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SUBSTITUTED HETEROCYCLES AS BROMODOMAIN INHIBITORS

20170360756 · 2017-12-21

    Inventors

    Cpc classification

    International classification

    Abstract

    The present application relates to substituted heterocycles compound of Formula I and pharmaceutical compositions thereof useful for the inhibition of BET protein function by binding to bromodomains (Formula I).

    ##STR00001##

    Claims

    1-76. (canceled)

    77. A compound of Formula I: ##STR00070## or a stereoisomer, tautomer, pharmaceutically acceptable salt, or hydrate thereof, wherein: X is selected from CH and N; Y is selected from —NH, —N—R.sub.1b and oxygen; Z is selected from N, and —CH—; R.sub.1a and R.sub.1b are independently selected from alkyl (C.sub.1-C.sub.6), carbocycle (C.sub.3-C.sub.10), heterocycle (C.sub.2-C.sub.10) optionally substituted with 1 to 3 groups selected from R.sub.4; R.sub.2 is selected from aryl (C.sub.5-C.sub.10) and heteroaryl (C.sub.5-C.sub.10) optionally substituted with 1 to 5 groups selected from R.sub.5; R.sub.3 is selected from carbocycle (C.sub.3-C.sub.10) and heterocycle (C.sub.2-C.sub.10) optionally substituted with 1 to 5 groups selected from R.sub.5; each R.sub.4 is independently selected from deuterium, alkyl (C.sub.1-C.sub.6), cycloalkyl (C.sub.3-C.sub.8), alkoxy (C.sub.1-C.sub.6), amino, —NHC(O)NH-alkyl(C.sub.1-C.sub.6), halogen, amide, —CF.sub.3, —CN, —N.sub.3, ketone, —S(O)-alkyl(C.sub.1-C.sub.4), —SO.sub.2-alkyl(C.sub.1-C.sub.6), thioalkyl(C.sub.1-C.sub.6), —COOH, and ester, each of which may be optionally substituted with hydrogen, F, Cl, Br, —OH, —NH.sub.2, —NHMe, —OMe, —SMe, oxo, and/or thio-oxo; and each R.sub.5 is independently selected from deuterium, alkyl (C.sub.1-C.sub.6), alkoxy (C.sub.1-C.sub.6), amino, —NHC(O)NH-alkyl(C.sub.1-C.sub.6), halogen, amide, —CF.sub.3, —CN, —N.sub.3, ketone, —S(O)-alkyl(C.sub.1-C.sub.4), —SO.sub.2-alkyl(C.sub.1-C.sub.6), thioalkyl(C.sub.1-C.sub.6), —COOH, and ester, each of which may be optionally substituted with hydrogen, F, Cl, Br, —OH, —NH.sub.2, —NHMe, —OMe, —SMe, oxo, and/or thio-oxo.

    78. A compound according to claim 77, wherein the compound is a compound of Formula Ia: ##STR00071## or a stereoisomer, tautomer, pharmaceutically acceptable salt, or hydrate thereof, wherein: R.sub.1a is selected from alkyl (C.sub.1-C.sub.6), carbocycle (C.sub.3-C.sub.10), heterocycle (C.sub.2-C.sub.10) optionally substituted with 1 to 3 groups selected from R.sub.4; R.sub.2 is selected from aryl (C.sub.5-C.sub.10) and heteroaryl (C.sub.5-C.sub.10) optionally substituted with 1 to 5 groups selected from R.sub.5; R.sub.3 is selected from carbocycle (C.sub.3-C.sub.10) and heterocycle (C.sub.2-C.sub.10) optionally substituted with 1 to 5 groups selected from R.sub.5; each R.sub.4 is independently selected from deuterium, alkyl (C.sub.1-C.sub.6), cycloalkyl (C.sub.3-C.sub.8), alkoxy (C.sub.1-C.sub.6), amino, —NHC(O)NH-alkyl(C.sub.1-C.sub.6), halogen, amide, —CF.sub.3, —CN, —N.sub.3, ketone, —S(O)-alkyl(C.sub.1-C.sub.4), —SO.sub.2-alkyl(C.sub.1-C.sub.6), thioalkyl(C.sub.1-C.sub.6), —COOH, and ester, each of which may be optionally substituted with hydrogen, F, Cl, Br, —OH, —NH.sub.2, —NHMe, —OMe, —SMe, oxo, and/or thio-oxo; and each R.sub.5 is independently selected from deuterium, alkyl (C.sub.1-C.sub.6), alkoxy (C.sub.1-C.sub.6), amino, —NHC(O)NH-alkyl(C.sub.1-C.sub.6), halogen, amide, —CF.sub.3, —CN, —N.sub.3, ketone (C.sub.1-C.sub.6), —S(O)-alkyl(C.sub.1-C.sub.4), —SO.sub.2-alkyl(C.sub.1-C.sub.6), thioalkyl(C.sub.1-C.sub.6), —COOH, and ester, each of which may be optionally substituted with hydrogen, F, Cl, Br, —OH, —NH.sub.2, —NHMe, —OMe, —SMe, oxo, and/or thio-oxo.

    79. The compound according to claim 77, wherein Y is —NR.sub.1b.

    80. The compound according to claim 78, wherein R.sub.1a is selected from the following structures, which may be optionally substituted with 1 to 3 groups selected from R.sub.4: ##STR00072##

    81. The compound according to claim 80, wherein R.sub.1a is selected from the following structures, which may be optionally substituted with 1 to 3 groups selected from R.sub.4: ##STR00073##

    82. The compound according to claim 81, wherein R.sub.1a is ##STR00074##

    83. The compound according to claim 78, wherein R.sub.1a is selected from the following structures, which may be optionally substituted with 1 to 3 groups selected from R.sub.4: ##STR00075##

    84. The compound according to claim 79, wherein R.sub.1b is alkyl (C.sub.1-C.sub.6).

    85. The compound according to claim 84, wherein R.sub.1b is selected from methyl and ethyl.

    86. The compound according to claim 78, wherein R.sub.2 is selected from bicyclic aryl (C.sub.5-C.sub.10) and bicyclic heteroaryl (C.sub.5-C.sub.10) groups optionally substituted with 1 to 5 R.sub.5 groups.

    87. The compound according to claim 78, wherein R.sub.2 is selected from the following: ##STR00076## each of which may be optionally substituted with 1 to 2 groups selected from alkyl (C.sub.1-C.sub.6), halogen, ketone, amide, and ester.

    88. The compound according to claim 87, wherein R.sub.2 is ##STR00077##

    89. The compound according to claim 78, wherein R.sub.3 is selected from the following structures: ##STR00078## each of which may be optionally substituted with 1 to 5 groups selected from R.sub.5.

    90. The compound according to claim 89, wherein R.sub.3 is selected from the following structures: ##STR00079## each of which may be optionally substituted with 1 to 5 groups selected from R.sub.5.

    91. The compound according to claim 90, wherein R.sub.3 is an unsubstituted phenyl group.

    92. The compound according to claim 77, wherein the compound of Formula I is selected from: 3-(5-(3-Acetylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one; Methyl 3-(4-(4-cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl)-4-methylbenzoate; 3-(4-(4-Cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl)-N-methylbenzamide; 3-(4-(4-Cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl)benzamide; 4-Cyclopropyl-3-(5-(3-oxo-2,3-dihydro-1H-inden-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(2-phenyl-5-(3-propionylphenyl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 3-(5-(3-Acetyl-4-fluorophenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one; 3-(5-(3-Acetyl-5-fluorophenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 3-(5-(5-acetyl-2-methylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one; 3-(5-(5-acetylpyridin-3-yl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one formate; 4-cyclopropyl-3-(5-(1,3-dimethyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-(Cyclopropylmethyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Isopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-(tert-Butyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopentyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclobutyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclohexyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Isobutyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 3-(5-(1-Methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-4-neopentyl-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-1-methyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-1-ethyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-1-(cyclopropylmethyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-1-(2-methoxyethyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(3-methylbenzo[d]isoxazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-benzo[d][1,2,3]triazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(3-methyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-3-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 3-(2-(1H-Indol-6-yl)-5-(1-methyl-1H-indazol-6-yl)-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(1H-pyrazol-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(1-methyl-1H-pyrazol-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one; and stereoisomers, tautomers, pharmaceutically acceptable salts, and hydrates thereof.

    93. The compound of claim 77, wherein the compound of Formula I is selected from: 3-(5-(3-acetylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one hydrochloride; methyl 3-(4-(4-cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl)-4-methyl benzoate hydrochloride; 3-(4-(4-cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl)-N-methyl benzamide hydrochloride; 3-(4-(4-cyclopropyl-5-oxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)-2-phenyl-1H-imidazol-5-yl) benzamide hydrochloride; 4-cyclopropyl-3-(5-(3-oxo-2,3-dihydro-1H-inden-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(2-phenyl-5-(3-propionylphenyl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 3-(5-(3-acetyl-4-fluorophenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one hydrochloride; 3-(5-(3-acetyl-5-fluorophenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 3-(5-(5-acetyl-2-methylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(3-methyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-benzo[d][1,2,3]triazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1,3-dimethyl-1H-indazol-5-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one dihydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-3-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one dihydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(1-methyl-1H-pyrazol-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(1H-pyrazol-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one 6hydrochloride; 4-cyclopropyl-1-methyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride; and stereoisomers, tautomers, and hydrates thereof.

    94. A pharmaceutical composition comprising the compound of claim 77, and a pharmaceutically acceptable carrier.

    95. A method for inhibition of BET protein function comprising administering a therapeutically effective amount of the compound of claim 77.

    96. A method of treating a disease or disorder selected from: autoimmune diseases or disorders; inflammatory diseases or disorders; acute non-autoimmune inflammatory diseases or disorders; and chronic non-autoimmune inflammatory diseases or disorders; comprising administering a therapeutically effective amount of the compound of claim 77.

    97. The method of claim 96, wherein: the autoimmune or inflammatory disorder is selected from Acute Disseminated Encephalomyelitis, Agammaglobulinemia, Allergic Disease, Ankylosing spondylitis, Anti-GBM/Anti-TBM nephritis, Anti-phospholipid syndrome, Autoimmune aplastic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune myocarditis, Autoimmune pancreatitis, Autoimmune retinopathy, Autoimmune thrombocytopenic purpura, Behcet's Disease, Bullous pemphigoid, Castleman's Disease, Celiac Disease, Churg-Strauss syndrome, Crohn's Disease, Cogan's syndrome, Dry eye syndrome, Essential mixed cryoglobulinemia, Dermatomyositis, Devic's Disease, Encephalitis, Eosinophlic esophagitis, Eosinophilic fasciitis, Erythema nodosum, Giant cell arteritis, Glomerulonephritis, Goodpasture's syndrome, Granulomatosis with Polyangiitis (Wegener's), Graves' Disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, Hemolytic anemia, Henoch-Schonlein purpura, idiopathic pulmonary fibrosis, IgA nephropathy, Inclusion body myositis, Type I diabetes, Interstitial cystitis, Kawasaki's Disease, Leukocytoclastic vasculitis, Lichen planus, Lupus (SLE), Microscopic polyangitis, Multiple sclerosis, Myasthenia gravis, myositis, Optic neuritis, Pemphigus, POEMS syndrome, Polyarteritis nodosa, Primary biliary cirrhosis, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum, Relapsing polychondritis, Rheumatoid arthritis, Sarcoidosis, Scleroderma, Sjogren's syndrome, Takayasu's arteritis, Transverse myelitis, Ulcerative colitis, Uveitis, and Vitiligo; and the acute or chronic non-autoimmune inflammatory disorder is selected from sinusitis, pneumonitis, osteomyelitis, gastritis, enteritis, gingivitis, appendicitis, irritable bowel syndrome, tissue graft rejection, chronic obstructive pulmonary disease (COPD), septic shock, osteoarthritis, acute gout, acute lung injury, acute renal failure, burns, Herxheimer reaction, and SIRS associated with viral infections.

    98. A method of treating cancer comprising administering a therapeutically effective amount of the compound according to claim 77.

    99. The method of claim 98, wherein the cancer is selected from Burkitt's lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, diffuse large B-cell lymphoma, follicular lymphoma, multiple myeloma, bladder cancer, breast cancer, colon cancer, melanoma, ovarian cancer, prostate cancer, small cell lung carcinoma, non-small cell lung cancer, NUT midline carcinoma, acute B-cell lymphoma, and head and neck squamous cell carcinoma.

    100. The method of claim 98, wherein the cancer is selected from: a cancer associated with overexpression, translocation, amplification, or rearrangement of a myc family oncoprotein; a cancer associated with overexpression, translocation, amplification, or rearrangement of BET proteins; a cancer that relies on pTEFb (Cdk9/cyclin T) and BET proteins to regulate oncogenes; a cancer associated with upregulation of BET responsive genes selected from CDK6, Bcl2, TYRO3, MYB, and hTERT; a cancer that is sensitive to effects of BET inhibition; a cancer associated with a virus; and a cancer associated with a gene regulated by a super enhancer.

    101. The method of claim 100, wherein: the cancer associated with overexpression, translocation, amplification, or rearrangement of a myc family oncoprotein is selected from B-acute lymphocytic leukemia, Burkitt's lymphoma, Diffuse large B-cell lymphoma, Multiple myeloma, Primary plasma cell leukemia, Atypical carcinoid lung cancer, Bladder cancer, Breast cancer, Cervix cancer, Colon cancer, Gastric cancer, Glioblastoma, Hepatocellular carcinoma, Large cell neuroendocrine carcinoma, Medulloblastoma, Melanoma, nodular, Melanoma, superficial spreading, Neuroblastoma, esophageal squamous cell carcinoma, Osteosarcoma, Ovarian cancer, Prostate cancer, Renal clear cell carcinoma, Retinoblastoma, Rhabdomyosarcoma, and Small cell lung carcinoma; the cancer associated with overexpression, translocation, amplification, or rearrangement of BET proteins is selected from NUT midline carcinoma, B-cell lymphoma, non-small cell lung cancer, esophageal cancer, head and neck squamous cell carcinoma, breast cancer, prostate cancer, and colon cancer; the cancer that relies on pTEFb (Cdk9/cyclin T) and BET proteins to regulate oncogenes is selected from chronic lymphocytic leukemia, multiple myeloma, follicular lymphoma, diffuse large B-cell lymphoma, Burkitt's lymphoma, Hodgkin's lymphoma, anaplastic large cell lymphoma, neuroblastoma and primary neuroectodermal tumor, rhabdomyosarcoma, prostate cancer, and breast cancer; the cancer associated with upregulation of BET responsive genes CDK6, Bcl2, TYRO3, MYB, and hTERT is selected from pancreatic cancer, breast cancer, colon cancer, glioblastoma, adenoid cystic carcinoma, T-cell prolymphocytic leukemia, malignant glioma, bladder cancer, medulloblastoma, thyroid cancer, melanoma, multiple myeloma, Barret's adenocarcinoma, hepatoma, prostate cancer, pro-myelocytic leukemia, chronic lymphocytic leukemia, mantle cell lymphoma, diffuse large B-cell lymphoma, small cell lung cancer, and renal carcinoma; the cancer sensitive to effects of BET inhibition is selected from NUT-midline carcinoma (NMV), acute myeloid leukemia (AML), acute B lymphoblastic leukemia (B-ALL), Burkitt's Lymphoma, acute B-cell Lymphoma, Melanoma, mixed lineage leukemia, multiple myeloma, pro-myelocytic leukemia (PML), non-Hodgkin's lymphoma, Neuroblastoma, Medulloblastoma, lung carcinoma (NSCLC, SCLC), breast cancer, prostate cancer, and colon carcinoma; and the cancer associated with a virus is associated with a virus selected from Epstein-Barr Virus (EBV), hepatitis B virus (HBV), hepatitis C virus (HCV), Kaposi's sarcoma associated virus (KSHV), human papilloma virus (HPV), Merkel cell polyomavirus, and human cytomegalovirus (CMV).

    102. The method of claim 100, wherein the gene regulated by a superenhancer is the MYC oncogene.

    103. The method of claim 98, wherein the cancer is resistant to treatment with immunotherapy, hormone-deprivation therapy, and/or chemotherapy.

    104. The method of claim 103, wherein administration of a therapeutically effective amount of said compound: restores sensitivity to immunotherapy, hormone-deprivation therapy, and/or chemotherapy; and/or inhibits proliferation of cancer cells, and/or induces cancer cell death or senescence.

    105. The method of claim 98, wherein said compound is combined with other therapies, chemotherapeutic agents, or antiproliferative agents.

    106. A method of treating a benign proliferative or fibrotic disorder selected from benign soft tissue tumors, bone tumors, brain and spinal tumors, eyelid and orbital tumors, granuloma, lipoma, meningioma, multiple endocrine neoplasia, nasal polyps, pituitary tumors, prolactinoma, pseudotumor cerebri, seborrheic keratoses, stomach polyps, thyroid nodules, cystic neoplasms of the pancreas, hemangiomas, vocal cord nodules, polyps, and cysts, Castleman disease, chronic pilonidal disease, dermatofibroma, pilar cyst, pyogenic granuloma, juvenile polyposis syndrome, idiopathic pulmonary fibrosis, renal fibrosis, post-operative stricture, keloid formation, scleroderma, and cardiac fibrosis, comprising administering a therapeutically effective amount of the compound of claim 77.

    107. A method of treating a disease or disorder selected from: a disease or disorder that benefits from up-regulation or ApoA-I transcription and protein expression; a metabolic disease or disorder; and a neurological disease or disorder; comprising administering a therapeutically effective amount of the compound of claim 77.

    108. The method of claim 107, wherein: the disease or disorder that benefits from up-regulation or ApoA-I transcription and protein expression is selected from cardiovascular disease, dyslipidemia, atherosclerosis, hypercholesterolemia, metabolic syndrome, and Alzheimer's disease; the metabolic disease or disorder is selected from obesity-associated inflammation, type II diabetes, and insulin resistance; and the neurological disease or disorder is selected from Alzheimer's disease, Parkinson's disease, Huntington disease, bipolar disorder, schizophrenia, Rubinstein-Taybi syndrome, and epilepsy.

    Description

    EXAMPLES

    [0379] General Methods.

    [0380] Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. Proton nuclear magnetic resonance spectra were obtained on a Bruker 400 MHz spectrometer. Spectra are given in ppm (δ) and coupling constants, J values, are reported in hertz (Hz). Mass spectra analyses were performed on Shimadzu 2020 Mass Spectrometer in ESI or APCI mode when appropriate.

    Abbreviations

    [0381] DCM: dichloromethane; DMF: dimethylformamide; EtOAc: ethyl acetate; NBS: N-bromosuccinimide; MeOH: methanol; PE: petroleum ether; SEM-CI: (2-(chloromethoxy)ethyl)trimethylsilane; TEA: triethylamine; TFA: trifluoroacetic acid; THF: tetrahydrofuran; TLC: thin layer chromatography.

    General Procedure A

    Example 1: Preparation of 3-(5-(3-Acetylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one

    [0382] ##STR00037##

    [0383] A solution of compound 1 (25.0 g, 184 mmol, 1.00 eq), ethyl prop-2-ynoate (18.0 g, 184 mmol, 1.0 eq) in MeOH (200 mL) was heated at 65° C. for 16 h. After cooling to 25° C., MeOH was removed under reduced pressure and the residue was purified by column chromatography (PE:EtOAc=3:1) to give an oil. The oil was dissolved in phenyl ether (200 mL) and heated at 200° C. for 30 min. TLC indicated conversion to a new spot. The reaction mixture was cooled to 25° C. and poured into petroleum ether (500 mL), extracted with 1N HCl (2×200 mL). The combined aqueous layers were carefully diluted with sodium bicarbonate to basic pH and extracted with ethyl acetate (5×200 mL). The combined organic layers were dried and concentrated. The residue was purified by column chromatography (3:1 PE:EtOAc) and washed with DCM (200 mL) to afford 2 (18.0 g, 83.2 mmol, 45% yield) as a yellow solid: .sup.1H NMR (400 MHz, CDCl.sub.3) δ 1.30 (t, J=7.03 Hz, 3H), 4.26 (q, J=6.94 Hz, 2H), 7.36-7.54 (m, 3H), 7.92-8.02 (m, 2H), 8.13 (br s, 1H).

    [0384] To a solution of compound 2 (18.0 g, 83.2 mmol, 1.0 eq) in EtOH (200 mL) was added hydrazine monohydrate (16.7 g, 333 mmol, 4.0 eq) in one portion at 25° C. The mixture was stirred at 80° C. for 16 h. TLC showed the reaction was completed. The mixture was cooled to 25° C. and concentrated under reduced pressure at 40° C. The residue was purified by silica gel chromatography (1:10 MeOH/EtOAc) to afford 3 (14.0 g, 69.2 mmol, 83% yield) as a yellow solid: .sup.1H NMR (400 MHz, DMSO-d6) δ 7.35-7.42 (m, 1H), 7.43-7.51 (m, 2H), 7.73 (s, 1H), 7.99-8.06 (m, 1H), 8.97 (br s, 1H).

    [0385] To a solution of compound 3 (14.0 g, 69.2 mmol, 1.0 eq) in THF (120 mL) was added a solution of isocyanatocyclopropane (28.8 g, 346 mmol, 5.0 eq) in DCM (500 mL) dropwise at 25° C. The mixture was stirred at 25° C. for 2 h. The resulting yellow precipitate was filtered and washed with EtOAc (200 mL) to afford 4 (14.0 g, 49.1 mmol, 70% yield): ESI m/z 286.1[M+1]+.

    [0386] To a solution of compound 4 (14 g, 49.07 mmol, 1.0 eq) in H.sub.2O (300 mL) was added NaOH (5.89 g, 147.21 mmol, 3.0 eq) in one portion at 25° C. Then the reaction mixture was heated at 100° C. After 40 h, TLC showed the reaction was completed and the mixture was cooled to 25° C. 3N HCl was added and at pH 5-6, a crystalline solid formed. The solid was isolated by filtration, washed with water (30 mL) and dried at 50° C. to afford 5 (8 g, 29.93 mmol, 61% yield): .sup.1H NMR (400 MHz, DMSO-d6) δ 0.80-0.90 (m, 2H), 0.93-1.04 (m, 2H), 3.13 (dt, J=6.90, 3.33 Hz, 1H), 7.42-7.61 (m, 2H), 7.91 (s, 1H), 8.10 (d, J=7.40 Hz, 2H).

    [0387] To a solution of compound 5 (8.00 g, 29.9 mmol, 1.0 eq) in DMF (60 mL) was added NBS (6.39 g, 35.9 mmol, 1.2 eq) in portions at 0° C. under N.sub.2 atmosphere. The resulting mixture was stirred at 0-10° C. for 30 min. TLC showed the reaction was complete. The reaction mixture was quenched with water (50 mL) and the aqueous phase was extracted with ethyl acetate (6×100 mL). The combined organic layers were washed with sat. sodium thiosulfate (100 mL), dried with anhydrous Na.sub.2SO.sub.4, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography (PE/EtOAc, 50-100%) to afford 6 (5.50 g, 15.9 mmol, 53% yield) as a yellow solid: .sup.1H NMR (400 MHz, DMSO-d6) δ 0.62 (br s, 2H), 0.78-0.87 (m, 2H), 2.97 (br s, 1H), 7.43-7.57 (m, 3H), 7.97 (d, J=7.15 Hz, 2H).

    [0388] Compound 6 (50 mg, 0.14 mmol, 1.0 eq), (3-acetylphenyl)boronic acid (45.9 mg, 0.280 mmol, 2.0 eq), K.sub.2CO.sub.3 (40 mg, 0.28 mmol, 2.0 eq) and Pd(dppf)Cl.sub.2.CH.sub.2Cl.sub.2 (24 mg, 0.03 mmol, 0.20 eq) in dioxane (2 mL) and H.sub.2O (0.5 mL) was degassed with nitrogen and then heated at 90° C. for 3 h under nitrogen atmosphere. LCMS showed the starting material was consumed completely. The reaction mixture was cooled to 25° C. and the solvent was removed in vacuo. The residue was dissolved in MeOH (10 mL) and filtered. The resulting filtrate was concentrated and purified by prep-HPLC (HCl buffer) to afford 3-(5-(3-acetylphenyl)-2-phenyl-1H-imidazol-4-yl)-4-cyclopropyl-1H-1,2,4-triazol-5(4H)-one hydrochloride (Example 1) (12 mg, 31.1 umol, 22% yield) as a white solid: .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.63-0.83 (m, 4H), 2.47 (dt, J=7.15, 3.33 Hz, 1H), 2.67 (s, 3H), 7.60-7.74 (m, 4H), 7.87-7.97 (m, 1H), 8.03-8.17 (m, 3H), 8.37 (t, J=1.57 Hz, 1H); ESI m/z 386.2[M+1].sup.+.

    General Procedure B

    Example 17: 4-(Cyclopropylmethyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one)

    [0389] ##STR00038## ##STR00039##

    [0390] To a solution of compound 2 (2.00 g, 9.25 mmol, 1.00 eq) in DMF (20 mL), was added NBS (1.98 g, 11.10 mmol, 1.20 eq) in portions at 0° C. under nitrogen. The mixture was stirred at 0-10° C. for 30 min. The mixture was diluted with water and the aqueous phase was extracted with EtOAc (3×80 mL). The combined organic fractions were washed with a solution of saturated sodium thiosulfate (30 mL), dried with anhydrous sodium sulfate and concentrated under vacuum to afford 7 (1.83 g, 6.20 mmol, 67% yield) as a yellow solid: .sup.1H NMR (400 MHz, DMSO-d6) δ 1.34 (t, J=7.16 Hz, 3H) 4.33 (q, J=6.97 Hz, 2H) 7.49 (d, J=6.41 Hz, 3H) 8.09 (d, J=6.40 Hz, 2H) 13.51-13.85 (m, 1H).

    [0391] To a mixture of compound 7 (800 mg, 2.71 mmol, 1.00 eq) and (1-methylindazol-6-yl)boronic acid (477 mg, 2.71 mmol, 1.00 eq) in dioxane (10 mL) was added KOAc (266.03 mg, 2.71 mmol, 1.00 eq) and Pd(dppf)Cl.sub.2 (198 mg, 271 umol, 0.10 eq) in one portion at 25° C. under nitrogen. Then the mixture was heated to 90° C. and stirred for 16 hr. The mixture was concentrated under reduced pressure and the residue was filtered through silica gel (0-100% EtOAc in PE). The crude product was purified by preparative HPLC to afford 8 (480 mg, 1.39 mmol, 51% yield) as a yellow solid: ESI m/z 347.1[M+1].

    [0392] To a mixture of compound 8 (450 mg, 1.30 mmol, 1.00 eq) in EtOH (5 mL) was added hydrazine hydrate (976 mg, 19.5 mmol, 15.0 eq) in one portion at 25° C. under nitrogen. Then the reaction mixture was stirred at 80° C. for 30 hours. The mixture was concentrated and the residue was taken up in EtOAc (10 mL). The suspension was stirred for 10 min and filtered to afford 9 (300 mg, 902 umol, 69% yield) as a gray solid: .sup.1H NMR (400 MHz, DMSO-d6) δ 4.09 (s, 3H) 4.51 (br. s., 2H) 7.42-7.55 (m, 3H) 7.64 (d, J=8.67 Hz, 1H) 7.76-7.84 (m, 1H) 8.07-8.23 (m, 4H) 9.15 (br. s., 1H) 12.94 (br. s., 1H).

    [0393] To a mixture of compound 9 (50 mg, 150 umol, 1.00 eq) in DMF (1.0 mL) was added carbonyldiimidazole (29 mg, 180 umol, 1.2 eq) in one portion at 25° C. under nitrogen. The reaction mixture was stirred at 25° C. for 16 hr and was concentrated in vacuum. The residue was taken up in water (5 mL) and stirred for 15 min. The precipitate was filtered and dried under vacuum to afford 10 (35 mg, 97 umol, 64% yield) as a gray solid: .sup.1H NMR (400 MHz, DMSO-d6) δ 4.10 (s, 3H) 7.03 (s, 2H) 7.42-7.57 (m, 4H) 7.86 (br. s., 1H) 8.11 (d, J=13.43 Hz, 4H) 13.08-13.36 (m, 1H).

    [0394] Compound 10 (35 mg, 97 umol, 1.00 eq) and cyclopropylmethanamine (69 mg, 976 umol, 10.0 eq) were taken up in MeOH (2.0 mL) in a microwave tube. The sealed tube was irradiated in the microwave at 90° C. for 1.5 hr. The reaction mixture was concentrated under vacuum to afford 11 (30 mg, 70 umol, 72% yield) as yellow solid: ESI m/z 430.2 [M+1]+.

    [0395] To a mixture of compound 11 (20 mg, 47 umol, 1.0 eq) in EtOH (1.0 mL) was added NaOH (3M, 16 uL, 1.0 eq) in one portion at 25° C. under nitrogen. The reaction mixture was stirred at 90° C. for 16 hr. 3N HCl was added to lower the pH to 5-6 which caused a precipitate to form. The solid was isolated by filtration and taken up in DMF (5 mL) for purification by preparative HPLC. 4-(Cyclopropylmethyl)-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one (Example 17) (13 mg, 32 umol, 68% yield) was isolated as a white solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.08-0.41 (m, 4H) 3.67 (br. s., 2H) 4.08 (s, 3H) 7.33-7.49 (m, 2H) 7.51-7.57 (m, 2H) 7.80 (d, J=8.41 Hz, 1H) 8.03-8.09 (m, 2H) 8.10-8.14 (m, 2H) 11.79 (br. s., 1H); ESI m/z 412.1[M+1].sup.+.

    General Procedure C

    Example 23: 4-Cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one

    [0396] ##STR00040## ##STR00041##

    [0397] A solution of compound 12 (18.0 g, 142.7 mmol, 1.0 eq) and hydrazine hydrate (35.7 g, 713.7 mmol, 5.0 eq) in EtOH (200 mL) was stirred at 90° C. for 15 h. After cooling to room temperature, a precipitate formed. The solid was filtered, and dried under vacuum to afford compound 13 (15.0 g, 119 mmol, 83% yield) as a white crystalline solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 4.35 (br. s., 2H) 7.61 (s, 1H) 7.72 (d, J=0.88 Hz, 1H) 9.10 (br. s., 1H) 12.49 (br. s., 1H).

    [0398] Carbonyldiimidazole (17.7 g, 109.4 mmol, 1.2 eq) was added to a suspension of compound 13 (11.5 g, 91.2 mmol, 1.0 eq) in DMF (110 mL) at 0° C. Initially, the solution cleared, but after stirring at 15° C. for 3 hr, a precipitate formed. The solid was filtered, washed with DCM (100 mL) and dried under vacuum to afford compound 14 (10.4 g, 67.7 mmol, 74% yield) as an off-white solid: ESI m/z 153.0[M+1].sup.+.

    [0399] A clear solution of compound 14 (5.00 g, 32.87 mmol, 1.00 eq) and cyclopropanamine (9.38 g, 164.4 mmol, 5.0 eq) in MeOH (5.00 mL) was stirred under microwave irradiation at 80° C. for 2 hr. After cooling, a precipitate formed. The reaction mixture was concentrated and the residue was triturated in ethyl acetate (40 mL). The solid was filtered and dried under vacuum to afford compound 15 (4.50 g, 21.5 mmol, 65% yield) as a light-yellow solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.34-0.42 (m, 2H) 0.52-0.61 (m, 2H) 2.42-2.49 (m, 1H) 6.49 (br. s., 1H) 7.71 (s, 1H) 7.75 (s, 2H) 9.34 (br. s., 1H) 12.53 (br. s., 1H).

    [0400] A clear solution of compound 15 (4.70 g, 22.5 mmol, 1.0 eq) in 2N NaOH (13 mL) was stirred under microwave irradiation at 140° C. for 4 hr. The reaction mixture was cooled to 0° C., and conc. HCl was added to adjust the pH to 8. The precipitate was filtered, washed with water (100 mL) and dried under vacuum to afford compound 16 (2.90 g, 14.6 mmol, 65.1% yield, 96.4% purity) as a white solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.75-0.90 (m, 4H) 3.01-3.09 (m, 1H) 7.64 (s, 1H) 7.80 (s, 1H) 11.52 (s, 1H) 12.50 (br. s., 1H); ESI m/z 192.1 [M+1].sup.+.

    [0401] N-Bromosuccinimide (5.36 g, 30.1 mmol, 1.2 eq) was added in portions to a 0° C. solution of compound 16 (4.80 g, 25.1 mmol, 1.0 eq) in DMF (40 mL) over 1 hr. The reaction mixture was stirred at 0° C. for 3 h. The reaction mixture was purified by p-HPLC directly to afford compound 17 (4.60 g, 12.0 mmol, 48% yield) as a white crystalline TFA salt: .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.51-0.61 (m, 2H) 0.73-0.83 (m, 2H) 2.85-2.95 (m, 1H) 7.94 (s, 1H) 11.97 (s, 1H); ESI m/z 270.0, 272.0 [M+1].sup.+.

    [0402] A solution of compound 17 (4.50 g, 11.7 mmol, 1.0 eq) in DMF (2 mL) was added dropwise to another solution of SEM-CI (5.86 g, 35.2 mmol, 3.0 eq) in THF (50 mL) at 0° C. Then the solution was stirred at 10° C. for 1 hr at which time a precipitate formed. Sodium hydride (1.88 g, 46.9 mmol, 4.0 eq) was added to the mixture in portions. Initially, the reaction mixture turn red, but the color faded as it was stirred for 15 hr. The reaction was quenched by the dropwise addition of water (10 mL) at 0° C. Additional water (100 mL) was added and the the pH was adjusted to 7 with aq. HCl. The reaction mixture was extracted with ethyl acetate (2×100 mL). The combined organic fractions were washed with brine, dried over anhydrous sodium sulfate and concentrated. The residue was purified by column chromatography (10-50% EA in PE) to afford compound 18 (2.90 g, 5.47 mmol, 46.7% yield) as a colorless oil: .sup.1H NMR (400 MHz, CDCl.sub.3) δ 0.00 (s, 9H) 0.02 (s, 9H) 0.63-0.78 (m, 2H) 0.78-0.94 (m, 4H) 0.94-1.02 (m, 2H) 3.03-3.11 (m, 1H) 3.39-3.48 (m, 2H) 3.63-3.74 (m, 2H) 5.21 (s, 2H) 5.33 (br. s., 2H) 7.68 (s, 1H); ESI m/z 532.1, 530.1 [M+1].sup.+.

    [0403] Compound 18 (1.50 g, 2.83 mmol, 1.0 eq), (1-methylindazol-6-yl)boronic acid (697 mg, 3.96 mmol, 1.40 eq), K.sub.2CO.sub.3 (782 mg, 5.66 mmol, 2.0 eq) and Pd(dppf)Cl.sub.2 (207 mg, 283 umol, 0.10 eq) were combined in a mixed solvent of dioxane (20 mL) and water (5 mL). The reaction mixture was degassed with nitrogen and then heated at 90° C. for 15 hr under nitrogen atmosphere. The reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (0-100% EA in DCM) to afford compound 19 (1.46 g, 80% purity) as a brown oil: ESI m/z 582.3 [M+1].sup.+.

    [0404] n-BuLi (2.5 M, 1.81 mL, 1.80 eq) was added to a solution of compound 19 (1.46 g, 2.51 mmol, 1.00 eq) in THF (33 mL) at −78° C. under a nitrogen atmosphere. The resulting slurry was stirred at −78° C. for 5 min, then a solution of 1,1,1,2,2,2-hexachloroethane (832 mg, 3.51 mmol, 1.40 eq) in THF (3 mL) was added in portions. The reaction mixture was stirred for 30 min at −78° C. as the color turned from brown to red. The reaction was quenched by the addition of water (40 mL) at 0° C. and the reaction mixture was extracted with ethyl acetate (2×100 mL). The combined organic fractions were washed with brine (50 mL), concentrated and the residue was purified by column chromatography (20%-100% EA in PE) to afford compound 20 (1.05 g, 85% purity) as a red gum: ESI m/z 616.2 [M+1].sup.+.

    [0405] Compound 20 (100 mg, 162 umol, 1.0 eq), 4-pyridylboronic acid (40 mg, 325 umol, 2.0 eq), K.sub.2CO.sub.3 (45 mg, 325 umol, 2.0 eq) were combined in a mixed solvent of dioxane (3 mL) and water (600 uL). Pd(PPh.sub.3).sub.4(19 mg, 16 umol, 0.10 eq) was added under nitrogen atmosphere, and then the mixture was stirred at 95° C. for 15 h. The reaction mixture was concentrated under reduced pressure and the residue was purified by column chromatography (0-10% MeOH in DCM) to afford compound 21 (80 mg) as a yellow oil: ESI m/z 659.2 [M+1]+.

    [0406] A solution of compound 21 (80 mg) in 6M hydrochloric acid (10 mL) was heated to 100° C. for 3 hr. The reaction mixture was concentrated and the residue was triturated in methanol (2 mL). The solid was isolated by filtration to give 4-cyclopropyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-(pyridin-4-yl)-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one dihydrochloride (Example 23) (23 mg, 58 umol, 48% yield) as a yellow solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 0.59-0.74 (m, 4H) 4.09 (s, 3H) 7.34 (d, J=8.67 Hz, 1H) 7.44-7.68 (m, 1H) 7.84 (d, J=8.48 Hz, 1H) 8.04-8.21 (m, 2H) 8.54 (d, J=5.46 Hz, 2H) 8.94 (d, J=6.40 Hz, 2H) 11.94 (br. s., 1H); ESI m/z 399.1 [M+1]+.

    General Procedure D

    Example 27: 4-Cyclopropyl-2-methyl-5-[4-(1-methylindazol-6-yl)-2-phenyl-1H-imidazol-5-yl]-1,2,4-triazol-3-one

    [0407] ##STR00042##

    [0408] SEM-CI (188 mg, 1.13 mmol, 1.95 eq) was added to a solution of compound 6 (200 mg, 578 umol, 1.0 eq) and TEA (146 mg, 1.44 mmol, 2.5 eq) in DMF (6 mL). The reaction mixture was purged with nitrogen and heated at 80° C. for 2 h. The yellow mixture was cooled to 25° C. and concentrated under reduced pressure. The residue was purified by silica gel chromatography (20-50% EA in PE) to afford 22 (161 mg) as yellow solid: ESI m/z 477.9[M+1].sup.+.

    [0409] Compound 22 (50 mg, 105 umol, 1.0 eq) and (1-methylindazol-6-yl)boronic acid (26.04 mg, 147.98 umol, 1.41 eq) were combined in 1,4-dioxane (2 mL) and water (400 uL). K.sub.2CO.sub.3 (30 mg, 217 umol, 2.1 eq) and Pd(PPh.sub.3).sub.4(18 mg, 16 umol, 0.15 eq) were added and the mixture was purged with nitrogen. The reaction mixture was heated at 90° C. for 20 h. The residue was purified by prep-TLC (5% MeOH in DCM) to afford 23 (38 mg, 67 umol, 64% yield) as yellow solid: ESI m/z 528.2[M+1].sup.+.

    [0410] Sodium hydride (4 mg, 100 umol, 1.5 eq) was added to a solution of compound 23 (38 mg, 67 umol, 1.0 eq) in THF (3 mL) at −10° C. and the reaction was stirred for 1 hr at −10° C. Iodomethane (610 mg, 4.30 mmol, 64.1 eq) was added dropwise and the reaction mixture was purged with nitrogen. After stirring for 16 hr at 15° C., the mixture was cooled to 0° C. and quenched with ice-water (15 mL). The reaction mixture was stirred for another 0.5 h and was extracted with DCM (2×10 mL). The combined organic fractions were washed with brine (2×10 mL), dried with anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was purified by prep-TLC (10% MeOH in DCM) to afford 24 (36 mg) as yellow solid: ESI m/z 542.2[M+1].sup.+.

    [0411] To a solution of compound 24 (20 mg, 37 umol, 1.0 eq) in DCM (10 mL) was added TFA (3 mL) at 14° C. The reaction mixture was heated at 40° C. for 1 hr and concentrated under reduced pressure. The residue was taken up in dioxane (3 mL), then ammonium hydroxide (1.29 mg, 36.9 umol, 1.0 eq) was added dropwise. The mixture was stirred at 14° C. for 0.5 h and was concentrated under reduced pressure. The residue was purified by preparative HPLC to afford 4-cyclopropyl-1-methyl-3-(5-(1-methyl-1H-indazol-6-yl)-2-phenyl-1H-imidazol-4-yl)-1H-1,2,4-triazol-5(4H)-one hydrochloride (Example 27) (3 mg, 7.3 umol, 20% yield) as light yellow solid: .sup.1H-NMR (DMSO-d.sub.6, 300 MHz): δ 8.17 (d, J=7.3 Hz, 2H), 8.09 (d, J=9.2 Hz, 2H), 7.77-7.85 (m, 1H), 7.44-7.62 (m, 3H), 7.27-7.34 (m, 1H), 4.08 (s, 3H), 3.39 (s, 3H), 1.12-1.27 (m, 1H), 0.63 ppm (d, J=4.5 Hz, 4H); ESI m/z 412.0 [M+1].sup.+.

    TABLE-US-00002 TABLE 2 Example Compounds General Example Chemical Name Structure Procedure Characterization Yield 1 3-(5-(3-acetylphenyl)-2- phenyl-1H-imidazol-4- yl)-4-cyclopropyl-1H- 1,2,4-triazol-5(4H)-one hydrochloride [00043]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.63-0.83 (m, 4H), 2.47 (dt, J = 7.15, 3.33 Hz, 1H), 2.67 (s, 3H), 7.60-7.74 (m, 4H), 7.87-7.97 (m, 1H), 8.03- 8.17 (m, 3H), 8.37 (t, J = 1.57 Hz, 1H); ESI m/z 386.2 [M + 1].sup.+. 22% 2 methyl 3-(4-(4- cyclopropyl-5-oxo-4,5- dihydro-1H-1,2,4- triazol-3-yl)-2-phenyl- 1H-imidazol-5-yl)-4- methyl benzoate hydrochloride [00044]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.72-0.79 (m, 2H), 0.80-0.87 (m, 2H), 2.20-2.29 (m, 1H), 2.41 (s, 3H), 3.93 (s, 3H), 7.57 (d, J = 8.03 Hz, 1H), 7.65- 7.74 (m, 3H), 8.06 (dd, J = 7.53, 2.01 Hz, 2H), 8.10 (dd, J = 8.03, 1.76 Hz, 1H), 8.16 (d, J = 1.63 Hz, 1H); ESI m/z 416.1 [M + 1].sup.+. 22% 3 3-(4-(4-cyclopropyl-5- oxo-4,5-dihydro-1H- 1,2,4-triazol-3-yl)-2- phenyl-1H-imidazol-5- yl)-N-methyl benzamide hydrochloride [00045]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.64- 0.84 (m, 4H), 2.22-2.36 (m, H), 2.97 (s, 3H), 7.63-7.80 (m, 4H), 7.83- 7.89 (m, 1H), 8.00 (d, J = 7.91 Hz, 1H), 8.10 (dd, J = 8.09, 1.44 Hz, 2H), 8.23 (t, J = 1.57 Hz, 1H); ESI m/z 401.1 [M + 1].sup.+. 28% 4 3-(4-(4-cyclopropyl-5- oxo-4,5-dihydro-1H- 1,2,4-triazol-3-yl)-2- phenyl-1H-imidazol-5- yl) benzamide hydrochloride [00046]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.65-0.82 (m, 4H), 2.24-2.36 (m, 1H), 3.33 (dt, J = 3.26, 1.63 Hz, 4H), 7.63-7.82 (m, 4H), 7.85-7.95 (m, 1H), 8.04-8.19 (m, 3H), 8.29 (t, J = 1.57 Hz, 1H); ESI m/z 387.1 [M + 1].sup.+. 22% 5 4-cyclopropyl-3-(5-(3- oxo-2,3-dihydro-1H- inden-5-yl)-2-phenyl- 1H-imidazol-4-yl)-1H- 1,2,4-triazol-5(4H)-one hydrochloride [00047]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.59-0.88 (m, 4H), 2.35 (dt, J = 7.03, 3.39 Hz, 1H), 2.72-2.88 (m, 2H), 3.23-3.31 (m, 2H), 7.66-7.83 (m, 4H), 7.96 (dd, J = 7.97, 1.82 Hz, 1H), 8.03-8.17 (m, 3H); ESI m/z 397.1 [M + 1].sup.+. 16% 6 4-cyclopropyl-3-(2- phenyl-5-(3- propionylphenyl)-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one hydrochloride [00048]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.64-0.82 (m, 4H), 1.22 (t, J = 7.15 Hz, 3H), 2.50 (tt, J = 7.07, 3.65 Hz, 1H) 3.11 (q, J = 7.15 Hz, 2H), 7.58- 7.70 (m, 4H), 7.90 (d, J = 7.78 Hz, 1H), 8.01- 8.15 (m, 3H), 8.35 (s, 1H); ESI m/z 400 [M + 1].sup.+ 63% 7 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one hydrochloride [00049]embedded image A .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.61-0.70 (m, 4H) 4.10 (s, 3H) 7.30 (dd, J = 8.41, 1.13 Hz, 1H) 7.51-7.69 (m, 3H) 7.83 (d, J = 8.53 Hz, 1H) 8.07- 8.20 (m, 2H) 8.26 (d, J = 6.90 Hz, 2H) 12.08 (s, 1H); ESI m/z 398.2 [M + H].sup.+.  4% 8 3-(5-(3-acetyl-4- fluorophenyl)-2-phenyl- 1H-imidazol-4-yl)-4- cyclopropyl-1H-1,2,4- triazol-5(4H)-one hydrochloride [00050]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.67-0.74 (m, 2H), 0.76-0.84 (m, 2H), 2.45 (dt, J = 6.93, 3.37 Hz, 1H), 2.69 (d, J = 4.64 Hz, 3H), 7.50 (dd, J = 10.67, 8.66 Hz, 1H), 7.69-7.80 (m, 4H), 7.90- 8.01 (m, 1H), 8.09 (d, J = 6.78 Hz, 2H), 8.27 (dd, J = 6.71, 2.45 Hz, 1H); ESI m/z 404 [M + 1].sup.+. 29% 9 3-(5-(3-acetyl-5- fluorophenyl)-2-phenyl- 1H-imidazol-4-yl)-4- cyclopropyl-1H-1,2,4- triazol-5(4H)-one hydrochloride [00051]embedded image A .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.61-0.73 (m, 4H) 2.62 (s, 3H) 2.81 (dt, J = 7.15, 3.33 Hz, 1H) 7.45-7.59 (m, 3H) 7.73 (dt, J = 9.25, 1.90 Hz, 1H) 7.86-7.91 (m, 1H) 8.11- 8.19 (m, 3H) 11.98- 12.10 (m, 1H); ESI m/z 404.1 [M + 1].sup.+ 13% 10 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-5-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one hydrochloride [00052]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.63-0.76 (m, 4H) 2.03-2.16 (m, 1H) 4.15 (s, 3H) 7.67-7.86 (m, 5H) 8.08-8.15 (m, 2H) 8.19 (s, 2H); ESI m/z 398.0 [M + 1].sup.+ 54% 11 3-(5-(5-acetyl-2- methylphenyl)-2- phenyl-1H-imidazol-4- yl)-4-cyclopropyl-1H- 1,2,4-triazol-5(4H)-one hydrochloride [00053]embedded image A .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ 0.64-0.84 (m, 4H) 2.28 (s, 3 H) 2.58 (s, 3 H) 7.45-7.61 (m, 4 H) 7.90-8.00 (m, 2 H) 8.11 (d, J = 7.28 Hz, 2 H) 11.73 (br. s., 1 H) ESI m/z 400.0 [M + 1].sup.+ 43% 12 3-(5-(5-acetylpyridin-3- yl)-2-phenyl-1H- imidazol-4-yl)-4- cyclopropyl-1H-1,2,4- triazol-5(4H)-one formate [00054]embedded image A .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 0.61-0.77 (m, 1H) 2.67 (s, 3 H) 2.79-2.96 (m, 1 H) 7.41- 7.59 (m, 3 H) 8.11 (d, J = 7.16 Hz, 2 H) 8.37 (br. s., 1 H) 8.62 (s, 1 H) 8.96- 9.16 (m, 2 H) 11.89 (br. s., 1 H); ESI m/z 387.0 [M + 1].sup.+ 22% 13 4-cyclopropyl-3-(5-(3- methyl-1H-indazol-5-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one hydrochloride [00055]embedded image A .sup.1HNMR (400 MHz, DMSO-d.sub.6) δ 0.63 (d, J = 4.52 Hz, 4H) 2.60- 2.66 (m, 1H) 7.45-7.59 (m, 5H) 8.15 (s, 1H) 8.22 (d, J = 7.40 Hz, 2H) 11.94 (br. s., 1H) 12.87 (br. s., 1H); ESI m/z 398.4 [M + 1].sup.+ 35% 14 4-cyclopropyl-3-(5-(1- methyl-1H- benzo[d][1,2,3]triazol-6- yl)-2-phenyl-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one hydrochloride [00056]embedded image A .sup.1H-NMR (DMSO-d.sub.6, 300 MHz): δ 11.90-12.05 (m, 1H), 8.22-8.33 (m, 1H), 8.11-8.21 (m, 2H), 7.99- 8.11 (m, 1H), 7.56 (t, J = 7.4 Hz, 4H), 4.34 (br. s., 3H), 2.63-2.68 (m, 1H), 0.64 ppm (br. s., 4H); ESI m/z 399.1 [M + 1].sup.+ 31% 15 4-cyclopropyl-3-(5-(1,3- dimethyl-1H-indazol-5- yl)-2-phenyl-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one hydrochloride [00057]embedded image A .sup.1H NMR (400 MHz, CD.sub.3OD): δ 8.16-8.23 (m, 1H), 8.08-8.15 (m, 2H), 7.70-7.81 (m, 3H), 7.61- 7.70 (m, 2H), 4.05 (s, 3H), 2.60 (s, 3H), 2.09- 2.20 (m, 1H), 0.62-0.75 (m, 4H); ESI m/z 412.0 [M + 1].sup.+ 13% 16 4-cyclopropyl-3-(5-(3- methylbenzo[d]isoxazol- 5-yl)-2-phenyl-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one [00058]embedded image A .sup.1H NMR (400 MHz, CDCl.sub.3) δ 8.10-8.15 (m, 1H), 8.00-8.07 (m, 2H), 7.81-7.86 (m, 1H), 7.63- 7.70 (m, 1H), 7.44-7.58 (m, 3H), 2.61 (s, 4H), 0.62-0.80 (m, 4H); ESI m/z 399.1 [M + 1].sup.+  2% 17 4-(cyclopropylmethyl)- 3-(5-(1-methyl-1H- indazol-6-yl)-2-phenyl- 1H-imidazol-4-yl)-1H- 1,2,4-triazol-5(4H)-one [00059]embedded image B .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.08-0.41 (m, 4H) 3.67 (br. s., 2H) 4.08 (s, 3H) 7.33-7.49 (m, 2H) 7.51-7.57 (m, 2H) 7.80 (d, J = 8.41 Hz, 1H) 8.03-8.09 (m, 2H) 8.10-8.14 (m, 2H) 11.79 (br. s., 1H); ESI m/z 412.1 [M + 1].sup.+ 68% 18 4-isobutyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one [00060]embedded image B .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 0.71 (d, J = 6.65 Hz, 6H) 1.87- 2.07 (m, 1H) 3.66 (br. s., 2H) 4.00 (s, 3 H) 7.12 (br. s., 1H) 7.31 (br, s., 2H) 7.48-7.70 (m, 2H) 7.90 (br. s., 1H) 8.02- 8.11 (m, 3H) 11.26- 11.47 (m, 1H); ESI m/z 414.2 [M + 1].sup.+ 44% 19 4-cyclobutyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one [00061]embedded image B .sup.1H NMR (400 MHz, DMSO-d.sub.6) δ 1.40-1.62 (m, 2H) 1.83 (br. s., 2H) 2.79 (br. s., 2H) 4.03 (s, 3H) 4.51 (br. s., 1H) 7.19- 7.53 (m, 4H) 7.67 (d, J = 8.16 Hz, 1H) 7.88- 8.04 (m, 2H) 8.10 (d, J = 7.40 Hz, 2H) 11.67 (br. s., 1H) 13.21 (s, 1H); ESI m/z 412.2 [M + H].sup.+ 50% 20 3-(5-(1-methyl-1H- indazol-6-yl)-2-phenyl- 1H-imidazol-4-yl)-4- neopentyl-1H-1,2,4- triazol-5(4H)-one [00062]embedded image B .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.72 (s, 9H) 3.62 (br. s., 2H) 4.13 (s, 3H) 7.42 (dd, J = 8.47, 1.19 Hz, 1H) 7.50 (d, J = 7.15 Hz, 1H) 7.52- 7.58 (m, 2H) 7.83 (d, J = 8.53 Hz, 1H) 8.01 (s, 1H) 8.04-8.09 (m, 3H); ESI m/z 428.3 [M + 1].sup.+ 17% 21 4-cyclopentyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one [00063]embedded image B .sup.1H NMR (400 MHz, CD.sub.3OD) δ 1.37-1.50 (m, 2H) 1.61 (br. s., 2H) 1.72 (br. s., 2H) 2.03 (dd, J = 12.86, 8.22 Hz, 2H) 4.11 (s, 3H) 4.24 (t, J = 8.78 Hz, 1H) 7.32 (d, J = 8.41 Hz, 1H) 7.47- 7.59 (m, 3H) 7.80-7.89 (m, 2H) 8.03-8.10 (m, 3H); ESI m/z 426 [M + 1].sup.+  7% 22 4-isopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-phenyl-1H-imidazol-4- yl)-1H-1,2,4-triazol- 5(4H)-one [00064]embedded image B .sup.1H NMR (400 MHz, CD.sub.3OD) δ 1.28 (d, J = 6.78 Hz, 6H) 4.12 (s, 3H) 4.14- 4.19 (m, 1H) 7.34 (dd, J = 8.41, 1.38 Hz, 1H) 7.47-7.58 (m, 3H) 7.83 (d, J = 8.78 Hz, 1H) 7.91 (s, 1H) 8.02-8.10 (m, 3 H); ESI m/z 400.3 [M + 1].sup.+ 10% 23 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-(pyridin-4-yl)-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one dihydrochloride [00065]embedded image C .sup.1H NMR (300 MHz, DMSO-d.sub.6) δ 0.59-0.74 (m, 4H) 4.09 (s, 3H) 7.34 (d, J = 8.67 Hz, 1H) 7.44- 7.68 (m, 1H) 7.84 (d, J = 8.48 Hz, 1H) 8.04- 8.21 (m, 2H) 8.54 (d, J = 5.46 Hz, 2H) 8.94 (d, J = 6.40 Hz, 2H) 11.94 (br. s., 1H); ESI m/z 399.1 [M + 1].sup.+ 48% 24 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-(pyridin-3-yl)-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one dihydrochloride [00066]embedded image C .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.64-0.79 (m, 4H) 2.36-2.54 (m, 1H) 4.16 (s, 3H) 7.42 (dd, J = 8.41, 1.25 Hz, 1H) 7.91 (d, J = 8.28 Hz, 1H) 8.10 (s, 1H) 8.13 (s, 1H) 8.31 (dd, J = 8.22, 5.83 Hz, 1H) 9.00 (d, J = 5.27 Hz, 1H) 9.28 (d, J = 8.28 Hz, 1H) 9.61 (s, 1H); ESI m/z 399.2 [M + 1].sup.+ 42% 25 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-(1-methyl-1H-pyrazol- 4-yl)-1H-imidazol-4-yl)- 1H-1,2,4-triazol-5(4H)- one hydrochloride [00067]embedded image C .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.63-0.73 (m, 4H) 2.08-2.24 (m, 1H) 4.09 (s, 3H) 4.16 (s, 3H) 7.36 (d, J = 8.41 Hz, 1H) 7.96 (d, J = 8.41 Hz, 1H) 8.05 (s, 1H) 8.13 (s, 1H) 8.27 (s, 1H) 8.54 (s, 1H); ESI m/z 402.2 [M + 1].sup.+ 38% 26 4-cyclopropyl-3-(5-(1- methyl-1H-indazol-6-yl)- 2-(1H-pyrazol-4-yl)-1H- imidazol-4-yl)-1H-1,2,4- triazol-5(4H)-one hydrochloride [00068]embedded image C .sup.1H NMR (400 MHz, CD.sub.3OD) δ 0.60-0.76 (m, 4H) 2.07-2.18 (m, 1H) 4.16 (s, 3H) 7.34-7.40 (m, 1H) 7.97 (d, J = 8.41 Hz, 1H) 8.02 (s, 1H) 8.14 (d, J = 0.75 Hz, 1H) 8.47 (s, 2H); ESI m/z 388.0 [M + 1].sup.+ 36% 27 4-cyclopropyl-1-methyl- 3-(5-(1-methyl-1H- indazol-6-yl)-2-phenyl- 1H-imidazol-4-yl)-1H- 1,2,4-triazol-5(4H)-one hydrochloride [00069]embedded image D .sup.1H-NMR(DMSO-d.sub.6, 300 MHz): δ 8.17 (d, J = 7.3 Hz, 2H), 8.09 (d, J = 9.2 Hz, 2H), 7.77-7.85 (m, 1H), 7.44-7.62 (m, 3H), 7.27-7.34 (m, 1H), 4.08 (s, 3H), 3.39 (s, 3H), 1.12-1.27 (m, 1H), 0.63 ppm (d, J = 4.5 Hz, 4H); ESI m/z 412.0 [M + 1].sup.+ 20%

    Example 28: Inhibition of Tetra-Acetylated histoneH4 Binding Individual BET Bromodomains

    [0412] Proteins were cloned and overexpressed with a N-terminal 6×His tag, then purified by nickel affinity followed by size exclusion chromatography. Briefly, E. coli BL21(DE3) cells were transformed with a recombinant expression vector encoding N-terminally Nickel affinity tagged bromodomains from Brd2, Brd3, Brd4. Cell cultures were incubated at 37° C. with shaking to the appropriate density and induced overnight with IPTG. The supernatant of lysed cells was loaded onto Ni-IDA column for purification. Eluted protein was pooled, concentrated and further purified by size exclusion chromatography. Fractions representing monomeric protein were pooled, concentrated, aliquoted, and frozen at −80° C. for use in subsequent experiments.

    [0413] Binding of tetra-acetylated histone H4 peptide (Millipore) and BET bromodomains was confirmed by Amplified Luminescent Proximity Homogenous Assay (AlphaScreen). N-terminally His-tagged bromodomains (BRD4(1) at 20 nM and BRD4(2) at 100 nM) and biotinylated tetra-acetylated histone H4 (10-25 nM) were incubated in the presence of nickel chelate acceptor beads and streptavidin donor beads (PerkinAlmer, 6760000K) added to a final concentration of 2 μg/ml under green light in a white 96 well microtiter plate (Greiner). For inhibition assays, serially diluted compounds were added to the reaction mixtures in a 0.1% final concentrations of DMSO. Final buffer concentrations were 50 mM HEPES, 100 mM NaCl and 0.1% BSA buffer, pH 7.4 and optimized to 30 min incubation time. Assay plates were read at 570 nM on a Synergy H4 Plate Reader (Biotek). IC.sub.50 values were determined from a dose response curve.

    [0414] Compounds with an IC.sub.50 value less than or equal to 0.3 μM were deemed to be highly active (+++); compounds with an IC.sub.50 value between 0.3 and 3 μM were deemed to be very active (++); compounds with an IC.sub.50 value between 3 and 30 μM were deemed to be active (+).

    TABLE-US-00003 TABLE 3 Inhibition of Tetra-acetylated Histone H4 Binding to Brd4 bromodomain 1 (BRD4(1)) and Brd 4 bromodomain 2 (BRD4(2)) Alpha Alpha Alpha Alpha Alpha Alpha Example Screen Screen Example Screen Screen Example Screen Screen Number BRD4(1) BRD4(2) Number BRD4(1) BRD4(2) Number BRD4(1) BRD4(2) 1 +++ + 2 + + 3 + + 4 + Not Active 5 Not Active Not Active 6 ++ + 7 +++ Not Active 8 +++ Not Active 9 ++ Not Active 10 + ++ 11 + Not Active 12 + ++ 13 ++ ++ 14 +++ +++ 15 ++ + 16 +++ ++ 17 Not Active Not Active 18 ++ + 19 ++ + 20 ++ + 21 + Not Active 22 + + 23 +++ + 24 + + 25 ++ + 26 + + 27 + Not Active

    Example 29: Inhibition of cMYC Expression in Cancer Cell Lines

    [0415] MV4-11 cells (CRL-9591) were plated at a density of 2.5×10.sup.4 cells per well in 96 well U-bottom plates and treated with increasing concentrations of test compound or DMSO (0.1%) in IMDM media containing 10% FBS and penicillin/streptomycin, and incubated for 3 h at 37° C. Triplicate wells were used for each concentration. Cells were pelleted by centrifugation and harvested using the mRNA Catcher PLUS kit according to manufacturer's instructions. The eluted mRNA isolated was then used in a one-step quantitative real-time PCR reaction, using components of the RNA UltraSense™ One-Step Kit (Life Technologies) together with Applied Biosystems TaqMan® primer-probes for cMYC and Cyclophilin. Real-time PCR plates were run on a ViiA™7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for cMYC to an internal control, prior to determining the fold expression of each sample, relative to the control.

    [0416] Compounds with an IC.sub.50 value less than or equal to 0.3 μM were deemed to be highly active (+++); compounds with an IC.sub.50 value between 0.3 and 3 μM were deemed to be very active (++); compounds with an IC.sub.50 value between 3 and 30 μM were deemed to be active (+).

    TABLE-US-00004 TABLE 4 Inhibition of c-myc Activity in Human AML MV4-11 cells Example c-myc Example c-myc Example c-myc Example c-myc Number activity Number activity Number activity Number activity 1 + 2 Not Active 4 + 6 + 7 + 8 Not Active 9 + 10 Not Active 13 + 14 + 18 Not Active 19 +

    Example 30: Inhibition of Cell Proliferation in Cancer Cell Lines

    [0417] MV4-11 cells (CRL-9591) were plated at a density of 5×10.sup.4 cells per well in 96 well flat bottom plates and treated with increasing concentrations of test compound or DMSO (0.1%) in IMDM media containing 10% FBS and penicillin/streptomycin. Triplicate wells were used for each concentration and a well containing only media was used as a control. Plates were incubated at 37° C., 5% CO.sub.2 for 72 h before adding 20 μL of the Cell Titer Aqueous One Solution (Promega) to each well and incubated at 37° C., 5% CO.sub.2 for an additional 3-4 h. The absorbance was read at 490 nm in a spectrophotometer and the percentage of cell titer relative to DMSO-treated cells was calculated after correcting for background by subtracting the blank well's signal. IC.sub.50 values were calculated using the GraphPad Prism software.

    [0418] Compounds with an IC.sub.50 value less than or equal to 0.3 μM were deemed to be highly active (+++); compounds with an IC.sub.50 value between 0.3 and 3 μM were deemed to be very active (++); compounds with an IC.sub.50 value between 3 and 30 μM were deemed to be active (+).

    TABLE-US-00005 TABLE 5 Inhibition of Cell Proliferation in Human AML MV-4-11 cells Cell Cell Cell Cell Example Proliferation Example Proliferation Example Proliferation Example Proliferation Number activity Number activity Number activity Number activity 1 + 2 Not Active 4 + 6 + 7 + 8 Not Active 9 + 10 Not Active 13 + 14 + − − − −

    Example 31: Inhibition of hIL-6 mRNA Transcription

    [0419] Human leukemic monocyte lymphoma U937 cells (CRL-1593.2) were plated at a density of 3.2×104 cells per well in a 96-well plate in 100 μL RPMI-1640 containing 10% FBS and penicillin/streptomycin, and differentiated into macrophages for 3 days in 60 ng/mL PMA (phorbol-13-myristate-12-acetate) at 37° C. in 5% CO.sub.2 prior to the addition of compound. The cells were pretreated for 1 h with increasing concentrations of test compound in 0.1% DMSO prior to stimulation with 1 ug/mL lipopolysaccharide from Escherichia coli. Triplicate wells were used for each concentration. The cells were incubated at 37° C., 5% CO.sub.2 for 3 h before the cells were harvested. At time of harvest, media was removed and cells were rinsed in 200 μL PBS. Cells were harvested using the mRNA Catcher PLUS kit according to manufacturer's instructions. The eluted mRNA was then used in a one-step quantitative real-time PCR reaction using components of the RNA UltraSense™ One-Step Kit (Life Technologies) together with Applied Biosystems TaqMan® primer-probes for hIL-6 and Cyclophilin. Real-time PCR plates were run on a ViiA™7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hIL-6 to an internal control, prior to determining the fold expression of each sample, relative to the control.

    [0420] Compounds with an IC.sub.50 value less than or equal to 0.3 μM were deemed to be highly active (+++); compounds with an IC.sub.50 value between 0.3 and 3 μM were deemed to be very active (++); compounds with an IC.sub.50 value between 3 and 30 μM were deemed to be active J+).

    TABLE-US-00006 TABLE 6 Inhibition of hIL-6 mRNA Transcription Example IL-6 Example IL-6 Example IL-6 Example IL-6 Number activity Number activity Number activity Number activity 1 + 7 + 8 + 9 + 14 + − − − − − −

    Examples 32: In Vivo Efficacy in Athymic Nude Mouse Strain of an Acute Myeloid Leukemia Xenograft Model Using MV4-11 Cells

    [0421] MV4-11 cells (ATCC) are grown under standard cell culture conditions and (NCr) nu/nu fisol strain of female mice age 6-7 weeks are injected with 5×10.sup.6 cells/animal in 100 μL PBS+100 μL Matrigel in the lower left abdominal flank. By approximately day 18-21 after MV4-11 cells injection, mice are randomized based on tumor volume (L×W×H)/2) of average ˜100-300 mm.sup.3. Mice are dosed orally with compound at 5 to 120 mg/kg b.i.d and/or q.d. on a continuous dosing schedule and at 2.5 to 85 mg/kg q.d. on a 5 day on 2 day off, 100 mg/kg q.d. on a 4 day on and 3 day off, 135 mg/kg q.d. on a 3 day on and 4 day off, 180 mg/kg on a 2 day on and 5 day off and 240 mg/kg on a 1 day on and 6 days off dosing schedules in EA006 formulation at 10 mL/kg body weight dose volume. Tumor measurements are taken with electronic micro calipers and body weights measured on alternate days beginning from dosing period. The average tumor volumes, percent Tumor Growth Inhibition (TGI) and % change in body weights are compared relative to Vehicle control animals. The means, statistical analysis and the comparison between groups are calculated using Student's t-test in Excel.

    [0422] Other embodiments of the present disclosure will be apparent to those skilled in the art from consideration of the specification and practice of the present disclosure disclosed herein. It is intended that the specification and examples be considered as exemplary only, with a true scope and spirit of the present disclosure being indicated by the following claims.