Apabetalone

Apabetalone (RVX-208) is an inhibitor of BET transcriptional regulators with selectivity for the second bromodomain. The IC50s are 87±10 μM and 0.51±0.041 μM for BD1 and BD2, respectively.

Compound 10A (3.26 g, 10 mmol) was added to a 100 mL single-mouth bottle.Compound 11 is ethylene carbonate (0.88 g, 10 mmol), Add 30 mL of N, N-dimethylformamide to dissolve it.At the same time, potassium carbonate (1.38 g, 10 mmol) was added.Warm the system to 110 ° C, The reaction was refluxed for 12 hours.After the reaction is over, Add 20 mL of water to the reaction system.Extract three times with dichloromethane (3*20 mL), Combine the organic phase, Dry with anhydrous sodium sulfate, filter, Distilling under reduced pressure to obtain a mixture, Purified by column chromatography (the eluent is petroleum ether / ethyl acetate, Volume ratio 2:1), A yellow solid RVX-208 (3.51 g, yield 95percent) was obtained.Intermediate 2 (58.74 kg), ?/, ?/-dimethylacetamide (280 kg), and starting material 3 (56.00 kg) were combined and p-toluenesulfonic acid monohydrate (5.90 kg) and 1/3 of the required sodium bisulfite (24.1 kg) were added. The mixture was heated to 115 0C and stirred for 90-105 minutes before the second 1/3 of the required sodium bisulfite (24.1 kg) was added. The remaining sodium bisulfite (24.1 kg) was added after another 90-105 minutes. The reaction mixture was stirred at 115 0C until the reaction was complete as determined by HPLC (approximately 1 hour, less than 4percent of intermediate 2 remaining). The reaction mixture was cooled to 25 0C and added to water (1770 kg). The mixture was stirred at 20 0C for 6 hours to complete the crystallization. The crude material was isolated by filtration, washed with water (234 kg) and dried under vacuum to constant weight. The crude material was dissolved in ?/, ?/-dimethylacetamide (252 kg) at 80 0C until all material had dissolved. The solution was cooled to 60 0C and heptane (918 kg) was slowly added over a period of 1 hour, maintaining a temperature above 35 0C.The solution was cooled to 35 0C and stirred at 35 0C for a minimum of 1 hour. The solid was isolated by filtration, washed with heptane (250 kg) and dried to constantweight under vacuum. Yield: 92.5percent; purity: 98.6percent. The dry solid (83.1 kg) was added to a 1 :1 mixture of ethanol and water (1V/1V; 1670 kg), and the mixture was heated to approximately 840C (reflux) until all material was in solution. The solution was cooled to 70 0C and polish-filtered, and then cooled to 30 0C over 2 hours. The solution was cooled to 0 0C. The mixture was stirred at 0 0C for at least 1 hour, before the material was isolated by filtration, washed with ethanol/water (1V/1V; 33 kg) and dried under vacuum to constant weight. The material was passed through a 60-mesh screen to afford 2-(4-(2-hydroxyethoxy)-3, 5-dimethylphenyl)-5, 7- dimethoxyquinazolin-4(3H)-one (4). Yield: 66.4 kg; 79.9percent.The intermediate H30-3 (15.2 g, 77.5 mol), H30-5 (15.06 g, 77.5 mmol), NAHSO3 (8.9 g, 85.3 mmol), P-TSA(1.34 g, 7.75 mmol), and NMP (140 mL) were added into a flask in the presence of nitrogen gas. The mixture was stirredfor 3 h at 130 °C. After the completion of the reaction detected by TLC, the mixture was extracted by adding water (450mL) and DCM (500 mL). The separated aqueous layer was extracted with DCM (4 3 400 mL). The resulting organiclayers were combined, washed with water (3 3 400 mL), dried by adding sodium sulfate, and filtered. The resultingfiltrate was distilled under reduced pressure. The resulting crude product was purified by silica gel column chromatography(eluent: dichloromethane: methanol = 80: 1) to give the intermediate H130: 2-(4-(2-hydroxyethoxy)-3, 5-dimethylphenyl)-5, 7-dimethoxyquinazoline-4(3H)-one (18 g, yield 62.7percent).A solution of 2-amino-4, 6-dimethoxybenzamide (0.60 g, 3.06 mmol) and 4-[2-(tert-butyidimethylsilanoxy)ethoxy]-3, 5-dimethylbenzaldehyde (0.856 g, 2.78 mmol) in N, N-dimethyl formamide (20 mL) was stirred at 70° C. for 1 h. Iodine (0.846 g, 3.33 mmol) and potassium carbonate (0.384 g, 2.78 mmol) were added and the reaction mixture was stirred at 70° C. for 16 h. The reaction mixture was poured into ice, and extracted with ethyl acetate. The organic layer was washed with water, brine, and dried over anhydrous NaTo a solution of 2-amino-4, 6-dimethoxy-benzamide (33.45 g, 0.170 mol) and 4-(2-hydroxy ethoxy)-3, 5-dimethyl benzaldehyde (33.3 g, 0.170 mol) in N, N-dimethyl acetamide (300 mL), NaHSOAt 70deg.] C 2-Amino-4, 6-dimethoxy-benzamide(0.60g, 3.06mmol) and 4- [2- (tert-butyldimethylsilyloxy)ethoxy]-3, 5-dimethyl-benzaldehyde(0.856g, 2.78mmol)in N, N- dimethylformamide(20 mL) was stirred for 1hour. Was added iodine (0.846g, 3.33mmol)and potassium carbonate (0.384g, 2.78mmol), thereaction mixture was at 70 stirred for 16 hours. The reactionmixture was poured onto ice, extracted with ethyl acetate. With water, theorganic layer was washed with brine, dried over anhydrous Na 2SO 4dry. The solvent was removed to give acrude product, which was purified by column chromatography to give a white solid of 2- (4- (2-hydroxyethoxy) -3, 5-dimethylphenyl) -5, 7-methoxy-quinazolin -4 (3H) - one (444mg, 39percent). Selected data: 229-231 . Alternatively, 2- (4-(2-hydroxyethoxy)-3, 5-dimethyl-phenyl)-5, 7-dimethoxy-quinazolin -4 (3H) - onecan be synthesized by the following method. In ethanol (350 mL of) of 3, 5-dimethyl-4-hydroxy-benzaldehyde(26.9g, 0.179mol) disposed dried 2L round bottomed flask with refluxcondenser and magnetic stirrer. 2-chloro-ethanol(87.6g, 1.074mol) and K 2CO 3(99g, 0.716mol), the reaction mixture was heated at reflux for 24 hours. The reaction mixture was cooled toroom temperature and filtered. The solvent was removed under reduced pressure.The crude product was diluted with ethyl acetate, and the organic layer waswashed with water, brine, Na 2SO 4dry. 45g crude product obtained after removalof the solvent. The crude product was purified by column chromatography (silicagel 230-400 mesh; with 50percent hexanes in ethyl acetate as eluent) to afford 33.3g (95percent) of product. 2-amino-4, 6-dimethoxy - benzamide (33.45g, 0.170mol)and 4- (2-hydroxyethoxy) -3, 5-dimethyl benzaldehyde(33.3g, 0.170 mol) was added NaHSO3(33.3g, 0.187mol) in N, N- dimethylacetamide (300mL) solution of and p-TSA(3.2g, 17.1mmol), at 150 reaction mixturewas heated for 14 hours.The reaction was cooled to room temperature. The solvent was removed underreduced pressure. The residue was diluted with water, followed by stirring at room temperature for30 minutes. The solid was isolated by filtration and dried to give the crudeproduct. The crude product was purified by column chromatography (silica gel230-400 mesh; used in CH 2Cl 25percent methanol as eluent) to afford 2- (4- (2-hydroxyethoxy) -3, 5-dimethyl-phenyl)-5, 7-dimethoxy-quinazoline -4 (3H) - one (33g, 52percent).A solution of 2-amino-4, 6-dimethoxyben2amide (0.60 g, 3.06 mmol) and 4-t2-(tert-butyldimethylsflanoxy)ethoxy]-3, 5-dfmethylbenzaldehyde (0.856 g, 2.78 mmol) in λ/, /V-dimethyl formamide (20 mL) was stirred at 70°C for 1 h. Iodine (0.846 g, 3.33 mmol) and potassium carbonate (0.384 g, 2.78 mmol) were added and the reaction mixture was stirred at 70°C for 16 h, The reaction mixture was poured into ice, and extracted with ethyl acetate. The organic layer was washed with water, brine, and dried over anhydrous NaTake 2-bromo-4, 6-dimethoxybenzamide (C-4) (388.5 mg, 1.50 mmol), Cuprous bromide (21.5 mg, 0.15 mmol), cesium carbonate (977.5 mg, 3.00 mmol), L-valine (34.5mg, 0.30mmol) in a thick-walled pressure tube, Add 5 ml of DMSO, intermediate B-3 (930 mg, 3.00 mmol), 1.90 ml of 26percent aqueous ammonia, Seal the pressure tube. The mixture was stirred at 100 ° C for 12 h, and then open for 12 h. Quenched with 6ml saturated ammonium chloride solution, Add 50 ml of ethyl acetate and 50 ml of distilled water.The mixture was separated and the aqueous layer was extracted with ethyl acetate (50ml×3).The organic layer was washed with distilled water (30 ml × 2) and saturated brine (30 ml).Dry over anhydrous sodium sulfate. Column chromatography, eluting with dichloromethane/methanol (v/v, 40:1) afforded 267 mg of Apabetalone.The yield was 48percent.2-Amino-4, 6-dimethoxybenzonitrile 8.9 g (0.05 mol) and 4-(2-hydroxyethoxy)-3, 5-dimethylbenzaldehyde 14.6 g were sequentially added to the reaction flask. (0.075mol), 50ml of tetrahydrofuran, 0.05g of sodium hydroxide, stirred and refluxed for 3.5h, After the reaction is completed, ethyl acetate and water are successively added, and the organic phase is separated and washed with water.Drying, recovering ethyl acetate under reduced pressure, and the residue was crystallized from ethanol.15.1 g of 2-[4-(2-hydroxyethoxy)-3, 5-dimethylphenyl]-5, 7-dimethoxyquinazolin-4(3H)-one, The yield is 81.5percent.The purity is 99.7percent (area normalization method).A solution of 2-amino-4, 6-dimethoxybenzamide (0.60 g, 3.06 mmol) and 4-[2-(tert-butyidimethylsilanoxy)ethoxy]-3, 5-dimethylbenzaldehyde (0.856 g, 2.78 mmol) in N, N-dimethyl formamide (20 mL) was stirred at 70° C. for 1 h. Iodine (0.846 g, 3.33 mmol) and potassium carbonate (0.384 g, 2.78 mmol) were added and the reaction mixture was stirred at 70° C. for 16 h. The reaction mixture was poured into ice, and extracted with ethyl acetate. The organic layer was washed with water, brine, and dried over anhydrous Na2-(4-(2-hydroxyethoxy)-3, 5-dimethylphenyl)-5, 7-dimethoxyquinazolin-4(3H)-one [0476] A solution of 2-amino-4, 6-dimethoxybenzamide (0.60 g, 3.06 mmol) and 4-[2-(tert-butyldimethylsilanoxy)ethoxy]-3, 5-dimethylbenzaldehyde (0.856 g, 2.78 mmol) in N, N-dimethyl formamide (20 mL) was stirred at 70° C. for 1 h. Iodine (0.846 g, 3.33 mmol) and potassium carbonate (0.384 g, 2.78 mmol) were added and the reaction mixture was stirred at 70° C. for 16 h. The reaction mixture was poured into ice, and extracted with ethyl acetate. The organic layer was washed with water, brine, and dried over anhydrous NaAt 70deg.] C 2-Amino-4, 6-dimethoxy-benzamide(0.60g, 3.06mmol) and 4- [2- (tert-butyldimethylsilyloxy)ethoxy]-3, 5-dimethyl-benzaldehyde(0.856g, 2.78mmol)in N, N- dimethylformamide(20 mL) was stirred for 1hour. Was added iodine (0.846g, 3.33mmol)and potassium carbonate (0.384g, 2.78mmol), thereaction mixture was at 70 stirred for 16 hours. The reactionmixture was poured onto ice, extracted with ethyl acetate. With water, theorganic layer was washed with brine, dried over anhydrous Na 2SO 4dry. The solvent was removed to give acrude product, which was purified by column chromatography to give a white solid of 2- (4- (2-hydroxyethoxy) -3, 5-dimethylphenyl) -5, 7-methoxy-quinazolin -4 (3H) - one (444mg, 39percent). Selected data: 229-231 . Alternatively, 2- (4-(2-hydroxyethoxy)-3, 5-dimethyl-phenyl)-5, 7-dimethoxy-quinazolin -4 (3H) - onecan be synthesized by the following method. In ethanol (350 mL of) of 3, 5-dimethyl-4-hydroxy-benzaldehyde(26.9g, 0.179mol) disposed dried 2L round bottomed flask with refluxcondenser and magnetic stirrer. 2-chloro-ethanol(87.6g, 1.074mol) and K 2CO 3(99g, 0.716mol), the reaction mixture was heated at reflux for 24 hours. The reaction mixture was cooled toroom temperature and filtered. The solvent was removed under reduced pressure.The crude product was diluted with ethyl acetate, and the organic layer waswashed with water, brine, Na 2SO 4dry. 45g crude product obtained after removalof the solvent. The crude product was purified by column chromatography (silicagel 230-400 mesh; with 50percent hexanes in ethyl acetate as eluent) to afford 33.3g (95percent) of product. 2-amino-4, 6-dimethoxy - benzamide (33.45g, 0.170mol)and 4- (2-hydroxyethoxy) -3, 5-dimethyl benzaldehyde(33.3g, 0.170 mol) was added NaHSO3(33.3g, 0.187mol) in N, N- dimethylacetamide (300mL) solution of and p-TSA(3.2g, 17.1mmol), at 150 reaction mixturewas heated for 14 hours.The reaction was cooled to room temperature. The solvent was removed underreduced pressure. The residue was diluted with water, followed by stirring at room temperature for30 minutes. The solid was isolated by filtration and dried to give the crudeproduct. The crude product was purified by column chromatography (silica gel230-400 mesh; used in CH 2Cl 25percent methanol as eluent) to afford 2- (4- (2-hydroxyethoxy) -3, 5-dimethyl-phenyl)-5, 7-dimethoxy-quinazoline -4 (3H) - one (33g, 52percent).