC varies for each compound, but can be estimated, for example, from in vitro data and animal experiments. The dosage required to achieve the MEC depends on individual characteristics and the route of administration. In the case of local administration or selective uptake, the effective local concentration of the drug may not be related to the plasma concentration.

[0336] The amount of compound or composition administered will 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.

[0337] The compounds and compositions may, if desired, be presented in a pack or dispenser device containing one or more unit dosage forms containing the active ingredient. Such a pack or device may, for example, comprise a metal or plastic foil, such as a blister pack, or a glass and rubber stopper, such as that found in vials. The pack or dispenser device may also be accompanied by instructions for administration. Compositions containing the compounds of the present invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for the treatment of the indicated condition.

[0338] 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. [Example]

[0339] Synthesis of exemplary compounds Example 1: Preparation of Compound 1 6-Chloronicotinic acid tert-butyl ester [ka]

[0340] 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).

[0341] 6-Hydrazinylnicotinic acid tert-butyl ester [ka]

[0342] 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-hydrazinylnicotinate (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).

[0343] Ethyl (E)-3-(dimethylamino)-2-(p-tolyl)acrylate [ka]

[0344] 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 mixture was stirred at 100° C. for 3 hours 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 ethyl (E)-3-(dimethylamino)-2-(p-tolyl)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 sufficiently pure to be used directly in the next step.

[0345] 6-(5-hydroxy-4-(p-tolyl)-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0346] To a solution of (E)-3-(dimethylamino)-2-(p-tolyl)ethyl acrylate (50.00 mg, 2.14 mmol) and tert-butyl 6-hydrazinylnicotinate (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 afford tert-butyl 6-(5-hydroxy-4-(p-toyl)-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 min (Method A). 1 HNMR (400MHz, DMSO-d6) 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).

[0347] Example 2: Preparation of Compound 2 (E)-2-(4-bromophenyl)-3-(dimethylamino)acrylate ethyl ester [ka]

[0348] To a solution of ethyl 2-(4-bromophenyl)acetate (1.01 g, 4.15 mmol) in 20.0 mL of dimethylformamide was added 2.47 g of dimethylformamide diethyl acetal (20.7 mmol). The mixture was stirred at 100° C. for 16 hours 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 ethyl (E)-2-(4-bromophenyl)-3-(dimethylamino)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 sufficiently pure to be used directly in the next step.

[0349] 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0350] To a solution of (E)-ethyl 2-(4-bromophenyl)-3-(dimethylamino)acrylate (300.0 mg, 1.01 mmol) and tert-butyl 6-hydrazinylnicotinate (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). 1HNMR (400MHz, DMSO-d6) δ 13.13 (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).

[0351] Example 3: Preparation of Compound 3 6-(4-(4-bromophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0352] 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 allowed to stir 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 (400MHz, DMSO-d6) 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).

[0353] Example 4: Preparation of Compound 4 Ethyl (E)-3-(dimethylamino)-2-(4-chlorophenyl)acrylate [ka]

[0354] 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 ethyl (E)-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 sufficiently pure to be used directly in the next step.

[0355] 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0356] To a solution of (E)-3-(dimethylamino)-2-(4-chlorophenyl)ethyl acrylate (0.83 mg, 3.95 mmol) and tert-butyl 6-hydrazinylnicotinate (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 (400MHz, DMSO-d6) 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).

[0357] Example 5: Preparation of Compound 5 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0358] 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 min (Method A). 1 HNMR (400MHz, DMSO-d6) 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).

[0359] Example 6: Preparation of Compound 6 Ethyl (E)-3-(dimethylamino)-2-(4-fluorophenyl)acrylate [ka]

[0360] To a solution of ethyl 2-(4-fluorophenyl)acetate (2.0 g, 10.98 mmol) in 5.0 mL of dimethylformamide was added 8.07 g of dimethylformamide diethyl acetal (5.0 mL). 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 ethyl (E)-3-(dimethylamino)-2-(4-fluorophenyl)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 min (Method B). The product was sufficiently pure to be used directly in the next step.

[0361] 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0362] To a solution of (E)-3-(dimethylamino)-2-(4-fluorophenyl)ethyl acrylate (567.0 mg, 2.39 mmol) and tert-butyl 6-hydrazinylnicotinate (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 (400MHz, DMSO-d6) δ8.91 (s, 1H), 8.41 (s, 3H), 7.93 (s, 2H), 7.19 (s, 2H), 1.57 (s, 9H).

[0363] Example 7: Preparation of Compound 7 6-(4-(4-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0364] 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 min (Method A). 1 HNMR (400MHz, DMSO-d6) δ8.95 (s, 1H), 8.45 (s, 3H), 7.94 (s, 2H), 7.20-7.17 (m, 2H).

[0365] Example 8: Preparation of Compound 8 2-(4-cyano-2-methylphenyl)acetic acid ethyl ester [ka]

[0366] 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 h. 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) + .

[0367] (E)-2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate ethyl ester [ka]

[0368] To a solution of ethyl 2-(4-cyano-2-methylphenyl)acetate (1.0 g, 5.0 mmol) in 10.0 mL of dimethylformamide was added 2.9 g of dimethylformamide diethyl acetal (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 ethyl (E)-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) + .

[0369] 6-Chloronicotinic acid tert-butyl ester [ka]

[0370] 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) + .

[0371] 6-Hydrazinylnicotinic acid tert-butyl ester [ka]

[0372] 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-hydrazinylnicotinate (5.0 g, 92.76% yield) as a yellow solid. LC-MS: m / z 210.0 (M+H) + .

[0373] 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0374] To a solution of (E)-ethyl 2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate (260 mg, 1.0 mmol) and tert-butyl 6-hydrazinylnicotinate (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) + .

[0375] Example 9: Preparation of Compound 9 Ethyl-3-acetoxy-2-(4-cyanophenyl)but-2-enoate [ka]

[0376] Under a N2 atmosphere, LHMDS (28.00 mL, 27.75 mmol) was added dropwise over 20 minutes to a mixture of ethyl 2-(4-cyanophenyl)acetate (3.50 g, 18.50 mmol) in anhydrous THF (100 mL) at -30 °C. The reaction was warmed to 0 °C and stirred at 0 °C for 30 minutes. The reaction was then cooled again to -30 °C, and a solution of acetyl chloride (2.18 g, 27.75 mmol) in THF (8 mL) was added dropwise over 15 minutes. The reaction was gradually warmed to room temperature and stirred for 4 hours. After the reaction was complete by TLC, the mixture was quenched with cold aqueous NH4Cl (50 mL) and extracted with EtOAc (3 x 80 mL). The combined organic layers were dried over anhydrous Na2SO4 (30 g), 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. 1H NMR δ ppm(300MHz,CDCl3)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).

[0377] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0378] To a mixture of ethyl 2-(4-cyanophenyl)-3-oxobutanoate (0.30 g, 1.38 mmol) in HOAc (24 ml) was added 4-hydrazinylbenzoic acid (1.06 g, 6.91 mmol). The mixture was stirred at 100° C. overnight. After completion of the reaction by TLC, the reaction was quenched with water (20 mL), causing a large amount of solid to precipitate. After filtration, the solid was dissolved in TEA (5 eq.), water (20 mL), and MeOH (20 mL). The solution was extracted with EtOAc (3×10 mL), and the pH of the aqueous phase was adjusted to 3 with diluted HCl solution. A large amount of solid precipitated. The solid was collected by filtration and dried to give the desired product (128 mg) as a yellow solid. LC-MS (ESI+): m / z 321 (M+H) + ;HPLC purity is 97.3%; 1 H NMR(300MHz,DMSO-d6)δ ppm 13.25(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).

[0379] Example 10: Preparation of Compound 10 6-Hydrazinylnicotinic acid tert-butyl ester [ka]

[0380] To a solution of tert-butyl 6-chloronicotinate (0.20 g, 0.94 mmol) in 1,4-dioxane (10 mL) was added hydrazine (0.35 g, 5.62 mmol) in one portion. The mixture was stirred at 80° C. overnight. After the reaction was complete by TLC analysis, the mixture was concentrated in vacuo. The residue was purified by preparative HPLC to give 180 mg of the title compound (180 mg) as an oil. 1 H NMR(300MHz, CDCl3)δ ppm 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).

[0381] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0382] 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-hydrazinylnicotinate. LC-MS (ESI-): m / z 375 (MH) - ;HPLC purity 96.3%; 1 H NMR(300MHz,CDCl3)δ ppm 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).

[0383] Example 11: Preparation of Compound 11 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid methyl ester [ka]

[0384] 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 methyl 6-hydrazinylnicotinate. LC-MS: m / z= 335 (M+H) + ;1H NMR(300MHz,DMSO-d6)δ ppm 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).

[0385] Example 12: Preparation of Compound 12 Di-tert-butyl 2-(4-cyano-2,5-difluorophenyl)malonate [ka]

[0386] Under a nitrogen atmosphere, di-tert-butyl malonate (8.26 g, 31.19 mmol) was added dropwise over 15 minutes to a solution of sodium hydride (60% dispersion in mineral oil) (5.09 g, 127.31 mmol) in anhydrous DMF (100 mL) at 0 °C. After stirring the reaction 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, followed by stirring at 60 °C overnight. After the reaction was complete by TLC analysis, the mixture was quenched with saturated aqueous NH4Cl (800 mL) and extracted with EtOAc (200 mL × 3). The combined organic phase was washed with water (100 mL × 2), dried over anhydrous Na2SO4 (50 g), 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)+ .

[0387] 2-(4-cyano-2,5-difluorophenyl)acetic acid [ka]

[0388] 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 TFA (35 mL) in one portion. The reaction was stirred at room temperature overnight. After the reaction was complete 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 the crude title compound was obtained as a white solid, which was used in the next step without further purification. LC-MS (ESI+): m / z 216 (M+H+H2O). + .

[0389] 2-(4-cyano-2,5-difluorophenyl)acetic acid methyl ester [ka]

[0390] 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 (4.0 mL) over 5 minutes at room temperature. The reaction was stirred at 70° C. for 3 hours. After the reaction was complete 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×3). The combined organic phases were dried over anhydrous NaSO (50 g), 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+H0) + .

[0391] (E)-2-(4-cyano-2,5-difluorophenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0392] The compound was synthesized according to the procedure for the preparation of (E)-ethyl 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) + .

[0393] 6-(4-(4-cyano-2,5-difluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0394] The compound was synthesized according to the procedure for the preparation of 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 is 95.4%, 1 H NMR(300MHz,DMSO-d6)δ ppm 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).

[0395] Example 13: Preparation of Compound 13 2-(3-fluoro-4-hydroxyphenyl)acetic acid methyl ester [ka]

[0396] To a mixture of 2-(3-fluoro-4-hydroxyphenyl)acetic acid (2.0 g, 11.76 mmol) in MeOH (20 ml) was added concentrated HSO (0.2 mL). The reaction was stirred at 65 °C for 2 hours. After the reaction was complete by TLC, the mixture was quenched with water (40 mL) and extracted with EtOAc (3 x 50 mL). The combined organic phases were dried, filtered, and directly concentrated to give the title product (2.49 g) as an oil. 1 H NMR (300MHz, CDCl3) δ ppm 7.03 (d, J = 1.5 Hz, 1H), 6.91 (dd, J = 3.9, 1.5 Hz, 2H), 3.70 (s, 3H), 3.54 (s, 2H).

[0397] 2-(3-fluoro-4-(((trifluoromethyl)sulfonyl)oxy)phenyl)methyl acetate [ka]

[0398] To a mixture of methyl 2-(3-fluoro-4-hydroxyphenyl)acetate (2.49 g, 13.50 mmol) in DCM (52 mL) under a N atmosphere was added trifluoromethanesulfonic anhydride (5.71 g, 20.25 mmol) dropwise over 15 minutes at 0 °C. 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 complete by TLC, the mixture was quenched with aqueous NaHCO (30 mL) and extracted with DCM (3 × 50 mL). The combined organic layers were dried over anhydrous NaSO (50 g), 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(300MHz, CDCl3)δ ppm 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).

[0399] 2-(4-cyano-3-fluorophenyl)acetic acid methyl ester [ka]

[0400] 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) under a N atmosphere, Zn(CN) (0.49 g, 4.14 mmol) and Pd(PPh) (0.80 g, 0.69 mmol) were added. The reaction was stirred at 80 °C for 4 h. After the reaction was complete by TLC, the mixture was quenched with water (50 mL) and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with water (2 × 20 mL), dried over anhydrous NaSO (50 g), 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 (300MHz, CDCl3) δ ppm 7.59 (t, J = 7.5 Hz, 1H), 7.20 (d, J = 8.7 Hz, 2H), 3.73 (s, 3H), 3.70 (s, 2H).

[0401] (E)-2-(4-cyano-3-fluorophenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0402] The compound was synthesized according to the procedure for the preparation of (E)-ethyl 2-(4-cyano-2-methylphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-cyano-3-fluorophenyl)acetate. LC-MS (ESI+): m / z 249 (M+H) + .

[0403] 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0404] The compound was synthesized according to the procedure for the preparation of 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(300MHz,DMSO-d6)δ ppm 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).

[0405] Example 14: Preparation of Compound 14 5-Fluoro-6-hydrazinyl nicotinic acid [ka]

[0406] To a solution of 6-chloro-5-fluoronicotinic acid (0.46 g, 2.59 mmol) in THF (22 mL) was added N2H4H2O (0.81 g, 12.95 mmol) dropwise over 1 min. The mixture was stirred at 65 °C overnight, after which a large amount of solid precipitated. The suspension was filtered, and the solid was slurried in MeOH (3 mL) for 1 h. After filtration, 440 mg of the desired product was obtained as a white solid. LC-MS (ESI+): m / z 172 (M+H) + .

[0407] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)-5-fluoronicotinic acid [ka]

[0408] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using 5-fluoro-6-hydrazinylnicotinic acid and (E)-2-(4-cyanophenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 325 (M+H) + ; 1 H-NMR(300MHz,DMSO-d6)δ ppm 8.78(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).

[0409] Example 15: Preparation of Compound 15 2-(4-chlorophenyl)-3-oxobutanoic acid methyl ester [ka]

[0410] Under a nitrogen atmosphere, LHMDS (8.1 mL, 8.11 mmol) was added dropwise over 10 minutes to a solution of methyl 2-(4-chlorophenyl)acetate (1.00 g, 5.41 mmol) in anhydrous THF (50 mL) at −40° C. The resulting mixture was stirred at −40° C. for 1 hour, and then 1-(1H-imidazol-1-yl)ethan-1-one (0.89 g, 8.11 mmol) was added portionwise over 10 minutes. After the addition, the mixture was warmed to room temperature and stirred for 1 hour. After the reaction was complete as indicated by TLC analysis, the residue was diluted with water (50 mL) and extracted with ethyl acetate (50 mL × 3). The combined organic phases were dried over NaSO (30 g), filtered, and concentrated to dryness to give the desired product (1.6 g) as an oil. LC-MS (ESI+): m / z 249 (M+Na). + .

[0411] 6-(4-(4-chlorophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0412] 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 methyl 2-(4-chlorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 330 (M+H) + ; 1 H-NMR(300MHz,DMSO-d6)δ ppm 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).

[0413] Example 16: Preparation of Compound 16 2-(4-(benzyloxy)-3-methylphenyl)acetic acid methyl ester [ka]

[0414] 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 Cs2CO3 (1.66 g, 5.10 mmol). The mixture was stirred at room temperature for 1 hour. After the reaction was complete by TLC analysis, the mixture was quenched with water (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were dried over anhydrous Na2SO4 (30 g), filtered, and concentrated in vacuo. The residue was purified by silica column chromatography (PE: EtOAc = 100:1 to 50:1) to give 498 mg of the title compound. 1H NMR(300MHz, CDCl3)δ ppm 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).

[0415] (Z)-2-(4-(benzyloxy)-3-methylphenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0416] The compound was synthesized according to the procedure for the preparation of 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 + ).

[0417] 6-(4-(4-(benzyloxy)-3-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0418] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(4-(benzyloxy)-3-methylphenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 402 (M+H + ).

[0419] 6-(5-hydroxy-4-(4-hydroxy-3-methylphenyl)-1H-pyrazol-1-yl)nicotinic acid [ka]

[0420] 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 under a hydrogen atmosphere from a balloon for 7 h. After the reaction was complete as indicated by TLC analysis, the suspension was filtered through a package of Celite 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 H NMR(300MHz,CD3OD)δ ppm 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).

[0421] Example 17: Preparation of Compound 17 (Z)-2-(6-chloropyridin-3-yl)-3-(dimethylamino)acrylate ethyl ester [ka]

[0422] The compound was synthesized according to the procedure for the preparation of 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 (300MHz, CDCl3)ppm8.18(d,J=2.4Hz,1H),7.66(s,1H),7.51(dd,J=2.4,8.1Hz,1H),7.26(d,J=8.4Hz,1H),3.64(s,3H),2.75(s,6H).

[0423] 6-(4-(6-chloropyridin-3-yl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0424] The compound was synthesized according to the procedure for the preparation of 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).

[0425] Example 18: Preparation of Compound 18 6-Hydrazinyl-4-methylnicotinic acid [ka]

[0426] 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 the reaction was complete as shown by LC-MS, the mixture was diluted with ethanol (6 mL), causing a large amount of solid to precipitate. After filtration, 725 mg of the desired product was obtained. LC-MS (ESI+): m / z 168 (M+H + ), 1 H NMR(300MHz,DMSO-d6)δ8.44(s,1H),7.56(brs,1H),6.43(s,1H),5.90(brs,2H),2.44(s,3H)

[0427] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid [ka]

[0428] To a solution of 6-hydrazinyl-4-methylnicotinic acid (0.30 g, 1.80 mmol) in acetic acid (20 ml) was added ethyl 2-(4-cyanophenyl)-3-oxobutanoate (0.43 g, 1.98 mmol). The reaction was stirred at 100° C. overnight. After the reaction was complete as indicated by LC-MS analysis, the mixture was cooled to room temperature, causing a large amount of solid to precipitate. After filtration, the filter cake was further purified by preparative HPLC to give 36 mg of the desired product. LC-MS (ESI+): m / z 335 (M+H + ), 1 H-NMR (300MHz, DMSO-d6) 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).

[0429] Example 19: Preparation of Compound 19 6-Hydrazinyl-2-methylnicotinic acid [ka]

[0430] The compound was synthesized using 6-fluoro-2-methylnicotinic acid according to the procedure for the preparation of 6-hydrazinyl-4-methylnicotinic acid. 1 H-NMR (300MHz, D2O) δ7.63(d,J=8.7Hz,1H),6.55(d,J=8.7Hz,1H),2.37(s,3H).

[0431] 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-2-methylnicotinic acid [ka]

[0432] The compound was synthesized using 6-hydrazinyl-2-methylnicotinic acid according to the procedure for the preparation of 6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)-4-methylnicotinic acid. LC-MS (ESI+): m / z 335 (M+H) + ; 1 H-NMR (300MHz, DMSO-d6) 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).

[0433] Example 20: Preparation of Compound 20 Ethyl 2-(5-chloropyridin-2-yl)-3-oxobutanoate [ka]

[0434] To a mixture of ethyl 2-(5-chloropyridin-2-yl)acetate (500 mg, 2.5 mmol) in anhydrous THF (15 mL) under a N atmosphere was added LHMDS (5.0 mL, 5.0 mmol) dropwise over 20 minutes at −55° C. After the reaction was stirred at 0° C. for 1 hour, a solution of 1-(1H-imidazol-1-yl)ethan-1-one (412 mg, 3.75 mmol) in THF (15 mL) was added dropwise over 15 minutes at −55° C. The reaction was slowly warmed to room temperature and stirred at room temperature for 1 hour. After the reaction was complete as indicated by TLC analysis, the mixture was quenched with aqueous NH4Cl solution (50 mL) and extracted with EtOAc (3×20 mL). The combined organic phase was dried over anhydrous Na2SO4 (20 g), filtered, and concentrated in vacuo. The residue was purified by silica column chromatography (Et0Ac:Hex=1:100) to give the title compound (152 mg) as a solid. LC-MS (ESI+): m / z 242 (M+H) + .

[0435] 6-(4-(5-chloropyridin-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0436] 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 ethyl 2-(5-chloropyridin-2-yl)-3-oxobutanoate. LC-MS (ESI+): m / z 331 (M+H) + ; 1 H NMR(300MHz,DMSO-d6)δ13.20(brs,2H),8.95(d,J=1.8Hz,1H),8.53-8.61(m,1H),8.52(s,1H),8.4 1(dd,J=8.7Hz,J=2.1Hz,1H),8.32(d,J=5.1Hz,1H),7.87(dd,J=8.7Hz,J=2.7Hz,1H),2.62(s,3H).

[0437] Example 21: Preparation of Compound 21 2-(4-bromo-2-methoxyphenyl)acetic acid methyl ester [ka]

[0438] To a solution of 2-(4-bromo-2-methoxyphenyl)acetic acid (2.00 g, 8.16 mmol) in MeOH (35 ml) was added SOCl (6 mL, 82.71 mmol) dropwise over 5 minutes at room temperature. After the addition, the mixture was stirred at 55° C. overnight. After the reaction was complete as indicated by TLC analysis, the reaction mixture was directly concentrated to dryness. The residue was diluted with aqueous NaHCO (50 mL) and extracted with EtOAc (100 mL×3). The combined organic phases were dried over NaSO, filtered, and concentrated to give the desired product (2.05 g) as a yellow oil. LC-MS (ESI+): m / z 281 (M+Na) + ;1 H-NMR (300MHz, CDCl3) δ7.02-7.08(m,2H),7.00(s,1H),3.93(s,3H),3.81(s,3H),3.57(s,2H).

[0439] 2-(4-cyano-2-methoxyphenyl)acetic acid methyl ester [ka]

[0440] Under nitrogen protection, to a solution of methyl 2-(4-bromo-2-methoxyphenyl)acetate (3.02 g, 11.66 mmol) in DMF (20 ml) was added Zn(CN) (2.74 g, 23.32 mmol), Pd(dba) (0.11 g, 0.12 mmol), and S-Phos (0.48 g, 1.17 mmol). The reaction was stirred at 110 °C for 4 h. After the reaction was complete as indicated by TLC analysis, the reaction was filtered to remove undissolved solids. The filtrate was directly concentrated to dryness. The residue was purified by flash silica chromatography (EtOAc / PE = 1 / 50 to 1 / 10) to give the desired product (2.04 g) as a white solid. LC-MS (ESI+): m / z 206 (M+H) + ; 1 H-NMR (300MHz, CDCl3) δ7.23-7.30 (m, 3H), 3.86 (s, 3H), 3.71 (s, 3H), 3.67 (s, 2H).

[0441] (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0442] 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 the reaction was complete as indicated by TLC analysis, the mixture was diluted with ice water (80 mL) and extracted with EtOAc (20 mL×3). The combined organic phases were dried over Na2SO4, filtered, and concentrated to give the desired product (1.23 g) as a brown oil. LC-MS (ESI+): m / z 261 (M+H) + ;

[0443] 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0444] To a suspension of (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)methyl acrylate (0.32 g, 1.24 mmol) in i-PrOH (6 mL) was added 6-hydrazinylnicotinic acid (0.19 g, 1.24 mmol) and HCl (1.25 mL, 1 M, 1.24 mmol). The reaction was stirred at room temperature for 6 hours, after which a large amount of solid 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. Dilute HCl solution (1 M, 5 mL) was added to the reaction, and the reaction was continued to stir at room temperature for an additional 20 minutes. A large amount of solid 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 (300MHz, DMSO-d6) 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).

[0445] 6-(4-(4-cyano-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0446] Under nitrogen protection, 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) was added in one portion to EtSNa (0.11 g, 1.34 mmol). The reaction was stirred at 150° C. for 48 hours. After the reaction was complete as indicated by TLC analysis, the mixture was diluted with water (80 mL) and the pH was adjusted to 2 with diluted HCl solution. A large amount of solid precipitated. After filtration, the solid was purified by preparative HPLC purification to give 20 mg of the title compound as a yellow solid. LC-MS (ESI+): m / z 321 (M−H) - ; 1 H-NMR(300MHz,CD3OD)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)

[0447] Example 22: Preparation of Compound 22 1-chloro-2-methoxy-4-methylbenzene [ka]

[0448] To a solution of 2-chloro-5-methylphenol (1 g, 6.99 mmol) in DMF (4 mL) was added K2CO3 (2.4 g, 17.48 mmol) and MeI (1.04 g, 7.34 mmol). The reaction was stirred at room temperature for approximately 6 hours. After the reaction was complete as indicated by TLC analysis, the reaction was diluted with water (20 mL) and extracted with EtOAc (20 mL x 3). The combined organic phase was washed with water (10 mL), dried over Na2SO4, and concentrated to dryness to give 1.05 g of the desired crude product. GC-MS (EI+): 156; 1H-NMR (300MHz, CDCl3) δ7.22 (d, J=8.1Hz, 1H), 6.69-6.74 (m, 2H), 3.88 (s, 3H), 2.33 (s, 3H).

[0449] 4-(Bromomethyl)-1-chloro-2-methoxybenzene [ka]

[0450] To a solution of 1-chloro-2-methoxy-4-methylbenzene (20.98 g, 0.13 mol) in CCl4 (200 mL) was added NBS (26.24 g, 0.15 mol) and BPO (1.62 g, 6.7 mmol). The resulting mixture was stirred at 80 °C overnight. After the reaction was complete by TLC analysis, the mixture was filtered, and the filtrate was diluted with diluted HCl solution (20 mL, 1N) and extracted with DCM (200 mL × 3). The combined organic layers were washed with saturated NaHCO3 solution (40 mL), dried over Na2SO4 (60 g), filtered, and concentrated to give 41.44 g of crude product. The crude product was further purified by silica column chromatography (PE / EtOAc = 100 / :1 to 60 / 1) to give 32.6 g of the title compound. 1 H-NMR (300MHz, CDCl3) δ7.32 (d, J=8.1Hz, 1H), 6.90-6.96 (m, 2H), 4.54 (s, 2H), 3.93 (s, 3H).

[0451] 2-(4-chloro-3-methoxyphenyl)acetonitrile [ka]

[0452] To a solution of 4-(bromomethyl)-1-chloro-2-methoxybenzene (10 g, 42.7 mmol) in EtOH (50 mL) and HO (10 mL), NaCN (3.14 g, 64.05 mmol) was added in one portion. The resulting mixture was stirred at 80 °C overnight. After the reaction was complete by TLC analysis, the reaction was diluted with water (100 mL) and extracted with DCM (200 mL × 2). The combined organic phase was washed with saturated NaHCO solution (50 mL) and brine (50 mL), dried, and concentrated to give 7.58 g of 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, CDCl3) δ7.34(d,J=8.1Hz,1H),6.83-6.88(m,2H),3.91(s,3H),3.73(s,2H).

[0453] 2-(4-chloro-3-methoxyphenyl)acetic acid [ka]

[0454] To a solution of 2-(4-chloro-3-methoxyphenyl)acetonitrile (5.58 g, 31 mmol) in ethanol (120 mL) and water (40 mL), KOH (8.68 g, 155 mol) was added in one portion. The reaction was stirred at reflux for approximately 3 hours. After the reaction was complete, as indicated by TLC analysis, the reaction was concentrated to remove most of the ethanol. The residue was adjusted to pH 3 with dilute HCl solution and extracted with EtOAc (200 mL x 3). The combined organic phase was washed with water (30 mL), dried, and concentrated to dryness to give 7.29 g of crude product. 1 H-NMR (300MHz, DMSO-d6) 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).

[0455] 2-(4-chloro-3-methoxyphenyl)acetic acid methyl ester [ka]

[0456] To a solution of 2-(4-chloro-3-methoxyphenyl)acetic acid (7.29 g, 36.45 mmol) in methanol (100 mL) was added concentrated H2SO4 (3.578 g, 36.45 mmol) in one portion. The reaction was stirred at 70 °C for 2 h. After the reaction was complete, as indicated by TLC analysis, the reaction was cooled to room temperature and concentrated to remove most of the methanol. The residue was diluted with water (20 mL) and extracted with EtOAc (50 mL × 2). The combined organic layers were dried and concentrated. The residue was purified by column chromatography (PE / EtOAc = 60 / 1 to 20 / 1) to give 4.68 g of the title compound. 1 H-NMR (300MHz, CDCl3) 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).

[0457] (Z)-2-(4-chloro-3-methoxyphenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0458] The compound was synthesized according to the procedure for the preparation of 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) + ;

[0459] 6-(4-(4-chloro-3-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0460] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(4-chloro-3-methoxyphenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 346 (M+H) + ;

[0461] 6-(4-(4-chloro-3-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0462] The compound was synthesized according to the procedure for the preparation of 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, CDCl3) δ 13.30 (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).

[0463] Example 23: Preparation of Compound 23 2-(4-bromo-3-fluorophenyl)acetic acid methyl ester [ka]

[0464] 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. 1H-NMR (300MHz, CDCl3) 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).

[0465] 2-(4-cyano-3-fluorophenyl)acetic acid methyl ester [ka]

[0466] 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, CDCl3) δ7.59(t,J=8.1Hz,1H),7.19(d,J=8.7Hz,2H),3.73(s,3H),3.70(s,2H).

[0467] 2-(4-cyano-3-fluorophenyl)-3-oxobutanoic acid methyl ester [ka]

[0468] Under nitrogen protection, to a solution of methyl 2-(4-cyano-3-fluorophenyl)acetate (363 mg, 1.88 mmol) in THF (15 mL) was added LiHMDS (2.9 mmol, 2.9 mL) dropwise over 10 minutes at −78° C. After the resulting mixture was stirred at −78° C. for 30 minutes, a solution of 1-acetylimidazole (249 mg, 2.25 mmol) in THF (5 mL) was added dropwise over 10 minutes to the reaction. The resulting mixture was continued to stir at −78° C. for 1 hour. After the reaction was complete as indicated by TLC analysis, the reaction was quenched with saturated NH4Cl solution (20 mL) and extracted with EtOAc (30 mL × 2). The combined organic phases were dried and concentrated to dryness to give 410 mg of crude title compound, which was used in the next step without further purification. LC-MS (ESI-): m / z 234 (M−H) -;

[0469] 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0470] The compound was synthesized according to the procedure for the preparation of 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 (300MHz, DMSO-d6) 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).

[0471] Example 24: Preparation of Compound 24 3-Methyl-4-vinylbenzonitrile [ka]

[0472] Under nitrogen protection, to a solution of 4-bromo-3-methylbenzonitrile (5 g, 25.51 mmol) and potassium vinyltrifluoroborate (6.84 g, 51.02 mmol) in THF / water (250 mL / 25 mL) was added CsCO (33.3 g, 102.04 mmol) and Pd(dppf)Cl (1.8 g, 2.55 mmol). The resulting mixture was stirred at 70 °C overnight. After completion of the reaction as indicated by TLC analysis, the reaction was quenched with water (100 mL) and extracted with EtOAc (50 mL × 2). The combined organic layers were dried and concentrated. The residue was purified by column chromatography (PE: EtOAc = 20 / 1 to 10 / 1) to give 3.2 g of the title compound as a pale yellow oil. GC-MS (EI+): m / z 143 (M + ).

[0473] 2-(4-cyano-2-methylphenyl)acetic acid [ka]

[0474] To a solution of 3-methyl-4-vinylbenzonitrile (3.2 g, 22.38 mmol) in DME (200 mL) and water (48 mL) was added I2 (0.57 g, 2.24 mmol) in one portion. To the resulting mixture at room temperature, Oxone (27.52 g, 44.76 mmol) was added portionwise over 10 minutes. After the addition, the reaction was stirred overnight at room temperature. After the reaction was complete as indicated by TLC analysis, 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 diluted with saturated Na2SO3 (50 mL) solution and extracted with EtOAc (50 mL × 3). The combined organic layers were dried and concentrated. The residue was slurried in n-hexane (30 mL) for 30 minutes and filtered. After repeating the n-hexane slurry and filtration purification twice, 3.56 g of crude product was obtained, which was used directly in the next step without further purification.

[0475] 2-(4-cyano-2-methylphenyl)acetic acid methyl ester [ka]

[0476] 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 (300MHz, CDCl3) 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).

[0477] 2-(4-cyano-2-methylphenyl)-3-oxobutanoic acid methyl ester [ka]

[0478] The compound was synthesized using methyl 2-(4-cyano-2-methylphenyl)acetate according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 232 (M+H) + ;

[0479] 6-(4-(4-cyano-2-methylphenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0480] The compound was synthesized according to the procedure for the preparation of 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) + ; 1H-NMR (300MHz, DMSO-d6) 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).

[0481] Example 25: Preparation of Compound 25 2-(4-Bromo-2-fluorophenyl)acetic acid methyl ester [ka]

[0482] 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 (300MHz, CDCl3) δ7.24-7.27(m,2H),7.14(t,J=8.4Hz,1H),3.71(s,3H),3.63(s,2H).

[0483] 2-(4-cyano-2-fluorophenyl)acetic acid methyl ester [ka]

[0484] 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 (300MHz, CDCl3) 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).

[0485] 2-(4-cyano-2-fluorophenyl)-3-oxobutanoic acid methyl ester [ka]

[0486] The compound was synthesized using methyl 2-(4-cyano-2-fluorophenyl)acetate according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 258 (M+Na). + .

[0487] 6-(4-(4-cyano-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0488] The compound was synthesized according to the procedure for the preparation of 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 H NMR(300MHz,DMSO-d6)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).

[0489] Example 26: Preparation of Compound 26 2-(4-chloro-3-fluorophenyl)acetic acid methyl ester [ka]

[0490] 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. 1H NMR (300MHz, CDCl3) 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).

[0491] 2-(4-chloro-3-fluorophenyl)-3-oxobutanoic acid methyl ester [ka]

[0492] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate using methyl 2-(4-chloro-3-fluorophenyl)acetate. LC-MS (ESI+): m / z 267 (M+Na) + ;

[0493] 6-(4-(4-chloro-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0494] The compound was synthesized according to the procedure for the preparation of 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(300MHz,DMSO-d6)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).

[0495] Example 27: Preparation of Compound 27 4-Amino-6-hydrazinylnicotinic acid [ka]

[0496] The compound was synthesized using 4-amino-6-fluoronicotinic acid according to the procedure for the preparation of 6-hydrazinyl-4-methylnicotinic acid. LC-MS (ESI+): m / z 169 (M+H) + ;

[0497] 4-Amino-6-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0498] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using 4-amino-6-hydrazinylnicotinic acid. LC-MS (ESI+): m / z 336 (M+H) + ; 1 H-NMR (300MHz, DMSO-d6) 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).

[0499] Example 28: Preparation of Compound 28 6-(4-(4-chloro-2-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0500] The compound was synthesized according to the procedure for the preparation of 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(300MHz,DMSO-d6)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).

[0501] Example 29: Preparation of Compound 29 2-Methyl-4-vinylbenzonitrile [ka]

[0502] The compound was synthesized using 4-bromo-2-methylbenzonitrile according to the procedure for the preparation of 3-methyl-4-vinylbenzonitrile. 1 H NMR (300MHz, CDCl3) 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).

[0503] 2-(4-cyano-3-methylphenyl)acetic acid methyl ester [ka]

[0504] 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 (300MHz, CDCl3) 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).

[0505] 2-(4-cyano-3-methylphenyl)-3-oxobutanoic acid methyl ester [ka]

[0506] The compound was synthesized using methyl 2-(4-cyano-3-methylphenyl)acetate according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate. LC-MS (ESI-): m / z 230 (MH) - ;

[0507] 6-(4-(4-cyano-3-methylphenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0508] The compound was synthesized according to the procedure for the preparation of 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, CD3OD) 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).

[0509] Example 30: Preparation of Compound 30 2-(4-bromo-2-chlorophenyl)acetic acid methyl ester [ka]

[0510] 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, CDCl3) 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).

[0511] 2-(2-chloro-4-cyanophenyl)acetic acid methyl ester [ka]

[0512] The compound was synthesized using methyl 2-(4-bromo-2-chlorophenyl)acetate according to the procedure for the preparation of methyl 2-(4-cyano-2-fluorophenyl)acetate. GC-MS (EI+): m / z 209.

[0513] (Z)-2-(2-chloro-4-cyanophenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0514] The compound was synthesized according to the procedure for the preparation of 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) +

[0515] 6-(4-(2-chloro-4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0516] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(2-chloro-4-cyanophenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 341 (M+H) + ; 1 H-NMR (300MHz, DMSO-d6) 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).

[0517] Example 31: Preparation of Compound 31 3,5-Dimethyl-4-vinylbenzonitrile [ka]

[0518] The compound was synthesized using 4-bromo-3,5-dimethylbenzonitrile according to the procedure for the preparation of 3-methyl-4-vinylbenzonitrile. 1 H-NMR (300MHz, CDCl3) 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).

[0519] 2-(4-cyano-2,6-dimethylphenyl)acetic acid methyl ester [ka]

[0520] 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, CDCl3) δ7.33 (s, 2H), 3.72 (s, 2H), 3.70 (s, 3H), 2.35 (s, 6H).

[0521] (Z)-2-(4-cyano-2,6-dimethylphenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0522] The compound was synthesized according to the procedure for the preparation of methyl (Z)-2-(4-cyano-2-methoxyphenyl)-3-(dimethylamino)acrylate using methyl 2-(4-cyano-2,6-dimethylphenyl)acetate. 1 H-NMR (300MHz, CDCl3) δ7.62 (s, 1H), 7.28 (s, 2H), 3.59 (s, 3H), 2.64 (s, 6H), 2.20 (s, 6H).

[0523] 6-(4-(4-cyano-2,6-dimethylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0524] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(4-cyano-2,6-dimethylphenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 335 (M+H) + ; 1 H-NMR (300MHz, CD3OD) δ8.03 (s, 1H), 8.52 (m, 2H), 7.75 (s, 1H), 7.40-7.47 (m, 2H), 2.30 (s, 6H).

[0525] Example 32: Preparation of Compound 32 2-(4-Methoxy-3-methylphenyl)acetic acid methyl ester [ka]

[0526] 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, CD3OD) 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).

[0527] 2-(4-Methoxy-3-methylphenyl)-3-oxobutanoic acid methyl ester [ka]

[0528] The compound was synthesized according to the procedure for the preparation of methyl 2-(4-cyano-3-fluorophenyl)-3-oxobutanoate using methyl 2-(4-methoxy-3-methylphenyl)acetate. LC-MS (ESI+): m / z 259 (M+Na) + ;

[0529] 6-(5-hydroxy-4-(4-methoxy-3-methylphenyl)-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0530] The compound was synthesized according to the procedure for the preparation of 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) + ; 1H-NMR(300MHz,DMSO-d6)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).

[0531] Example 33: Preparation of Compound 33 Diethyl 2-(2-bromo-4-cyanophenyl)malonate [ka]

[0532] Under nitrogen protection, NaH (600 mg, 15 mmol) was added portionwise to a solution of diethyl malonate (2 g, 12.5 mmol) in DMF over 5 minutes at 0°C. After stirring the reaction at 0°C for 30 minutes, 3-bromo-4-fluorobenzonitrile (2.08 g, 10 mmol) was added to the reaction in one portion, and the reaction was stirred at 80°C for 1.5 hours. After the reaction was complete as indicated by TLC analysis, the reaction was quenched with water (20 mL) and the pH was adjusted to 5 with diluted HCl solution. The resulting mixture was then extracted with EtOAc (50 mL x 2). The combined organic phase was washed with water (20 mL) and brine (20 mL), dried, and concentrated to dryness. The residue was purified by column chromatography (PE / EtOAc = 20 / 1 to 10 / 1) to give 1.8 g of the title compound. LC-MS(ESI+):m / z 340,342(M+H) + . 1 H-NMR (300MHz, CDCl3) 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).

[0533] 2-(2-bromo-4-cyanophenyl)acetic acid ethyl ester [ka]

[0534] To a solution of diethyl 2-(2-bromo-4-cyanophenyl)malonate (1.8 g, 5.3 mmol) and water (96 mg, 5.3 mmol) in DMSO (20 mL) was added LiCl (339 mg, 8 mmol) in one portion. The resulting mixture was stirred at 120 °C overnight. After completion of the reaction as indicated by TLC analysis, the reaction was quenched with water (20 mL), the pH was adjusted to 6 with diluted HCl solution, and extracted with EtOAc (20 mL × 2). The combined organic phase was washed with water (20 mL) and brine (20 mL), dried, and concentrated. The residue was purified by column chromatography (PE / EtOAc = 20 / 1 to 10 / 1) to give 780 mg of the title compound. 1 H-NMR(300MHz,CDCl3)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)

[0535] 2-(4-cyano-2-vinylphenyl)acetic acid ethyl ester [ka]

[0536] 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(300MHz,CDCl3)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=17.4Hz,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).

[0537] 2-(4-cyano-2-ethylphenyl)acetic acid ethyl ester [ka]

[0538] To a solution of methyl 2-(4-cyano-2-vinylphenyl)acetate (560 mg, 2.6 mmol) in methanol (10 mL) was added Pd / C (56 mg). The suspension was stirred under a hydrogen atmosphere from a balloon for about 1 hour. After the reaction was complete as indicated by TLC analysis, the suspension was filtered through a package of Celite, and the filter cake was washed with methanol (10 mL). The filtrate was concentrated to dryness to give 540 mg of the crude title compound. 1 H-NMR(300MHz,CDCl3)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).

[0539] (Z)-2-(4-cyano-2-ethylphenyl)-3-(dimethylamino)acrylate ethyl ester [ka]

[0540] 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,CDCl3)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)

[0541] 6-(4-(4-cyano-2-ethylphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0542] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(4-cyano-2-ethylphenyl)-3-(dimethylamino)ethyl acrylate. LC-MS (ESI+): m / z 335 (M+H) + ; 1 H-NMR(300MHz,DMSO-d6)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)

[0543] Example 34: Preparation of Compound 34 5-Hydrazinylpyrazine-2-carboxylic acid [ka]

[0544] The compound was synthesized according to the procedure for the preparation of 6-hydrazinyl-4-methylnicotinic acid using 5-fluoropyrazine-2-carboxylic acid. LC-MS (ESI+): m / z 155 (M+H) + ;

[0545] 5-(4-(4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)pyrazine-2-carboxylic acid [ka]

[0546] The compound was synthesized using 5-hydrazinylpyrazine-2-carboxylic acid according to the procedure for the preparation of 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid. LC-MS (ESI+): m / z 322 (M+H) + ; 1H-NMR (300MHz, DMSO-d6) 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).

[0547] Example 35: Preparation of Compound 35 2-(2-Methoxypyridin-4-yl)acetic acid ethyl ester [ka]

[0548] To a solution of 2-methoxy-4-methylpyridine (2.0 g, 16.2 mmol) in anhydrous tetrahydrofuran (50.0 mL) was added lithium diisopropylamide (16.0 mL, 32.0 mmol, 2.0 M in n-heptane) at −78° C. under nitrogen. The mixture was stirred at −78° C. for 10 minutes, and diethyl carbonate (3.78 g, 32.0 mmol) was added. The mixture was warmed 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 afford 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).

[0549] Ethyl (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)acrylate [ka]

[0550] To a solution of ethyl 2-(2-methoxypyridin-4-yl)acetate (1.95 g, 10 mmol) in 3.0 mL of dimethylformamide was added 5.95 g of dimethylformamide diethyl acetal (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 give ethyl (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)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).

[0551] 6-(5-hydroxy-4-(2-methoxypyridin-4-yl)-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0552] To a solution of (E)-3-(dimethylamino)-2-(2-methoxypyridin-4-yl)ethyl acrylate (0.25 g, 1.0 mmol) and tert-butyl 6-hydrazinylnicotinate (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 min (Method A). 1HNMR(400MHz,DMSO-d6)δ 13.53(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).

[0553] Example 36: Preparation of Compound 36 6-Hydrazinylnicotinic acid ethyl ester [ka]

[0554] 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 was obtained as a yellow oil (600 mg, 3.31 mmol, 61.4% yield). LC-MS: m / z=182.1 (M+H). + , retention time 0.37 min (Method A). The product was sufficiently pure to be used directly in the next step.

[0555] Ethyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate [ka]

[0556] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (300 mg, 1.30 mmol) and ethyl 6-hydrazinylnicotinate (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 give 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 (400MHz, DMSO-d6) 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).

[0557] Example 37: Preparation of Compound 37 6-Chloronicotinic acid isopropyl ester [ka]

[0558] To a solution of 6-chloronicotinic acid (3.0 g, 19.2 mmol) in dichloromethane (50.0 mL) was added carbonyldiimidazole (3.42 g, 21.1 mmol) at room temperature. The mixture was stirred for 1.0 h, and isopropyl alcohol (3.78 g, 32.0 mmol) was added. Dichloromethane was removed in vacuo. A catalytic amount of sodium isopropoxide (164 mg, 2.0 mmol) was added, and the mixture was heated at 90° C. for 1.0 h. The solution was concentrated in vacuo. 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 isopropyl 6-chloronicotinate (3.4 g, 17.1 mmol, 89% yield) as a yellow solid. LC-MS: m / z=200.0 [M+H] +, retention time 2.04 min (Method A). The product was sufficiently pure to be used directly in the next step.

[0559] 6-Hydrazinylnicotinic acid isopropyl ester [ka]

[0560] 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-hydrazinylnicotinate (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).

[0561] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid isopropyl ester [ka]

[0562] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (300 mg, 1.30 mmol) and isopropyl 6-hydrazinylnicotinate (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 at 90° C. overnight and cooled to precipitate a solid. The solid was filtered, washed with ethanol, and dried to give isopropyl 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinate (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(400MHz,DMSO-d6)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).

[0563] Example 38: Preparation of Compound 38 6-Chloronicotinic acid tert-butyl ester [ka]

[0564] 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).

[0565] 6-Hydrazinylnicotinic acid tert-butyl ester [ka]

[0566] 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 h. The mixture was cooled and concentrated to dryness. The residue was partitioned between ethyl acetate and water. The organic phase was washed with brine, dried over sodium sulfate, and concentrated. The residue was triturated with petroleum ether and filtered to give tert-butyl 6-hydrazinylnicotinate (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).

[0567] 2-(4-cyanophenyl)acetic acid methyl ester [ka]

[0568] To a mixture of 2-(4-cyanophenyl)acetic acid (5.0 g, 31.0 mmol) in methanol (10.0 mL) was added hydrochloric acid in methanol (20.0 mL, 3.0 M) at 0° C. The mixture was stirred at 70° C. for 3.0 h 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).

[0569] (E)-2-(4-cyanophenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0570] 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)acrymethyl (5.20 g, 25.4 mmol, 89% yield) as a yellow solid. LC-MS: m / z=231.0 [M+H] + , retention time 1.70 min (Method A). The product was sufficiently pure to be used directly in the next step.

[0571] 6-(4-(4-cyanophenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid tert-butyl ester [ka]

[0572] To a solution of (E)-methyl 2-(4-cyanophenyl)-3-(dimethylamino)acrylate (2.0 g, 8.70 mmol) and tert-butyl 6-hydrazinylnicotinate (1.82 g, 8.70 mmol) in ethanol (20 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 give 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). 1HNMR(400MHz,DMSO-d6)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).

[0573] Example 39: Preparation of Compound 39 2-(4-chloro-2-methoxyphenyl)acetic acid methyl ester [ka]

[0574] The compound was synthesized according to the procedure for the preparation of 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, CDCl3) δ7.10 (d, J = 8.1 Hz, 1H), 6.85-6.92 (m, 2H), 3.86 (s, 3H), 3.71 (s, 3H), 3.67 (s, 2H).

[0575] (Z)-2-(4-chloro-2-methoxyphenyl)-3-(dimethylamino)acrylate methyl ester [ka]

[0576] The compound was synthesized according to the procedure for the preparation of 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) + .

[0577] 6-(4-(4-chloro-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0578] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-2-methoxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid using (Z)-2-(4-chloro-2-methoxyphenyl)-3-(dimethylamino)methyl acrylate. LC-MS (ESI+): m / z 346 (M+H) + ;

[0579] 6-(4-(4-chloro-2-hydroxyphenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0580] The compound was synthesized according to the procedure for the preparation of 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 (300MHz, DMSO-d6) 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).

[0581] Example 40: Preparation of Compound 40 Diethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)malonate [ka]

[0582] Under nitrogen protection, a solution of ethyl 3-(ethylperoxy)-3-oxopropanoate (10.20 g, 63.50 mmol) in DMF (100 ml) was added to CsCO (45.50 g, 139.70 mmol). After stirring the reaction at 70 °C for 10 minutes, 4-fluoro-3-(trifluoromethyl)benzonitrile (12.00 g, 63.50 mmol) was added to the reaction. The resulting mixture was stirred at 70 °C for 2 hours. After the reaction was complete as indicated by TLC, the reaction was quenched with water (300 mL) and extracted with EtOAc (500 mL × 2). The combined organic phase was washed with water (50 mL), dried over NaSO, filtered, and concentrated to give the desired product (20.8 g) as an oil. 1 H-NMR (300MHz, CDCl3) 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).

[0583] 2-(4-cyano-2-(trifluoromethyl)phenyl)acetic acid ethyl ester [ka]

[0584] To a solution of diethyl 2-(4-cyano-2-(trifluoromethyl)phenyl)malonate (15.00 g, 45.60 mmol) in DMSO (150 ml) was added LiCl (2.90 g, 68.40 mmol) and water (0.80 mL, 45.6 mmol). The mixture was stirred at 120° C. overnight. After the reaction was completed as indicated by TLC, the reaction was quenched with water (300 mL) and extracted with EtOAc (150 mL×3). The combined organic layer was washed with water (100 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by silica chromatography (EA / PE=1 / 25) to give the desired product (8.28 g) as a white solid. 1H-NMR (300MHz, CDCl3) 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).

[0585] (Z)-2-(4-cyano-2-(trifluoromethyl)phenyl)-3-(dimethylamino)acrylate ethyl ester [ka]

[0586] 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 at 150° C. overnight. After the reaction was complete as indicated by TLC, the mixture was diluted with water (100 mL) and extracted with EtOAc (150 mL×3). The combined organic layers were dried over NaSO, filtered, and concentrated. The residue was purified by silica chromatography (EA / PE=1 / 9) to give the desired product (3.2 g) as a brown oil. 1 H-NMR(300MHz,CDCl3)δ7.94(s,1H),7.75(d,J=7.8Hz,1H),7.59(s,1H),7.4 6(d,J=7.8Hz,1H),3.95-4.08(m,2H),2.66(brs,6H),1.10(t,J=7.2Hz,3H).

[0587] 6-(4-(4-cyano-2-(trifluoromethyl)phenyl)-5-hydroxy-1H-pyrazol-1-yl)nicotinic acid [ka]

[0588] To a solution of (Z)-ethyl 2-(4-cyano-2-trifluoromethyl)phenyl)-3-(dimethylamino)acrylate (1.00 g, 3.20 mmol) in i-PrOH (38 mL) was added 6-hydrazinylnicotinic 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 the reaction was complete as indicated by LCMS analysis, DIEA (15 equiv.) was added and the mixture was stirred at room temperature for 2 days. The mixture was adjusted to pH 3, after which a large amount of solid precipitated. After filtration and slurry purification, 90 mg of the desired product was obtained as a solid. The HPLC purity was 93.1%. LC-MS (ESI+): m / z 375 (M+H) + ; 1 H-NMR (300MHz, DMSO-d6) 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).

[0589] Example 41: Preparation of Compound 41 Ethyl 3-oxo-2-(pyridin-4-yl)butanoate [ka]

[0590] The compound was synthesized using ethyl 3-oxo-2-(pyridin-4-yl)butanoate according to the procedure for the preparation of methyl 2-(4-cyano-2-fluorophenyl)-3-oxobutanoate. LC-MS (ESI+): m / z 208 (M+H) +

[0591] 6-(5-hydroxy-3-methyl-4-(pyridin-4-yl)-1H-pyrazol-1-yl)nicotinic acid [ka]

[0592] The compound was synthesized according to the procedure for the preparation of 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,CD3OD)δ9.00(s,1H),8.24-8.33(m,4H),7.84(d,J=6.0Hz,2H),2.44(s,3H)

[0593] 4-(1-(5-carboxypyridin-2-yl)-5-hydroxy-3-methyl-1H-pyrazol-4-yl)pyridine 1-oxide [ka]

[0594] 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 m-CPBA (166 mg, 0.96 mmol) in one portion at room temperature. The resulting mixture was stirred at room temperature for about 2 hours. After the reaction was complete as indicated by HPLC analysis, 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, CD3OD) 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).

[0595] Example 42: Preparation of Compound 42 2-(3-bromo-4-chlorophenyl)acetic acid methyl ester [ka]

[0596] 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, CDCl3) 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).

[0597] 2-(4-chloro-3-cyanophenyl)acetic acid methyl ester [ka]

[0598] 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, CDCl3) δ7.61 (s, 1H), 7.48 (s, 2H), 3.73 (s, 3H), 3.64 (s, 2H).

[0599] 2-(4-chloro-3-cyanophenyl)-3-oxobutanoic acid methyl ester [ka]

[0600] 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.

[0601] 6-(4-(4-chloro-3-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0602] The compound was synthesized according to the procedure for the preparation of 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 (300MHz, DMSO-d6) 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).

[0603] Example 43: Preparation of Compound 43 2-(6-oxo-1,6-dihydropyridin-3-yl)acetic acid: [ka]

[0604] 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 give 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.

[0605] 2-(6-oxo-1,6-dihydropyridin-3-yl)methyl acetate: [ka]

[0606] 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 partitioned 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).

[0607] 2-(6-Methoxypyridin-3yl)acetic acid methyl ester [ka]

[0608] To a solution of methyl 2-(6-oxo-1,6-dihydropyridin-3-yl)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 reaction solution was diluted with ethyl acetate and water. The organic layer was washed with brine, dried over sodium sulfate, and concentrated. The residue was purified by flash chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a mixture of methyl 2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)acetate and methyl 2-(6-methoxypyridin-3-yl)acetate (400 mg, 2.21 mmol, 82.1% yield) as a yellow oil. LC-MS: m / z = 182.1 (M+H). + , retention time 1.33 min (Method A).

[0609] Methyl (E)-3-(dimethylamino)-2-(6-methoxypyridin-3-yl)acrylate [ka]

[0610] To a solution of methyl 2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)acetate and methyl 2-(6-methoxypyridin-3-yl)acetate (400 mg, 2.21 mmol) in N,N-dimethylformamide (5.0 mL) was added N,N-dimethylformamide diethyl acetal (1.63 g, 11.1 mmol). 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 give a mixture of methyl (E)-3-(dimethylamino)-2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)acrylate and methyl (E)-3-(dimethylamino)-2-(6-methoxypyridin-3-yl)acrylate (400 mg, 1.69 mmol, 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 in the next step.

[0611] tert-Butyl 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinate [ka]

[0612] A mixture of methyl (E)-3-(dimethylamino)-2-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)acrylate and methyl (E)-3-(dimethylamino)-2-(6-methoxypyridin-3-yl)acrylate (350 mg, 1.48 mmol) in ethanol (5.0 mL) was stirred in a sealed tube at 90° C. for 16.0 h and cooled. The insoluble solid was filtered, and the filtrate was concentrated to dryness. The two isomers were separated by reverse-phase preparative HPLC as a white solid. tert-Butyl 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)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(500MHz,DMSO-d6)δ12.97(br,1H),8.91(d,J=1.5Hz,1H),8.70(s,1H),8.4 7-8.39(m,2H),8.19-8.14(m,1H),6.85-6.83(m,1H),3.85(s,3H),1.58(s,9H).

[0613] 6-(5-hydroxy-4-(6-methoxypyridin-3-yl)-1H-pyrazol-1-yl)nicotinic acid [ka]

[0614] 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 overnight at room temperature 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 minutes (Method A). 1 HNMR (400MHz, DMSO-d6) 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).

[0615] Example 44: Preparation of Compound 44 Diethyl 2-(3-chloro-4-cyanophenyl)malonate [ka]

[0616] 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,CDCl3)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).

[0617] 2-(3-chloro-4-cyanophenyl)acetic acid ethyl ester [ka]

[0618] 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,CDCl3)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).

[0619] Ethyl (E)-3-acetoxy-2-(3-chloro-4-cyanophenyl)but-2-enoate [ka]

[0620] The compound was synthesized according to the procedure for the preparation of 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) + .

[0621] 6-(4-(3-chloro-4-cyanophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid [ka]

[0622] The compound was synthesized according to the procedure for the preparation of 6-(4-(4-cyano-3-fluorophenyl)-5-hydroxy-3-methyl-1H-pyrazol-1-yl)nicotinic acid using ethyl (E)-3-acetoxy-2-(3-chloro-4-cyanophenyl)but-2-enoate. LC-MS (ESI+): m / z 355 (M+H) + ; 1 H-NMR(300MHz,DMSO-d6)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).

[0623] Example 45: In vitro assay demonstrates PHD inhibition Enzymatic half-maximal inhibitory concentration (IC 50 ) values ​​were determined on selected compounds of the present invention.

[0624] Time-resolved fluorescence resonance energy transfer (TR-FRET) assays were utilized to determine the enzymatic half-maximal inhibitory concentrations (IC ) of PHD inhibitors against full-length human prolyl-4-hydroxylase domain (PHD) enzymes, PHD1, PHD2, and PHD3. 50The TR-FRET assay was developed based on the specific binding of hydroxylated HIF-1α peptide to a complex formed by VHL, EloB, and EloC (VBC), generating a fluorescent signal. The TR-FRET terbium (Tb)-donor (anti-monoclonal antibody, anti-6His-Tb-cryptate gold) and D2-acceptor (streptavidin [SA]-D2) are linked to the VBC complex and HIF-1α peptide, respectively. The VBC complex specifically binds to the HIF-1α peptide when hydroxylated, allowing energy transfer from the TR-FRET donor to the acceptor (Figure 1).

[0625] material and method Unless otherwise noted, all chemicals and materials were of standard laboratory grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).

[0626] reagent TR-FRET Reagents 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).

[0627] An N-terminally biotinylated HIF-1α C35 synthetic peptide representing amino acids 547–581 and containing the proline 564 PHD2 hydroxylation site was purchased from California Peptide Research (Salt Lake City, UT, USA).

[0628] Recombinant proteins VBC complex His-tagged recombinant VHL protein, EloB, and EloC complex (His-VBC) was provided by Axxam (Milan, Italy). Recombinant human VHL (National Center for Biotechnology Information [NCBI] accession number NP_00542.1) contained a C-terminal His tag at amino acids 55–213 and was designated VHL-His. VHL-His was coexpressed with full-length human EloB (NCBI accession number Q15370.1) and full-length human EloC (NCBI accession number Q15369.1) in E. coli and purified as a His-VBC complex by affinity chromatography on a nickel-nitrilotriacetic acid (Ni-NTA) column. Purity (approximately 80%) was assessed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).

[0629] PHD1 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 (NCBI accession no. NP_542770.2) with an N-terminal FLAG tag (molecular weight 44.9 kDa). Purity (>90%) was assessed by SDS-PAGE.

[0630] PHD2 Full-length human PHD2 enzyme was produced using a baculovirus-infected insect cell (BIIC) expression system from Beryllium (Bedford, MA, USA). The PHD2 construct contained amino acids 1–426 of PHD2 (UniProt Knowledgebase [UniProtKB] / Swiss-Prot accession number Q9GZT9.1) and an N-terminal His tag and tobacco etch virus (TEV) protease cleavage site. The construct was expressed in Sf9 insect cells, purified through a 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.

[0631] PHD3 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 (NCBI accession number NP_071356.1) with an N-terminal 6-His tag (catalog number 81033, lot number 24417001). Purity was assessed by SDS-PAGE and found to be >75% pure.

[0632] PHD inhibitors. Small molecule PHD inhibitors were synthesized and their identities confirmed as described herein.

[0633] TR-FRET assay procedure PHD inhibitor compounds were pre-incubated with PHD enzymes in a 10 μL reaction volume in a white 384-well Optiplate microplate (catalog no. 6007290, Perkin Elmer, Waltham, MA, USA). 5 μL of PHD inhibitor compound was serially diluted in 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 mix (60 nM PHD1, 20 nM PHD2, 140 nM PHD3) prepared as a 4x concentrate in dilution buffer containing PHD enzymes, 40 μM ferrous ammonium sulfate (FAS), and 4 mM sodium ascorbate (Na). The plate was incubated for 30 minutes at room temperature without rotation.

[0634] Next, 5 microliters of VBC / anti-6His-Tb-cryptate gold mix (prepared as a 4-fold concentrate in dilution buffer containing 20 nM His-VBC, 1.32 nM monoclonal antibody, and anti-6His-Tb-cryptate gold) was added. This step was immediately followed by the addition of 5 μL of HIF-1α C35 substrate mix (prepared as a 4-fold concentrate in dilution buffer containing 120 nM biotin-labeled HIF-1α C35, 132 nM SA-D2, and 4 μM 2-oxoglutarate (2-OG)) to reach a final reaction volume of 20 μL.

[0635] The final assay reaction contained 50 mM HEPES, pH 7.5, 50 mM NaCl, 1 μM 2-OG, 10 μM FAS, 1 mM Na-ascorbate, 0.01% Tween-20, 0.01% purified BSA, 30 nM biotin-labeled 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) along with diluted compound.

[0636] IC of PHD inhibitor compounds 50 For IC measurements, reactions were incubated at room temperature for 10 minutes and then read on a Perkin Elmer EnVision (Waltham, MA, USA) at an excitation wavelength of 340 nm and emission wavelengths of 615 and 665 nm. Data represent the quotient of the signal intensities at 665 nm and 615 nm, calculated automatically by Envision Manager software (Perkin Elmer, Waltham, MA, USA). 50 Values ​​(mean, standard deviation, standard error of the mean, geometric mean, and 95% confidence interval) were determined using a four-parameter curve fit using GraphPad Prism 7.0 (GraphPad, La Jolla, CA, USA) and represent compound concentration plotted against the calculated ratio of 665 nm and 615 nm. TR-FRET assays were performed in triplicate at each compound concentration, and assays were repeated three times independently.

[0637] Ki is calculated based on the following Cheng-Prusoff equation: 50 It was calculated from. Ki = IC50 / (1+[2-OG] / Km)

[0638] The final concentration of 2-OG in both the PHD1 and PHD2 assays is 1 uM. The Km of 2-OG for PHD1 was determined to be 12.7 nM, while the Km of 2-OG for PHD2 was determined to be 22.6 nM. Exemplary Compounds [Table 4] TIFF0007768890000277.tif245170TIFF0007768890000278.tif244170TIFF0007768890 000279.tif245170TIFF0007768890000280.tif245170TIFF0007768890000281.tif59170

[0639] From the present description, those skilled in the art can easily ascertain the essential characteristics of the present invention, and can make various changes and modifications to the present invention to adapt it to various uses and conditions without departing from its spirit and scope.

[0640] All U.S. or foreign references, patents, or applications cited in this application are hereby incorporated by reference in their entirety as if set forth herein. In the event of any conflict, the material literally disclosed herein will control. The present application provides the following aspects of the invention. (Aspect 1) A compound of formula A, (chemical 1) TIFF0007768890000282.tif25170 or a pharmaceutically acceptable salt thereof, wherein: Ar 1 is phenyl or a 6-membered nitrogen-containing heteroaryl, wherein said phenyl or heteroaryl is selected from the group consisting of halogen, CN, OH, C optionally substituted with one or more halogens, 1-3 Alkyl, or C 1-3 optionally substituted with alkoxy; R 2 is H or C 1 - 3 is alkyl, Ar 2 is a halogen, OH, amine, or C 1-3 a 6-membered nitrogen-containing heteroaryl optionally substituted with alkyl; R 4 is hydrogen or C 1-4 is alkyl, Formula (A) represents the following compound: (Case 2) TIFF0007768890000283.tif108170 or a pharmaceutically acceptable salt thereof, excluding: (Aspect 2) Ar 1 teeth, (3) TIFF0007768890000284.tif19170 where: X, Y, and Z are independently CH or N, wherein N is optionally oxidized; Each R 1 are independently hydrogen, halogen, CN, OH, C optionally substituted with one or more halogens 1-3 Alkyl, and C 1-3 alkoxy; The compound of embodiment 1, wherein m is 1, 2, 3, or 4. (Aspect 3) Ar 1 at least one R is CN or halogen 1 The compound according to any one of the preceding aspects, wherein R is phenyl substituted by (Aspect 4) Ar 1 C optionally substituted with one or more halogens 1-3 one or two R independently selected from alkyl, halogen, CN, or OH; 1 The compound of embodiment 3, wherein the compound is substituted with a group. (Aspect 5) Ar 1 is a pyridyl N-oxide, or C 1-3 At least one R is alkoxy or halogen 1 The compound according to any one of embodiments 1 to 2, wherein the compound is pyridyl optionally substituted by: (Aspect 6) Ar 2 but, (C4) TIFF0007768890000285.tif24170 where: A and B are independently CH or N, wherein N is optionally oxidized; Each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 is selected from the group consisting of alkyl, The compound according to any one of embodiments 1 to 5, wherein n is 0, 1, or 2. (Aspect 7) Ar 2 is a pyridyl or pyrazinyl group, said group being unsubstituted or containing halogen, C 1-3 A compound according to any one of embodiments 1 to 6, comprising a substituent which is alkyl, or OH. (Aspect 8) R 2 is H or CH 3 A compound according to any one of aspects 1 to 7, wherein (Aspect 9) R 4 The compound according to any one of embodiments 1 to 8, wherein is H. (Aspect 10) R 4 But C 1-4 The compound according to any one of embodiments 1 to 8, wherein the compound is alkyl. (Aspect 11) A structure according to formula (I): (C5) TIFF0007768890000286.tif28170 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 are independently 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 is alkyl, Each R 3 are independently hydrogen, halogen, OH, amine, and C 1-3 is selected from the group consisting of alkyl, R 4 is hydrogen or C 1-4 The compound according to embodiment 1, wherein the aryl group is alkyl. (Aspect 12) The structure of formula (Ia): (6) TIFF0007768890000287.tif29170 or a pharmaceutically acceptable salt thereof. (Aspect 13) A structure of formula (Ib): (7) TIFF0007768890000288.tif33170 or a pharmaceutically acceptable salt thereof. (Aspect 14) The structure of formula (Ic): (8) TIFF0007768890000289.tif33170 or a pharmaceutically acceptable salt thereof. (Aspect 15) A structure according to formula (II): (9) TIFF0007768890000290.tif37170 or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3, or 4; n is 0, 1, or 2; R 1 are independently each time 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 is alkyl, R 3 are each independently hydrogen, halogen, OH, amine, or C 1-3 is selected from the group consisting of alkyl, R 4 is hydrogen or C 1-4 The compound according to embodiment 1, wherein the aryl group is alkyl. (Aspect 16) The structure of formula (IIa): (C10) TIFF0007768890000291.tif41170 or a pharmaceutically acceptable salt thereof. (Aspect 17) The structure of formula (IIb): (Chem.11) TIFF0007768890000292.tif36170 or a pharmaceutically acceptable salt thereof. (Aspect 18) The structure of formula (IIc): (C12) TIFF0007768890000293.tif33170 or a pharmaceutically acceptable salt thereof. (Aspect 19) The structure of formula (IId): (C13) TIFF0007768890000294.tif33170 or a pharmaceutically acceptable salt thereof. (Aspect 20) The structure of formula (IIe): (C14) TIFF0007768890000295.tif33170 or a pharmaceutically acceptable salt thereof. (Aspect 21) The structure of formula (IIf): (C15) TIFF0007768890000296.tif30170 or a pharmaceutically acceptable salt thereof. (Aspect 22) The structure of formula (IIg): (C16) TIFF0007768890000297.tif32170 or a pharmaceutically acceptable salt thereof. (Aspect 23) A structure according to formula III: (C17) TIFF0007768890000298.tif32170 or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3, or 4; n is 0, 1, or 2; R 1 independently each time hydrogen, halogen, OH, and C optionally substituted with one or more halogens 1-3 is selected from the group consisting of alkyl, R 2 is hydrogen or C 1 - 3 is alkyl, R 3 are each independently hydrogen, halogen, OH, amine, and C 1-3 is selected from the group consisting of alkyl, R 4 is hydrogen or C 1-4 is alkyl, R 5 is CN or halogen. (Aspect 24) The structure of formula (IIIa): (C18) TIFF0007768890000299.tif41170 or a pharmaceutically acceptable salt thereof. (Aspect 25) The structure of formula (IIIb): (C19) TIFF0007768890000300.tif42170 or a pharmaceutically acceptable salt thereof. (Aspect 26) The structure of formula (IIIc): (20) TIFF0007768890000301.tif42170 or a pharmaceutically acceptable salt thereof. (Aspect 27) The structure of formula (IIId): (21) TIFF0007768890000302.tif40170 or a pharmaceutically acceptable salt thereof. (Aspect 28) The structure of formula (IIIe): (22) TIFF0007768890000303.tif36170 or a pharmaceutically acceptable salt thereof. (Aspect 29) A structure of formula (IIIf): (23) TIFF0007768890000304.tif35170 or a pharmaceutically acceptable salt thereof. (Aspect 30) A structure of formula III(g): (24) TIFF0007768890000305.tif29170 or a pharmaceutically acceptable salt thereof. (Aspect 31) The compound of embodiment 11, 12, or 14, wherein X is CH. (Aspect 32) The compound of embodiment 11, 12, or 14, wherein X is N. (Aspect 33) 33. The compound according to embodiment 32, wherein N is optionally oxidized. (Aspect 34) The compound of embodiment 11, 12, or 14, wherein Y is CH. (Aspect 35) The compound of embodiment 11, 12, or 14, wherein Y is N. (Aspect 36) The compound of embodiment 11, 12, or 14, wherein Z is CH. (Aspect 37) The compound of embodiment 11, 12, or 14, wherein Z is N. (Aspect 38) The compound of embodiment 11, 12, or 14, wherein A is CH. (Aspect 39) The compound of embodiment 11, 12, or 14, wherein A is N. (Aspect 40) The compound of embodiment 11, 12, or 14, wherein B is CH. (Aspect 41) The compound of embodiment 11, 12, or 14, wherein B is N. (Aspect 42) The compound according to any one of embodiments 11 to 30, wherein m is 1. (Aspect 43) The compound according to any one of embodiments 11 to 30, wherein m is 2. (Aspect 44) The compound according to any one of embodiments 11 to 30, wherein m is 3. (Aspect 45) A compound according to any one of aspects 11 to 22, wherein m is 4. (Aspect 46) The compound according to any one of embodiments 11 to 16, 19, 22 to 24, 27, and 30, wherein n is 0. (Aspect 47) The compound according to any one of embodiments 11 to 16, 19, 22 to 24, 27, and 30, wherein n is 1. (Aspect 48) The compound according to any one of embodiments 11 to 16, 19, 22 to 24, 27, and 30, wherein n is 2. (Aspect 49) R 1 The compound according to any one of embodiments 11 to 30, wherein is hydrogen. (Aspect 50) R 1 The compound according to any one of embodiments 11 to 30, wherein is halogen. (Aspect 51) R 1 is F. (Aspect 52) R 1 51. The compound according to embodiment 50, wherein is Cl. (Aspect 53) R 1 51. The compound according to embodiment 50, wherein is Br. (Aspect 54) R 1 The compound of any one of embodiments 11-30, wherein is CN. (Aspect 55) R 1 The compound according to any one of embodiments 11 to 30, wherein is OH. (Aspect 56) R 1 C optionally substituted with one or more halogens 1-3 The compound according to any one of embodiments 11 to 30, wherein the aryl group is alkyl. (Aspect 57) R 1 But C 1-3 57. The compound according to embodiment 56, wherein the aryl group is alkyl. (Aspect 58) R 1 57. The compound according to embodiment 56, wherein is methyl. (Aspect 59) R 1 57. The compound according to embodiment 56, wherein is ethyl. (Aspect 60) R 1 CF 3 57. The compound of embodiment 56, wherein (Aspect 61) R 1 But C 1-3 The compound according to any one of embodiments 11 to 30, which is alkoxy. (Aspect 62) R 1 62. The compound according to embodiment 61, wherein is methoxy. (Aspect 63) R 2 The compound according to any one of embodiments 11-18 and 23-26, wherein is hydrogen. (Aspect 64) R 2 But C 1-3 The compound according to any one of embodiments 11 to 18 and 23 to 26, wherein is alkyl. (Aspect 65) R 2 65. The compound according to embodiment 64, wherein is methyl. (Aspect 66) R 3 The compound according to any one of embodiments 11-16, 19, 22-24, 27, and 30, wherein is hydrogen. (Aspect 67) R 3 The compound of any one of embodiments 11-16, 19, 22-24, 27, and 30, wherein is halogen. (Aspect 68) R 3 57. The compound according to embodiment 56, wherein (Aspect 69) R 3 The compound according to any one of embodiments 11-16, 19, 22-24, 27, and 30, wherein is OH. (Aspect 70) R 3 The compound of any one of embodiments 11-16, 19, 22-24, 27, and 30, wherein is an amine. (Aspect 71) R 3 NH 2 71. The compound of embodiment 70, wherein (Aspect 72) R 3 But C 1-3 The compound according to any one of embodiments 11 to 16, 19, 22 to 24, 27, and 30, wherein is alkyl. (Aspect 73) R 3 73. The compound of embodiment 72, wherein is methyl. (Aspect 74) R 4 The compound of any one of embodiments 11-15, 17, 20, 22, 25, 28, and 30, wherein is hydrogen. (Aspect 75) R 4 But C 1-4 The compound of any one of embodiments 11 to 15, 17, 20, 22, 25, and 30, wherein is alkyl. (Aspect 76) R 4 76. The compound according to embodiment 75, wherein is methyl. (Aspect 77) R 4 76. The compound according to embodiment 75, wherein is ethyl. (Aspect 78) R 4 76. The compound according to embodiment 75, wherein is isopropyl. (Aspect 79) R 4 76. The compound according to embodiment 75, wherein is tert-butyl. (Aspect 80) R 5 The compound according to any one of aspects 23 to 30, wherein is F. (Aspect 81) R 5 The compound according to any one of aspects 23 to 30, wherein is Cl. (Aspect 82) R 5 The compound according to any one of embodiments 23 to 30, wherein is Br. (Aspect 83) R 5 The compound of any one of embodiments 23-30, wherein is CN. (Aspect 84) (Table 1) TIFF0007768890000306.tif238170TIFF0007768890000307.tif239170TIFF0007768890000308.tif97170 or a pharmaceutically acceptable salt thereof. (Aspect 85) Aspect 85. A compound according to any one of aspects 1 to 84, or a pharmaceutically acceptable salt thereof, wherein at least one hydrogen atom is replaced with a deuterium atom. (Aspect 86) A pharmaceutical composition comprising a compound according to any one of aspects 1 to 85, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. (Aspect 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 embodiments 1-85. (Aspect 88) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is ischemia-reperfusion injury. (Aspect 89) 89. The method of embodiment 88, wherein said ischemia-reperfusion injury is selected from stroke, myocardial infarction, and acute kidney injury. (Aspect 90) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is inflammatory bowel disease. (Aspect 91) 91. The method of embodiment 90, wherein the inflammatory bowel disease is ulcerative colitis. (Aspect 92) 91. The method of embodiment 90, wherein the inflammatory bowel disease is Crohn's disease. (Aspect 93) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is cancer. (Aspect 94) 94. The method of embodiment 93, wherein the cancer is colorectal cancer. (Aspect 95) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is a liver disease. (Aspect 96) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is atherosclerosis. (Aspect 97) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is a cardiovascular disease. (Aspect 98) 88. The method of embodiment 87, wherein the disease mediated by PHD activity is an ocular disease or condition. (Aspect 99) 99. The method of embodiment 98, wherein said disease or condition of the eye is selected from radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia. (Aspect 100) 88. The method of embodiment 87, wherein the disease is anemia. (Aspect 101) 101. The method of embodiment 100, wherein the anemia is anemia associated with chronic kidney disease. (Aspect 102) 88. The method of embodiment 87, wherein the disease is chronic kidney disease. (Aspect 103) 88. The method of embodiment 87, wherein the disease is associated with hyperoxia. (Aspect 104) 104. The method of embodiment 103, wherein the disease is retinopathy of prematurity. (Aspect 105) 104. The method of embodiment 103, wherein the disease is bronchopulmonary dysplasia (BPD). (Aspect 106) 88. The method of embodiment 87, wherein said 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 liver cirrhosis.

Claims

1. A compound of formula A, 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, wherein: Ar １ is phenyl, wherein the phenyl is a C optionally substituted with halogen, CN, OH, one or more halogens; １－３ Alkyl, or C １－３ substituted with alkoxy, R ２ is H or C １ - ３ is alkyl, Ar ２ is a halogen, OH, amine, or C １－３ a 6-membered nitrogen-containing heteroaryl optionally substituted with alkyl; R ４ is hydrogen or C １－４ alkyl, and Formula (A) represents the following compound: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof, excluding:

2. Ar １ at least one R is CN or halogen １ 2. The compound of claim 1, wherein the phenyl is substituted by:

3. Ar １ C optionally substituted with one or more halogens １－３ one or two R independently selected from alkyl, halogen, CN, or OH; １ The compound of claim 2 , substituted with a group.

4. Ar ２ but, 【Transformation 3】 where: A and B are independently CH or N, wherein N is optionally oxidized; Each R ３ are independently hydrogen, halogen, OH, amine, and C １－３ alkyl, and The compound of any one of claims 1 to 3, wherein n is 0, 1, or 2.

5. Ar ２ is a pyridyl or pyrazinyl group, said group being unsubstituted or selected from the group consisting of halogen, C １－３ The compound of any one of claims 1 to 4, which comprises a substituent that is alkyl, or OH.

6. R ２ is H or CH ３ The compound according to any one of claims 1 to 5,

7. R ４ The compound of any one of claims 1 to 6, wherein is H.

8. R ４ But C １－４ The compound of any one of claims 1 to 6, which is alkyl.

9. The structure of formula (Ib): 【Chemistry 4】 or a pharmaceutically acceptable salt thereof, wherein: R 1 is independently selected at each occurrence from the group consisting of halogen, CN, OH, C 1-3 alkyl optionally substituted with one or more halogens, and C 1-3 alkoxy; A and B are independently CH or N, wherein N is optionally oxidized; and 2. The compound of claim 1, wherein each R 3 is independently selected from the group consisting of hydrogen, halogen, OH, amine, and C 1-3 alkyl.

10. A structure according to formula (II): 【Transformation 5】 or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3, or 4; n is 0, 1, or 2; R １ independently each time, 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 is alkyl, R ３ are each independently hydrogen, halogen, OH, amine, or C １－３ alkyl, and R ４ is hydrogen or C １－４ The compound of claim 1 , wherein the aryl group is alkyl.

11. The structure of formula (IIa): 【Transformation 6】 or a pharmaceutically acceptable salt thereof.

12. The structure of formula (IIb): 【Transformation 7】 or a pharmaceutically acceptable salt thereof.

13. The structure of formula (IIc): 【Transformation 8】 or a pharmaceutically acceptable salt thereof.

14. The structure of formula (IId): 【Chemistry 9】 or a pharmaceutically acceptable salt thereof.

15. The structure of formula (IIe): 【Chemistry 10】 or a pharmaceutically acceptable salt thereof.

16. A structure of formula (IIf): 【Chemistry 11】 or a pharmaceutically acceptable salt thereof.

17. The structure of formula (IIg): 【Chemistry 12】 or a pharmaceutically acceptable salt thereof.

18. A structure according to formula III: 【Chemistry 13】 or a pharmaceutically acceptable salt thereof, wherein: m is 1, 2, 3, or 4; n is 0, 1, or 2; R １ independently at each occurrence, halogen, OH, and C optionally substituted with one or more halogens. １－３ is selected from the group consisting of alkyl, R ２ is hydrogen or C １ - ３ is alkyl, R ３ are each independently hydrogen, halogen, OH, amine, and C １－３ is selected from the group consisting of alkyl, R ４ is hydrogen or C １－４ is alkyl, R ５ is CN or a halogen.

19. The structure of formula (IIIa): 【Chemistry 14】 19. The compound of claim 18, wherein:

20. The structure of formula (IIIb): 【Chemistry 15】 19. The compound of claim 18, wherein:

21. The structure of formula (IIIc): 【Chemistry 16】 19. The compound of claim 18, wherein:

22. The structure of formula (IIId): 【Chemistry 17】 19. The compound of claim 18, wherein:

23. The structure of formula (IIIe): [Chemistry 18] 19. The compound of claim 18, wherein:

24. A structure of formula (IIIf): 【Chemistry 19】 19. The compound of claim 18, wherein:

25. The structure of formula III(g): 【Chemistry 20】 19. The compound of claim 18, wherein:

26. The compound of any one of claims 9 to 25, wherein m is 1.

27. The compound of any one of claims 9 to 25, wherein m is 2.

28. The compound of any one of claims 9 to 25, wherein m is 3.

29. The compound according to any one of claims 9 to 17, wherein m is 4.

30. 26. The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein n is 0.

31. 26. The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein n is 1.

32. 26. The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein n is 2.

33. R １ The compound of any one of claims 9 to 25, wherein is halogen.

34. R １ 34. The compound of claim 33, wherein is F.

35. R １ 34. The compound of claim 33, wherein is Cl.

36. R １ 34. The compound of claim 33, wherein is Br.

37. R １ The compound of any one of claims 9 to 25, wherein is CN.

38. R １ The compound of any one of claims 9 to 25, wherein is OH.

39. R １ C optionally substituted with one or more halogens １－３ The compound of any one of claims 9 to 25, which is alkyl.

40. R １ But C １－３ 40. The compound of claim 39, which is alkyl.

41. R １ 40. The compound of claim 39, wherein is methyl.

42. R １ 40. The compound of claim 39, wherein is ethyl.

43. R １ is CF ３ 40. The compound of claim 39, wherein:

44. R １ But C １－３ The compound of any one of claims 9 to 25, which is alkoxy.

45. R １ 45. The compound of claim 44, wherein is methoxy.

46. R ２ The compound of any one of claims 9 to 13 and 18 to 21, wherein is hydrogen.

47. R ２ But C １－３ The compound of any one of claims 9 to 13 and 18 to 21, which is alkyl.

48. R ２ 48. The compound of claim 47, wherein is methyl.

49. R ３ The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein is hydrogen.

50. R ３ The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein is halogen.

51. R ３ 40. The compound of claim 39, wherein is F.

52. R ３ The compound of any one of claims 9 to 11, 14, 17 to 19, 22 and 25, wherein is OH.

53. R ３ The compound of any one of claims 11, 14, 17-19, 22 and 25, wherein is an amine.

54. R ３ NH ２ 54. The compound of claim 53, wherein:

55. R ３ But C １－３ The compound according to any one of claims 9 to 11, 14, 17 to 19, 22 and 25, which is alkyl.

56. R ３ 56. The compound of claim 55, wherein is methyl.

57. R ４ 26. The compound of any one of claims 9, 10, 12, 15, 17, 20, 23, and 25, wherein is hydrogen.

58. R ４ But C １－４ 26. The compound of any one of claims 9, 10, 12, 15, 17, 20, 23, and 25, wherein the compound is alkyl.

59. R ４ 59. The compound of claim 58, wherein is methyl.

60. R ４ 59. The compound of claim 58, wherein is ethyl.

61. R ４ 59. The compound of claim 58, wherein is isopropyl.

62. R ４ 59. The compound of claim 58, wherein is tert-butyl.

63. R ５ The compound of any one of claims 18 to 25, wherein is F.

64. R ５ The compound of any one of claims 18 to 25, wherein is Cl.

65. R ５ The compound of any one of claims 18 to 25, wherein is Br.

66. R ５ The compound of any one of claims 18 to 25, wherein is CN. 【Request Item 67】 【Table 1】 or a pharmaceutically acceptable salt thereof. 【Request Item 68】 【Table 2】 or a pharmaceutically acceptable salt thereof. 【Request Item 69】 【Chemistry 21】 or a pharmaceutically acceptable salt thereof. 【Request Item 70】 【Chemistry 22】 or a pharmaceutically acceptable salt thereof. 【Request Item 71】 【Chemistry 23】 or a pharmaceutically acceptable salt thereof. 【Request Item 72】 【Chemistry 24】 or a pharmaceutically acceptable salt thereof. 【Request Item 73】 【Chemistry 25】 or a pharmaceutically acceptable salt thereof. 【Request Item 74】 【Chemistry 26】 or a pharmaceutically acceptable salt thereof. 【Request Item 75】 【Chemistry 27】 or a pharmaceutically acceptable salt thereof. 【Request Item 76】 【Table 3】 or a pharmaceutically acceptable salt thereof.

77. 77. The compound according to any one of claims 1 to 76, or a pharmaceutically acceptable salt thereof, wherein at least one hydrogen atom is replaced with a deuterium atom.

78. 78. A pharmaceutical composition comprising a compound according to any one of claims 1 to 77, or a pharmaceutically acceptable salt thereof.

79. A pharmaceutical composition for treating a disease mediated by PHD activity, comprising a compound according to any one of claims 1 to 77.

80. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is ischemia-reperfusion injury.

81. 81. The pharmaceutical composition of claim 80, wherein the ischemia-reperfusion injury is selected from stroke, myocardial infarction, and acute kidney injury.

82. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is inflammatory bowel disease.

83. 83. The pharmaceutical composition of claim 82, wherein the inflammatory bowel disease is ulcerative colitis.

84. 83. The pharmaceutical composition of claim 82, wherein the inflammatory bowel disease is Crohn's disease.

85. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is cancer.

86. 86. The pharmaceutical composition of claim 85, wherein the cancer is colorectal cancer.

87. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is a liver disease.

88. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is atherosclerosis.

89. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is a cardiovascular disease.

90. 80. The pharmaceutical composition of claim 79, wherein the disease mediated by PHD activity is an ocular disease or condition.

91. 91. The pharmaceutical composition of claim 90, wherein the disease or condition of the eye is selected from radiation retinopathy, retinopathy of prematurity, diabetic retinopathy, age-related macular degeneration, and ocular ischemia.

92. 80. The pharmaceutical composition of claim 79, wherein the disease is anemia.

93. 93. The pharmaceutical composition of claim 92, wherein the anemia is anemia associated with chronic kidney disease.

94. 80. The pharmaceutical composition of claim 79, wherein the disease is chronic kidney disease.

95. 80. The pharmaceutical composition of claim 79, wherein the disease is associated with hyperoxia.

96. 96. The pharmaceutical composition of claim 95, wherein the disease is retinopathy of prematurity.

97. 96. The pharmaceutical composition of claim 95, wherein the disease is bronchopulmonary dysplasia (BPD).

98. 80. The pharmaceutical composition of claim 79, 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 liver cirrhosis.

Images