PPARG inverse agonists and uses thereof

Abstract

Compounds of formula (I), as well as pharma- ceutically acceptable salts and compositions thereof, are provided that are useful for treating a variety of conditions associated with PPARG. TIFF2024529534000052.tif3258

Description

[Technical field]

[0001] Related Applications

[0001] This application claims the benefit of priority to U.S. Provisional Application No. 63 / 229,861, filed August 5, 2021, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] PPAR gamma (PPARG) is a type II ligand-dependent nuclear hormone receptor (belonging to the PPAR nuclear receptor subfamily) that functions as an obligate heterodimer with the retinoid X receptor (RXR). PPARG is expressed primarily in adipose tissue, colon, macrophages and the luminal layer of the urothelium. PPARG is known as a master regulator of adipogenesis and functions to regulate adipocyte differentiation, fatty acid storage and glucose metabolism. PPARG has also been shown to play an important role in macrophage metabolism and inflammation induced by IL4, controlling glutamine metabolism. In normal urothelium, PPARG is essential for its homeostasis and renewal.

[0003] The role of PPARG in cancer was originally inferred from genomic studies that identified PAX8-PPARG chromosomal rearrangements in follicular thyroid carcinoma. More recently, PPARG was found to be overexpressed and genetically altered in the luminal subtype of urothelial carcinoma. This is consistent with reports that long-term use of PPARG agonists is associated with increased incidence of urothelial carcinoma. The majority of urothelial carcinomas are urothelial carcinomas, which are classified as either non-muscle invasive urothelial carcinoma (NMIUC, 70%), muscle invasive urothelial carcinoma (MIUC, 25%) or metastatic urothelial carcinoma (MUC, 5%). MIUC is usually newly diagnosed, but 10-20% of NMIUC cases may eventually progress. MIUC is a heterogeneous and aggressive disease associated with a 5-year survival rate of 60% for patients with localized disease and less than 10% for patients with distant metastases. The molecular understanding of NMIUC and MIUC has improved significantly, including the relationship between molecular subtypes and urothelial differentiation. Several molecular classes of MIUC have been proposed, with an activated PPARG signature prominent in the luminal subtype. First-line treatment is chemotherapy, and although there are several options for chemotherapy-ineligible or second-line patients, options are limited and overall survival is poor.

[0004]

[0004] There is a need to develop effective PPARG modulators for treating cancers such as NMIUC, MIUC and MUC, and related conditions. Summary of the Invention [Means for solving the problem]

[0005] As used herein, the compound of formula I

[0006] [ka]

[0007] and pharma- ceutically acceptable salts and compositions thereof, 1 , R2 , R 3 , R 4 , R 5 , R 6 , R 7 , q and r are as described herein. In one embodiment, the disclosed compounds of formula I and their pharma- ceutically acceptable salts modulate PPARG (e.g., as agonists, such as inverse agonists) and are useful in various therapeutic applications, such as in the treatment of cancer. Thus, their use for treating diseases that respond to the inhibition of PPARG is included.

[0008] Also included are pharmaceutical compositions comprising the disclosed compounds of Formula I and pharma ceutically acceptable salts thereof, as well as methods for their preparation. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] 1. Overview of the compound In a first embodiment, a compound of formula I

[0010] [ka]

[0011] or a pharma- ceutically acceptable salt thereof (In the formula, X, Y and Z each independently represent N or -CR 4 and at least one of X, Y or Z is N; R 1 is hydrogen, halo, (C1-C4) alkyl or hydroxyl; R 2 ,Halo,-SR g , -SOR g , -SO2R g -OR g and R 3 is cyano or nitro, R 4is hydrogen, halo, (C1-C4) alkyl, (C1-C4) alkoxy or hydroxyl; R 5 is halo, halo(C1-C4)alkyl or cyano; R 6 is halo, halo(C1-C4)alkyl, (C1-C4)alkyl or cyano; R 7 is halo, (C1-C4) alkyl, (C1-C4) alkoxy, halo(C1-C4) alkyl, halo(C1-C4) alkoxy, -(C1-C4) alkyl OR a , -(C1-C4) alkylC(O)R a , -(C1-C4) alkyl C(O)OR a , -C(O)NR a R b , -(C1-C4) alkylC(O)NR a R b , -C(O)R a , -C(O)OR a , -NR a R b , -(C1-C4) alkylNR a R b , -C(O)NR a SO3H, -NR a C(O)R b , -NR a C(O)OR b , -NR a C(S)OR b , -NR c C(O)N a R b , -NR c C(S)NR a R b , -NR c S(O)2NR a R b , -C(S)R a , -S(O)2R a , -S(O)R a , -C(S)OR a , -C(S)NR a R b , -NR a C(S)R b , -SR a, phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl, each of said phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl being optionally and independently selected from R 8 is substituted with 1 to 3 groups selected from R 8 is halo, (C1-C4) alkyl, halo(C1-C4) alkyl, (C1-C4) alkoxy, halo(C1-C4) alkoxy, nitro, oxo, cyano, -(C1-C4) alkyl OR d , -(C1-C4) alkylC(O)R d , -(C1-C4) alkyl C(O)OR d , -C(O)NR d R e , -(C1-C4) alkylC(O)NR d R e , -C(O)R d , -C(O)OR d , -NR d R e , -(C1-C4) alkylNR d R e , -C(O)NR d SO3H, -NR d C(O)R e , -NR d C(O)OR e , -NR d C(S)OR e , -NR f C(O)N d R e , -NR f C(S)NR d R e , -NR f S(O)2NR d R e , -C(S)R d , -S(O)2R d , -S(O)R d , -C(S)OR d , -C(S)NR d R e , -NR d C(S)R e and -SR d is selected from R a , Rb , R c , R d , R e , R f and R g are each independently hydrogen or (C1-C4)alkyl optionally substituted with 1 or 2 -NR'R'' groups, where R' and R'' are each independently selected from hydrogen, (C1-C4)alkyl and halo(C1-C4)alkyl; q and r are each independently 0 or 1. is provided. 2.Definition When used in connection with describing a chemical group that may have multiple points of attachment, a hyphen (-) refers to the point of attachment of the group to the variable for which it is defined. For example, -NR b C(O)OR c and -NR b C(S)OR c means that the point of attachment for this group occurs on the nitrogen atom.

[0012]

[0009] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), and iodine (iodo, -I).

[0013] The term "alkyl", whether used alone or as part of a larger moiety such as "haloalkyl", means a saturated straight or branched chain monovalent hydrocarbon radical.

[0011] "Alkoxy" refers to an alkyl group bonded through an oxygen linking atom, represented by -O-alkyl. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, proproxy, and butoxy.

[0014] The term "haloalkyl" includes mono-, poly- and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine and iodine.

[0013] "Haloalkoxy" is a haloalkyl group that is attached to another moiety through an oxygen atom, for example, --OCHF2 or --OCF3.

[0015] The term oxo means the group ═O.

[0015] The term "5-7 membered heteroaryl" used alone or as part of a larger moiety refers to a 5-7 membered aromatic group containing 1-4 heteroatoms selected from N, O and S. Monocyclic heteroaryls include, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, triazinyl, tetrazinyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, etc. Optional substituents on the heteroaryl group may be present at any substitutable position, including, for example, the position at which the heteroaryl is attached.

[0016] The term "4-6 membered heterocyclyl" refers to a 4-6 membered saturated or partially unsaturated heterocycle containing 1-4 heteroatoms independently selected from N, O and S. The heterocyclyl ring can be attached to its pendant group at any heteroatom or carbon atom resulting in a stable structure. Examples of monocyclic saturated or partially unsaturated heterocyclic groups include, but are not limited to, tetrahydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, pyrrolidinyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, oxetanyl, dioxolanyl, morpholinyl, thiomorpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl and tetrahydropyrimidinyl. Optional substituents on the heterocyclyl group can be present at any substitutable position, including the position at which the heterocyclyl is attached.

[0017]

[0017] Compounds with one or more chiral centers can exist in various stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial configuration. Stereoisomers include all diastereomeric, optical, and epimeric forms, and racemates, as well as mixtures thereof. When the stereochemical configuration at a chiral center in a compound with one or more chiral centers is indicated by its chemical name (e.g., when the configuration is indicated by a chemical name with "R" or "S") or structure (e.g., when the configuration is indicated by a "wedge-shaped" bond), the enrichment of the indicated configuration relative to the opposite configuration is greater than 50%, 60%, 70%, 80%, 90%, 99% or 99.9%. The "enrichment of the indicated configuration relative to the opposite configuration" is a mole percentage and is determined by dividing the number of compounds with the indicated stereochemical configuration at the chiral center by the total number of all compounds with the same or opposite stereochemical configuration in the mixture.

[0018]

[0018] When a disclosed compound is named or depicted by structure without indicating stereochemistry, it is understood that the name or structure encompasses one of the possible stereoisomers or geometric isomers, without the other, or a mixture of the stereoisomers or geometric isomers encompassed.

[0019] The disclosed compounds may exist in one or more tautomeric forms, such as those below, and are included herein.

[0020] [ka]

[0021]

[0020] The terms "subject" and "patient" may be used interchangeably and refer to a mammal in need of treatment, such as companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, the subject is a human in need of treatment.

[0022]

[0021] The terms "inhibit," "inhibition," or "inhibiting" include a reduction in the baseline activity of a biological activity or process.

[0022] As used herein, the terms "treatment", "treat" and "treating" refer to reversing, alleviating, delaying the onset of, or preventing the progression of a disease or disorder as described herein, or one or more symptoms thereof. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other embodiments, treatment may be administered asymptomatically. For example, treatment may be administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of symptoms and / or exposure to a particular organism or other susceptibility factor), i.e., prophylactic treatment. Treatment may also be continued after symptoms have disappeared, e.g., to delay their recurrence.

[0023] The term "pharmaceutical acceptable carrier" refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants or vehicles that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and wool fat.

[0024]

[0024] For use in medicine, the salts of the compounds described herein refer to non-toxic "pharmaceutical acceptable salts". Pharmaceutically acceptable salt forms include pharmaceutical acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutical acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and organic acids (e.g., acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). Compounds of the present teachings having an acidic group, such as a carboxylic acid, can form pharmaceutical acceptable salts with a pharmaceutical acceptable base(s). Suitable pharmaceutical acceptable base salts include, for example, ammonium salts, alkali metal salts (e.g., sodium and potassium salts), and alkaline earth metal salts (e.g., magnesium and calcium salts). Compounds having a quaternary ammonium group also include counterions such as chloride, bromide, iodide, acetate, perchlorate, and the like. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0025]

[0025] The term "effective amount" or "therapeutically effective amount" refers to an amount of a compound described herein that elicits a desired or beneficial biological or medical response in a subject, for example, a dosage of between 0.01 and 100 mg / kg body weight / day. 3.Compound

[0026] In a second embodiment, the compound of formula I is represented by formula II

[0026] [ka]

[0027] or a pharma- ceutically acceptable salt thereof, wherein the variables are as described above for formula I. In a third embodiment, the compound of formula I is represented by formula III

[0028] [ka]

[0029] or a pharma- ceutically acceptable salt thereof, wherein the variables are as described above for formula I. In a fourth embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 2 is halo, -S(C1-C4)alkyl, -SO(C1-C4)alkyl, -SO2(C1-C4)alkyl, or -O(C1-C4)alkylN[(C1-C4)alkyl]2, and the remaining variables are as described above for Formula I. Alternatively, as part of a fourth embodiment, R in a compound of Formula I, II, or III, or a pharma- ceutically acceptable salt thereof, 2 is chloro, -SCH3, -SOCH3, -SO2CH3, or -O(CH2)2N(CH3)2, and the remaining variables are as described above for Formula I. Alternatively, as part of the fourth embodiment, R in a compound of Formula I, II, or III, or a pharma- ceutically acceptable salt thereof, 2 is chloro, and the remainder of the variables are as described above for Formula I.

[0030] In a fifth embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 3 is cyano, and the remainder of the variables are as described above for Formula I or the fourth embodiment.

[0031] In a sixth embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 1 is hydrogen, fluoro, hydroxyl, or methyl, and the remaining variables are as described above for Formula I, or the fourth or fifth embodiment. Alternatively, as part of the sixth embodiment, R in a compound of Formula I, II, or III, or a pharma- ceutically acceptable salt thereof, 1 is hydrogen, and the remaining variables are as described above for Formula I, or the fourth or fifth embodiment.

[0032] In a seventh embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 5 is halo or cyano, and the remaining variables are as described above for Formula I, or the fourth, fifth or sixth embodiment. Alternatively, as part of the seventh embodiment, R in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, 5 is halo, and the remaining variables are as described above for Formula I or the fourth, fifth or sixth embodiment.

[0033]

[0032] In an eighth embodiment, q in a compound of Formula I, II or III or a pharma- ceutically acceptable salt thereof is 1, and the remaining variables are as described above for Formula I or the fourth, fifth, sixth or seventh embodiment.

[0034] In a ninth embodiment, r in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is 1, and the remaining variables are as described above for Formula I or the fourth, fifth, sixth, seventh or eighth embodiment. Alternatively, as part of the ninth embodiment, r in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is 0, and the remaining variables are as described above for Formula I or the fourth, fifth, sixth, seventh or eighth embodiment.

[0035] In a tenth embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 6 is halo, and the remaining variables are as described above for Formula I or the fourth, fifth, sixth, seventh or eighth embodiment.

[0036] In an eleventh embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 7 is halo, halo(C1-C4)alkyl, (C1-C4)alkyl, (C1-C4)alkoxy, -(C1-C4)alkylOR a, -C(O)NR a R b , phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl, each of said phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl being optionally and independently selected from R 8 and the remaining variables are as described above for Formula I, or the fourth, fifth, sixth, seventh, eighth, ninth or tenth embodiment. Alternatively, as part of the eleventh embodiment, R in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is 7 is halo, halo(C1-C4)alkyl, (C1-C4)alkyl, (C1-C4)alkoxy, -(C1-C4)alkylOR a , -C(O)NR a R b , phenyl, pyridinyl, piperazinyl, piperidinyl, pyrrolidinyl, thiomorpholinyl, pyrazolyl, and oxetanyl, each of said phenyl, pyridinyl, pyrazolyl, pyrrolidinyl, piperazinyl, thiomorpholinyl, piperidinyl, and oxetanyl optionally and independently selected from R 8 and the remaining variables are as described above for Formula I, or the fourth, fifth, sixth, seventh, eighth, ninth or tenth embodiment. Alternatively, as part of the eleventh embodiment, R in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is 7 is halo, halo(C1-C4)alkyl, (C1-C4)alkyl, (C1-C4)alkoxy, -(C1-C4)alkylOR a , -C(O)NR a R b , phenyl, pyridinyl, pyrazolyl, and oxetanyl, each of said phenyl, pyridinyl, pyrazolyl, and oxetanyl optionally and independently being R 8 and the remaining variables are as described above for Formula I or the fourth, fifth, sixth, seventh, eighth, ninth or tenth embodiment.

[0037] In a twelfth embodiment, R in a compound of formula I, II or III or a pharma- ceutically acceptable salt thereof is 8 is halo, -C(O)NR d R e , (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, oxo, and cyano, with the remaining variables being as described above for Formula I or the third, fourth, fifth, sixth, seventh, eighth, ninth, tenth or eleventh embodiment. Alternatively, as part of the twelfth embodiment, R in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is selected from the group consisting of aryl, aryloxy ... 8 is selected from halo, (C1-C4)alkyl, halo(C1-C4)alkyl, (C1-C4)alkoxy, halo(C1-C4)alkoxy, oxo, and cyano, and the remaining variables are as described above for Formula I or the third, fourth, fifth, sixth, seventh, eighth, ninth, tenth or eleventh embodiment. Alternatively, as part of the twelfth embodiment, R in a compound of Formula I, II or III, or a pharma- ceutically acceptable salt thereof, is 8 is -C(O)NR d R e , (C1-C4)alkyl, and oxo, with the remaining variables being as described above for Formula I or the third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, or eleventh embodiment. Alternatively, as part of the twelfth embodiment, R in a compound of Formula I, II, or III, or a pharma- ceutically acceptable salt thereof, is selected from 8 is selected from -C(O)N(CH), CH, and oxo, and the remaining variables are as described above for Formula I or the third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, or eleventh embodiment. Alternatively, as part of the twelfth embodiment, R in a compound of Formula I, II, or III, or a pharma- ceutically acceptable salt thereof, is 8 is halo(C1-C4)alkyl, and the remaining variables are as described above for Formula I or the third, fourth, fifth, sixth, seventh, eighth, ninth, tenth or eleventh embodiment.

[0038] Compounds having formula I, II, or III are further disclosed in the examples and are included in this disclosure, including pharma- ceutically acceptable salts and neutral forms thereof. 4. Use, Formulation and Administration

[0038] The compounds and compositions described herein are generally useful for modulating the activity of PPARG. In some embodiments, the compounds, pharma- ceutically acceptable salts, and pharmaceutical compositions described herein inhibit the activity of PPARG. In some embodiments, the compounds and pharma- ceutically acceptable salts disclosed herein are agonists of PPARG. In some embodiments, the compounds and pharma- ceutically acceptable salts disclosed herein are agonists of PPARG. In some embodiments, the compounds and pharma- ceutical acceptable salts disclosed herein are inverse agonists of PPARG. In one embodiment, an "inverse agonist" refers to an agent that binds to the same receptor binding site as an agonist (e.g., the binding site of a nuclear receptor such as PPARG) and exerts the opposite effect by not only antagonizing the effect of the agonist, but also suppressing spontaneous receptor signaling (if present).

[0039] In some aspects, the compounds and pharma- ceutically acceptable salts disclosed herein overcome the activation state of PPARG function resulting from altered PPARG activity (mutation, amplification or overexpression) or RXRA activating mutation. In some aspects, the compounds and pharma- ceutically acceptable salts disclosed herein increase the repression state (NCOR1 recruitment) to a higher degree than previously disclosed PPARG regulators, such as previous inverse agonists. Such results occur even in the context of mutants. For example, see the table in the Examples section that qualitatively evaluates NCOR1 recruitment and repression of PPARG target genes in HT1197.

[0040] In some aspects, the compounds and pharmaceutical compositions described herein are useful for treating disorders related to PPARG function.Therefore, provided herein is a method for treating disorders related to PPARG function, comprising administering to a subject in need of treatment of the disorder a therapeutically effective amount of the compounds described herein or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising the disclosed compounds or a pharma- ceutically acceptable salt thereof.

[0041]

[0041] Also provided is the use of a compound described herein or a pharma- ceutically acceptable salt thereof, or a pharmaceutical composition comprising a disclosed compound or a pharma- ceutically acceptable salt thereof, for the preparation of a medicament for treating a disorder associated with PPARG function.Also provided is a pharmaceutical composition comprising a compound described herein or a pharma- ceutically acceptable salt thereof, or a disclosed compound or a pharma- ceutically acceptable salt thereof, for use in treating a disorder associated with PPARG.

[0042] In one aspect, the PPARG-associated disorder is cancer. In some aspects, the cancer is associated with an upregulated peroxisome proliferator-activated receptor (PPAR) signaling pathway. In some aspects, the upregulated PPAR signaling pathway is associated with increased expression of one or more genes selected from uroplakin 1A (UPK1A), uroplakin IB (UPK1B), uroplakin (UPK2), keratin 20 (KRT20), GATA binding protein 3 (GAT A3), nuclear receptor corepressor 1 (NCORl), nuclear receptor corepressor 2 (NCOR2), fatty acid binding protein 4 (FABP4), forkhead box A1 (FOXA1), CD36 molecule (CD36), acyl-CoA oxidase 1 (ACOX1), 3-hydroxy-3-methylglutaryl-CoA synthase 2 (HMGCS2), acyl-CoA synthase long chain family member 5 (ACSL5), arachidonate 5-lipoxygenase (ALOX5), acyl-CoA synthase long chain family member 1 (ACSL1), and angiopoietin-like 4 (ANGPTL4).

[0043] In some embodiments, the cancer treated by the compounds described herein, their pharmaceutically acceptable salts and pharmaceutical compositions is selected from breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, renal cancer, bladder cancer, testicular cancer, urothelial cancer (e.g., non-muscle invasive urothelial cancer, muscle invasive urothelial cancer, metastatic urothelial cancer), skin cancer, melanoma, colon cancer, kidney cancer, brain cancer and hematopoietic cancer (e.g., lymphoma, multiple myeloma and leukemia). In one embodiment, the cancer treated by the compounds described herein, their pharmaceutically acceptable salts and pharmaceutical compositions is urothelial cancer, e.g., non-muscle invasive urothelial cancer, muscle invasive urothelial cancer and metastatic urothelial cancer.

[0044]

[0044] Other uses besides cancer are contemplated, such as angiogenesis and neovascularization in metabolic diseases (e.g. osteoporosis, rickets, arthropathy, obesity, type 1 and type 2 diabetes), lipid metabolism disorders, pancreatitis, glucose metabolism disorders, diabetic nephropathy, diabetic complications, hyperuricemia, osteoporosis, rickets, arthropathy, inflammatory diseases (e.g. inflammatory skin diseases such as psoriasis, atopic dermatitis, eczema, acne vulgaris, other dermatitis and pruritus), pulmonary disorders (e.g. asthma and chronic obstructive pulmonary disease), autoimmune diseases, neurodegenerative diseases (e.g. multiple sclerosis, Alzheimer's disease and Parkinson's disease), cardiovascular diseases (e.g. selected from atherosclerosis, venous and arterial occlusive diseases), stenosis after invasive procedures, cardiomyopathy, myocardial fibrosis, congestive heart failure, neoplastic diseases and renal diseases.

[0045] In certain aspects, the pharmaceutical compositions described herein are formulated for administration to a patient in need of such compositions. The pharmaceutical compositions described herein can be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally or via a subcutaneously implanted reservoir. The term "parenterally" as used herein includes subcutaneous, intravenous, intramuscular, intraarterial, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the pharmaceutical compositions described herein may be aqueous or oleaginous suspensions. These suspensions may be formulated by techniques well known in the art using suitable dispersing or wetting agents and suspending agents.

[0046]

[0046] In some aspects, the pharmaceutical composition is administered orally.

[0047] The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound employed, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, as well as the judgment of the treating physician and the severity of the particular disease being treated. The amount of the compounds described herein in the composition will also depend on the particular compound in the pharmaceutical composition. Example chemical synthesis

[0048] The following representative examples are intended to help illustrate the disclosure, but they are not intended, and should not be construed, to limit the scope of the invention.

[0047]

[0049] Typical starting materials used were obtained from commercial sources or prepared in other instances unless otherwise noted. Preparation of compounds

[0050] The compounds claimed herein were prepared according to the procedures outlined in the following schemes.

[0048]

[0051]

[0049] [ka]

[0050]

[0052] Quinolones such as S4 can be prepared by the general synthetic methodology shown in Scheme 1. Compounds of formula S3 can be prepared from coupling of acid chloride S1 with 2-ketoaniline S2 with or without the addition of a base. Treatment of amide S3 with a base such as sodium hydroxide or lithium hydroxide in an ether solvent such as dioxane or 2-methyl-tetrahydrofuran at elevated temperature affords the target quinolone compound S4.

[0051]

[0053]

[0052] [ka]

[0053]

[0054] Quinolones such as S5 can be prepared by treating a halo-quinolone such as S4 with a base and a nucleophile.

[0055] Preparation of starting materials

[0054] [ka]

[0055]

[0056] 3-Chloro-6-cyanopicolinic acid: To a solution of ethyl 3-chloro-6-cyanopicolinate in DCE (10 mL) at 20° C. was added hydroxy(trimethyl)stannane (859 mg, 4.75 mmol, 2 equiv.). The mixture was stirred at 50° C. for 16 h. The mixture was diluted with water (15 mL) and extracted with ethyl acetate (2×10 mL). The combined organic layers were washed with brine (15 mL×2), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give the title compound as a white solid (268 mg, 49% yield, 80% purity). 1H NMR (400 MHz, CD3OD) δ 8.20 (d, J = 8.6 Hz, 1H), 8.00 - 7.95 (m, 1H).LCMS[M+1]=183.0 / 185.0.

[0056] [ka]

[0057] 3-Chloro-6-cyanopicolinoyl chloride: 3-Chloro-6-cyanopicolinic acid (120 mg, 657 μmol, 1 equiv.) was added to a flask containing SOCl2 (2 mL) at 20° C. The mixture was then stirred at 80° C. for 3 h under N2. The reaction mixture was concentrated under reduced pressure to give the title compound as a yellow solid (130 mg, crude). The crude product was used directly in the next step without further purification.

[0058] [ka]

[0059] 5-Chloro-2-cyanoisonicotinyl chloride: 5-Chloro-2-cyanoisonicotinic acid (200 mg, 1.10 mmol, 1 equiv.) was added to a flask containing SOCl2 (2 mL) at 20°C. The mixture was then stirred at 80°C for 1 h under N2. The reaction mixture was concentrated under reduced pressure to give the title compound as a white solid (220 mg, crude). The crude product was used directly in the next step without further purification.

[0060] [ka]

[0061] 6-Cyano-3-(methylthio)picolinoyl chloride Step 1, 6-cyano-3-(methylthio)picolinic acid: To a solution of ethyl 3-chloro-6-cyanopicolinate (2 g, 9.5 mmol, 1.0 equiv) in DMF (3 mL) was added sodium methanethiolate (1.5 g, 21.4 mmol, 2.2 equiv). The mixture was stirred at 0° C. for 2 h. The reaction mixture was poured into ice water (15 mL) and stirred for 10 min. The pH of the solution was adjusted to pH=5-6 with aqueous 1 M HCl. The white solid that precipitated during the pH adjustment was filtered off and washed with H2O (5 mL). The filter cake was dried under vacuum to give the title compound as a white solid (800 mg, 43% yield). 1 H NMR (400 MHz, DMSO-d6) δ 8.13 - 8.08 (m, 1H), 8.05 - 7.99 (m, 1H), 2.49 (br s, 3H). Step 2, 6-Cyano-3-(methylthio)picolinoyl chloride: A solution of 6-cyano-3-(methylthio)picolinic acid (300 mg, 1.5 mmol, 1.0 equiv) in SOCl2 (5 mL) was stirred at 80° C. for 1 h. The reaction mixture was concentrated under reduced pressure to give the title compound as a white solid (350 mg, crude). The product was used in the next step without further purification.

[0062] [ka]

[0063] 2-Cyano-5-(methylthio)isonicotinoyl chloride Step 1, 2-cyano-5-methylsulfanyl-pyridine-4-carboxylic acid: To a solution of sodium methanethiolate (17.83 g, 254 mmol, 2.5 equiv) in DMF (130 mL) was added a solution of methyl 5-chloro-2-cyano-pyridine-4-carboxylate (20 g, 102 mmol, 1.0 equiv) in DMF (260 mL) at 0 °C. The mixture was stirred at 0 °C for 0.5 h under N2. The pH of the reaction mixture was adjusted to pH = 1 with concentrated HCl. The white solid that precipitated during the pH adjustment was filtered off and washed with H2O (5 mL). The filter cake was dried under vacuum to give the title compound as a white solid (18.0 g, 91% yield). 1 H NMR (400 MHz, DMSO-d6) δ 8.75 (s, 1H), 8.21 (s, 1H), 2.62 (s, 3H). Step 2, 2-cyano-5-(methylthio)isonicotinoyl chloride: A solution of 2-cyano-5-(methylthio)isonicotinic acid (115 mg, 592 μmol, 1.0 equiv) in SOCl2 (4 mL) was stirred at 100° C. for 2 h under N2. The mixture was concentrated in vacuo to give the title compound as a yellow solid (125 mg, crude). The product was used in the next step without further purification.

[0064] [ka]

[0065] 1-(2-amino-4-chloro-5-(4-methylpiperazin-1-yl)phenyl)ethanone Step 1, 1-(5-bromo-2-chloro-4-nitro-phenyl)-4-methyl-piperazine: To a solution of 1-bromo-4-chloro-5-fluoro-2-nitro-benzene (2.0 g, 7.86 mmol, 1.0 equiv.) in DMF (20 mL), 1-methylpiperazine (787 mg, 7.86 mmol, 1.0 equiv.) and K2CO3 (1.09 g, 7.86 mmol, 1.0 equiv.) were added. The mixture was stirred at room temperature for 4 h. The mixture was added to water (100 mL). The precipitate that formed was filtered off and the material was washed with water (3 x 50 mL). The material was purified by silica gel column chromatography (10:1 to 3:1 petroleum ether:ethyl acetate + 10% DCM) to give the title compound as an orange solid (2.4 g, 91% yield). LCMS: [M+1]=334.0. 1 H NMR (400 MHz, chloroform-d) δ 8.05 (s, 1H), 7.24 (s, 1H), 3.29 - 3.17 (m, 4H), 2.68 - 2.55 (m, 4H), 2.38 (s, 3H).

[0066] Step 2, 2-Bromo-5-chloro-4-(4-methylpiperazin-1-yl)aniline: A mixture of 1-(5-bromo-2-chloro-4-nitro-phenyl)-4-methyl-piperazine (2.4 g, 7.17 mmol, 1.0 equiv), Fe (2.00 g, 35.8 mmol, 5.0 equiv) and NH4Cl (1.92 g, 35.8 mmol, 5.0 equiv) in MeOH (20 mL) and H2O (5 mL) was purged with N2, then the mixture was stirred at 80 °C for 16 h under N2. The suspension was filtered through a Celite pad and the pad cake was washed with MeOH (3 x 30 mL). The filtrate was concentrated under reduced pressure. The residue was diluted with water (20 mL). The pH of the solution was adjusted to pH = 7-8 with saturated aqueous sodium bicarbonate solution. The aqueous phase was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were washed with brine (10 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound as an orange solid (1.7 g, crude). LCMS: [M+1]=304.0. 1H NMR (400 MHz, DMSO-d6) δ 7.11 (s, 1H), 6.87 (s, 1H), 5.29 - 5.16 (m, 2H), 2.80 (br t, J = 4.4 Hz, 4H), 2.42 (br s, 4H), 2.20 (s, 3H).

[0067] Step 3, 5-chloro-2-(1-ethoxyvinyl)-4-(4-methylpiperazin-1-yl)aniline: To a solution of 2-bromo-5-chloro-4-(4-methylpiperazin-1-yl)aniline (300 mg, 985 μmol, 1 equiv.) and tributyl(1-ethoxyvinyl)stannane (399 uL, 1.18 mmol, 1.2 equiv.) in toluene (5 mL) was added Pd(PPh3)4 (113.8 mg, 98.4 μmol, 0.1 equiv.). The mixture was stirred at 120 °C for 16 h under N2. The reaction mixture was cooled to room temperature and quenched with an aqueous solution of KF (20 mL). The aqueous phase was extracted with ethyl acetate (2×30 mL) and the combined organic layers were washed with saturated aqueous sodium bicarbonate, brine, dried over anhydrous Na2SO4, filtered and concentrated to give the title compound as a brown solid (250 mg, crude). LCMS [M+1]=296.1. 1 H NMR (400 MHz, chloroform-d) δ 7.39 (s, 1H), 6.74 (s, 1H), 6.11 (br s, 2H), 3.00 (br s, 4H), 2.62 (br s, 4H), 2.59 - 2.52 (m, 3H), 2.41 - 2.31 (m, 3H), 1.30 - 1.23 (m, 2H).

[0068] Step 4, 1-(2-amino-4-chloro-5-(4-methylpiperazin-1-yl)phenyl)ethanone: A solution of 5-chloro-2-(1-ethoxyvinyl)-4-(4-methylpiperazin-1-yl)aniline (250 mg, 845 μmol, 1.0 equiv) in aqueous HCl (1M, 2.54 mL, 3.0 equiv) was stirred at 20 °C for 2 h. The reaction mixture was diluted with H2O (10 mL) and extracted with EtOAc (3 × 10 mL). The pH of the aqueous layer was adjusted to pH = 8 with saturated aqueous NaHCO3. The aqueous layer was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (50:1 to 10:1 DCM:MeOH) to afford the title compound as a yellow solid (140 mg, 62% yield). 1 H NMR (400 MHz, chloroform-d) δ 7.39 (s, 1H), 6.74 (s, 1H), 6.11 (br s, 2H), 3.00 (br s, 4H), 2.69 - 2.57 (m, 4H), 2.57 - 2.54 (m, 3H), 2.43 - 2.31 (m, 3H).

[0066] [ka]

[0067]

[0069] 1-(6-amino-2,4-difluoro-3-(1-methylpiperidin-4-yl)phenyl)ethan-1-one

[0070] Step 1, 1-(6-amino-2,4-difluoro-3-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethan-1-one: 1-(6-amino-3-bromo-2,4-difluorophenyl)ethan-1-one (1.3 g, 5.3 mmol, 1 equiv.) and 1-methyl-4-(4,4,5,6-tetrahydropyridin-4-yl)phenyl)ethan-1-one in THF (1.2 mL) and HO (0.3 mL). To a mixture of 5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine (1.8 g, 8.0 mmol, 1.5 equiv.) at room temperature was added K3PO4 (2.5 g, 11.7 mmol, 2.2 equiv.) and di-tert-butyl(cyclopentyl)phosphane-dichloropalladium;iron (347 mg, 532 μmol, 0.1 equiv.) under N2. The mixture was stirred at 80 °C for 2 h. The reaction mixture was filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (10:1 to 2:1 petroleum ether:ethyl acetate) to give the title compound as a yellow oil (1.4 g, 97% yield). 1 H NMR (400 MHz, methanol-d4) δ 6.29 (dd, J = 1.8, 12.2 Hz, 1H), 5.73 (br s, 1H), 3.13 (q, J = 2.8 Hz, 2H), 2.70 (t, J = 5.8 Hz, 2H), 2.51 (d, J = 8.8 Hz, 3H), 2.43 (br d, J = 1.8 Hz, 2H), 2.38 (s, 3H).

[0071] Step 2, 1-(6-amino-2,4-difluoro-3-(1-methylpiperidin-4-yl)phenyl)ethan-1-one: To a solution of 1-(6-amino-2,4-difluoro-3-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)ethan-1-one (1.2 g, 4.4 mmol, 1 equiv.) in MeOH (12 mL) was added 20% Pd(OH)2 on carbon (500 mg) under N2. The suspension was degassed under vacuum and purged with H2 several times. The mixture was mixed under H2 (50 Psi) at 70 °C for 3 h. The reaction mixture was filtered and concentrated under reduced pressure to give the title compound as a pale yellow solid (1.0 g, 84% yield). 1H NMR (400 MHz, methanol-d4) δ 6.27 (dd, J = 1.8, 13.2 Hz, 1H), 2.99 (br d, J = 6.8 Hz, 2H), 2.92 - 2.83 (m, 1H), 2.51 (d, J = 9.2 Hz, 3H), 2.30 (s, 3H), 2.19 - 2.07 (m, 4H), 1.72 - 1.65 (m, 2H).

[0068] [ka]

[0069]

[0072] 1-(2-amino-4-chloro-6-fluorophenyl)ethanone: To a solution of 3-chloro-5-fluoroaniline (20 g, 137 mmol, 1.0 equiv) in p-xylene (40 mL) at 0-5 °C was added BCl3 (1 M, 182.7 mL, 1.3 equiv) over 2 h. The mixture was allowed to warm to room temperature within 0.5 h and stirred at room temperature for 10 min. Acetonitrile (57.8 mL, 1.10 mol, 8 equiv) was then added dropwise over 20 min at room temperature. The mixture was stirred at room temperature for 10 min and then p-xylene (45 mL) was added. AlCl3 (10.2 g, 76.9 mmol, 0.5 equiv) was then added and the reaction mixture was stirred at room temperature for 1 h and then at 75-77 °C for an additional 12 h. Aqueous HCl (4 N, 200 mL) was then added to the mixture and the mixture was stirred at 80 °C for 4 h. The mixture was poured into water (800 mL) and extracted with ethyl acetate (1.5 L). The organic phase was washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50:1 to 20:1 petroleum ether:ethyl acetate) to give the title compound as a pale yellow solid (4.6 g, 18% yield). LCMS [M+1] = 188.0 / 190.0. 1 H NMR (400 MHz, chloroform-d) δ 6.53 - 6.41 (m, 2H), 6.36 (dd, J = 2.0, 11.8 Hz, 2H), 2.58 (d, J = 8.4 Hz, 3H).

[0070] [ka]

[0071]

[0073] 1-(3-acetyl-4-amino-2,6-difluorophenyl)-N,N-dimethylpiperidine-3-carboxamide: The title compound was prepared following the synthetic steps described for 1-(2-amino-4-chloro-5-(4-methylpiperazin-1-yl)phenyl)ethenone using 2-bromo-3,4,5-trifluoro-1-nitrobenzene and N,N-dimethylpiperidine-3-carboxamide as starting materials. LCMS [M-1]=324.1. 1 H NMR (400 MHz, chloroform-d) δ 6.41 - 6.18 (m, 2H), 6.12 (dd, J = 1.6, 12.6 Hz, 1H), 3.34 - 3.20 (m, 1H), 3.10 (s, 3H), 3.03 (br d, J = 16.0 Hz, 2H), 2.94 (s, 3H), 2.57 (d, J = 8.8 Hz, 3H), 1.95 - 1.84 (m, 1H), 1.82 - 1.53 (m, 5H).

[0074] Example 1

[0072] [ka]

[0073]

[0075] 5-Chloro-6-(6,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile

[0076] Scheme 1, Step 1. N-(2-Acetyl-4,5-difluorophenyl)-3-chloro-6-cyanopicolinamide: To a solution of 1-(2-amino-4,5-difluorophenyl)ethan-1-one (100 mg, 584 μmol, 1 equiv.) in THF (2 mL) was added NaH (25.7 mg, 642 μmol, 60% purity, 1.1 equiv.) at 0° C. The mixture was stirred at 0° C. for 5 min under N2. Then, a solution of 3-chloro-6-cyanopicolinoyl chloride (129 mg, 642 μmol, 1 equiv.) in THF (1 mL) was added dropwise to the mixture at 0° C. and the mixture was mixed at 20° C. for 16 h. The reaction mixture was quenched by the addition of saturated aqueous NH4Cl (20 mL) and the mixture was extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The crude product was triturated with ethyl acetate (3 mL) at 20° C. for 20 min. The product was then filtered off and the precipitate was washed with ethyl acetate (3×0.3 mL). The solid was dried under reduced pressure to give the title compound as a white solid (130 mg, 66% yield). 1 H NMR (400 MHz, DMSO-d6) δ 13.03 (s, 1H), 8.68 (dd, J = 7.8, 13.8 Hz, 1H), 8.45 (d, J = 8.4 Hz, 1H), 8.37 - 8.21 (m, 2H), 2.75 - 2.64 (m, 3H).

[0077] Scheme 1, step 2. 5-Chloro-6-(6,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile: To a solution of N-(2-acetyl-4,5-difluorophenyl)-3-chloro-6-cyanopicolinamide (100 mg, 298 μmol, 1 equiv.) in dioxane (5 mL) was added LiOH (7.1 mg, 297 μmol, 1 equiv.) at 20° C. The mixture was stirred at 110° C. for 16 h under N2. The pH of the mixture was adjusted to 6 with aqueous 1 M HCl. The mixture was then diluted with water (15 mL) and extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Waters Xbridge BEH C18 100mm x 30mm x 10um; mobile phase: 10-40% acetonitrile in water (+NH4HCO3)) to give the title compound as a yellow solid (21mg, 21% yield). 1 H NMR (400 MHz, DMSO-d6) δ 8.49 (d, J = 8.4 Hz, 1H), 8.28 (d, J = 8.4 Hz, 1H), 8.00 (dd, J = 9.0, 11.0 Hz, 1H), 7.72 (dd, J = 7.0, 11.4 Hz, 1H), 6.53 (s, 1H).LCMS[M+1]=318.0 / 320.0.

[0074]

[0078] Example 2

[0075] [ka]

[0076]

[0079] 5-Chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile

[0080] Scheme 1, Step 1. N-(2-Acetyl-3,5-difluorophenyl)-5-chloro-2-cyanoisonicotinamide: To a solution of 1-(2-amino-4,6-difluorophenyl)ethan-1-one (170 mg, 993 μmol, 1 equiv.) in THF (1 mL) was added NaH (43.7 mg, 1.09 mmol, 60% purity, 1.1 equiv.) at 0° C. Then a solution of 5-chloro-2-cyanoisonicotinoyl chloride (220 mg, 1.09 mmol, 1.1 equiv.) in THF (1 mL) was added dropwise to the mixture. The mixture was stirred at 20° C. for 16 h. The reaction mixture was quenched by the addition of saturated aqueous NH4Cl (15 mL) and then extracted with ethyl acetate (2×30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was purified by preparative TLC (petroleum ether:ethyl acetate=3:1) to give the title compound as a yellow solid (84 mg, 25% yield). 1 H NMR (400 MHz, CDCl3) δ 12.56 (br s, 1H), 8.84 (s, 1H), 8.52 - 8.45 (m, 1H), 7.91 (s, 1H), 6.77 - 6.74 (dd, J = 2.6, 8.2 Hz, 1H), 2.70 (d, J = 8.4 Hz, 3H).

[0081] Scheme 1, step 2. 5-Chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile: To a solution of N-(2-acetyl-3,5-difluorophenyl)-5-chloro-2-cyanoisonicotinamide (53 mg, 158 μmol, 1 equiv.) in dioxane (1 mL) was added LiOH (7.6 mg, 316 μmol, 2 equiv.) at 20 °C under N2. The mixture was stirred at 110 °C for 1 h. The reaction mixture was concentrated under reduced pressure and the pH was adjusted to 6-7 with aqueous 1 M HCl. The mixture was filtered. The filtrate was then washed with water and dried under reduced pressure to give the title compound as a white solid (31 mg, 58% yield). 1H NMR (400 MHz, DMSO-d6) δ 12.23 (br s, 1H), 9.10 (s, 1H), 8.49 (s, 1H), 7.24 - 7.10 (m, 2H), 6.18 (s, 1H).LCMS[M+1]=318.0 / 320.0.

[0077]

[0082] Example 3

[0078] [ka]

[0079]

[0083] 5-Chloro-6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile

[0084] Scheme 1, Step 1. N-(2-Acetyl-3,5-difluorophenyl)-3-chloro-6-cyanopicolinamide: NaH (20.6 mg, 514 μmol, 60% purity, 1.1 equiv.) was added to a solution of 1-(2-amino-4,6-difluorophenyl)ethan-1-one in THF (0.8 mL) at 0° C. Then, a solution of 3-chloro-6-cyanopicolinoyl chloride (1.1 equiv.) in THF (0.4 mL) was added to the solution at 0° C. The mixture was stirred at 20° C. for 16 h. The mixture was diluted with water (15 mL) and extracted with ethyl acetate (2×10 mL). The combined organic layers were washed with brine (2×15 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was triturated with ethyl acetate (1.5 mL) at 20° C. to give the title compound as a white solid (102 mg, 57% yield, 87% purity). 1 H NMR (400 MHz, DMSO-d6) δ 12.38 (s, 1H), 8.45 (d, J = 8.4 Hz, 1H), 8.33 (d, J = 8.4 Hz, 1H), 8.06 - 8.01 (m, 1H), 7.30 (ddd, J = 2.4, 9.0, 11.6 Hz, 1H), 2.61 (d, J = 6.4 Hz, 3H).LCMS[M+1]=336.0 / 338.0.

[0080]

[0085] Scheme 1, step 2. 5-Chloro-6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile: To a solution of N-(2-acetyl-3,5-difluorophenyl)-3-chloro-6-cyanopicolinamide in dioxane (1 mL) was added LiOH (5.0 mg, 209 μmol, 1 equiv.) at 20 °C. The mixture was stirred at 110 °C for 2 h. Then, another portion of LiOH (5.0 mg, 209 μmol, 1 equiv.) was added to the reaction mixture at 20 °C, and the mixture was stirred at 110 °C for another 2 h. The pH of the reaction mixture was adjusted to 5-6 with aqueous 1 M HCl. The mixture was extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC (column: Phenomenex Luna 80 mm x 30 mm x 3 um; mobile phase: water (HCl)-acetonitrile) to give the title compound as a white solid (28.6 mg, 43% yield). 1 H NMR (400 MHz, CD3OD) δ 8.35 (d, J = 8.6 Hz, 1H), 8.08 (s, 1H), 7.29 (br d, J = 9.6 Hz, 1H), 7.00-7.11 (m, 1H), 6.75 ppm (s, 1H).LCMS[M+1]=317.9 / 319.9.

[0081]

[0086] The compounds in Table 1 were prepared according to Scheme 1 using procedures similar to those described in Examples 1, 2 and 3.

[0082] [Table 1]

[0083]

[0087] Example 5

[0084] [ka]

[0085]

[0088] 5-Chloro-6-(4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile

[0089] Scheme 1, Step 1. N-(2-Acetylphenyl)-3-chloro-6-cyanopicolinamide: To a solution of 1-(2-aminophenyl)ethan-1-one (200 mg, 1.48 mmol, 1 equiv.) in DCM (2.0 mL) was added triethylamine (618 μL, 4.44 mmol, 3 equiv.). Then, a solution of 3-chloro-6-cyanopicolinoyl chloride (360 mg) in DCM (2.0 mL) was added dropwise to the mixture at 25° C. The mixture was stirred at 25° C. for 0.5 h. The reaction mixture was partitioned between water (5 mL) and DCM (15 mL). The organic phase was separated, dried over Na2SO4, filtered, and concentrated under reduced pressure. The resulting residue was triturated with methanol (3 mL) and dried under reduced pressure to give the title compound as a white solid (140 mg, 32% yield). 1 H NMR (400 MHz, DMSO-d6) δ 12.85 (br s, 1H), 8.75 - 8.61 (m, 1H), 8.50 - 8.41 (m, 1H), 8.32 - 8.11 (m, 2H), 7.77 - 7.63 (m, 1H), 7.36 - 7.26 (m, 1H), 2.75 - 2.67 (m, 3H).LCMS[M+1]=300.1 / 302.1.

[0086]

[0090] Scheme 1, Step 2. 5-Chloro-6-(4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile: To a solution of LiOH (12.8 mg, 534 μmol, 2 equiv.) in dioxane (1.00 mL) at 25° C. was added N-(2-acetylphenyl)-3-chloro-6-cyanopicolinamide (80 mg, 267 μmol, 1 equiv.). The mixture was stirred at 110° C. for 6 h. The reaction mixture was quenched by the addition of saturated aqueous NH4Cl solution (3.00 mL) and the mixture was extracted with ethyl acetate (3×5 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex luna C18 80 mm x 40 mm x 3 μm; mobile phase: 20-40% acetonitrile (+HCl) in water) to give the title compound as a white solid (20 mg, 27% yield). 1 H NMR (400 MHz, CD3OD) δ 8.49 (d, J = 8.4 Hz, 1H), 8.44 (d, J = 8.4 Hz, 1H), 8.17 (d, J = 8.4 Hz, 1H), 8.08 (d, J = 3.6 Hz, 2H), 7.82 (td, J = 4.2, 8.4 Hz, 1H), 7.44 - 7.41 (m, 1H).LCMS[M+1]=281.9 / 284.0.

[0087]

[0091] The compounds in Table 2 were prepared according to Scheme 1 using procedures similar to those described in Example 5.

[0088] [Table 2]

[0089]

[0092] Example 7

[0090] [ka]

[0091]

[0093] 6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-methoxypicolinonitrile

[0094] Scheme 2, step 1. 6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-methoxypicolinonitrile: NaOMe (85.0 mg, 1.5 mmol, 5 equiv.) was added to a solution of 5-chloro-6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile (100 mg, 315 μmol, 1 equiv.) in DMF (1 mL) cooled to 0 °C. The reaction mixture was then stirred at 20 °C for 16 h. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm × 30 mm × 3 um; mobile phase: 40-70% acetonitrile (+ formic acid) in water) to give the title compound as a white solid (20 mg, 20% yield). 1 H NMR (400 MHz, DMSO-d6) δ 11.77 (br s, 1H), 8.26 (d, J = 8.8 Hz, 1H), 7.93 (d, J = 8.8 Hz, 1H), 7.50 (br d, J = 10.4 Hz, 1H), 7.10 (br t, J = 9.8 Hz, 1H), 6.68 (s, 1H), 4.04 (s, 3H).LCMS[M+1]=314.0.

[0092]

[0095] Example 8

[0093] [ka]

[0094]

[0096] 6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylthio)picolinonitrile

[0097] Scheme 2, step 1. 6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylthio)picolinonitrile: NaSMe (56.0 mg, 799 μmol, 2.5 equiv.) was added to a solution of 5-chloro-6-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile (100 mg, 315 μmol, 1 equiv.) in DMF (1 mL) cooled to 0° C., and the mixture was mixed for 1 h at 0° C. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm×30 mm×3 um; mobile phase: 20%-60% acetonitrile (+ formic acid) in water) to give the title compound as a white solid (57 mg, 55% yield). 1 H NMR (400 MHz, CD3OD) δ 8.04 (d, J = 8.6 Hz, 1H), 7.97 - 7.91 (m, 1H), 7.24 (br d, J = 9.4 Hz, 1H), 6.99 (ddd, J = 2.4, 9.4, 11.6 Hz, 1H), 6.63 (br s, 1H), 2.60 (s, 3H).LCMS[M+1]=330.0.

[0095]

[0098] Example 9

[0096] [ka]

[0097]

[0099] 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-methoxypicolinonitrile

[0100] Scheme 2, step 1. 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-methoxypicolinonitrile: To a solution of 5-chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile (100 mg, 315 μmol, 1 equiv.) in DMF (1.5 mL) was added NaOMe (85.0 mg, 1.5 mmol, 5 equiv.) at 0 °C. The mixture was stirred at 20 °C for 16 h. The reaction solution was concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Luna 80 mm × 30 mm × 3 um; mobile phase: 15-45% methanol in water (+HCl)) to give the title compound as a yellow solid (4.6 mg, 4.5% yield). 1 H NMR (400 MHz, DMSO-d6): δ 8.79 (s, 1H), 8.25-8.29 (m, 1H), 7.11-7.25 (m, 2H), 6.24-6.37 (m, 1H), 4.08 (s, 3H).LCMS[M+1]=314.0.

[0098]

[0101] Example 10

[0099] [ka]

[0100]

[0102] 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylthio)picolinonitrile Scheme 2, step 1. 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylthio)picolinonitrile: To a solution of 5-chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile (80 mg, 252 μmol, 1 equiv.) in DMF (1.5 mL) was added NaSMe (44.8 mg, 640 μmol, 2.54 equiv.) at 0 °C. The mixture was stirred at 0 °C for 1 h. The reaction mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75 mm × 30 mm × 3 um; mobile phase: 30-60% acetonitrile (+ formic acid) in water) to give the title compound as a white solid (16 mg, 19% yield). 1 H NMR (400 MHz, DMSO-d6): δ 11.96-12.26 (m, 1H), 8.81 (s, 1H), 8.19 (s, 1H), 7.11-7.25 (m, 2H), 6.11-6.26 (m, 1H), 2.66-2.66 (m, 3H).LCMS[M+1]=329.9.

[0101]

[0103] The compounds in Table 3 were prepared according to Scheme 1 using procedures similar to those described in Example 7.

[0102] [Table 3]

[0103]

[0104] Example 12

[0104] [ka]

[0105] 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(ethylthio)picolinonitrile

[0106] Scheme 2, step 1. 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(ethylthio)picolinonitrile: To a solution of ethanethiol (29.1 μL, 393 μmol, 2.5 equiv.) in DMF (2 mL) was added NaH (15.7 mg, 393 μmol, 2.5 equiv.; 60% dispersion in oil) at 0° C. and the mixture was mixed under N2 for 30 min. 5-Chloro-4-(5,7-difluoro-4-oxo-1H-quinolin-2-yl)pyridine-2-carbonitrile (50 mg, 157 μmol, 1 equiv.) was then added to the mixture in one portion. The mixture was stirred at room temperature for 16 h under N2. The reaction mixture was poured into saturated aqueous NH4Cl (5 mL) and stirred for 2 min. The aqueous phase was extracted with ethyl acetate (2×5 mL). The combined organic layers were washed with brine (2×5 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75×30 mm×3 um; mobile phase: 1-40% acetonitrile in water (+ formic acid modifier) ​​to give the title compound as a white solid (51.7 mg, 91% yield). LCMS: [M+1]=344.0. 1 H NMR (400 MHz, DMSO-d6) δ 12.11 (br s, 1 H) 8.85 (s, 1 H) 8.21 (s, 1 H) 7.05 - 7.24 (m, 2 H) 6.06 (br s, 1 H) 3.23 (br d, J = 7.2 Hz, 2 H) 1.26 (t, J = 7.2 Hz, 3 H).

[0107] Example 13

[0106] [ka]

[0107]

[0108] 5-Chloro-6-(5,7-difluoro-6-(4-methylpiperazin-1-yl)-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile

[0109] Step 1, 1-(3-Bromo-2,6-difluoro-4-nitro-phenyl)-4-methyl-piperazine: To a solution of 1-methylpiperazine (1.73 mL, 15.6 mmol, 1.0 equiv) and K2CO3 (3.24 g, 23.4 mmol, 1.5 equiv) in DMSO (40 mL) was added 4-bromo-1,2,3-trifluoro-5-nitro-benzene (4 g, 15.63 mmol, 1.0 equiv). The mixture was stirred at 20 °C for 16 h. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (3 x 50 mL). The combined organic layers were washed with brine (2 x 100 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound as a brown solid (4.73 g, 90% yield). LCMS [M+1] = 336.0. 1 H NMR (400 MHz, chloroform-d) δ 7.66 (dd, J = 1.8, 11.8 Hz, 1H), 3.40 (br t, J = 4.6 Hz, 4H), 2.58 - 2.51 (m, 4H), 2.36 (s, 3H).

[0108]

[0110] Step 2, 2-Bromo-3,5-difluoro-4-(4-methylpiperazin-1-yl)aniline: A solution of 1-(3-bromo-2,6-difluoro-4-nitro-phenyl)-4-methyl-piperazine (2.5 g, 7.44 mmol, 1.0 equiv) in EtOH (20 mL) was added to a solution of NH4Cl (1.99 g, 37.2 mmol, 5.0 equiv) and Fe (2.08 g, 37.2 mmol, 5.0 equiv) in H2O (10 mL) at room temperature. The solution was then stirred at 80 °C for 1.5 h under N2. The reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under reduced pressure. The residue was then diluted with water (50 mL) and extracted with DCM (3 x 40 mL). The combined organic layers were washed with brine (70 mL), dried over Na2SO4, filtered and concentrated under reduced pressure to give the title compound as a brown solid (2.08 g, 91% yield). LCMS [M+1] = 306.0. 1H NMR (400 MHz, chloroform-d) δ 6.32 (dd, J = 2.2, 12.6 Hz, 1H), 4.14 (br s, 2H), 3.16 - 3.09 (m, 4H), 2.59 - 2.48 (m, 4H), 2.35 (s, 3H).

[0109]

[0111] Step 3, 1-[6-amino-2,4-difluoro-3-(4-methylpiperazin-1-yl)phenyl]ethenone: To a solution of 2-bromo-3,5-difluoro-4-(4-methylpiperazin-1-yl)aniline (1 g, 3.27 mmol, 1.0 equiv.) in toluene (10 mL) was added tributyl(1-ethoxyvinyl)stannane (3.31 mL, 9.80 mmol, 3.0 equiv.) and Pd(PPh3)4 (377 mg, 327 μmol, 0.1 equiv.) under N2. The mixture was stirred at 120° C. for 16 h under N2. The reaction mixture was cooled to room temperature and the reaction was quenched by the addition of an aqueous solution of KF (30 mL). The mixture was stirred at room temperature for 2 h, diluted with water (10 mL) and extracted with EtOAc (3×20 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was diluted in dioxane (15 mL) and 4N HCl in dioxane (2.08 mL, 1 equiv.) was added. The mixture was stirred at room temperature for 1 h. The reaction mixture was quenched by addition of saturated aqueous NaHCO3 (50 mL), diluted with water (50 mL) and extracted with DCM (3×30 mL). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (1:0-1:3 petroleum ether:ethyl acetate) to give impure material, which was re-purified by preparative HPLC (column: Phenomenex luna C18 (250×70 mm×15 um); mobile phase: 15-35% acetonitrile in water (+ formic acid modifier)) to give the title compound as a yellow solid (190 mg, 8.5% yield). LCMS[M+1]=270.2.

[0110]

[0112] Step 4, N-[2-acetyl-3,5-difluoro-4-(4-methylpiperazin-1-yl)phenyl]-3-chloro-6-cyano-pyridine-2-carboxamide: A mixture of 3-chloro-6-cyano-pyridine-2-carboxylic acid (71 mg, 389 μmol, 1.0 equiv) in SOCl2 (2 mL) was purged with N2, and then the mixture was stirred under N2 for 2 h at 100° C. The reaction mixture was concentrated under reduced pressure to give 3-chloro-6-cyano-pyridine-2-carbonyl chloride (80 mg, crude) as a white solid, which was used in the next step of this step without further purification. A mixture of 3-chloro-6-cyano-pyridine-2-carbonyl chloride (77.6 mg, 386 μmol, 1.3 equiv.), 1-[6-amino-2,4-difluoro-3-(4-methylpiperazin-1-yl)phenyl]ethanone (80 mg, 297 μmol, 1.0 equiv.) in isopropyl acetate (3.5 mL) was purged with N2, and then the mixture was stirred at 80° C. for 16 h under N2. The reaction mixture was concentrated under reduced pressure, and the residue was purified by preparative TLC (10:1 DCM:MeOH) to give the title compound as a yellow solid (55 mg, 43% yield). LCMS [M+1]=434.1. 1 H NMR (400 MHz, methanol-d4) δ 8.26 (dd, J = 1.8, 14.2 Hz, 1H), 8.22 (d, J = 8.4 Hz, 1H), 8.02 (d, J = 8.4 Hz, 1H), 4.55 (br s, 4H), 3.38 - 3.34 (m, 4H), 2.69 (s, 3H), 2.63 (d, J = 7.6 Hz, 3H).

[0111]

[0113] Step 5, 5-chloro-6-(5,7-difluoro-6-(4-methylpiperazin-1-yl)-4-oxo-1,4-dihydroquinolin-2-yl)picolinonitrile: To a solution of N-[2-acetyl-3,5-difluoro-4-(4-methylpiperazin-1-yl)phenyl]-3-chloro-6-cyano-pyridine-2-carboxamide (85 mg, 196 μmol, 1.0 equiv.) in 2-MeTHF (2 mL), LiOH (9.4 mg, 392 μmol, 2.0 equiv.) was added. The reaction mixture was stirred at 90 °C for 12 h. The reaction mixture was cooled to room temperature and then the pH was adjusted to pH = 2-3 with aqueous 1 M HCl. The aqueous phase was extracted with ethyl acetate (5 mL). The aqueous phase was purified by preparative HPLC (column: Phenomenex Luna C18 75×30 mm×3 um; mobile phase: 1-40% acetonitrile in water (+ formic acid modifier)) to give the title compound as a yellow solid (28.7 mg, 35% yield). LCMS [M+1]=416.1. 1 H NMR (400 MHz, methanol-d4) δ 8.32 (d, J = 8.4 Hz, 1H), 8.05 (d, J = 8.4 Hz, 1H), 7.28 (dd, J = 1.8, 11.8 Hz, 1H), 6.65 (s, 1H), 3.39 (br t, J = 4.6 Hz, 4H), 2.95 (br s, 4H), 2.62 (s, 3H).

[0114] The compounds in Table 4 were prepared according to Scheme 1 using procedures similar to those described in Example 13.

[0112] [Table 4-1]

[0113] [Table 4-2]

[0114] [Table 4-3]

[0115] [Table 4-4]

[0116] Example 27

[0117] [ka]

[0118] 6-Chloro-5-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)nicotinonitrile Step 1, 2-hydroxy-5-iodonicotinic acid: To a solution of methyl 2-chloro-5-iodonicotinate (3.0 g, 10.7 mmol, 1.0 equiv) in THF (20 mL) and HO (7 mL) was added LiOH·HO (1.3 g, 32.2 mmol, 3.0 equiv). The mixture was stirred at 50° C. for 16 h. The mixture was concentrated under reduced pressure to remove THF. The pH of the mixture was adjusted to pH=6 with aqueous 1M HCl. During the pH adjustment, a white solid precipitated and was collected by filtration. The filter cake was washed with HO (10 mL) and concentrated in vacuo to give 2-hydroxy-5-iodonicotinic acid as a white solid. (2.5 g, 91% yield) LCMS [M+1]=265.9. 1 H NMR (400 MHz, DMSO-d6) δ 8.28 (d, J = 2.4 Hz, 1H), 8.21 (d, J = 2.4 Hz, 1H). Step 3, N-(2-acetyl-3,5-difluorophenyl)-2-chloro-5-iodonicotinamide: A solution of 2-hydroxy-5-iodonicotinic acid (2.5 g, 9.7 mmol, 1.0 equiv) in POCl3 (15 mL) was stirred for 5 h at 90° C. The reaction mixture was concentrated under reduced pressure to remove volatiles to give the title compound as a white solid (3 g, crude), which was used in the next step of the reaction without further purification.

[0119] To a solution of 1-(2-amino-4,6-difluorophenyl)ethanone (1.4 g, 8.7 mmol, 1.0 equiv.) in THF (15 mL) was added NaH (384.2 mg, 9.6 mmol, 1.1 equiv.; 60% dispersion in oil). Then, 2-chloro-5-iodonicotinoyl chloride (2.9 g, 9.61 mmol, 1.1 equiv.) was added to the mixture. The mixture was stirred at room temperature for 16 h. The reaction mixture was quenched by the addition of a saturated aqueous solution of NH4Cl (10 mL). The aqueous layer was extracted with ethyl acetate (2×40 mL) and the combined organic layers were washed with brine (15 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with ethyl acetate at 25° C. for 0.5 h to give the title compound as a white solid (3.0 g, 79% yield). LCMS[M+1]=436.9. 1 H NMR (400 MHz, DMSO-d6) δ 11.34 (s, 1H), 8.80 (d, J = 2.4 Hz, 1H), 8.36 (d, J = 2.4 Hz, 1H), 7.50 - 7.40 (m, 1H), 7.30 (ddd, J = 2.6, 8.8, 11.2 Hz, 1H), 2.54 (d, J = 3.8 Hz, 3H).

[0120] Step 4, 2-(2-chloro-5-iodopyridin-3-yl)-5,7-difluoroquinolin-4(1H)-one: To a solution of N-(2-acetyl-3,5-difluorophenyl)-2-chloro-5-iodonicotinamide (740 mg, 1.6 mmol, 1.0 equiv.) in dioxane (7 mL) was added LiOH (40.6 mg, 1.6 mmol, 1.0 equiv.). The mixture was stirred at 110° C. for 16 h. The reaction mixture was cooled to room temperature, diluted with H2O (5 mL), and extracted with ethyl acetate (3×10 mL). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was triturated with acetonitrile at room temperature for 0.5 h to give the title compound as a white solid (200 mg, 28% yield). LCMS [M+1]=418.9. 1H NMR (400 MHz, DMSO-d6) δ 12.11 (br s, 1H), 8.85 (d, J = 2.0 Hz, 1H), 8.52 (d, J = 1.6 Hz, 1H), 7.12 (br d, J = 8.2 Hz, 2H), 6.10 (br s, 1H).

[0121] Step 5, 6-chloro-5-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)nicotinonitrile: To a solution of 2-(2-chloro-5-iodopyridin-3-yl)-5,7-difluoroquinolin-4(1H)-one (150 mg, 358 μmol, 1.0 equiv.) in DMA (3 mL) was added Zn(CN)2 (25.2 mg, 215 μmol, 0.6 equiv.) and Pd(PPh3)4 (41.4 mg, 35.8 μmol, 0.1 equiv.) under N2. The mixture was stirred at 120° C. for 3 h under N2. The reaction mixture was cooled to room temperature and then poured into water (5 mL). The aqueous phase was extracted with ethyl acetate (2×10 mL). The combined organic layers were washed with brine (2×10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex C18 75×30 mm×3 um; mobile phase: 10-40% acetonitrile in water (+NH4HCO3 modifier)) to give the title compound as a white solid (52.1 mg, 44% yield). LCMS[M+1]=317.9. 1 H NMR (400 MHz, DMSO-d6) δ 12.18 (br d, J = 2.2 Hz, 1H), 9.11 (d, J = 2.2 Hz, 1H), 8.73 (d, J = 2.2 Hz, 1H), 7.16 (br d, J = 10.6 Hz, 2H), 6.35 - 5.92 (m, 1H).

[0122] Example 28

[0120] [ka]

[0121]

[0123] 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfinyl)picolinonitrile 4-(5,7-Difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfinyl)picolinonitrile: To a mixture of 4-(5,7-difluoro-4-oxo-1H-quinolin-2-yl)-5-methylsulfanyl-pyridine-2-carbonitrile (60 mg, 182 μmol, 1 equiv.) in DCM (1 mL) was added m-CPBA (55.4 mg, 273 μmol, 85% purity, 1.5 equiv.) at room temperature under N2. The mixture was stirred at room temperature for 16 h. The reaction mixture was poured into a saturated aqueous solution of Na2SO3 (15 mL). The aqueous phase was extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine (2×20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified twice by preparative HPLC (1-column: Waters Xbridge BEH C18 100×30 mm×10 um; mobile phase: 10-35% acetonitrile in water (+NH4HCO3 modifier)-ACN]; and 2-column: Phenomenex Luna C18 150×30 mm×5 um; mobile phase: 1-35% acetonitrile in water (+formic acid modifier) ​​to give the title compound as an off-white solid (8.4 mg, 13% yield). LCMS [M+1]=346.0. 1 H NMR (400 MHz, DMSO-d6) δ 9.37 (s, 1H), 8.60 (s, 1H), 7.73 - 7.51 (m, 1H), 7.48 - 7.20 (m, 2H), 2.52 (br d, J = 1.8 Hz, 3H).

[0122]

[0124] Example 29

[0123] [ka]

[0124]

[0125] 4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfonyl)picolinonitrile 4-(5,7-Difluoro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfonyl)picolinonitrile: To a mixture of 4-(5,7-difluoro-4-oxo-1H-quinolin-2-yl)-5-methylsulfanyl-pyridine-2-carbonitrile (110 mg, 334 μmol, 1.0 equiv.) in acetone (1 mL), water (0.6 mL), methanol (0.7 mL) and THF (0.7 mL) was added oxone (616.03 mg, 1.00 mmol, 3.0 equiv.). The mixture was stirred at 40° C. for 32 h. The reaction mixture was poured into H2O (15 mL). The aqueous phase was extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine (2×20 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75×30 mm×3 um; mobile phase: 20-50% acetonitrile in water (+ formic acid modifier) ​​to give the title compound as an off-white solid (16.8 mg, 14% yield). LCMS [M+1]=361.9. 1 H NMR (400 MHz, DMSO-d6) δ 11.96 (br s, 1H), 9.35 (br s, 1H), 8.43 (br s, 1H), 7.27 - 6.88 (m, 2H), 6.18 (br s, 1H), 3.75 - 3.38 (m, 3H).

[0125]

[0126] Example 30

[0126] [ka]

[0127] 4-(7-chloro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfonyl)picolinonitrile

[0128] Step 1, N-(2-acetyl-5-chloro-phenyl)-2-cyano-5-methylsulfanyl-pyridine-4-carboxamide: To a solution of 1-(2-amino-4-chloro-phenyl)ethanone (262 mg, 1.5 mmol, 1.0 equiv) in isopropyl acetate (3 mL) was added 2-cyano-5-methylsulfanyl-pyridine-4-carbonyl chloride (328.5 mg, 1.54 mmol, 1.0 equiv). The mixture was stirred at 80° C. for 16 h. The reaction mixture was cooled to room temperature and the volatiles were removed under reduced pressure. The residue was triturated with MTBE at room temperature for 30 min. The mixture was filtered and the filter cake was concentrated under reduced pressure to give the title compound as a white solid (440 mg, 82% yield). LCMS [M+1]=346.0. 1 HNMR (400 MHz, DMSO-d6) δ 11.90 (s, 1H), 8.82 (s, 1H), 8.38 (d, J = 2.2 Hz, 1H), 8.23 ​​(s, 1H), 8.08 (d, J = 8.6 Hz, 1H), 7.44 (dd, J = 2.2, 8.6 Hz, 1H), 2.65 (d, J = 7.8 Hz, 6H).

[0128]

[0129] Step 2, 4-(7-chloro-4-oxo-1H-quinolin-2-yl)-5-methylsulfanyl-pyridine-2-carbonitrile: To a solution of N-(2-acetyl-5-chloro-phenyl)-2-cyano-5-methylsulfanyl-pyridine-4-carboxamide (345 mg, 998 μmol, 1.0 equiv.) in dioxane (3 mL) was added LiOH (23.8 mg, 998 μmol, 1.0 equiv.). The mixture was stirred at 110° C. for 12 h. The pH of the mixture was adjusted to pH=3-4 with aqueous 1M HCl. The mixture was then diluted with water (3 mL) and a precipitate formed. The mixture was filtered and concentrated under reduced pressure to give a residue. The crude product was triturated with acetonitrile at room temperature for 30 min to give the title compound as a white solid (235 mg, 72% yield). LCMS[M+1]=328.1. 1H NMR (400 MHz, DMSO-d6) δ 12.08 (br s, 1H), 8.81 (s, 1H), 8.21 (s, 1H), 8.12 (d, J = 8.6 Hz, 1H), 7.58 (d, J = 1.6 Hz, 1H), 7.42 (dd, J = 1.8, 8.6 Hz, 1H), 6.17 (s, 1H), 2.74 - 2.60 (m, 3H).

[0129]

[0130] Step 3, 4-(7-chloro-4-oxo-1,4-dihydroquinolin-2-yl)-5-(methylsulfonyl)-picolinonitrile: To a solution of 4-(7-chloro-4-oxo-1H-quinolin-2-yl)-5-methylsulfanyl-pyridine-2-carbonitrile (288 mg, 879 μmol, 1.0 equiv.) in acetone (0.35 mL), HO (0.2 mL), MeOH (0.25 mL), THF (0.25 mL) was added oxone (1.6 g, 2.6 mmol, 3.0 equiv.). The mixture was stirred at 40° C. for 16 h. The mixture was quenched by the addition of saturated aqueous Na2SO3 (111 mg, 879 μmol, 1.0 equiv.). The mixture was then diluted with water (5 mL) and filtered. The filtrate was concentrated in vacuo and the residue was purified by preparative HPLC (column: Phenomenex Luna C18 200×40 mm×10 um; mobile phase: 20-50% acetonitrile in water (+ formic acid modifier)) to give the title compound as a white solid (74.7 mg, 24% yield). LCMS [M+1]=360.8. 1 H NMR (400 MHz, DMSO-d6) δ 12.77 - 11.60 (m, 1H), 9.35 (s, 1H), 8.54 (br s, 1H), 8.18 (br d, J = 8.8 Hz, 1H), 8.00 - 7.29 (m, 2H), 7.08 - 5.99 (m, 1H), 3.54 (br s, 3H).

[0130]

[0131] The compounds in Table 5 were prepared according to Scheme 1 using procedures similar to those described for Example 30.

[0131] [Table 5]

[0132] Example 33

[0133] [ka]

[0134] 5-Chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)pyrimidine-2-carbonitrile Step 1, N-(2-acetyl-3,5-difluoro-phenyl)-5-chloro-2-methylsulfanyl-pyrimidine-4-carboxamide: A mixture of 5-chloro-2-methylsulfanyl-pyrimidine-4-carboxylic acid (1.4 g, 6.84 mmol, 1.0 equiv) in SOCl2 (20 mL) was purged with N2, and then the mixture was stirred at 80° C. for 0.5 h under N2. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to give 5-chloro-2-methylsulfanyl-pyrimidine-4-carbonyl chloride (1.45 g, crude) as a black solid. The product was used directly in the next step of the reaction without further purification.

[0135] To a solution of 1-(2-amino-4,6-difluoro-phenyl)ethanone (1.0 g, 5.8 mmol, 1.0 equiv) in isopropyl acetate (20 mL) was added 5-chloro-2-methylsulfanyl-pyrimidine-4-carbonyl chloride (1.4 g, 6.4 mmol, 1.1 equiv). The mixture was purged with N2 and then stirred at 80° C. for 2 h under N2. The mixture was cooled to room temperature and then diluted with H2O (5 mL), filtered, and concentrated under reduced pressure. The residue was triturated with MTBE at room temperature for 5 min to give the title compound as a white solid (1.9 g, 90% yield). LCMS [M+1]=358.0. 1H NMR (400 MHz, chloroform-d) δ 13.16 (br s, 1H), 8.69 (s, 1H), 8.61 - 8.52 (m, 1H), 6.70 (ddd, J = 2.6, 8.2, 11.8 Hz, 1H), 2.80 (s, 3H), 2.68 (d, J = 8.0 Hz, 3H).

[0136] Step 2, 2-(5-chloro-2-methylsulfanyl-pyrimidin-4-yl)-5,7-difluoro-1H-quinolin-4-one: To a solution of N-(2-acetyl-3,5-difluoro-phenyl)-5-chloro-2-methylsulfanyl-pyrimidine-4-carboxamide (1.0 g, 2.8 mmol, 1.0 equiv.) in dioxane (10 mL) was added LiOH (134 mg, 5.6 mmol, 2.0 equiv.). The mixture was purged with N2, and then the mixture was stirred at 110° C. for 6 h under N2. The pH of the reaction mixture was adjusted to pH=6 with aqueous 1M HCl and diluted with H2O (5 mL). The precipitate was filtered off, and the filter cake was triturated with MTBE at room temperature for 5 min to give the title compound as a white solid (720 mg, 76% yield). LCMS[M+1]=340.0. 1 H NMR (400 MHz, methanol-d4) δ 8.84 (s, 1H), 7.26 (br d, J = 9.4 Hz, 1H), 7.05 - 6.95 (m, 1H), 6.73 (br s, 1H), 2.63 (s, 3H).

[0137] Step 3, 2-(5-chloro-2-methylsulfonyl-pyrimidin-4-yl)-5,7-difluoro-1H-quinolin-4-one: To a solution of 2-(5-chloro-2-methylsulfanyl-pyrimidin-4-yl)-5,7-difluoro-1H-quinolin-4-one (300 mg, 883 μmol, 1.0 equiv.) in acetone (15 mL), HO (8 mL), MeOH (1 mL) and THF (1 mL) was added oxone (1.6 g, 2.6 mmol, 3.0 equiv.). The mixture was purged with N2 and then stirred at room temperature for 2 h under N2. The reaction mixture was diluted with HO (20 mL) and the aqueous layer was extracted with ethyl acetate (5×15 mL). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (3:1 to 0:1 petroleum ether:ethyl acetate) to give the title compound as a yellow solid (185 mg, 52% yield). LCMS [M+1] = 371.9. 1 H NMR (400 MHz, methanol-d4) δ 9.31 (s, 1H), 7.29 (br d, J = 7.8 Hz, 1H), 7.03 (br t, J = 10.0 Hz, 1H), 6.90 (br s, 1H), 3.47 (s, 3H).

[0138] Step 4, 5-chloro-4-(5,7-difluoro-4-oxo-1,4-dihydroquinolin-2-yl)pyrimidine-2-carbonitrile: To a solution of 2-(5-chloro-2-(methylsulfonyl)pyrimidin-4-yl)-5,7-difluoroquinolin-4(1H)-one (50.0 mg, 135 μmol, 1.0 equiv.) in DMSO (2.5 mL) cooled to 10° C., NaCN (16.5 mg, 336 μmol, 2.5 equiv.) was added and the mixture was stirred at 10° C. for 10 min. The reaction was diluted with H2O (10 mL) and the aqueous phase was extracted with ethyl acetate (10×15 mL). The combined organic phase was washed with brine (2×10 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative HPLC (column: Phenomenex Luna C18 75×30 mm×3 um; mobile phase: 10-50% acetonitrile in water (+ formic acid modifier)) to give the title compound as an off-white solid (18.2 mg, 42% yield). LCMS [M+1]=318.9. 1 H NMR (400 MHz, DMSO-d6) δ 12.58 - 11.84 (m, 1H), 9.42 (s, 1H), 7.43 (br d, J = 9.8 Hz, 1H), 7.27 (br s, 1H), 6.96 - 6.49 (m, 1H).

[0139] Example 34

[0140] [ka]

[0141] (S)-1-(2-(2-cyano-5-(methylsulfonyl)pyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide Step 1, N-[2-acetyl-4-[3-(dimethylcarbamoyl)-1-piperidyl]-3,5-difluoro-phenyl]-5-chloro-2-cyano-pyridine-4-carboxamide: To a solution of 1-(3-acetyl-4-amino-2,6-difluoro-phenyl)-N,N-dimethyl-piperidine-3-carboxamide (300 mg, 922 μmol, 1.0 equiv.) in isopropyl acetate (5 mL) was added 5-chloro-2-cyano-pyridine-4-carbonyl chloride (222 mg, 1.1 mmol, 1.2 equiv.) at room temperature under N2. The mixture was stirred at 80° C. for 2 hours. The reaction mixture was cooled to room temperature and poured into water (20 mL). The aqueous phase was extracted with ethyl acetate (3×20 mL). The combined organic layers were washed with brine, dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (50:1 to 1:1 petroleum ether:ethyl acetate) to give the title compound as a yellow solid (360 mg, 80% yield). LCMS [M+1] = 490.2. 1 H NMR (400 MHz, chloroform-d) δ 12.13 (s, 1H), 8.82 (s, 1H), 8.39 (dd, J = 1.8, 13.6 Hz, 1H), 7.90 (s, 1H), 3.41 - 3.31 (m, 1H), 3.24 - 3.14 (m, 3H), 3.14 - 3.10 (m, 3H), 2.96 (s, 4H), 2.71 - 2.63 (m, 3H), 2.00 - 1.88 (m, 1H), 1.87 - 1.66 (m, 3H).

[0142] Step 2, 1-[2-(5-chloro-2-cyano-4-pyridyl)-5,7-difluoro-4-oxo-1H-quinolin-6-yl]-N,N-dimethyl-piperidine-3-carboxamide: To a mixture of N-[2-acetyl-4-[3-(dimethylcarbamoyl)-1-piperidyl]-3,5-difluoro-phenyl]-5-chloro-2-cyano-pyridine-4-carboxamide (360 mg, 735 μmol, 1.0 equiv.) in 2-MeTHF (5 mL), LiOH (52.8 mg, 2.2 mmol, 3.0 equiv.) was added at room temperature under N2. The mixture was stirred at 80° C. for 16 h. The pH of the reaction mixture was adjusted to pH=5-6 with aqueous 1N HCl. The mixture was poured into ice water (70 mL) and the aqueous phase was extracted with ethyl acetate (2 x 70 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (100:1 to 30:1 DCM:MeOH) to give the title compound as a yellow solid (185 mg, 53% yield). LCMS [M+1] = 472.2. 1 H NMR (400 MHz, DMSO-d6) δ 12.13 - 11.89 (m, 1H), 9.15 - 8.94 (m, 1H), 8.46 (s, 1H), 7.21 - 7.06 (m, 1H), 6.08 (s, 1H), 3.11 (br d, J = 6.4 Hz, 3H), 3.07 - 2.97 (m, 4H), 2.97 - 2.84 (m, 1H), 2.80 (s, 3H), 1.85 (br d, J = 11.8 Hz, 1H), 1.78 - 1.62 (m, 2H), 1.58 - 1.39 (m, 1H).

[0143] Step 3, (S)-1-(2-(2-cyano-5-(methylthio)pyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide: To a mixture of 1-(2-(5-chloro-2-cyanopyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide (182 mg, 386 μmol, 1.0 equiv) in DMF (3 mL) was added NaSMe (67.6 mg, 964 μmol, 2.5 equiv) in one portion at 0° C. under N2. The mixture was stirred at 0° C. for 1 h. The reaction mixture was poured into ice water (20 mL) and stirred for 10 min. The aqueous phase was extracted with ethyl acetate (2 x 20 mL). The combined organic layers were washed with brine (3 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. Further material was extracted from the aqueous phase by adjusting the pH to 5-6 with aqueous 1M HCl and collecting the precipitate that formed. The combined crude products were triturated with acetonitrile. The crude racemic title compound was resolved into enantiomers by SFC (column: DAICEL CHIRALCEL OJ (250 mm x 30 mm x 10 um); mobile phase: 25% MeOH in CO2 (+0.1% NH3H2O) to give (R)-1-(2-(2-cyano-5-(methylthio)pyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide (early-eluting enantiomer) (35.0 mg, 19% yield) and the title compound (late-eluting enantiomer) (35.0 mg, 19% yield) as yellow solids. LCMS [M+1] = 484.0. 1H NMR (400 MHz, DMSO-d6) δ 12.52 - 11.50 (m, 1H), 8.78 (s, 1H), 8.14 (s, 1H), 7.48 - 7.01 (m, 1H), 6.46 - 5.96 (m, 1H), 3.14 (br d, J = 6.2 Hz, 3H), 3.05 (s, 4H), 2.95 - 2.85 (m, 2H), 2.81 (s, 3H), 2.65 (s, 3H), 1.85 (br d, J = 11.0 Hz, 1H), 1.79 - 1.61 (m, 2H), 1.57 - 1.40 (m, 1H).

[0144] Step 4, (S)-1-(2-(2-cyano-5-(methylsulfonyl)pyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide: To a solution of (S)-1-(2-(2-cyano-5-(methylthio)pyridin-4-yl)-5,7-difluoro-4-oxo-1,4-dihydroquinolin-6-yl)-N,N-dimethylpiperidine-3-carboxamide (31.1 mg, 64.2 μmol, 1.0 equiv.) in DCM (0.5 mL) cooled to 0° C. was added m-CPBA (39.1 mg, 193 μmol, 3.0 equiv.; 85% purity). The mixture was stirred at room temperature for 3 h. The reaction was quenched by the addition of a saturated aqueous solution of Na2SO3. The mixture was concentrated in vacuo. The residue was diluted with DMF (2 mL) and filtered to remove solids. The filtrate was purified by preparative HPLC (column: Phenomenex Luna C18 75 x 30 mm x 3 um; mobile phase: 30-60% acetonitrile in water (+ formic acid modifier) ​​to give the title compound as a yellow solid (4.0 mg, 7.5 μmol, 12% yield). LCMS [M+1] = 516.1. 1H NMR (400 MHz, methanol-d4) δ 9.40 (s, 1H), 8.29 (br s, 1H), 7.26 - 6.82 (m, 1H), 6.47 - 6.12 (m, 1H), 3.28-3.05 (m, 10H), 3.14 - 3.05 (m, 1H), 3.01 (s, 3H), 1.98 (br dd, J = 3.2, 12.8 Hz, 1H), 1.85 (br s, 2H), 1.74 - 1.58 (m, 1H).

[0145] Biochemical and cellular assays PPARγ-NCOR1 recruitment assay:

[0146] Compound potency (EC 50 ) and the maximal extent of NCOR1 recruitment to PPARG was assessed with a TR-FRET binding assay measuring the association of a biotinylated NCOR1 ID2 peptide (biotin-GHSFADPASNLGLEDIIRKALMG-amide) to the PPARG / RXRA LBD heterodimer. Specifically, 20 microliters of TR-FRET master mix consisting of 2 nM WT PPARG LBD (E. coli expressed, His-TEV-Q203-Y477; Uniprot ID P37231-2), 2 nM WT RXRA LBD or mutant S427F RXRA LBD (E. coli expressed, Flag-TEV-E228-T462; P19793-1), 50 nM NCOR1, 80 nM rosiglitazone, 25 nM streptavidin-d2 (Cisbio) and 0.3 nM anti-His Tb (Cisbio) in 25 mM MOPS pH 7.4, 25 mM KCl, 1 mM EDTA, 0.01% BSA, 0.01% Tween-20 and 1 mM TCEP was added to 60 μL of compound (0.3% fc) in DMSO. Ten-point dose response titrations of DMSO (v / v) were added to 384-well plates in duplicate. The mixtures were incubated for 3 hours and read on an EnVision plate reader (Perkin Elmer) at Ex / Em 615 / 665. Potency (EC50 To determine the extent of NCOR1 recruitment, TR-FRET ratios were normalized to the average ratio of DMSO control wells (0%) and to the average maximum ratio of a positive control compound (T0070907 (2-chloro-5-nitro-N-4-pyridinyl-benzamide); defined as 100%) in the CDD Vault and analyzed using the Levenberg-Marquardt algorithm.

[0146]

[0147] PPARγ-MED1 Blockade Assay:

[0148] Compound potency (IC 50 ) and the maximal extent of MED1 repulsion towards PPARG was assessed in a TR-FRET binding assay measuring the association of biotinylated MED1 LxxLL peptide (biotin- VSSMAGNTKNHPMLMNLLKDNPAQ-amide) to the PPARG / RXRA LBD heterodimer. Specifically, 20 microliters of TR-FRET master mix consisting of 2 nM WT PPARG LBD (E. coli expressed, His-TEV-Q203-Y477; Uniprot ID P37231-2), 2 nM WT RXRA LBD (E. coli expressed, Flag-TEV-E228-T462; P19793-1), 350 nM NCOR1, 80 nM rosiglitazone, 175 nM streptavidin-d2 (Cisbio) and 0.3 nM anti-His Tb (Cisbio) in 25 mM MOPS pH 7.4, 25 mM KCl, 1 mM EDTA, 0.01% BSA, 0.01% Tween-20 and 1 mM TCEP was added to 60 μL of the compounds in DMSO (0.3% DMSO). Ten-point reaction titrations of fc (v / v) were added to a 384-well plate containing duplicates. The mixtures were incubated for 3 hours and read on an EnVision plate reader (Perkin Elmer) at Ex / Em 615 / 665. The potency of MED1 repulsion (IC 50To determine the degree of FRET concentration, TR-FRET ratios were normalized to the average ratio of DMSO control wells (0%) and to the average minimum ratio for a positive control compound (GW9662 (2-chloro-5-nitrobenzanilide); defined as 100%) in the CDD Vault and analyzed using the Levenberg-Marquardt algorithm.

[0147]

[0149] Bladder Cancer Pharmacodynamic Assay

[0150] 5637 (PPARG amplified) and HT1197 (RXRA S427F mutant) cells were used for evaluation of PPARG target gene regulation using quantitative PCR. Cells were treated with PPARG inverse agonists for 24 hours, after which FABP4 (IDT, Cat. No. Hs.PT 58.20106818) and ANGPTL4 (IDT, Cat. No. Hs.PT 58.25480012) expression was analyzed, using expression of the housekeeping gene TBP (IDT, Cat. No. Hs.PT 58v.39858774) to normalize expression across samples. Quantitative PCR was performed using an ABI QuantStudio 7 Flex reaction system. Data were analyzed and reported relative to DMSO controls using the comparative Ct method (ΔΔCt).

[0148] Table 6

[0151] For PPARG-NCOR recruitment assay, EC 50 The concentrations were expressed as follows: A: <10 nM, B: 10-100 nM, C: 100-1,000 nM, D: 1,000-10,000 nM, E: >10,000 nM. The percentage of NCOR recruitment was expressed as follows: A: >100% (> control compound, T907), B: <100% (< control compound, T907).

[0149]

[0152] For PPARG-MED1 recruitment assay, EC 50were expressed as follows: A: <10 nM, B: 10-100 nM, C: 100-1,000 nM, D: 1,000-10,000 nM, E: >10,000 nM. The % of MED1 blockade was expressed as follows: A: >100% (> control compound, GW9662), B: <100% (< control compound, GW9662).

[0150]

[0153] For HT1197 cell assay, EC 50 are expressed as follows: A: <10 nM, B: 10-100 nM, C: 100-1,000 nM, D: 1,000-10,000 nM, E: >10,000 nM, ND: not determined. The % inhibition of ANGPTL4, PPARG target genes at a compound concentration of 100 nM is expressed as a percentage of the DMSO control experiment.

[0151] [Table 6]

[0152]

[0154] While a number of embodiments have been described, it will be apparent that our basic examples can be modified to provide other embodiments that utilize the compounds and methods of the present invention. It will therefore be appreciated that the scope of the present invention is defined by the scope of the appended claims rather than by the specific embodiments shown by way of example.

[0153]

[0155] The contents of all references cited throughout this application (including literature references, issued patents, published patent applications, and co-pending patent applications) are expressly incorporated herein by reference in their entirety. Unless otherwise defined, all technical and scientific terms used herein are given the meaning commonly known to those of ordinary skill in the art.

Claims

1. Formula I 【Chemical 1】 or a pharmaceutically acceptable salt thereof (In the formula, X, Y and Z are each independently N or -CR ４ and at least one of X, Y or Z is N; R １ is hydrogen, halo, (C １ ~C ４ ) alkyl or hydroxyl; R ２ is halo, -SR ｇ , -SOR ｇ , -SO ２ R ｇ or -OR ｇ and R ３ is cyano or nitro, R ４ is hydrogen, halo, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy or hydroxyl; R ５ Ha, halo, halo (C １ ~C ４ ) alkyl or cyano; R ６ Ha, halo, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkyl or cyano; R ７ Halo, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkyl OR ａ , -(C １ ~C ４ ) alkylC(O)R ａ , -(C １ ~C ４ ) alkylC(O)OR ａ , —C(O)NR ａ R ｂ , -(C １ ~C ４ ) alkylC(O)NR ａ R ｂ , -C(O)R ａ , -C(O)OR ａ , -NR ａ R ｂ , -(C １ ~C ４ ) alkylNR ａ R ｂ , —C(O)NR ａ SO ３ H, -NR ａ C(O)R ｂ , -NR ａ C(O)OR ｂ , -NR ａ C(S)OR ｂ , -NR ｃ C(O)N ａ R ｂ , -NR ｃ C(S)NR ａ R ｂ , -NR ｃ S (O) ２ NR ａ R ｂ , -C(S)R ａ , -S(O) ２ R ａ , -S(O)R ａ , -C(S)OR ａ , -C(S)NR ａ R ｂ , -NR ａ C(S)R ｂ , -SR ａ , phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl, each of said phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl optionally and independently selected from R ８ is substituted with 1 to 3 groups selected from R ８ Halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ ) alkoxy, nitro, oxo, cyano, -(C １ ~C ４ ) alkyl OR ｄ , -(C １ ~C ４ ) alkylC(O)R ｄ , -(C １ ~C ４ ) alkylC(O)OR ｄ , —C(O)NR ｄ R ｅ , -(C １ ~C ４ ) alkylC(O)NR ｄ R ｅ , -C(O)R ｄ , -C(O)OR ｄ , -NR ｄ R ｅ , -(C １ ~C ４ ) alkylNR ｄ R ｅ , —C(O)NR ｄ SO ３ H, -NR ｄ C(O)R ｅ , -NR ｄ C(O)OR ｅ , -NR ｄ C(S)OR ｅ , -NR ｆ C(O)N ｄ R ｅ , -NR ｆ C(S)NR ｄ R ｅ , -NR ｆ S (O) ２ NR ｄ R ｅ , -C(S)R ｄ , -S(O) ２ R ｄ , -S(O)R ｄ , -C(S)OR ｄ , -C(S)NR ｄ R ｅ , -NR ｄ C(S)R ｅ and -SR ｄ is selected from R ａ , R ｂ , R ｃ , R ｄ , R ｅ , R ｆ and R ｇ are each independently substituted with hydrogen or optionally with one or two —NR′R″ groups (C １ ~C ４ ) alkyl, and R′ and R″ are each independently hydrogen, (C １ ~C ４ ) alkyl and halo(C １ ~C ４ ) alkyl; q and r are each independently 0 or 1.

2. The compound is of formula II 【Chemistry 2】 2. The compound of claim 1, wherein:

3. The compound is of formula II 【Chemistry 3】 3. The compound of claim 2, wherein:

4. R ２ But halo, -S (C １ ~C ４ ) alkyl, —SO(C １ ~C ４ ) alkyl, —SO ２ (C １ ~C ４ ) alkyl or —O(C １ ~C ４ ) alkylN[(C １ ~C ４ ) alkyl] ２ 4. The compound of claim 1, wherein:

5. R ２ But chloro, -SCH ３ , -SOCH ３ , -SO ２ CH ３ or -O(CH ２ ) ２ N (CH ３ ) ２ 4. The compound of claim 1, wherein:

6. R ２ 4. The compound of claim 1, wherein is chloro, or a pharmaceutically acceptable salt thereof.

7. R ３ 4. The compound of claim 1, wherein R is cyano, or a pharmaceutically acceptable salt thereof.

8. R １ 4. The compound of claim 1, wherein is hydrogen, fluoro, hydroxyl, or methyl, or a pharmaceutically acceptable salt thereof.

9. R １ 4. The compound of claim 1, wherein is hydrogen, or a pharmaceutically acceptable salt thereof.

10. R ５ 4. The compound of claim 1, wherein is halo or cyano, or a pharmaceutically acceptable salt thereof.

11. R ５ 4. The compound of claim 1, wherein is halo, or a pharmaceutically acceptable salt thereof.

12. 4. The compound of claim 1, wherein q is 1, or a pharmaceutically acceptable salt thereof.

13. 4. The compound of claim 1, wherein r is 1, or a pharmaceutically acceptable salt thereof.

14. 4. The compound of claim 1, wherein r is 0, or a pharmaceutically acceptable salt thereof.

15. R ６ 4. The compound of claim 1, wherein is halo, or a pharmaceutically acceptable salt thereof.

16. R ７ But, halo, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkyl OR ａ , —C(O)NR ａ R ｂ , phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl, each of said phenyl, 4- to 6-membered heterocyclyl, and 5- to 7-membered heteroaryl optionally and independently selected from R ８ 4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, substituted with 1 to 3 groups selected from:

17. R ７ But, halo, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkyl OR ａ , —C(O)NR ａ R ｂ , phenyl, pyridinyl, piperazinyl, piperidinyl, pyrrolidinyl, thiomorpholinyl, pyrazolyl, and oxetanyl, each of said phenyl, pyridinyl, pyrazolyl, pyrrolidinyl, piperazinyl, thiomorpholinyl, piperidinyl, and oxetanyl optionally and independently selected from R ８ 4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, substituted with 1 to 3 groups selected from:

18. R ７ But, halo, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, -(C １ ~C ４ ) alkyl OR ａ , —C(O)NR ａ R ｂ , phenyl, pyridinyl, pyrazolyl, and oxetanyl, each of said phenyl, pyridinyl, pyrazolyl, and oxetanyl optionally and independently selected from R ８ 4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, substituted with 1 to 3 groups selected from:

19. R ８ halo, -C(O)NR ｄ R ｅ , (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ 4. The compound of claim 1, wherein R is selected from alkoxy, oxo, and cyano, or a pharmaceutically acceptable salt thereof.

20. R ８ But, Halo, (C １ ~C ４ ) alkyl, halo (C １ ~C ４ ) alkyl, (C １ ~C ４ ) alkoxy, halo (C １ ~C ４ 4. The compound of claim 1, wherein R is selected from alkoxy, oxo, and cyano, or a pharmaceutically acceptable salt thereof.

21. R ８ is -C(O)NR ｄ R ｅ , (C １ ~C ４ 4. The compound of claim 1, wherein R is selected from the group consisting of aryl, aryl, aryl(s), ...

22. R ８ is -C(O)N(CH ３ ) ２ , C.H. ３ 4. The compound of claim 1, wherein the compound is selected from:

23. R ８ Halo (C １ ~C ４ 4. The compound of claim 1, wherein R is 1 or 2; or a pharmaceutically acceptable salt thereof.

24. If the compound has the structural formula: 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】 【Chemistry 4-4】 【Chemistry 4-5】 【Chemistry 4-6】 2. The compound of claim 1, which is: or a pharmaceutically acceptable salt thereof.

25. 4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

26. 26. The pharmaceutical composition of claim 25 for treating a cancer responsive to inhibition of PPARG in a subject.

27. 27. The pharmaceutical composition of claim 26, wherein the cancer is selected from breast cancer, pancreatic cancer, ovarian cancer, prostate cancer, renal cancer, bladder cancer, testicular cancer, urothelial cancer, skin cancer, melanoma, colon cancer, kidney cancer, brain cancer, and hematopoietic cancer.

28. 28. The pharmaceutical composition of claim 27, wherein the cancer is bladder cancer.

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