Substituted bicyclic compounds as bromodomain inhibitors

Abstract

The invention relates to substituted bicyclic compounds, which are useful for inhibition of BET protein function by binding to bromodomains, pharmaceutical compositions comprising these compounds, and use of the compounds and compositions in therapy.

Claims

1. A method for treating prostate cancer in a mammal, comprising administering a therapeutically effective amount of a compound of formula: ##STR00084## or a tautomer, pharmaceutically acceptable salt, or hydrate thereof, wherein: D.sub.1 is selected from isoxazole, optionally substituted with one or more groups independently selected from deuterium, alkyl(C.sub.1-C.sub.4), alkoxy(C.sub.1-C.sub.4), amino, halogen, amide, C(O)cycloamino, CF.sub.3, CN, N.sub.3, ketone (C.sub.1-C.sub.4), S(O)Alkyl(C.sub.1-C.sub.4), SO.sub.2alkyl(C.sub.1-C.sub.4), -thioalkyl(C.sub.1-C.sub.4), COOH, and ester, wherein said alkyl(C.sub.1-C.sub.4), alkoxy(C.sub.1-C.sub.4), amino, amide, ketone (C.sub.1-C.sub.4), S(O)Alkyl(C.sub.1-C.sub.4), SO.sub.2alkyl(C.sub.1-C.sub.4), -thioalkyl(C.sub.1-C.sub.4), and ester are optionally substituted with one or more groups independently selected from hydrogen, F, Cl, Br, OH, NH.sub.2, NHMe, OMe, SMe, oxo, and thio-oxo; X is present and selected from (NH), O, NHCR.sub.xR.sub.y, NHSO.sub.2, and CR.sub.xR.sub.yNH; Z.sub.1 is NR.sub.a; R.sub.a is selected from hydrogen, deuterium, and alkyl (C.sub.1-3); R.sub.3 is selected from isoxazole, pyrazole, pyridyl, thiazole, isothiazole, pyrimidine, phenyl, cyclohexene, benzo[d]oxazolyl, naphthyl, and quinolyl, optionally substituted with one or more groups independently selected from deuterium, alkyl(C.sub.1-C.sub.4), OH, alkoxy(C.sub.1-C.sub.4), amino, halogen, amide, CF.sub.3, CN, N.sub.3, ketone (C.sub.1-C.sub.4), S(O)Alkyl(C.sub.1-C.sub.4), SO.sub.2alkyl(C.sub.1-C.sub.4), -thioalkyl(C.sub.1-C.sub.4), carboxyl, and ester, wherein said alkyl(C.sub.1-C.sub.4), alkoxy(C.sub.1-C.sub.4), amino, amide, ketone (C.sub.1-C.sub.4), S(O)Alkyl(C.sub.1-C.sub.4), SO.sub.2alkyl(C.sub.1-C.sub.4), -thioalkyl(C.sub.1-C.sub.4), and ester are optionally substituted with one or more groups independently selected from hydrogen, F, Cl, Br, OH, NH.sub.2, NHMe, OMe, SMe, oxo, and/or thio-oxo; R.sub.1 and R.sub.2 are independently selected from hydrogen, deuterium, alkyl, OH, NH.sub.2, -thioalkyl, and alkoxy; and R.sub.x and R.sub.y are each independently selected from hydrogen, alkyl(C.sub.1-5), halogen, OH, CF.sub.3, deuterium, amino, and alkoxy(C.sub.1-5), or two substituents selected from R.sub.x, R.sub.y and R.sub.1 may be connected in a 5- or 6-membered ring to form a bicyclic carbocycle or bicyclic heterocycle.

2. The method of claim 1, wherein D.sub.1 is ##STR00085##

3. A method for treating prostate cancer in a mammal, comprising administering a therapeutically effective amount of a compound selected from: 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(2-(trifluoromethyl)phenyl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(2-(trifluoromethyl)pyridin-3-yl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(4-methylisothiazol-5-yl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(4-fluoro-2-(trifluoromethyl)phenyl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2-methoxy-5-methyl phenyl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2-methoxypyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 3-(6-(3,5-dimethylisoxazol-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-2-methylbenzonitrile; 4,6-bis(3,5-dimethylisoxazol-4-yl)-1,3-dimethyl-1H-benzo[d]imidazol-2(3H)-one; 3-(6-(3,5-dimethylisoxazol-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-4-methylbenzonitrile; 5-(3,5-dimethylisoxazol-4-yl)-7-(4-methoxypyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(5-fluoro-2-methoxyphenyl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(5-chloro-2-methylphenyl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(6-amino-2-methylpyridin-3-yl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(3,5-dimethyl-1H-pyrazol-4-yl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 6-(3,5-dimethylisoxazol-4-yl)-4-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one; 6-(3,5-dimethylisoxazol-4-yl)-4-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazole-2(3H)-thione; 6-(3,5-dimethylisoxazol-4-yl)-4-(4-methylpyridin-3-yl)-1H-benzo[d]imidazole-2-thiol; 3-(6-(3,5-dimethylisoxazol-4-yl)-2-thioxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-4-methylbenzonitrile; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-((1,3,5-trimethyl-1H-pyrazol-4-yl)amino)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-((2-methylpyridin-3-yl)amino)-1H-benzo[d]imidazol-2(3H)-one; 5-(5-(hydroxymethyl)-3-methylisoxazol-4-yl)-1-methyl-7-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one; 3-(6-(3,5-dimethylisoxazol-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-4-methylbenzamide; 3-(6-(3,5-dimethylisoxazol-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-2-methylbenzamide; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-((2-methylpyridin-3-yl)oxy)-1H-benzo[d]imidazol-2(3H)-one; 7-(3,5-dimethyl-1H-pyrazol-4-yl)-5-(5-(hydroxymethyl)-3-methylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-((3,5-dimethylisoxazol-4-yl)amino)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(3,5-dichloropyridin-4-yl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(2-chlorophenyl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(3-methylpyridin-4-yl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(o-tolyl)-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2-fluoro-5-methoxyphenyl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(5-chloro-2-methoxyphenyl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2-fluoro-3-methoxyphenyl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2,4-dimethylthiazol-5-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 5-(3,5-dimethylisoxazol-4-yl)-7-(2-methoxy-6-methyl pyridin-3-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(benzo[d]oxazol-5-yl)-5-(3,5-dimethyl isoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; 7-(cyclohex-1-en-1-yl)-5-(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one; and tautomers, pharmaceutically acceptable salts, and hydrates thereof.

4. The method of claim 1, 2, or 3, wherein the compound is administered in combination with another anticancer agent.

5. The method of claim 4, wherein the anticancer agent is selected from ABT-737, Azacitidine (Vidaza), AZD1152 (Barasertib), AZD2281 (Olaparib), AZD6244 (Selumetinib), BEZ235, Bleomycin Sulfate, Bortezomib (Velcade), Busulfan (Myleran), Camptothecin, Cisplatin, Cyclophosphamide (Clafen), CYT387, Cytarabine (Ara-C), Dacarbazine, DAPT (GSI-IX), Decitabine, Dexamethasone, Doxorubicin (Adriamycin), Etoposide, Everolimus (RAD001), Flavopiridol (Alvocidib), Ganetespib (STA-9090), Gefitinib (Iressa), Idarubicin, Ifosfamide (Mitoxana), IFNa2a (Roferon A), Melphalan (Alkeran), Methazolastone (temozolomide), Metformin, Mitoxantrone (Novantrone), Paclitaxel, Phenformin, PKC412 (Midostaurin), PLX4032 (Vemurafenib), Pomalidomide (CC-4047), Prednisone (Deltasone), Rapamycin, Revlimid (Lenalidomide), Ruxolitinib (INCB018424), Sorafenib (Nexavar), SU11248 (Sunitinib), SU11274, Vinblastine, Vincristine (Oncovin), Vinorelbine (Navelbine), Vorinostat (SAHA), and WP1130 (Degrasyn).

Description

EXAMPLES

(1) General Methods.

(2) Unless otherwise noted, reagents and solvents were used as received from commercial suppliers. Proton nuclear magnetic resonance spectra were obtained on a Bruker AVANCE 300 spectrometer at 300 MHz or Bruker AVANCE 500 spectrometer at 500 MHz. Spectra are given in ppm () and coupling constants, J values, are reported in hertz (Hz). Tetramethylsilane was used as an internal standard for .sup.1H nuclear magnetic resonance. Mass spectra analyses were performed on Waters Aquity UPLC Mass Spectrometer in ESI or APCI mode when appropriate, Agilent 6130A Mass Spectrometer in ESI, APCI, or MultiMode mode when appropriate or Applied Biosystems API-150EX Spectrometer in ESI or APCI mode when appropriate. Silica gel chromatography were in general performed on a Teledyne Isco CombiFlash Rf 200 system or a Teledyne Isco CombiFlash Companion system.

Preparation of 4,6-bis(3,5-dimethylisoxazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 1)

(3) ##STR00023##

(4) Step 1:

(5) To a solution of 1 (5.0 g, 16.9 mmol) in ethanol (35 mL) was added iron (4.7 g, 84.5 mmol) and acetic acid (15 mL). The reaction was heated at 85 C. for an hour. The reaction mixture was cooled to room temperature, diluted with methanol (150 mL) and neutralized with sodium carbonate. The organic layer was dried over sodium sulfate, filtered and concentrated. Purification by chromatography (silica gel, 0-20% ethyl acetate/hexanes) afforded 2 (3.15 g, 70%) as a brown solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 6.77 (d, J=2.1 Hz, 1H), 6.65 (d, J=2.1 Hz, 1H), 5.17 (s, 2H), 4.77 (s, 2H).

(6) Step 2:

(7) To a solution of 2 (3.15 g, 11.8 mmol) in 1,4-dioxane (50 mL) was added 1,1-carbonyldiimidazole (2.3 g, 14.2 mmol). The reaction was heated at 65 C. for 8 hours. The reaction mixture was cooled to room temperature and concentrated in vacuo. Purification by chromatography (silica gel, 0-10% methanol/ethyl acetate) and further trituration with methanol afforded 3 (2.9 g, 83%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.26 (s, 1H), 11.08 (s, 1H), 7.32 (d, J=1.8 Hz, 1H), 7.06 (d, J=1.8 Hz, 1H).

(8) Step 3:

(9) To a suspension of 3 (200 mg, 0.69 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was added 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (535 mg, 2.40 mmol), sodium carbonate (290 mg, 2.74 mmol) and tetrakis(triphenylphosphine)palladium (158 mg, 0.14 mmol). The reaction mixture was purged with nitrogen and was heated at 95 C. for 16 h. The mixture was diluted with methylene chloride (30 mL) and washed with brine (210 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. Purification by chromatography (silica gel, 0-100% ethyl acetate/hexanes) afforded Example Compound 1 (70 mg, 32%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 10.85 (s, 1H), 10.72 (s, 1H), 6.91 (s, 1H), 6.83 (d, J=1.5 Hz, 1H), 2.41 (s, 3H), 2.31 (s, 3H), 2.24 (s, 3H), 2.15 (s, 3H); ESI m/z 325 [M+H].sup.+.

Preparation of 5,7-bis(3,5-dimethylisoxazol-4-yl)-1-methyl-1H-benzo[d]imidazol-2(3H)-one (Example Compound 2)

(10) ##STR00024##

(11) Step 1:

(12) To a solution of 3 (400 mg, 1.37 mmol) in tetrahydrofuran (15 mL) was added di-t-butyl dicarbonate (299 mg, 1.37 mmol) and potassium carbonate (189 mg, 1.37 mmol). The reaction was stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate (30 mL) and washed with water and brine. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to afford 4 (550 mg, >100%) as an off-white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.83 (s, 1H), 7.72 (s, 1H), 7.52 (d, J=6.0 Hz, 1H), 1.57 (s, 9H).

(13) Step 2:

(14) To a solution of 4 (550 mg, 1.40 mmol) in tetrahydrofuran (10 mL) was added methyl iodide (0.12 mL, 1.96 mmol) and potassium carbonate (232 mg, 1.68 mmol). The reaction was stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate (30 mL) and washed with sat. sodium bicarbonate and brine. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to afford 5 (550 mg, 96%) as an off-white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.89 (d, J=1.8 Hz, 1H), 7.63 (d, J=1.8 Hz, 1H), 3.56 (s, 3H), 1.58 (s, 9H).

(15) Step 3:

(16) To a solution of 5 (550 mg, 1.40 mmol) in methylene chloride (10 mL) was added trifluoroacetic acid (3.40 mL) and the reaction was stirred at room temperature for 30 min. The reaction mixture was concentrated in vacuo, and the residue was then diluted with ethyl acetate (30 mL), and washed with sat. sodium bicarbonate and brine. The organic layer was separated, dried over sodium sulfate and concentrated in vacuo to afford 6 (440 mg, >100%) as an off-white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.37 (bs, 1H), 7.32 (d, J=1.8 Hz, 1H), 7.11 (d, J=1.8 Hz, 1H), 3.53 (s, 3H).

(17) Step 4:

(18) To a solution of 6 (430 mg, 1.41 mmol) in 1,4-dioxane (13 mL) and water (3 mL) was added 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (1.1 g, 4.92 mmol), sodium carbonate (598 mg, 5.64 mmol) and tetrakis(triphenylphosphine)palladium (0) (163 mg, 0.14 mmol). The reaction mixture was purged with nitrogen and then heated at 95 C. for 16 h. The mixture was diluted with methylene chloride (50 mL) and washed with brine (210 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. Purification by chromatography (silica gel, 0-50% ethyl acetate/methylene chloride) afforded Example Compound 2 (220 mg, 46%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 7.01 (d, J=1.8 Hz, 1H), 6.79 (d, J=1.8 Hz, 1H), 2.95 (s, 3H), 2.49 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H), 2.10 (s, 3H); ESI m/z 339 [M+H].sup.+.

Preparation of 5,7-bis(3,5-dimethylisoxazol-4-yl)benzo[d]oxazol-2(3H)-one (Example Compound 3)

(19) ##STR00025##

(20) Step 1:

(21) A solution of 7 (1.73 g, 6.48 mmol) and 1,1-carbonyldiimidazole (2.63 g, 16.23 mmol) in 1,4-dioxane (60 mL) was refluxed for 16 h. After cooling to room temperature, the reaction mixture was mixed with silica gel (10 g) and concentrated. The resulting residue was purified by chromatography (silica gel, 0-50% ethyl acetate/heptane) to afford 8 (1.62 g, 85%) as a light brown solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 12.16 (br s, 1H), 7.53 (d, J=1.8 Hz, 1H), 7.29 (d, J=1.8 Hz, 1H); MM m/z 292 [M+H].sup.+.

(22) Step 2:

(23) A mixture of 8 (322 mg, 1.10 mmol), potassium (3,5-dimethylisoxazol-4-yl)trifluoroborate (782 mg, 3.85 mmol), potassium phosphate (1.05 g, 4.95 mmol) and tetrakis(triphenylphosphine)palladium(0) (153 mg, 0.13 mmol) in toluene (15 mL)/water (0.5 mL) was purged with nitrogen for 5 minutes. Then the reaction mixture was heated for 16 h at 90 C. After cooling to room temperature, potassium (3,5-dimethylisoxazol-4-yl)trifluoroborate (220 mg, 1.08 mmol), tetrakis(triphenylphosphine)palladium(0) (50 mg, 0.043 mmol), 1,4-dioxane (3 mL)/water (2 mL) were added. The reaction mixture was purged with nitrogen for two minutes, and then heated for 16 h at 90 C. After cooling to room temperature, the reaction mixture was concentrated. The resulting residue was purified by chromatography (silica gel, 0-50% ethyl acetate/heptane) followed by trituration with methylene chloride/hexanes to afford Example Compound 3 (45 mg, 13%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.90 (br s, 1H), 7.15-7.08 (m, 2H), 2.43 (s, 3H), 2.41 (s, 3H), 2.26 (s, 3H), 2.24 (s, 3H); MM m/z 324 [MH].sup..

General Procedure A: 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(2-methylpyridin-3-yl)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 4)

(24) ##STR00026##

(25) Step 1:

(26) To a solution of 9 (1.00 g, 4.61 mmol) in 1,4-dioxane (40 mL) and water (4 mL) was added 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (1.23 g, 5.53 mmol), potassium carbonate (1.27 g, 9.22 mmol), and tetrakis(triphenylphosphine)palladium(0) (266 mg, 0.231 mmol). The reaction mixture was purged with nitrogen and heated at 90 C. for 16 h. The reaction mixture was cooled to room temperature, concentrated and purified by chromatography (silica gel, 0-30% ethyl acetate/hexanes) to give 10 (950 mg, 88%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 8.02 (d, J=2.1 Hz, 1H), 7.26 (dd, J=2.1 Hz, 8.5 Hz, 1H), 6.89 (d, J=8.6 Hz, 1H), 6.14 (s, 2H), 2.40 (s, 3H), 2.26 (s, 3H); ESI m/z 234 [M+H].sup.+.

(27) Step 2:

(28) To a solution of 10 (940 mg, 4.03 mmol) in acetic acid (15 mL) at 0 C. was added N-bromosuccinimide (753 mg, 4.23 mmol). The reaction was warmed to room temperature and stirred for 16 h. The mixture was concentrated in vacuo. The residue was suspended in hot MeOH, cooled to room temperature and was basified with 10% aq. NaHCO.sub.3. The mixture was diluted with water and filtered. The solid was washed with water and dried in vacuo to afford 11 (1.10 g, 87%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 8.04 (d, J=2.1 Hz, 1H), 7.61 (d, J=2.1 Hz, 1H), 6.69 (br.s, 2H), 2.40 (s, 3H), 2.26 (s, 3H); ESI m/z 312 [M+H].sup.+.

(29) Step 3:

(30) To a solution of 11 (1.00 g, 3.21 mmol) in DMF (10 mL) was added NaH (60% dispersion in mineral oil, 141 mg, 3.53 mmol) at room temperature under nitrogen. The mixture was stirred at room temperature for 30 min and iodomethane (410 mg, 2.98 mmol) was added. The reaction mixture was stirred at room temperature for 16 h. NH.sub.4Cl/H.sub.2O (10 mL) was added, the mixture was stirred for 30 min, concentrated and purified by chromatography (silica gel, 0-25% ethyl acetate/hexanes) to give 12 (370 mg, 35%) as an orange solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 7.75 (d, J=2.1 Hz, 1H), 7.57 (d, J=2.1 Hz, 1H), 6.25 (q, J=5.6 Hz, 1H), 3.06 (d, J=5.5 Hz, 3H), 2.40 (s, 3H), 2.26 (s, 3H).

(31) Step 4:

(32) To a solution of 12 (2.43 g, 7.45 mmol) in tetrahydrofuran (40 mL) was added sodium dithionite (7.78 g, 44.7 mmol) in water (40 mL). The reaction mixture was stirred at room temperature for 2 h and concentrated under vacuum. To the residue was added 2N HCl (30 mL), the mixture was heated to reflux for 1 min, and concentrated under vacuum. The residue was dissolved in MeOH, adjusted to pH 8 by saturated NaHCO.sub.3 (10% in water) and concentrated under vacuum. The residue was purified by chromatography (silica gel, 0-100% ethyl acetate/hexanes) to afford 13 (1.92 g, 87%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 6.79 (d, J=1.8 Hz, 1H), 6.50 (d, J=1.8 Hz, 1H), 4.08 (br.s, 2H), 3.29 (br.s, 1H), 2.71 (s, 3H), 2.38 (s, 3H), 2.25 (s, 3H); ESI m/z 296 [M+H].sup.+.

(33) Step 5:

(34) To a mixture of 13 (1.92 g, 6.49 mmol) in 1,4-dioxane (50 mL) was added 1,1-carbonyldiimidazole (2.10 g, 12.9 mmol) and DMAP (10 mg). The reaction was heated in a sealed tube at 100 C. for 16 h. The mixture was concentrated and purified by chromatography (silica gel, 0-100% ethyl acetate in hexanes) to afford 14 (2.03 g, 97%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 9.28 (s, 1H), 7.08 (d, J=1.4 Hz, 1H), 6.89 (d, J=1.4 Hz, 1H), 3.78 (s, 3H), 2.39 (s, 3H), 2.25 (s, 3H); ESI m/z 322 [M+H].sup.+.

(35) Step 6:

(36) To a solution of 14 (100 mg, 0.31 mmol) in 1,4-dioxane (3 mL) and water (0.5 mL) was added 2-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (88 mg, 0.40 mmol), sodium carbonate (66 mg, 0.62 mmol) and tetrakis(triphenylphosphine)palladium (0) (18 mg, 0.016 mmol). The reaction mixture was purged with nitrogen and then heated at 95 C. for 16 h. The mixture was diluted with methylene chloride (50 mL) and washed with brine (210 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. Purification by chromatography (silica gel, 0-5% methanol/methylene chloride) afforded Example Compound 4 (55 mg, 53%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 8.54 (dd, J=5.0, 1.7 Hz, 1H), 7.74 (dd, J=7.6, 1.8 Hz, 1H), 7.36-7.29 (m, 1H), 7.00 (d, J=1.8 Hz, 1H), 6.78 (d, J=1.5 Hz, 1H), 2.70 (s, 3H), 2.40 (s, 3H), 2.31 (s, 3H), 2.23 (s, 3H); ESI m/z 335 [M+H].sup.+.

Preparation of 6-(3,5-dimethylisoxazol-4-yl)-4-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 22)

(37) ##STR00027##

(38) Step 1:

(39) To a mixture of 11 (500 mg, 1.6 mmol), 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (454 mg, 1.92 mmol), potassium carbonate (443 mg, 3.20 mmol), water (2 mL) and 1,4-dioxane (9 mL) was added tetrakis(triphenylphosphine)palladium(0) (93 mg, 0.08 mmol). The suspension was heated at 90 C. for 17 h. After cooling to room temperature, methanol (20 mL) and silica gel (10 g) were added. The mixture was concentrated to dryness and the resulting powder was purified by flash chromatography (silica gel, 0-90% ethyl acetate/hexanes) affording 15 as a yellow solid (291 mg, 53%): .sup.1H NMR (500 MHz, CDCl.sub.3) 8.05 (d, J=2.0 Hz, 1H), 7.10 (d, J=2.5 Hz, 1H), 6.26 (br s, 2H), 3.82 (s, 3H), 2.43 (s, 3H), 2.29 (s, 3H), 2.14 (s, 3H), 2.13 (s, 3H).

(40) Step 2:

(41) To a solution of 15 (290 mg, 0.85 mmol) in THF (20 mL) was added a solution of sodium dithionite (887 mg, 5.10 mmol) in water (20 mL). The solution stirred at room temperature for 17 h. The reaction was concentrated to dryness and methanol (30 mL) was added. The suspension stirred at room temperature for 3 h and was filtered. The filtrate was concentrated to dryness and a solution of 2N aq. HCl (20 mL) was added. The solution was brought to reflux for 5 minutes and then cooled to room temperature. The solvent was removed under reduced pressure and silica gel (10 g) and methanol (20 mL) were added. The methanol was removed and the adsorbed silica mixture was subject to flash chromatography (silica gel, 0-50% CMA (CMA: 80% CH.sub.2Cl.sub.2, 18% methanol, 2% NH.sub.4OH) in CH.sub.2Cl.sub.2) affording 16 as a light brown solid (201 mg, 76%): .sup.1H NMR (500 MHz, CDCl.sub.3) 6.59 (d, J=2.0 Hz, 1H), 6.44 (d, J=2.0 Hz, 1H), 3.80 (s, 3H), 3.48 (br s, 4H), 2.39 (s, 3H), 2.27 (s, 3H), 2.16 (s, 3H), 2.14 (s, 3H).

(42) Step 3:

(43) To a solution of 16 (200 mg, 0.64 mmol) in anhydrous 1,4-dioxane (10 mL) at room temperature was added 1,1-carbonyldiimidazole (125 mg, 0.77 mmol). The mixture was heated at 65 C. for 17 h and then cooled to room temperature. After adding silica gel (10 g) and concentrating the mixture to dryness, the material was subject to flash chromatography (silica gel, 0-10% methanol in CH.sub.2Cl.sub.2) and the product fractions were concentrated to an off-white solid. The solid was triturated with ethyl acetate (20 mL) and the suspension was filtered. The solid collected was dried in a vacuum oven for 17 h affording the product Example Compound 22 (197 mg, 91%) as an off-white solid: .sup.1H NMR (500 MHz, DMSO-d.sub.6) 10.7 (s, 1H), 10.4 (s 1H), 6.82 (d, J=1.5 Hz, 1H), 6.68 (d, J=1.5 Hz, 1H), 3.70 (s, 3H), 2.40 (s, 3H), 2.23 (s, 3H), 2.12 (s, 3H), 2.05 (s, 3H); ESI m/z 338 [M+H].sup.+.

General Procedure B: Preparation of 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-((1,3,5-trimethyl-1H-pyrazol-4-yl)amino)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 26)

(44) ##STR00028##

(45) Step 1:

(46) To a solution of 14 (2.03 g, 6.30 mmol) in dichloromethane (100 mL) was added triethylamine (2.63 mL, 18.9 mmol) followed by trityl chloride (5.27 g, 18.9 mmol). The mixture was stirred at room temperature overnight. The mixture was concentrated, the residue was purified by chromatography (silica gel, 0-20% ethyl acetate/hexanes) to give 17 (1.55 g, 44%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.15 (m, 15H), 7.10 (d, J=1.3 Hz, 1H), 6.16 (d, J=1.3 Hz, 1H), 3.72 (s, 3H), 2.15 (s, 3H), 1.96 (s, 3H); ESI m/z 564 [M+H].sup.+.

(47) Step 2:

(48) To a solution of 17 (200 mg, 0.355 mmol) in toluene (10 mL) under nitrogen atmosphere was added 1,3,5-trimethyl-1H-pyrazol-4-amine (66 mg, 0.53 mmol), cesium carbonate (231 mg, 0.710 mmol), 2-dicyclohexylphosphino-2,4,6-tri-i-propyl-1,1-biphenyl (25 mg, 0.053 mmol), and tris(dibenzylideneacetone) dipalladium(0) (33 mg, 0.036 mmol). The reaction mixture was heated at 90 C. overnight, cooled to room temperature, and purified by chromatography (silica gel, 0-100% ethyl acetate/hexanes) to give 18 (140 mg, 67%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.47 (d, J=7.3 Hz, 6H), 7.24 (t, J=6.5 Hz, 6H), 7.18 (t, J=6.5 Hz, 3H), 6.28 (s, 1H), 5.85 (d, J=1.3 Hz, 1H), 5.65 (d, J=1.3 Hz, 1H), 3.71 (s, 3H), 3.70 (s, 3H), 2.10 (s, 3H), 2.03 (s, 3H), 2.00 (s, 3H), 1.84 (s, 3H).

(49) Step 3:

(50) A mixture of 18 (140 mg, 0.236 mmol) and TFA (2 mL) were stirred at room temperature overnight. The reaction mixture was concentrated under vacuum. The residue was dissolved in MeOH and basified using concentrated NH.sub.4OH. The mixture was concentrated under vacuum and purified by reverse phase HPLC on Polaris C.sub.18 column eluted with 10-90% CH.sub.3CN in H.sub.2O to give Example Compound 26 (24 mg, 28%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 5.45 (s, 1H), 5.89 (s, 1H), 3.77 (s, 3H), 3.72 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 2.06 (s, 3H); ESI m/z 367 [M+H].sup.+.

Preparation of 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-((2-methylpyridin-3-yl)oxy)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 31)

(51) ##STR00029##

(52) Step 1:

(53) To a solution of 17 (200 mg, 0.355 mmol) in DMSO (10 mL) under nitrogen atmosphere was added 2-methylpyridin-3-ol (58 mg, 0.53 mmol), K.sub.3PO.sub.4 (188 mg, 0.888 mmol), picolinic acid (9 mg, 0.07 mmol), and CuI (7 mg, 0.04 mmol). The reaction mixture was heated at 90 C. overnight, cooled to room temperature, and concentrated under vacuum. The residue was purified by chromatography (silica gel, 0-100% ethyl acetate/hexanes) to give 19 (130 mg, 62%) as a yellow solid: .sup.1H NMR (500 MHz, CDCl.sub.3) 8.30 (dd, J=1.5, 4.5 Hz, 1H), 7.46 (d, J=7.4 Hz, 6H), 7.33-7.20 (m, 9H), 7.18-7.10 (m, 2H), 6.24 (d, J=1.3 Hz, 1H), 5.55 (d, J=1.3 Hz, 1H), 3.51 (s, 3H), 2.58 (s, 3H), 2.05 (s, 3H), 1.91 (s, 3H); ESI m/z 593 [M+H].sup.+.

(54) Step 2:

(55) A mixture of 19 (130 mg, 0.220 mmol) and TFA (2 mL) were stirred at room temperature overnight. The reaction mixture was concentrated under vacuum. The residue was dissolved in MeOH and basified with concentrated NH.sub.4OH. The mixture was concentrated under vacuum and purified by reverse phase HPLC on a Polaris C.sub.18 column eluted with 10-90% CH.sub.3CN in H.sub.2O to give Example Compound 31 (35 mg, 46%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 8.21 (dd, J=1.5, 4.6 Hz, 1H), 7.31 (dd, J=1.5, 8.3 Hz, 1H), 7.27 (dd, J=4.6, 8.4 Hz, 1H), 6.87 (d, J=1.4 Hz, 1H), 6.49 (d, J=1.4 Hz, 1H), 3.51 (s, 3H), 2.59 (s, 3H), 2.34 (s, 3H), 2.17 (s, 3H); ESI m/z 351 [M+H].sup.+.

General Procedure C: 5-(5-(hydroxymethyl)-3-methylisoxazol-4-yl)-1-methyl-7-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 28)

(56) ##STR00030##

(57) Step 1:

(58) To a solution of 20 (3.20 g, 28.32 mmol) in AcOH (5 mL) was added N-bromosuccinimide (6.05 g, 33.98 mmol) and H.sub.2SO.sub.4 (0.1 mL). The reaction mixture was heated to 120 C. for 3 h. The reaction mixture was concentrated, the residue was dissolved in EtOAc (200 mL), washed with saturated NaHCO.sub.3 (100 mL), saturated Na.sub.2S.sub.2O.sub.3 (350 mL) and brine (100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated to give 21 (5.50 g, 83%) as a pale yellow solid: .sup.1H NMR (300 MHz, CDCl.sub.3) 5.16 (s, 2H), 2.31 (s, 3H), 2.13 (s, 3H).

(59) Step 2:

(60) To a solution of 22 (10.0 g, 37.9 mmol) in DMF (100 mL) at 0 C. was added NaH (60%, 1.97 g, 49.3 mmol). The mixture was stirred at 0 C. for 30 minutes, CH.sub.3I (3.54 mL, 56.9 mol) was added dropwise, the mixture was stirred at 0 C. for 1 h, then warmed to room temperature and stirred overnight. The reaction was quenched with saturated NH.sub.4Cl (100 mL) and extracted with EtOAc (3150 mL). The combined organic layer was washed with brine (3150 mL), dried over sodium sulfate, filtered and concentrated. The residue was triturated with EtOAc/hexanes to afford 23 (8.5 g, 80%) as an orange solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 8.39 (q, J=5.1 Hz, 1H), 8.35 (d, J=1.2 Hz, 1H), 7.72 (dd, J=8.7, 0.9 Hz, 1H), 6.98 (d, J=9.7 Hz, 1H), 2.97 (d, J=4.8 Hz, 3H), 1.29 (s, 12H).

(61) Step 3:

(62) A mixture of 21 (2.34 g, 10.0 mmol), 23 (4.0 g, 14.4 mmol) and potassium carbonate (4.14 g, 30.0 mmol) in 1,4-dioxane (60 mL) and water (10 mL) was purged with nitrogen for 10 minutes, PdCl.sub.2(dppf) (817 mg, 1.0 mmol) was then added. The reaction mixture was heated at 90 C. for 7 h, diluted with EtOAc (300 mL), washed with brine (2100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by chromatography (silica gel, 10-50% ethyl acetate/hexanes) to afford 24 (1.15 g, 37%) as an orange gum: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 8.30 (q, J=5.1 Hz, 1H), 8.08 (d, J=2.1 Hz, 1H), 7.62 (dd, J=9.0, 1.8 Hz, 1H), 7.11 (d, J=9.0 Hz, 1H), 5.14 (s, 2H), 3.00 (d, J=4.8 Hz, 3H), 2.26 (s, 3H), 2.05 (s, 3H).

(63) Step 4:

(64) A solution of 24 (1.15 g, 3.77 mmol) in CH.sub.3CN (50 mL) was cooled to 0 C. and N-bromosuccinimide (1.21 g, 6.79 mmol) was added portionwise. The reaction mixture was stirred at 0 C. for 30 minutes, then warmed to rt for 3 h. The reaction mixture was diluted with EtOAc (200 mL), then washed with saturated Na.sub.2S.sub.2O.sub.3 (350 mL) and brine (100 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was suspended in EtOAc/hexanes (1/1, 100 mL), sonicated and filtered, and the filtrate was concentrated to give 25 (1.31 g, 90%) as an orange solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.87 (d, J=2.1 Hz, 1H), 7.81 (d, J=2.1 Hz, 1H), 6.55 (q, J=5.1 Hz, 1H), 5.15 (s, 2H), 2.73 (d, J=5.4 Hz, 3H), 2.25 (s, 3H), 2.03 (s, 3H).

(65) Step 5:

(66) A mixture of 25 (95 mg, 0.243 mmol), 1,3,5-trimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (115 mg, 0.486 mmol) and Na.sub.2CO.sub.3 (77 mg, 0.729 mmol) in 1,4-dioxane (5 mL) and water (0.4 mL) was purged with nitrogen for 5 minutes, Pd(PPh.sub.3).sub.4 (28 mg, 0.024 mmol) was added and the reaction mixture was heated at 90 C. for 18 h. The reaction mixture was diluted with EtOAc (30 mL), filtered and concentrated. The residue was purified by chromatography (silica gel, 0-5% methanol/ethyl acetate) to afford 26 (38 mg, 38%) as an orange oil: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.91 (d, J=2.4 Hz, 1H), 7.26 (d, J=2.4 Hz, 1H), 6.99 (q, J=5.4 Hz, 1H), 5.17 (s, 2H), 3.71 (s, 3H), 2.45 (d, J=5.4 Hz, 3H), 2.28 (s, 3H), 2.09 (s, 3H), 2.02 (s, 3H), 2.01 (s, 3H).

(67) Step 6:

(68) To a solution of 26 (38 mg, 0.092 mmol) in tetrahydrofuran (5 mL) and water (4 mL) was added sodium dithionite (104 mg, 0.60 mmol). The reaction mixture was stirred at room temperature for 4 h, 2 N HCl (1 mL) was added, the mixture was heated to reflux for 15 minutes then cooled to rt. Na.sub.2CO.sub.3 was added slowly to adjust to pH 9. The mixture was extracted with CH.sub.2Cl.sub.2 (50 mL), the organic layer was washed with brine (30 mL), filtered and concentrated. The residue was dissolved in 1,4-dioxane (2 mL), 1,1-carbonyldiimidazole (19 mg, 0.12 mmol) was added and the mixture was heated to 100 C. for 18 h. The mixture was concentrated, the residue was dissolved in THF (3 mL), NaOH (1 N in water, 0.5 mL) was added and the reaction mixture was heated to 50 C. for 2 h. The mixture was diluted with EtOAc (15 mL), washed with brine (310 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. The residue was purified by chromatography (silica gel, 0-10% methanol/ethyl acetate) followed by trituration with EtOAc/hexanes to afford Example Compound 28 (9 mg, 24%) as an off-white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.08 (s, 1H), 7.03 (d, J=1.5 Hz, 1H), 6.74 (d, J=1.8 Hz, 1H), 5.65 (t, J=5.7 Hz, 1H), 4.49 (d, J=5.7 Hz, 2H), 3.73 (s, 3H), 2.88 (s, 3H), 2.27 (s, 3H), 2.09 (s, 3H), 1.97 (s, 3H); ESI m/z 368 [M+H].sup.+.

Preparation of 4,6-bis(3,5-dimethylisoxazol-4-yl)-1,3-dimethyl-1H-benzo[d]imidazol-2(3H)-one (Example Compound 15)

(69) ##STR00031##

(70) Step 1:

(71) To a solution of 3 (300 mg, 1.03 mmol) in tetrahydrofuran (6 mL) was added methyl iodide (0.16 mL, 2.57 mmol) and potassium carbonate (284 mg, 2.06 mmol). The reaction was stirred at room temperature for 16 h. The reaction mixture was diluted with ethyl acetate (30 mL) and washed with sat. sodium bicarbonate and brine. The organic layer was separated, dried over sodium sulfate and concentrated. The residue was triturated with EtOAc to afford 27 (150 mg, 46%) as an off-white solid: ESI m/z 320 [(M+2)+H].sup.+.

(72) Step 2:

(73) To a solution of 27 (150 mg, 0.47 mmol) in 1,4-dioxane (5 mL) and water (1 mL) was added 3,5-dimethyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoxazole (366 mg, 1.64 mmol), sodium carbonate (1998 mg, 1.88 mmol) and tetrakis(triphenylphosphine)palladium (0) (27 mg, 0.024 mmol). The reaction mixture was purged with nitrogen and then heated at 95 C. for 16 h. The mixture was diluted with methylene chloride (50 mL) and washed with brine (210 mL). The organic layer was dried over sodium sulfate, filtered and concentrated. Purification by chromatography (silica gel, 0-50% ethyl acetate/methylene chloride) afforded Example Compound 15 (48 mg, 29%) as a white solid: .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.26 (d, J=1.8 Hz, 1H), 6.84 (d, J=1.8 Hz, 1H), 3.40 (s, 3H), 3.00 (s, 3H), 2.43 (s, 3H), 2.29 (s, 3H), 2.26 (s, 3H), 2.09 (s, 3H); ESI m/z 353 [M+H].sup.+.

General Procedure D: Preparation of 5-(3,5-dimethylisoxazol-4-yl)-1-methyl-7-(3-methylpyridin-4-yl)-1H-benzo[d]imidazol-2(3H)-one (Example Compound 38)

(74) ##STR00032##

(75) Step 1:

(76) To a solution of 17 (500 mg, 0.887 mmol) in 1,4-dioxane (10 mL) was added 4,4,4,4,5,5,5,5-octamethyl-2,2-bi(1,3,2-dioxaborolane) (338 mg, 1.33 mmol), potassium acetate (174 mg, 1.77 mmol), and [1,1-bis(diphenylphosphino)ferrocene] dichloropalladium(II) (65 mg, 0.089 mmol). The reaction mixture was purged with nitrogen for 5 minutes and then heated at 100 C. for 16 h. The reaction mixture was cooled to room temperature, concentrated and purified by chromatography (silica gel, 0-50% ethyl acetate in hexanes) to afford 28 (310 mg, 57%) as a yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 7.50-7.40 (m, 6H), 7.30-7.18 (m, 10H), 6.27 (d, J=1.6 Hz, 1H), 3.51 (s, 3H), 2.13 (s, 3H), 1.95 (s, 3H), 1.39 (s, 12H); ESI m/z 612 [M+H].sup.+.

(77) Step 2:

(78) To a solution of 28 (100 mg, 0.164 mmol) in 1,4-dioxane (10 mL) and water (1 mL) was added 4-bromo-3-methylpyridine (57 mg, 0.33 mmol), potassium bicarbonate (68 mg, 0.49 mmol), and tetrakis(triphenylphosphine)palladium(0) (9 mg, 0.008 mmol). The reaction mixture was purged with nitrogen for 5 minutes and then heated at 90 C. for 16 h. The reaction mixture was cooled to room temperature and concentrated. The residue was dissolved in TFA (2 mL) and stirred at room temperature for 2 h. The mixture was concentrated. The residue was purified by chromatography (silica gel, 0-20% methanol/ethyl acetate). The product was further purified by reverse phase HPLC on a Polaris C.sub.18 column eluting with 10-90% CH.sub.3CN in H.sub.2O to give Example Compound 38 (28 mg, 51%) as an off-white solid: .sup.1H NMR (500 MHz, CD.sub.3OD) 8.53 (s, 1H), 8.47 (d, J=4.9 Hz, 1H), 7.42 (d, J=5.0 Hz, 1H), 7.09 (d, J=1.4 Hz, 1H), 6.79 (d, J=1.4 Hz, 1H), 2.88 (s, 3H), 2.41 (s, 3H), 2.25 (s, 3H), 2.19 (s, 3H); ESI m/z 335 [M+H].sup.+.

General Procedure E: Preparation of 3-(6-(3,5-dimethylisoxazol-4-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-4-methylbenzamide (Example Compound 29)

(79) ##STR00033##

(80) To a solution of Example 16 (35 mg, 0.10 mmol) in ethanol (2 mL) was added 2 N NaOH (0.49 mL). The reaction mixture was heated to 85 C. for 2 h. The reaction mixture was diluted in methylene chloride (70 mL), washed with brine (25 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by chromatography (silica gel, 0-10% methanol/methylene chloride) to afford Example 29 (34 mg, 92%) as white solid: .sup.1H NMR (500 MHz, DMSO-d.sub.6) 11.11 (br.s, 1H), 7.94 (br.s, 1H), 7.87 (dd, J=7.8, 2.0 Hz, 1H), 7.82 (d, J=2.0 Hz, 1H), 7.42 (d, J=8.0 Hz, 1H), 7.31 (br.s, 1H), 6.99 (d, J=2.0 Hz, 1H), 6.75 (d, J=1.5 Hz, 1H) 2.67 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.15 (s, 3H); ESI m/z 377 [M+H].sup.+.

Preparation of 6-(3,5-dimethylisoxazol-4-yl)-4-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-benzo[d]imidazole-2(3H)-thione (Example 23)

(81) ##STR00034##

(82) Lawesson's reagent (0.485 g, 1.20 mmol) was added to a solution of Example Compound 22 (0.337 g, 1.00 mmol) in 1,4-dioxane (2 mL). The reaction was stirred at 180 C. for 6 h under microwave heating conditions. The reaction was cooled to rt, concentrated under reduced pressure and quenched with water (75 mL) The resulting precipitate was collected by filtration, washed with water, then ethyl acetate (20 mL) and dried under vacuum. The residue was purified by flash column chromatography (silica gel, 0-5% methanol/dichloromethane) followed by prep. HPLC to afford Example 23 (0.066 g, 19%) as a white solid: .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.37 (br s, 2H), 7.01 (s, 1H), 6.87 (s, 1H), 3.71 (s, 3H), 2.42 (s, 3H), 2.24 (s, 3H), 2.11 (s, 3H), 2.05 (s, 3H); ESI MS m/z 352 [MH].sup..

Preparation of 6-(3,5-dimethylisoxazol-4-yl)-4-(4-methylpyridin-3-yl)-1H-benzo[d]imidazole-2(3H)-thione (Example Compound 24)

(83) ##STR00035##

(84) Step 1:

(85) To a degassed solution of 11 (6.24 g, 20 mmol), 4-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (6.57 g, 30 mmol) and K.sub.3PO.sub.4 (12.74 g, 60 mmol) in 1,4-dioxane (126 mL) and water (12.6 mL) was added Pd(PPh.sub.3).sub.4 (2.31 g, 2 mmol). The reaction was heated at 100 C. for 20 h under N.sub.2. The reaction was cooled to rt, dried over MgSO.sub.4, filtered through silica gel and concentrated under reduced pressure. The residue was purified by chromatography (silica gel, 80% CH.sub.2Cl.sub.2/ethyl acetate) to give an impure mixture that was dissolved in ethyl acetate (200 mL) and extracted with 2N HCl (22 mL) and water (420 mL). The combined aqueous extracts were washed with diethyl ether (250 mL) and basified with solid K.sub.2CO.sub.3 (about 7.3 g) to pH 9. The aqueous was extracted with chloroform (420 mL). The combined organics were dried over MgSO.sub.4, filtered and concentrated under reduced pressure to give 29 (4.48 g, 60%) as an orange solid: .sup.1H NMR (400 MHz, CDCl.sub.3) 8.61 (d, J=5.2 Hz, 1H), 8.46 (s, 1H), 8.14 (d, J=2.0 Hz, 1H), 7.33 (d, J=5.2 Hz, 1H), 7.16 (d, J=2.0 Hz, 1H), 6.06 (br.s, 2H), 2.44 (s, 3H), 2.29 (s, 3H), 2.24 (s, 3H).

(86) Step 2:

(87) Concentrated hydrochloric acid (20.7 mL, 249 mmol) was added in one portion to a stirred suspension of 29 (4.48 g, 13.8 mmol) and tin granules (4.92 g, 41.4 mmol) in ethanol (146 mL). The reaction was stirred at rt for 23 h. After that time the resulting precipitate was collected by filtration, washed with ethanol (250 mL), then Et.sub.2O (2100 mL) and dried under vacuum. The material was dissolved in water (100 mL) and the pH of the resulting solution was adjusted to 9 with solid K.sub.2CO.sub.3 (4.9 g). The aqueous solution was extracted with chloroform (620 mL). The combined organics were dried over MgSO.sub.4, filtered and concentrated under reduced pressure to 30 (3.27 g, 80%) as a yellow solid: .sup.1H NMR (400 MHz, CDCl.sub.3) 8.51 (d, J=5.2 Hz, 1H), 8.46 (s, 1H), 7.26 (d, J=5.2 Hz, 1H), 6.66 (d, J=2.0 Hz, 1H), 6.47 (d, J=2.0 Hz, 1H), 3.54 (br.s, 2H), 3.31 (br s, 2H), 2.40 (s, 3H), 2.27 (s, 3H), 2.23 (s, 3H).

(88) Step 3: 1,1-thiocarbonyldiimidazole (0.267 g, 1.5 mmol) was added in one portion to a stirred suspension of 30 (0.294 g, 1.0 mmol) in anhydrous THF (10 mL). The reaction was heated at reflux with stirring for 21 h. The reaction was cooled to rt and concentrated under reduced pressure. The residue was dissolved in chloroform (20 mL), washed with water (310 mL), dried over MgSO.sub.4, filtered and concentrated under reduced pressure. The residue was purified by flash column chromatography (silica gel, 0-2% methanol/chloroform) to give Example 2 Compound 4 (0.305 g, 91%) as a yellow solid: .sup.1H NMR (400 MHz, CDCl.sub.3) 12.58 (br.s, 1H), 11.33 (br.s, 1H), 8.45 (s, 1H), 8.23 (d, J=5.4 Hz, 1H), 7.22 (d, J=5.4 Hz, 1H), 7.15 (d, J=1.2 Hz, 1H), 6.91 (d, J=1.2 Hz, 1H), 2.41 (s, 3H), 2.28 (s, 3H), 2.27 (s, 3H); ESI MS m/z 337 [M+H].sup.+.

Preparation of 3-(6-(3,5-dimethylisoxazol-4-yl)-2-thioxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-4-methylbenzonitrile (Example Compound 25)

(89) ##STR00036##

(90) Starting with (5-cyano-2-methylphenyl)boronic acid, compound 31 was prepared using the method for Example Compound 22 step 1 to 2.

(91) A mixture of 31 (0.2 g, 0.63 mmol) and 1,1-thiocarbonyldiimidazole (0.17 g, 0.95 mmol) in THF (8.0 mL) was heated at reflux for 18 h. The reaction was cooled to rt, filtered and concentrated under reduced pressure. Ice-cold water (20 mL) was added to the residue and the product was extracted with chloroform (220 mL). The combined organics were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography (silica gel, 0-3% MeOH/dichloromethane) to give Example Compound 25 (0.19 g, 83.9%) as an off-white solid: .sup.1H NMR (400 MHz, CDCl.sub.3) 10.26 (br.s, 2H), 7.68 (d, J=8.2 Hz, 1H), 7.58 (s, 1H), 7.50 (d, J=7.8 Hz, 1H), 7.13 (s, 1H), 6.92 (s, 1H), 2.43 (s, 3H), 2.29 (s, 6H); ESI MS m/z 361 [M+H].sup.+.

(92) TABLE-US-00002 TABLE 1 Examples prepared using methods described above. Example General Purity Compound Chemical Name Structure procedure Characterization HPLC 1 4,6-bis(3,5- dimethylisoxazol- 4-yl)-1H- benzo[d]imidazol- 2(3H)-one none .sup.1H NMR (300 MHz, DMSO-d.sub.6) 10.85 (s, 1H), 10.72 (s, 1H), 6.91 (s, 1H), 6.83 (d, J = 1.5 Hz, 1H), 2.41 (s, 3H), 2.31 (s, 3H), 2.24 (s, 3H), 2.15 (s, 3H); ESI m/z 325 [M + H]+. 98.3 2 5,7-bis(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one none .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.79 (d, J = 1.8 Hz, 1H), 2.95 (s, 3H), 2.49 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H), 2.10 (s, 3H); ESI m/z 339 [M + H]+. >99 3 5,7-bis(3,5- dimethylisoxazol- 4-yl)benzo [d]oxazol- 2(3H)-one none .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.90 (br s, 1H), 7.15-7.08 (m, 2H), 2.43 (s, 3H), 2.41 (s, 3H), 2.26 (s, 3H), 2.24 (s, 3H); MM m/z 324 [M H]. >99 4 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(2- methylpyridin- 3-yl)-1H- benzo[d]imidazol- 2(3H)-one 0 A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 8.54 (dd, J = 5.0, 1.7 Hz, 1H), 7.74 (dd, J = 7.6, 1.8 Hz, 1H), 7.36-7.29 (m, 1H), 7.00 (d, J = 1.8 Hz, 1H), 6.78 (d, J = 1.5 Hz, 1H), 2.70 (s, 3H), 2.40 (s, 3H), 2.31 (s, 3H), 2.23 (s, 3H); ESI m/z 335 [M + H]+. >99 5 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(2- (trifluoromethyl) phenyl)-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 7.93-7.87 (m, 1H), 7.79-7.66 (m, 2H), 7.64-7.57 (m, 1H), 7.00 (d, J = 1.5 Hz, 1H), 6.79-6.76 (m, 1H), 2.62 (s, 3H), 2.38 (s, 3H), 2.20 (s, 3H); ESI m/z 388 [M + H]+. 98.3 6 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(4- methylpyridin- 3-yl)-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.17 (s, 1H), 8.54-8.47 (m, 2H), 7.39 (d, J = 4.8 Hz, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.78 (d, J = 1.5 Hz, 1H), 2.70 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H), 2.14 (s, 3H); ESI m/z 335 [M + H]+. >99 7 7-(1,3- dimethyl-1H- pyrazol-4-yl)- 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.06 (s, 1H), 7.76 (s, 1H), 6.92 (d, J = 1.5 Hz, 1H), 6.71 (d, J = 1.8 Hz, 1H), 3.82 (s, 3H), 2.95 (s, 3H), 2.39 (s, 3H), 2.21 (s, 3H), 2.06 (s, 3H); ESI m/z 338 [M + H]+. >99 8 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(2- (trifluoromethyl) pyridin-3- yl)-1H-benzo [d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.29 (s, 1H), 8.86 (d, J = 5.3 Hz, 1H), 8.10-8.07 (m, 1H), 7.91-7.87 (m, 1H), 7.08 (d, J = 1.7 Hz, 1H), 6.95 (d, J = 1.7 Hz, 1H), 2.87 (s, 3H), 2.49 (s, 3H), 2.22 (s, 3H); ESI m/z 389 [M + H]+. 96.8 9 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7- (1,3,5-trimethyl- 1H-pyrazol-4-yl)- 1H-benzo [d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.05 (s, 1H), 6.93 (d, J = 1.7 Hz, 1H), 6.65 (d, J = 1.7 Hz, 1H), 3.72 (s, 3H), 2.88 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.08 (s, 3H), 1.97 (s, 3H); ESI m/z 352 [M + H]+. >99 10 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(4- methylisothiazol- 5-yl)-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.23 (s, 1H), 9.15 (s, 1H), 7.03 (d, J = 1.7 Hz, 1H), 6.86 (d, J = 1.7 Hz, 1H), 2.98 (s, 3H), 2.40 (s, 3H), 2.25 (s, 3H), 2.22 (s, 3H); ESI m/z 341 [M + H]+. >99 11 5-(3,5- dimethylisoxazol- 4-yl)-7-(4- fluoro-2- (trifluoromethyl) phenyl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.18 (s, 1H), 7.81 (dd, J = 9.5, 2.4 Hz, 1H), 7.32-7.59 (m, 2H), 7.01 (d, J = 1.8 Hz, 1H), 6.79-6.77 (m, 1H), 2.66 (s, 3H), 2.37 (s, 3H), 2.20 (s, 3H); ESI m/z 406 [M + H]+. >99 12 5-(3,5- dimethylisoxazol- 4-yl)-7-(2- methoxy-5- methylphenyl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.04 (s, 1H), 7.76-7.18 (m, 1H), 7.12 (d, J = 2.2 Hz, 1H), 7.00 (d, J = 8.4 Hz, 1H), 6.93 (d, J = 1.8 Hz, 1H), 6.71 (d, J = 1.5 Hz, 1H), 3.70 (s, 3H), 2.80 (s, 3H), 2.40 (s, 3H), 2.29 (s, 3H), 2.22 (s, 3H); ESI m/z 364 [M + H]+. 99.0 13 5-(3,5- dimethylisoxazol- 4-yl)-7-(2- methoxypyridin- 3-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.12 (s, 1H), 8.27 (dd, J = 4.9, 1.9 Hz, 1H), 7.78 (dd, J = 7.4, 1.9 Hz, 1H), 7.17-7.10 (m, 1H), 6.98 (d, J = 1.8 Hz, 1H), 6.77 (d, J = 1.8 Hz, 1H), 3.85 (s, 3H), 2.82 (s, 3H), 2.40 (s, 3H), 2.23 (s, 3H); ESI m/z 351 [M + H]+. >99 14 3-(6-(3,5- dimethylisoxazol- 4-yl)-3- methyl-2-oxo- 2,3-dihydro- 1H- benzo[d]imidazol- 4-yl)-2- methylbenzonitrile 0 A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.19 (s, 1H), 7.90 (dd, J = 7.7, 1.2 Hz, 1H), 7.68 (dd, J = 8.1, 1.2 Hz, 1H), 7.49 (t, J = 8.0 Hz, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.76 (d, J = 1.8 Hz, 1H), 2.68 (s, 3H), 2.40 (s, 3H), 2.30 (s, 3H), 2.24 (s, 3H); ESI m/z 359 [M + H]+. >99 15 4,6-bis(3,5- dimethylisoxazol- 4-yl)-1,3- dimethyl-1H- benzo[d]imidazol- 2(3H)-one none .sup.1H NMR (300 MHz, DMSO-d.sub.6) 7.26 (d, J = 1.8 Hz, 1H), 6.84 (d, J = 1.8 Hz, 1H), 3.40 (s, 3H), 3.00 (s, 3H), 2.43 (s, 3H), 2.29 (s, 3H), 2.26 (s, 3H), 2.09 (s, 3H); ESI m/z 353 [M + H]+. >99 16 3-(6-(3,5- dimethylisoxazol- 4-yl)-3- methyl-2-oxo- 2,3-dihydro-1H- benzo[d]imidazol- 4-yl)-4- methylbenzonitrile A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.18 (s, 1H), 7.86-7.81 (m, 2H), 7.59-7.54 (m, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.76 (d, J = 1.8 Hz, 1H), 2.68 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.17 (s, 3H); ESI m/z 359 [M + H]+. >99 17 5-(3,5- dimethylisoxazol- 4-yl)-7-(4- methoxypyridin- 3-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.13 (s, 1H), 8.56 (d, J = 5.7 Hz, 1H), 8.40 (s, 1H), 7.20 (d, J = 5.7 Hz, 1H), 6.99 (d, J = 1.8 Hz, 1H), 6.77 (d, J = 1.8 Hz, 1H), 3.84 (s, 3H), 2.82 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H); ESI m/z 351 [M + H]+. >99 18 5-(3,5- dimethylisoxazol- 4-yl)-7-(5- fluoro-2- methoxyphenyl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.09 (s, 1H), 7.33-7.20 (m, 2H), 7.16-7.07 (m, 1H), 6.96 (d, J = 1.7 Hz, 1H), 6.75 (d, J = 1.7 Hz, 1H), 3.72 (s, 3H), 2.82 (s, 3H), 2.40 (s, 3H), 2.23 (s, 3H); ESI m/z 366 [M H]. 99.0 19 7-(5-chloro-2- methylphenyl)- 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (500 MHz, DMSO-d.sub.6) 11.11 (s, 1H), 7.42 (dd, J = 8.0, 2.5 Hz, 1H), 7.40 (d, J = 2.0 Hz, 1H), 7.37 (d, J = 8.5 Hz, 1H), 6.98 (d, J = 1.5 Hz, 1H), 6.73 (d, J = 2.0 Hz, 1H), 2.71 (s, 3H), 2.34 (s, 3H), 2.22 (s, 3H), 2.06 (s, 3H); ESI m/z 368 [M + H]+. 98.6 20 7-(6-amino-2- methylpyridin- 3-yl)-5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (500 MHz, DMSO-d.sub.6) 11.03 (s, 1H), 7.27 (d, J = 8.5 Hz, 1H), 6.92 (d, J = 1.5 Hz, 1H), 6.69 (d, J = 1.5 Hz, 1H), 6.35 (d, J = 8.5 Hz, 1H), 5.97 (s, 2H), 2.82 (s, 3H), 2.39 (s, 3H), 2.22 (s, 3H), 2.07 (s, 3H); ESI m/z 350 [M + H]+. >99 21 7-(3,5- dimethyl-1H- pyrazol-4-yl)- 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (500 MHz, CD3OD) 7.03 (d, J = 1.5 Hz, 1H), 7.35- 7.33 (m, 1H), 7.15 (d, J = 2.0 Hz, 1H), 6.76 (d, J = 1.5 Hz, 1H), 3.02 (s, 3H), 2.42 (s, 3H), 2.26 (s, 3H), 2.13 (br.s, 6H); ESI m/z 338 [M + H]+. >99 22 6-(3,5- dimethylisoxazol- 4-yl)-4-(1,3,5- trimethyl-1H- pyrazol-4-yl)-1H- benzo[d]imidazol- 2(3H)-one none .sup.1H NMR (500 MHz, DMSO-d.sub.6) 10.7 (s, 1H), 10.4 (s 1H), 6.82 (d, J = 1.5 Hz, 1H), 6.68 (d, J = 1.5 Hz, 1H), 3.70 (s, 3H), 2.40 (s, 3H), 2.23 (s, 3H), 2.12 (s, 3H), 2.05 (s, 3H); ESI m/z 338 [M + H]+. >99 23 6-(3,5- dimethylisoxazol- 4-yl)-4-(1,3,5- trimethyl-1H- pyrazol-4-yl)- 1H- benzo[d]imidazole- 2(3H)-thione No general procedure .sup.1H NMR (400 MHz, DMSO-d.sub.6) 12.37 (br.s, 2H), 7.01 (s, 1H), 6.87 (s, 1H), 3.71 (s, 3H), 2.42 (s, 3H), 2.24 (s, 3H), 2.11 (s, 3H), 2.05 (s, 3H); ESI m/z 352 [M H]. 98.5 24 6-(3,5- dimethylisoxazol- 4-yl)-4-(4- methylpyridin- 3-yl)-1H- benzo[d]imidazole- 2-thiol 0 No general procedure .sup.1H NMR (400 MHz, CDCl.sub.3) 12.58 (br.s, 1H), 11.33 (br.s, 1H), 8.45 (s, 1H), 8.23 (d, J = 5.4 Hz, 1H), 7.22 (d, J = 5.4 Hz, 1H), 7.15 (d, J = 1.2 Hz, 1H), 6.91 (d, J = 1.2 Hz, 1H), 2.41 (s, 3H), 2.28 (s, 3H), 2.27 (s, 3H); ESI MS m/z 337 [M + H]+. 98.7 25 3-(6-(3,5- dimethylisoxazol- 4-yl)-2- thioxo-2,3- dihydro-1H- benzo[d]imidazol- 4-yl)-4- methylbenzonitrile No general procedure .sup.1H NMR (400 MHz, CDCl.sub.3) 10.26 (br.s, 2H), 7.68 (d, J = 8.2 Hz, 1H), 7.58 (s, 1H), 7.50 (d, J = 7.8 Hz, 1H), 7.13 (s, 1H), 6.92 (s, 1H), 2.43 (s, 3H), 2.29 (s, 6H); ESI MS m/z 361 [M + H]+. 97.4 26 5-(3,5- dimethylisoxazol- 4-yl)-1-methyl-7- ((1,3,5- trimethyl-1H- pyrazol-4- yl)amino)-1H- benzo[d]imidazol- 2(3H)-one B .sup.1H NMR (500 MHz, CD.sub.3OD) 5.45 (s, 1H), 5.89 (s, 1H), 3.77 (s, 3H), 3.72 (s, 3H), 2.27 (s, 3H), 2.14 (s, 3H), 2.10 (s, 3H), 2.06 (s, 3H); ESI m/z 367 [M + H]+. 97.9 27 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-((2- methylpyridin- 3-yl)amino)-1H- benzo[d]imidazol- 2(3H)-one B .sup.1H NMR (500 MHz, CD.sub.3OD) 7.86 (dd, J = 1.3, 4.8 Hz, 1H), 7.08-7.03 (m, 1H), 6.93 (d, J = 1.5 Hz, 1H), 6.78 (d, J = 1.5 1H), 6.77 (dd, J = 1.2, 8.2 Hz, 1H), 3.35 (s, 3H), 2.55 (s, 3H), 2.39 (s, 3H), 2.23 (s, 3H); ESI m/z 350 [M + H]+. >99 28 5-(5-(hydroxyl methyl)-3- methylisoxazol- 4-yl)-1-methyl-7- (1,3,5- trimethyl-1H- pyrazol-4-yl)- 1H-benzo [d]imidazol- 2(3H)-one C .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.08 (s, 1H), 7.03 (d, J = 1.5 Hz, 1H), 6.74 (d, J = 1.8 Hz, 1H), 5.65 (t, J = 5.7 Hz, 1H), 4.49 (d, J = 5.7 Hz, 2H), 3.73 (s, 3H), 2.88 (s, 3H), 2.27 (s, 3H), 2.09 (s, 3H), 1.97 (s, 3H); ESI m/z 368 [M + H]+. 98.7 29 3-(6-(3,5- dimethylisoxazol- 4-yl)-3- methyl-2-oxo- 2,3-dihydro- 1H-benzo [d]imidazol-4- yl)-4-methyl benzamide E .sup.1H NMR (500 MHz, DMSO-d.sub.6) 11.11 (br.s, 1H), 7.94 (br.s, 1H), 7.87 (dd, J = 7.8, 2.0 Hz, 1H), 7.82 (d, J = 2.0 Hz, 1H), 7.42 (d, J = 8.0 Hz, 1H), 7.31 (br.s, 1H), 6.99 (d, J = 2.0 Hz, 1H), 6.75 (d, J = 1.5 Hz, 1H) 2.67 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.15 (s, 3H); ESI m/z 377 [M + H]+. >99 30 3-(6-(3,5- dimethylisoxazol- 4-yl)-3- methyl-2-oxo- 2,3-dihydro- 1H-benzo [d]imidazol-4- yl)-2-methyl benzamide E .sup.1H NMR (500 MHz, DMSO-d.sub.6) 11.10 (br.s, 1H), 7.79 (br.s, 1H), 7.45 (br.s, 1H), 7.41 (dd, J = 7.5, 1.5 Hz, 1H), 7.35 (dd, J = 7.5, 1.5 Hz, 1H), 7.30 (t, J = 7.5 Hz, 1H), 6.98 (d, J = 2.0 Hz, 1H), 6.68 (d, J = 1.5 Hz, 1H), 2.69 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.11 (s, 3H); ESI m/z 377 [M + H]+. >99 31 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-((2- methylpyridin- 3-yl)oxy)-1H- benzo[d]imidazol- 2(3H)-one No general procedure .sup.1H NMR (500 MHz, CD.sub.3OD) 8.21 (dd, J = 1.5, 4.6 Hz, 1H), 7.31 (dd, J = 1.5, 8.3 Hz, 1H), 7.27 (dd, J = 4.6, 8.4 Hz, 1H), 6.87 (d, J = 1.4 Hz, 1H), 6.49 (d, J = 1.4 Hz, 1H), 3.51 (s, 3H), 2.59 (s, 3H), 2.34 (s, 3H), 2.17 (s, 3H); HPLC 97.0%, tR = 8.5 min; ESI m/z 351 [M + H]+. 97.0 32 7-(3,5- dimethyl-1H- pyrazol-4-yl)- 5-(5-(hydroxyl methyl)-3- methylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one C .sup.1H NMR (300 MHz, DMSO-d.sub.6) 12.37 (br.s, 1H), 11.07 (s, 1H), 7.03 (d, J = 1.8 Hz, 1H), 6.76 (d, J = 1.5 Hz, 1H), 5.65 (t, J = 5.7 Hz, 1H), 4.49 (d, J = 5.7 Hz, 2H), 2.87 (s, 3H), 2.27 (s, 3H), 2.04 (br.s, 6H); ESI m/z 354 [M + H]+. 98.7 33 5-(3,5- dimethylisoxazol- 4-yl)-7- ((3,5-dimethyl isoxazol-4- yl)amino)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one B .sup.1H NMR (300 MHz, CD.sub.3OD) 6.57 (d, J = 1.5 Hz, 1H), 6.02 (d, J = 1.5 Hz, 1H), 3.75 (s, 3H), 2.03 (s, 3H), 2.29 (s, 3H), 2.13 (s, 3H), 2.11 (s, 3H); ESI m/z 354 [M + H]+. 95.7 34 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7- (naphthalen- 1-yl)-1H- benzo[d]imidazol- 2(3H)-one 0 A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 8.06 (s, 1H), 8.03 (s, 1H), 7.63-7.50 (m, 5H), 7.07 (d, J = 1.5 Hz, 1H), 6.86 (d, J = 1.8 Hz, 1H), 2.41 (s, 3H), 2.40 (s, 3H), 2.24 (s, 3H); ESI MS m/z 370 [M + H]+. 95.6 35 7-(3,5- dichloropyridin- 4-yl)-5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one D .sup.1H NMR (500 MHz, CD.sub.3OD) 8.71 (s, 2H), 7.14 (d, J = 1.6 Hz, 1H), 6.83 (d, J = 1.6 Hz, 1H), 2.97 (s, 3H), 2.42 (s, 3H), 2.26 (s, 3H); ESI m/z 389 [M + H]+. >99 36 5-(3,5- dimethylisoxazol- 4-yl)-1-methyl-7- (quinolin-3- yl)-1H-benzo [d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 9.06 (d, J = 2.4 Hz, 1H), 8.53 (d, J = 2.1 Hz, 1H), 8.10 (d, J = 7.8 Hz, 1H), 8.08 (d, J = 6.9 Hz, 1H), 7.86- 7.80 (m, 1H), 7.72- 7.69 (m, 1H), 7.07 (d, J = 1.5 Hz, 1H), 6.99 (d, J = 1.5 Hz, 1H), 2.87 (s, 3H), 2.43 (s, 3H), 2.25 (s, 3H); ESI MS m/z 371 [M + H]+. >99 37 7-(2-chloro phenyl)-5- (3,5-dimethyl isoxazol-4-yl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 7.63-7.43 (m, 4H), 7.01 (d, J = 1.5 Hz, 1H), 6.76 (d, J = 1.5 Hz, 1H), 2.76 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H); ESI MS m/z 354 [M + H]+. 97.8 38 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(3- methylpyridin- 4-yl)-1H- benzo[d]imidazol- 2(3H)-one D .sup.1H NMR (500 MHz, CD.sub.3OD) 8.53 (s, 1H), 8.47 (d, J = 4.9 Hz, 1H), 7.42 (d, J = 5.0 Hz, 1H), 7.09 (d, J = 1.4 Hz, 1H), 6.79 (d, J = 1.4 Hz, 1H), 2.88 (s, 3H), 2.41 (s, 3H), 2.25 (s, 3H), 2.19 (s, 3H); ESI m/z 335 [M + H]+. >99 39 5-(3,5- dimethylisoxazol- 4-yl)-7- (3,5-dimethyl pyridin-4-yl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one D .sup.1H NMR (500 MHz, CD.sub.3OD) 8.37 (s, 2H), 7.10 (d, J = 1.6 Hz, 1H), 6.72 (d, J = 1.6 Hz, 1H), 2.82 (s, 3H), 2.41 (s, 3H), 2.25 (s, 3H), 2.11 (s, 6H); ESI m/z 349 [M + H]+. >99 40 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-7-(o- tolyl)-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.1 (s, 1H), 7.35-7.29 (m, 4H), 6.96 (d, J = 1.8 Hz, 1H), 6.71 (d, J = 1.8 Hz, 1H), 2.67 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H), 2.09 (s, 3H); ESI MS m/z 334 [M + H]+. >99 41 5-(3,5- dimethylisoxazol- 4-yl)-7-(2- fluoro-5- methoxyphenyl)- 1-methyl- 1H-benzo [d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 7.30-7.24 (m, 1H), 7.07-7.01 (m, 3H), 6.85 (d, J = 1.5 Hz, 1H), 3.79 (s, 3H), 2.89 (s, 3H), 2.41 (s, 3H), 2.23 (s, 3H); ESI m/z 368 [M + H]+. >99 42 7-(5-chloro-2- methoxyphenyl)- 5-(3,5- dimethylisoxazol- 4-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.1 (s, 1H), 7.51-7.48 (m, 1H), 7.39 (d, J = 2.7 Hz, 1H), 7.15 (d, J = 9.0 Hz, 1H), 6.96 (d, J = 1.5 Hz, 1H), 6.75 (d, J = 1.5 Hz, 1H), 3.74 (s, 3H), 2.82 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H); ESI MS m/z 384 [M + H]+. 94.2 43 5-(3,5- dimethylisoxazol- 4-yl)-7-(2- fluoro-3- methoxyphenyl)- 1-methyl- 1H-benzo[d] imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 7.28-7.21 (m, 2H), 7.06-7.01 (m, 2H), 6.81 (d, J = 1.5 Hz, 1H), 3.89 (s, 3H), 2.85 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H); ESI MS m/z 368 [M + H]+. >99 44 5-(3,5- dimethylisoxazol- 4-yl)-7- (2,4-dimethyl thiazol-5-yl)-1- methyl-1H- benzo[d]imidazol- 2(3H)-one 0 A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 7.01 (d, J = 1.8 Hz, 1H), 6.83 (d, J = 1.5 Hz, 1H), 2.94 (s, 3H), 2.66 (s, 3H), 2.39 (s, 3H), 2.21 (s, 3H), 2.15 (s, 3H); ESI MS m/z 355 [M + H]+. >99 45 5-(3,5- dimethylisoxazol- 4-yl)-7-(2- methoxy-6- methylpyridin methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.1 (s, 1H), 7.65 (d, J = 7.2 Hz, 1H), 6.99 (s, 1H), 6.96 (d, J = 1.8 Hz, 1H), 6.73 (d, J = 1.8 Hz, 1H), 3.82 (s, 3H), 2.83 (s, 3H), 2.48 (s, 3H), 2.40 (s, 3H), 2.22 (s, 3H); ESI MS m/z 365 [M + H]+. >99 46 7-(benzo[d] oxazol-5-yl)-5- (3,5-dimethyl isoxazol-4-yl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d.sub.6) 11.2 (s, 1H), 8.85 (s, 1H), 7.92 (d, J = 1.2 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.56-7.53 (m, 1H), 7.00 (d, J = 1.5 Hz, 1H), 6.85 (d, J = 1.8 Hz, 1H), 2.79 (s, 3H), 2.42 (s, 3H), 2.24 (s, 3H); ESI MS m/z 361 [M + H]+. 91.4 47 7-(cyclohex-1- en-1-yl)-5- (3,5-dimethyl isoxazol-4-yl)- 1-methyl-1H- benzo[d]imidazol- 2(3H)-one A .sup.1H NMR (300 MHz, DMSO-d6) 11.0 (s, 1H), 6.82 (d, J = 1.8 Hz, 1H), 6.67 (d, J = 1.8 Hz, 1H), 5.68- 5.67 (m, 1H), 3.30 (s, 3H), 2.38 (s, 3H), 2.29-2.27 (m, 2H), 2.20 (s, 3H), 2.19- 2.18 (m, 2H), 1.77- 1.65 (m, 4H); ESI MS m/z 324 [M + H]+. >99

Example 1: Inhibition of Tetra-Acetylated Histone H4 Binding Individual BET Bromodomains

(93) Proteins were cloned and overexpressed with a N-terminal 6His 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.

(94) Binding of tetra-acetylated histone H4 and BET bromodomains was confirmed by a Homogenous Time Resolved Fluorescence Resonance Energy Transfer (HTRF) method. N-terminally His-tagged bromodomains (200 nM) and biotinylated tetra-acetylated histone H4 peptide (25-50 nM, Millipore) were incubated in the presence of Europium Cryptate-labeled streptavidin (Cisbio Cat. #610SAKLB) and XL665-labeled monoclonal anti-His antibody (Cisbio Cat. #61HISXLB) in a white 96 well microtiter plate (Greiner). For inhibition assays, serially diluted test compound was added to these reactions in a 0.2% final concentration of DMSO. Duplicate wells were used for each concentration tested. Final buffer concentrations were 30 mM HEPES pH 7.4, 30 mM NaCl, 0.3 mM CHAPS, 20 mM phosphate pH 7.0, 320 mM KF, 0.08% BSA. After a 2 h incubation at room temperature, fluorescence was measured at 665 and 620 nm with a SynergyH4 plate reader (Biotek). The binding inhibitory activity was shown by a decrease in 665 nm relative to 620 nm fluorescence. IC.sub.50 values were determined from a dose response curve.

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

(96) TABLE-US-00003 TABLE 2 Inhibition of Tetra-acetylated Histone H4 Binding to Brd4 bromodomain 1 (BRD4(1) as Measured by FRET Example FRET Example FRET FRET Com- activity Com- activity Example activity pound BRD4(1) pound BRD4(1) Compound BRD4(1) 1 +++ 2 +++ 3 +++ 4 +++ 5 +++ 6 +++ 7 +++ 8 +++ 9 +++ 10 +++ 11 ++ 12 +++ 13 +++ 14 +++ 15 +++ 16 +++ 17 +++ 18 +++ 19 +++ 20 +++ 21 +++ 22 +++ 23 +++ 24 +++ 25 +++ 26 +++ 27 +++ 28 +++ 29 +++ 30 +++ 31 +++ 32 +++ 33 +++ 34 +++ 35 +++ 36 ++ 37 ++ 38 +++ 39 +++ 40 +++ 41 +++ 42 +++ 43 ++ 44 ++ 45 ++ 46 +++

Example 2: Inhibition of c-MYC Expression in Cancer Cell Lines

(97) MV4-11 cells (CRL-9591) were plated at a density of 2.510.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 ViiA7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hMYC to an internal control, prior to determining the fold expression of each sample, relative to the control.

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

(99) TABLE-US-00004 TABLE 3 Inhibition of c-myc Activity in Human AML MV4-11 cells Example c-myc Example c-myc Example c-myc Example c-myc Compound activity Compound activity Compound activity Compound activity 1 ++ 2 +++ 3 + 4 +++ 5 +++ 6 +++ 7 +++ 8 ++ 9 +++ 10 +++ 11 ++ 13 +++ 14 +++ 15 ++ 16 +++ 17 +++ 18 ++ 19 +++ 20 ++ 21 +++ 22 +++ 23 +++ 24 +++ 25 +++ 26 +++ 27 +++ 28 ++ 29 +++ 30 +++ 31 ++ 32 +++ 33 +++ 34 +++ 35 NA 36 ++ 37 ++ 38 ++ 39 +++ 40 +++ 44 ++ 45 +++

Example 3: Inhibition of Cell Proliferation in Cancer Cell Lines

(100) In this example, cell titer in MV4-11 cells were quantitated to measure the inhibition of proliferation when treated with a compound of the present disclosure.

(101) MV4-11 cells (CRL-9591) were plated at a density of 510.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 L of the CellTiter 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 Graph Pad Prism software.

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

(103) TABLE-US-00005 TABLE 4 Inhibition of Cell Proliferation in Human AML MV-4-11 cells Cell Cell Cell Cell Example Proliferation Example Proliferation Example Proliferation Example Proliferation Compound activity Compound activity Compound activity Compound activity 1 +++ 2 ++ 3 + 4 ++ 5 +++ 6 ++ 7 +++ 8 ++ 9 +++ 10 +++ 11 ++ 12 ++ 13 ++ 14 ++ 15 ++ 16 ++ 17 ++ 18 ++ 19 ++ 20 +++ 21 +++ 22 +++ 23 +++ 24 ++ 25 ++ 26 ++ 27 ++ 28 ++ 29 ++ 30 +++ 31 ++ 32 ++ 33 ++ 34 ++ 35 Not 36 ++ available 37 ++ 38 ++ 39 ++ 40 ++ 44 + 45 ++

Example 4: Inhibition of hIL-6 mRNA Transcription

(104) In this example, hIL-6 mRNA in tissue culture cells were quantitated to measure the transcriptional inhibition of hIL-6 when treated with a compound of the present disclosure.

(105) Human leukemic monocyte lymphoma U937 cells (CRL-1593.2) were plated at a density of 3.2104 cells per well in a 96-well plate in 100 L 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% CO2 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% CO2 for 3 h before the cells were harvested. At time of harvest, media was removed and cells were rinsed in 200 L 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 ViiA7 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.

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

(107) TABLE-US-00006 TABLE 5 Inhibition of hIL-6 mRNA Transcription Example IL-6 Example IL-6 Example IL-6 Example IL-6 Compound activity Compound activity Compound activity Compound activity 1 +++ 2 +++ 3 ++ 4 +++ 5 +++ 6 +++ 7 +++ 8 ++ 9 +++ 10 +++ 11 ++ 12 ++ 13 ++ 16 ++ 18 +++ 19 ++ 20 +++ 21 +++ 22 +++ 23 +++ 26 ++ 27 +++ 28 ++ 29 ++ 30 +++ 31 +++ 32 +++ 33 ++ 34 +++ 35 ++ 38 ++ 39 +++ 40 ++ 44 ++ 45 ++

Example 5: Inhibition of hIL-17 mRNA Transcription

(108) In this example, hIL-17 mRNA in human peripheral blood mononuclear cells were quantitated to measure the transcriptional inhibition of hIL-17 when treated with a compound of the present disclosure.

(109) Human peripheral blood mononuclear cells were plated (2.010.sup.5 cells per well) in a 96-well plate in 45 L OpTimizer T Cell expansion media (Life Technologies) containing 20 ng/ml IL-2 and penicillin/streptomycin. The cells were treated with increasing concentrations of the test compound or DMSO (0.1%), and incubated at 37 C., 5% CO2 for 1 h before addition of 10 stock OKT3 antibody at 10 ug/ml in media. Triplicate wells were used for each concentration. Cells were incubated at 37 C., 5% CO2 for 6 h before the cells were harvested. At time of harvest, cells were pelleted by centrifugation at 800 rpm for 5 min. 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-17 and Cyclophilin. Real-time PCR plates were run on a ViiA7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hIL-17 to an internal control, prior to determining the fold induction of each unknown sample, relative to the control.

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

(111) TABLE-US-00007 TABLE 6 Inhibition of hIL-17 mRNA Transcription Example Compound IL-17 activity Example Compound IL-17 activity 1 +++ 2 +++ 4 ++ 5 ++ 7 ++ 9 +++ 10 +++ 11 +++ 13 ++ 18 +++ 20 ++ 21 +++ 22 +++ 28 ++ 30 +++

Example 6: Inhibition of hVCAM mRNA Transcription

(112) In this example, hVCAM mRNA in tissue culture cells is quantitated to measure the transcriptional inhibition of hVCAM when treated with a compound of the present disclosure.

(113) Human umbilical vein endothelial cells (HUVECs) are plated in a 96-well plate (4.010.sup.3 cells per well) in 100 L EGM media and incubated for 24 h prior to the addition of increasing concentrations of the compound of interest or DMSO (0.1%). Triplicate wells were used for each concentration. The cells are pretreated for 1 h with the test compound prior to stimulation with tumor necrosis factor- when they are incubated for an additional 24 h before the cells are harvested. At time of harvest, the spent media is removed and HUVECs are rinsed in 200 L 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 hVCAM and Cyclophilin. Real-time PCR plates were run on a ViiA7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hVCAM to an internal control, prior to determining the fold induction of each unknown sample, relative to the control.

Example 7: Inhibition of hMCP-1 mRNA Transcription

(114) In this example, hMCP-1 mRNA in human peripheral blood mononuclear cells is quantitated to measure the transcriptional inhibition of hMCP-1 when treated with a compound of the present disclosure.

(115) Human Peripheral Blood Mononuclear Cells are plated at a density of 1.010.sup.5 cells per well in a 96-well plate in RPMI-1640 containing 10% FBS and penicillin/streptomycin. The cells are treated with increasing concentrations of the compound or DMSO (0.1%), and incubated at 37 C., 5% CO2 for 3 h before the cells are harvested. At time of harvest, cells are transferred to V-bottom plates and pelleted by centrifugation at 800 rpm for 5 min. 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 hMCP-1 and Cyclophilin. Real-time PCR plates were run on a ViiA7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hMCP-1 to an internal control, prior to determining the fold induction of each unknown sample, relative to the control.

Example 8: Up-Regulation of hApoA-1 mRNA Transcription

(116) In this example, hApoA-I mRNA in tissue culture cells was quantitated to measure the transcriptional up-regulation of hApoA-I when treated with a compound of the present disclosure.

(117) Huh7 cells (2.510.sup.5 per well) were plated in a 96-well plate using 100 L DMEM per well, (Gibco DMEM supplemented with penicillin/streptomycin and 10% FBS), 72 h before the addition of the compound. The cells are treated with increasing concentrations of the compound or DMSO (0.1%), and incubated at 37 C., 5% CO2 for 48 h. Spent media was removed from the Huh-7 cells and placed on ice for immediate use with the LDH cytotoxicity assay Kit II from Abcam. The cells remaining in the plate were rinsed with 100 L 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 hApoA-I and Cyclophilin. Real-time PCR plates were run on a ViiA7 real time PCR machine (Applied Biosystems), data was analyzed, normalizing the Ct values for hApoA-1 to an internal control, prior to determining the fold induction of each unknown sample, relative to the control.

(118) Compounds with an EC.sub.170 value less than or equal to 0.3 M were deemed to be highly active (+++); compounds with an EC.sub.170 value between 0.3 and 3 M were deemed to be very active (++); compounds with an EC.sub.170 value between 3 and 30 M were deemed to be active (+).

(119) TABLE-US-00008 TABLE 7 Up-regulation of hApoA-1 mRNA Transcription. Example ApoA-1 Example ApoA-1 Example ApoA-1 Compound activity Compound activity Compound activity 1 +++ 4 +++ 7 +++ 9 +++ 13 +++

Example 9: In Vivo Efficacy in Athymic Nude Mouse Strain of an Acute Myeloid Leukemia Xenograft Model Using MV4-11 Cells

(120) MV4-11 cells (ATCC) were grown under standard cell culture conditions and (NCr) nu/nu fisol strain of female mice age 6-7 weeks were injected with 510.sup.6 cells/animal in 100 L PBS+100 L Matrigel in the lower left abdominal flank. By approximately day 18-21 after MV4-11 cells injection, mice were randomized based on tumor volume (LWH)/2) of average 100-300 mm.sup.3. Mice were 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 were 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 were compared relative to Vehicle control animals. The means, statistical analysis and the comparison between groups were calculated using Student's t-test in Excel.

(121) TABLE-US-00009 TABLE 8 In vivo efficacy in athymic nude mouse strain of an acute myeloid leukemia xenograft model Example Compound In vivo activity Example 9 Active

Example 10: In Vivo Efficacy in Athymic Nude Mouse Strain of an Acute Myeloid Leukemia Xenograft Model Using OCI-3 AML Cells

(122) OCI-3 AML cells (DMSZ) were grown under standard cell culture conditions and (NCr) nu/nu fisol strain of female mice age 6-7 weeks were injected with 1010.sup.6 cells/animal in 100 L PBS+100 L Matrigel in the lower left abdominal flank. By approximately day 18-21 after OCI-3 AML cells injection, mice were randomized based on tumor volume (LWH)/2) of average 100-300 mm.sup.3. Mice were dosed orally with compound at 30 mg/kg b.i.d on a continuous dosing schedule and at 2.5 to 45 mg/kg q.d. on a 5 day on and 2 day off dosing schedule in EA006 formulation at 10 mL/kg body weight dose volume. Tumor measurements were 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 were compared relative to Vehicle control animals. The means, statistical analysis and the comparison between groups were calculated using Student's t-test in Excel.

(123) TABLE-US-00010 TABLE 9 In vivo efficacy in athymic nude mouse strain of an acute myeloid leukemia xenograft model using OCI-3 AML cells Example Compound In vivo activity Example 9 Active

Example 11: Evaluation of Target Engagement

(124) MV4-11 and MM1.s cells (ATCC) were grown under standard cell culture conditions and (NCr) nu/nu fisol strain of female mice age 6-7 weeks were injected with 510.sup.6 cells/animal in 100 L PBS+100 L Matrigel in the lower left abdominal flank. By approximately day 28 after MV4-11 and MM1.s cells injection, mice were randomized based on tumor volume (LWH)/2) of average 500 mm.sup.3. Mice were dosed orally with compound in EA006 formulation at 10 mL/kg body weight dose volume and tumors harvested 3, 6, 12, 24 hrs post dose for Bcl2 and c-myc gene expression analysis as PD biomarkers.

(125) TABLE-US-00011 TABLE 10 Evaluation of Target Engagement. Example Compound In vivo activity Example 9 Active

Example 12: In Vivo Efficacy in Athymic Nude Mouse Strain of Multiple Myeloma Xenograft Model Using MM1.s Cells

(126) MM1.s cells (ATCC) were grown under standard cell culture conditions and SCID-Beige strain of female mice age 6-7 weeks were injected with 1010.sup.6 cells/animal in 100 L PBS+100 L Matrigel in the lower left abdominal flank. By approximately day 21 after MM1.s cells injection, mice were randomized based on tumor volume (LWH)/2) of average 120 mm.sup.3. Mice were dosed orally with compound at 25 to 90 mg/kg b.i.d and or q.d in EA006 formulation at 10 mL/kg body weight dose volume. Tumor measurements were 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 were compared relative to Vehicle control animals. The means, statistical analysis and the comparison between groups were calculated using Student's t-test in Excel.

(127) TABLE-US-00012 TABLE 11 In vivo efficacy in athymic nude mouse strain of multiple myeloma xenograft model using MM1.s cells Example Compound In vivo activity Example 9 Active

Example 13: In Vivo Efficacy in Mouse Endotoxemia Model Assay

(128) Sub lethal doses of Endotoxin (E. Coli bacterial lipopolysaccharide) are administered to animals to produce a generalized inflammatory response which is monitored by increases in secreted cytokines. Compounds are administered to C57/Bl6 mice at T=4 hours orally at 75 mg/kg dose to evaluate inhibition in IL-6 and IL-17 and MCP-1 cytokines post 3-h challenge with lipopolysaccharide (LPS) at T=0 hours at 0.5 mg/kg dose intraperitoneally.

Example 14: In Vivo Efficacy in Rat Collagen-Induced Arthritis

(129) Rat collagen-induced arthritis is an experimental model of polyarthritis that has been widely used for preclinical testing of numerous anti-arthritic agents. Following administration of collagen, this model establishes a measurable polyarticular inflammation, marked cartilage destruction in association with pannus formation and mild to moderate bone resorption and periosteal bone proliferation. In this model, collagen was administered to female Lewis strain of rats on Day 1 and 7 of study and dosed with compounds from Day 11 to Day 17. Test compounds were evaluated to assess the potential to inhibit the inflammation (including paw swelling), cartilage destruction and bone resorption in arthritic rats, using a model in which the treatment is administered after the disease has been established.

Example 15: In Vivo Efficacy in Experimental Autoimmune Encephalomyelitis (EAE) Model of MS

(130) Experimental autoimmune encephalomyelitis (EAE) is a T-cell-mediated autoimmune disease of the CNS which shares many clinical and histopathological features with human multiple sclerosis (MS). EAE is the most commonly used animal model of MS. T cells of both Th1 and Th17 lineage have been shown to induce EAE. Cytokines IL-23, IL-6 and IL-17, which are either critical for Th1 and Th17 differentiation or produced by these T cells, play a critical and non-redundant role in EAE development. Therefore, drugs targeting production of these cytokines are likely to have therapeutic potential in treatment of MS.

(131) Compounds of Formula I were administered to EAE mice to assess anti-inflammatory activity. In this model, EAE is induced by MOG.sub.35-55/CFA immunization and pertussis toxin injection in female C57Bl/6 mice.

Example 16: Ex Vivo Effects on T Cell Function from Splenocyte and Lymphocyte Cultures Stimulated with External MOG Stimulation

(132) Mice were immunized with MOG/CFA and simultaneously treated with the compound for 11 days on a b.i.d regimen. Inguinal Lymph node and spleen were harvested, cultures were set up for lymphocytes and splenocytes and stimulated with external antigen (MOG) for 72 hours. Supernatants from these cultures were analyzed for TH1, Th2 and Th17 cytokines using a Cytometric Bead Array assay.

(133) 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.