* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.
Stage #1: With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride In N,N-dimethyl-formamide at -20 - 25℃; Inert atmosphere; Large scale Stage #2: With N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at -30 - 25℃; Large scale
In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20 mi DMF (30.20 kg) was charged into the 50 L flask reactor. Then the stirrer was started. Maintaining the temperature at 15-25°C, ASYM-112394 (3.22 kg, 2.76 kg after corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. The mixture was cooled to -10 to -20°C and 1-hydroxybenzotriazole hydrate (2.10 kg) was added into the mixture at -10 to -20°C. Then EDCI (2.41 kg) was added into the mixture in five portions at an interval of about 5-10 mm. The mixture was cooled to -20 to -30°C and ASYM-111888 (Asymchem) (1.96 kg) was added into the mixture at -20 to -30°C. Then DIEA (1.77 kg) was added into the mixture at the rate of 3-4 kg/h. The mixture was heated to 15-25°C at the rate of 5-10°C/h. The mixture was reacted at 15-25°C. After 6-8 h,the mixture was sampled and analyzed by HPLC every 2-4 h until the content of ASYM-112394 was 2percent. The mixture was cooled to 0-10°C and the reaction mixture was quenched with a solution which was prepared from ethyl acetate (28.80 kg) in purified water (12.80 kg) at 0-10°C. The mixture was extracted three times with ethyl acetate (28.80 kg). For each extraction the mixture was stirred for 20-30 mm and settled for 20-30 mm before separation. The organic phases were combined and washed twice with purified water (12.80 kg). The mixture was stirred for 20-30 mm and settled for 20-30 mm before separation for each time. Then the obtained organic phase was filtered through an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor. The mixture was washed twice with a 5percent acetic acid solution, which was prepared from acetic acid (2.24 kg) in purified water (42.50 kg). The solution was added at the rate of 10-20 kg/h. The organic phase was washed twice with a sodium carbonate solution, which was prepared from sodium carbonate (9.41 kg) in purified water (48.00 kg). The organic phase was washed twice with a sodium chloride solution, which was prepared from sodium chloride (16.00 kg) in purified water (44.80 kg). The organic phase was transferred into a 300 L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added into the mixture and the mixture was stirred for 2-4 h at 15-30°C. The mixture was filtered with a nutsche filter, which was pre-loaded with about 1 cm thick silica gel (7.50 kg). The filter cake was soaked and washed with ethyl acetate (14.40 kg) before filtration. The filtrates were combined and the combined filtrate was added into a 72 L flask through an in-line fluid filter. The mixture was concentrated at T40°C under reduced pressure (P-0.08 MPa) until 3-4 L remained. Then MTBE (4.78 kg) was added into the mixture. The mixture was cooled to 0-10°C for crystallization with stirring. After 1 h, the mixture was sampled and analyzed by wtpercentevery 1-2 h until the wtpercent of the mother liquor was 5percent or the change of wtpercent between consecutive samples was 1percent. The mixture was filtered with a vacuum filter flask and the filter cake was dried in the tray drier under nitrogen at 30-40°C until KF0.5percent. 3.55 kg of product was recovered as an off-white solid at 100percent purity.2e(°)9.1 ±0.210.0 ± 0.210.2 ± 0.211.4± 0.212.5 ± 0.212.7 ± 0.213.3 ± 0.215.2 ± 0.215.4 ± 0.216.0 ± 0.217.1 ±0.217.6 ± 0.218.2 ± 0.218.8 ± 0.219.2 ± 0.219.5 ± 0.2d space (A)9.690 ± 0.2128.869 ± 0.1788.664 ± 0.1697.742 ± 0.1357.066±0.1126.956 ± 0.1096.637 ± 0.0995.833 ± 0.0765.769 ± 0.0755.531 ± 0.0695.173 ± 0.0605.038 ± 0.0574.876 ± 0.0534.723 ± 0.0504.624 ± 0.0484.556 ± 0.046Intensity (percent)12275258215469384212100[0234] The resulting 4-(5-Chloro-2-isopropylam inopyridin-4-yl)-1 H-pyrrole-2- carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was analyzed by XRPD (FIG. 1). Peaks shown in FIG. 1 are listed in Table 2, prominent peaks are listed in Table 3.Table 2: XRPD peaks observed for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxyl ic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide free base.20.3 ± 0.220.5 ± 0.221.4± 0.221.7 ± 0.221.9± 0.223.1 ±0.223.3 ± 0.223.6 ± 0.224.3 ± 0.225.2 ± 0.225.6 ± 0.226.6 ± 0.227.0 ± 0.227.7 ± 0.227.9 ± 0.228.2 ± 0.228.7 ± 0.228.9 ± 0.229.2 ± 0.230.2 ± 0.230.6 ± 0.24.381 ± 0.0434.327 ± 0.0424.145 ± 0.0384.102 ± 0.0374.057 ± 0.0373.847 ± 0.0333.812 ± 0.0323.774 ± 0.0323.653 ± 0.0303.530 ± 0.0283.476 ± 0.0273.355 ± 0.0253.297 ± 0.0243.214 ± 0.0233.191 ± 0.0223.159 ± 0.0223.106 ± 0.0213.083 ± 0.0213.057 ± 0.0202.957 ± 0.0192.923 ± 0.019141244111213252611923713103949149Table 3: Prominent XRPD peaks for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxyl ic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide free base.2e(°)9.1 ±0.212.5 ± 0.215.2 ± 0.215.4 ± 0.2d space (A)9.690 ± 0.2127.066±0.1125.833 ± 0.0765.769 ± 0.075Intensity (percent)1225154619.2 ± 0.219.5 ± 0.220.3 ± 0.220.5 ± 0.221.4± 0.221.7 ± 0.221.9± 0.223.1 ±0.223.3 ± 0.223.6 ± 0.224.3 ± 0.227.7 ± 0.227.9 ± 0.230.2 ± 0.24.624 ± 0.0484.556 ± 0.0464.381 ± 0.0434.327 ± 0.0424.145 ± 0.0384.102 ± 0.0374.057 ± 0.0373.847 ± 0.0333.812 ± 0.0323.774 ± 0.0323.653 ± 0.0303.214 ± 0.0233.191 ± 0.0222.957 ± 0.01912100141244111213252611131014[0235]FT-IR was performed onisopropylaminopyridin-4-yI)-1 H-pyrrole-2-carboxylica sample of 4-(5-Chloro-2-acid [1 -(3-chlorophenyl)-2-Position (cm1)681712748783807827857878Intensity0.01 740.00250.00140.00580.0010.00820.00450.00069hydroxyethyl]amide free base as described in Example 1 (FIG. 2). Observed peaks from FIG. 2 are listed in Table 4.Table 4: Observed FT-IR peaks for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxyl ic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide free base.Position (cm1) Intensity897 0.00067916 0.00056932 0.0008996 0.00041040 0.000741080 0.00691101 0.000811126 0.000961145 0.00141170 0.00271197 0.00111208 0.00281235 0.00131255 0.00151268 0.00211294 0.00131350 0.00181364 0.0021385 0.000771398 0.000771439 0.00171451 0.00141466 0.00191487 0.00891504 0.00331523 0.00651533 0.00631568 0.00211603 0.01081629 0.00622927 0.000242974 0.000283235 0.000523405 0.00026Position (cm1) Intensity; DSC was performed on a sample of 4-(5-Chloro-2- isopropylam inopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2- hydroxyethyl]amide free base as described in Example 1 (FIG. 3) and showed an endotherm having an onset temperature of approximately 184°C.
78.81%
With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In N,N-dimethyl-formamide at -30 - 25℃; Inert atmosphere; Large scale
[0151] In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20 mm. DMF (30.20 kg) was charged into the 50 L flask reactor. Then the stirrer was started. Maintaining the temperature at 15-25°C, ASYM-112394 (3.22 kg, 2.76 kgafter corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. The mixture was cooled to -10 to -20°C and 1-hydroxybenzotriazole hydrate (2.10 kg) was added into the mixture at -10 to -20°C. Then EDCI (2.41 kg) was added into the mixture in five portions at an interval of about 5-10 mm. The mixture was cooled to -20 to -30°C and ASYM-1 11888 (Asymchem) (1.96 kg) was added into the mixture at -20 to -30°C. Then DIEA (1 .77 kg) was added into the mixture at the rate of 3-4 kg/h. The mixture was heated to 15-25°C at the rate of 5-10°C/h. The mixture was reacted at 15-25°C. After 6-8 h, the mixture was sampled and analyzed by HPLC every 2-4 h until the content of ASYM-112394 was 2percent. The mixture was cooled to 0-10°C and the reaction mixture was quenched with a solution which was prepared from ethyl acetate (28.80 kg) in purified water (12.80 kg) at 0-10°C. The mixture was extracted three times with ethyl acetate (28.80 kg). For each extraction the mixture was stirred for 20-30 mm and settled for 20-30 mm before separation. The organic phases were combined and washed twice with purified water (12.80 kg). The mixture was stirred for 20-30 mm and settled for 20-30 mm before separation for each time. Then the obtained organic phase was filtered through an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor. The mixture was washed twice with a 5percent acetic acid solution, which was prepared from acetic acid (2.24 kg) in purified water (42.50 kg). The solution was added at the rate of 10-20 kg/h. The organic phase was washed twice with a sodium carbonate solution, which was prepared from sodium carbonate (9.41 kg) in purified water (48.00 kg). The organic phase was washed twice with a sodium chloride solution, which was prepared from sodium chloride (16.00 kg) in purified water (44.80 kg). The organic phase was transferred into a 300 L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added into the mixture and the mixture was stirred for 2-4 h at 15-30°C. The mixture was filtered with a nutsche filter, which was pre-loaded with about 1 cm thick silica gel (7.50 kg). The filter cake was soaked and washed with ethyl acetate (14.40 kg) before filtration. The filtrates were combined and the combined filtrate was added into a 72 L flask through an in-line fluid filter. The mixture was concentrated at T40°C under reduced pressure (P-0.08 MPa) until 3-4 L remained. Then MTBE (4.78 kg) was added into the mixture. The mixture was cooled to 0-10°C for crystallization with stirring. After 1 h, the mixture was sampled and analyzed by wtpercent every 1-2 h until the wtpercent of the mother liquor was 5percent or the change of wtpercent between consecutive samples was 1 percent. The mixture was filtered with a vacuum filter flask and the filter cake was dried in the tray drier under nitrogen at 30-40°C until KF0.5percent. 3.55 kg of product was recovered as an off-white solid at 100percent purity.
64%
With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine In DMF (N,N-dimethyl-formamide) at 20℃;
4-(5-Chloro- 2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid [1-(3- chlorophenyl) -2-hydroxyethyl] amide: To a suspension of 4-(5-chloro-2- isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid (1.93 g, 6.9 mmol, 1.0 equivalent) in DMF (5.0 mL) was added EDCI (1.45 g, 7.6 mmol, 1.1 equivalents), HOBt (0.94 g, 6.9 mmol, 1.0 equivalent) and (S)-3-chlorophenylglycynol (1.58 g, 7.6 mmol, 1.1 equivalents). Diisopropylethylamine (2.7 mL) was then added and the resulting mixture was stirred a room temperature overnight. The mixture was then poured into water and extracted with ethyl acetate. After drying over sodium sulfate, the solvent was removed and the crude was adsorbed on silica gel. Purification was effected by flash chromatography on silica, eluting with mixtures of hexanes/acetone (from 80: 20 to 60: 40) to afford the title compound as white solid (1.9 g, 64percent). Rt(min) 4.981s. FIA MS: 433.1 ES+; 431.2 ES-. (at)HNMR (CD30D) 8 1.31 (d, 6H), 3.85 (in, 3H), 5.15 (t, 1H), 7.01 (s, 1H), 7.25 (m, 3H), 7.4 (s, 1H), 7.45 (s, 1H), 7.7 (s, 1H), 7.95 (s, 1H).
With 1H-tetrazole; In dichloromethane; acetonitrile; at 20℃; for 16.0h;
di-tert-Butyl 4-(5-chloro-2-(isopropylamino )pyridine-4-yl)-N-( (8)-1-(3- chlorophenyl)-2-hydroxyetliyl)-1H pyrrole-2-carboxamide phosphate: Compound 1-9 (1 g, 2.3 mmol, 1.0 equivalent) and tetrazole (241 mg, 3.45 mmol, 1.5 equivalents) were dissolved in dichloromethane (5 mL) and acetonitrile (5 mL) under nitrogen at room temperature. Di-tert-butyl diisopropyl phosphoamidite (1.1 mL, 3.45 mmol, 1.5 equivalents) was added dropwise and the resulting mixture was stirred for 16 hours. The reaction mixture was then cooled on an ice bath, treated with a solution of 6 M tert-butylhydroxyperoxide (3 mL) and stirred for 20 minutes. The clear solution was diluted in dichloromethane and small amount of methanol, washed with Na2S203, water and dried over sodium sulfate. The crude oil was adsorbed on silica gel and was first purified by flash chromatography eluting with mixtures of hexane/acetone (from 90:10 to 60: 40) and then by reversed phase HPLC (acetonitrile/water/1 % TFA), yielding the di-t-butyl ether intermediate as a white solid (336 mg). HPLC Rt: 6.53 minutes, MS FIA: 625.0 ES+; 623.1ES-.
With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine; In DMF (N,N-dimethyl-formamide); at 20℃; for 16.0h;Product distribution / selectivity;
4-(5-chloro-2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid [1-(3- chlorophenyl)-2-hydroxyethyl] amide (1-9): 4-Bromo-2-chloro-5-N- isopropylpyridin-2-amine N-oxide (25 mg, 0.094 mmol, 1.0 equivalent) and 4- (4,4,5,5-tetraniethyl-[1 ,3,2]dioxaborolan-2-yl)-1 -(2,4,6-trimethylbenzensulfonyl)-lH- pyrrole-2-carboxylic acid methyl ester (39 mg, 0.094 mmol, 1.0 equivalent) were dissolved in benzene (5 mL) then aqueous 2M Na2C03 (1 mL) and Pd (PPh3)4 mg, 0.1 mmol, 0.2 equivalent) were added and the resulting suspension was heated at reflux at 80 C for 16 hours. The reaction mixture was diluted in ethyl acetate, washed with water and dried over anhydrous sodium sulfate to afford 4-(5-chloro-2- isopropylamino-pyridin-4-yl)-1-(2,4,6-trimethyl-benzenesulfonyl)-1H pyrrole-2- carboxylic acid methyl ester N-oxide (Rt(min) 6.859. MS FIA: 492.0 ES+) which was then treated with a 2 M solution of PC13 in dichloromethane (1 mL) at room temperature. After 10 minutes, the solvent was removed under a stream of nitrogen and the crude oil was dissolved in methanol (1 mL) and aqueous 1 M NaOH (1 mL). The resulting mixture was heated at reflux for 16 hours then the crude solution was acidified using aqueous 1 M HCI and the solvent was removed. The resulting 4-(5- chloro-2-isopropylainino-pyridin-4-yl)-lH-pyrrole-2-carboxylic acid (Rt(min) 3.527. MS FIA: 279.4 ES+; 278.2 Es-) was suspended in DMF (3 mL) together with EDCI (36 mg, 0.19 mmol, 2 equivalents), HOBt (26 mg, 0.19 mmol, 2 equivalents), (S)-3- chlorophenylglycinol HCI salt (59 mg, 0.28 mmol, 3 equivalents) and DIEA (0.12 mL, 0.75 mmol, 8 equivalents). The resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted in ethyl acetate, washed with water and dried over sodium sulfate. After removing the solvent under reduced pressure, the crude product was purified by reversed phase HPLC (acetonitrile/water/TFA 1 %) to afford the title compound as a white solid (4.8 mg, 8.1%).
In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20 mi DMF (30.20 kg) was charged into the 50 L flask reactor. Then the stirrer was started. Maintaining the temperature at 15-25C, ASYM-112394 (3.22 kg, 2.76 kg after corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. The mixture was cooled to -10 to -20C and 1-hydroxybenzotriazole hydrate (2.10 kg) was added into the mixture at -10 to -20C. Then EDCI (2.41 kg) was added into the mixture in five portions at an interval of about 5-10 mm. The mixture was cooled to -20 to -30C and ASYM-111888 (Asymchem) (1.96 kg) was added into the mixture at -20 to -30C. Then DIEA (1.77 kg) was added into the mixture at the rate of 3-4 kg/h. The mixture was heated to 15-25C at the rate of 5-10C/h. The mixture was reacted at 15-25C. After 6-8 h,the mixture was sampled and analyzed by HPLC every 2-4 h until the content of ASYM-112394 was 2%. The mixture was cooled to 0-10C and the reaction mixture was quenched with a solution which was prepared from ethyl acetate (28.80 kg) in purified water (12.80 kg) at 0-10C. The mixture was extracted three times with ethyl acetate (28.80 kg). For each extraction the mixture was stirred for 20-30 mm and settled for 20-30 mm before separation. The organic phases were combined and washed twice with purified water (12.80 kg). The mixture was stirred for 20-30 mm and settled for 20-30 mm before separation for each time. Then the obtained organic phase was filtered through an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor. The mixture was washed twice with a 5% acetic acid solution, which was prepared from acetic acid (2.24 kg) in purified water (42.50 kg). The solution was added at the rate of 10-20 kg/h. The organic phase was washed twice with a sodium carbonate solution, which was prepared from sodium carbonate (9.41 kg) in purified water (48.00 kg). The organic phase was washed twice with a sodium chloride solution, which was prepared from sodium chloride (16.00 kg) in purified water (44.80 kg). The organic phase was transferred into a 300 L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added into the mixture and the mixture was stirred for 2-4 h at 15-30C. The mixture was filtered with a nutsche filter, which was pre-loaded with about 1 cm thick silica gel (7.50 kg). The filter cake was soaked and washed with ethyl acetate (14.40 kg) before filtration. The filtrates were combined and the combined filtrate was added into a 72 L flask through an in-line fluid filter. The mixture was concentrated at T40C under reduced pressure (P-0.08 MPa) until 3-4 L remained. Then MTBE (4.78 kg) was added into the mixture. The mixture was cooled to 0-10C for crystallization with stirring. After 1 h, the mixture was sampled and analyzed by wt%every 1-2 h until the wt% of the mother liquor was 5% or the change of wt% between consecutive samples was 1%. The mixture was filtered with a vacuum filter flask and the filter cake was dried in the tray drier under nitrogen at 30-40C until KF0.5%. 3.55 kg of product was recovered as an off-white solid at 100% purity.2e()9.1 ±0.210.0 ± 0.210.2 ± 0.211.4± 0.212.5 ± 0.212.7 ± 0.213.3 ± 0.215.2 ± 0.215.4 ± 0.216.0 ± 0.217.1 ±0.217.6 ± 0.218.2 ± 0.218.8 ± 0.219.2 ± 0.219.5 ± 0.2d space (A)9.690 ± 0.2128.869 ± 0.1788.664 ± 0.1697.742 ± 0.1357.066±0.1126.956 ± 0.1096.637 ± 0.0995.833 ± 0.0765.769 ± 0.0755.531 ± 0.0695.173 ± 0.0605.038 ± 0.0574.876 ± 0.0534.723 ± 0.0504.624 ± 0.0484.556 ± 0.046Intensity (%)12275258215469384212100[0234] The resulting 4-(5-Chloro-2-isopropylam inopyridin-4-yl)-1 H-pyrrole-2- carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was analyzed by XRPD (FIG. 1). Peaks shown in FIG. 1 are listed in Table 2, prominent peaks are listed in Table 3.Table 2: XRPD peaks observed for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxyl ic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide free base.20.3 ± 0.220.5 ± 0.221.4± 0.221.7 ± 0.221.9± 0.223.1 ±0.223.3 ± 0.223.6 ± 0.224.3 ± 0.225.2 ± 0.225.6 ± 0.226.6 ± 0.227.0 ± 0.227.7 ± 0.227.9 ± 0.228.2 ± 0.228.7 ± 0.228.9 ± 0.229.2 ± 0.230.2 ± 0.230.6 ± 0.24.381 ± 0.0434.327 ± 0.0424.145 ± 0.0384.102 ± 0.0374.057 ± 0.0373.847 ± 0.0333.812 ± 0.0323.774 ± 0.0323.653 ± 0.0303.530 ± 0.0283.476 ± 0.0273.355 ± 0.0253.297 ± 0.0243.214 ± 0.0233.191 ± 0.0223.159 ± 0.0223.106 ± 0.0213.083 ± 0.0213.057 ± 0.0202.957 ± 0.0192.923 ± 0.019141244111213252611923713103949149Table 3: Prominent XRPD peaks for 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxyl ic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide free base.2e()9.1 ±0.212.5 ± 0.215.2 ± 0.215.4 ± 0.2d space (A)9.690 ± 0.2127.066±0.1125.833 ± 0.0765.769 ± 0.075Intensity (%)1225154619.2 ± 0.219.5 ± 0.220.3 ± 0.220.5 ± 0.221.4± 0.221.7 ± 0.221.9± 0.223.1 ±0.223.3 ± 0.223.6 ± 0.224.3 ± 0.227.7 ± 0.227.9 ± 0.230.2 ± 0.24.624 ± 0.0484.556 ± 0.0464.381 ± 0.0434.327 ± 0.0424.145 ...
78.81%
With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine; In N,N-dimethyl-formamide; at -30 - 25℃;Inert atmosphere; Large scale;
[0151] In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20 mm. DMF (30.20 kg) was charged into the 50 L flask reactor. Then the stirrer was started. Maintaining the temperature at 15-25C, ASYM-112394 (3.22 kg, 2.76 kgafter corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. The mixture was cooled to -10 to -20C and 1-hydroxybenzotriazole hydrate (2.10 kg) was added into the mixture at -10 to -20C. Then EDCI (2.41 kg) was added into the mixture in five portions at an interval of about 5-10 mm. The mixture was cooled to -20 to -30C and ASYM-1 11888 (Asymchem) (1.96 kg) was added into the mixture at -20 to -30C. Then DIEA (1 .77 kg) was added into the mixture at the rate of 3-4 kg/h. The mixture was heated to 15-25C at the rate of 5-10C/h. The mixture was reacted at 15-25C. After 6-8 h, the mixture was sampled and analyzed by HPLC every 2-4 h until the content of ASYM-112394 was 2%. The mixture was cooled to 0-10C and the reaction mixture was quenched with a solution which was prepared from ethyl acetate (28.80 kg) in purified water (12.80 kg) at 0-10C. The mixture was extracted three times with ethyl acetate (28.80 kg). For each extraction the mixture was stirred for 20-30 mm and settled for 20-30 mm before separation. The organic phases were combined and washed twice with purified water (12.80 kg). The mixture was stirred for 20-30 mm and settled for 20-30 mm before separation for each time. Then the obtained organic phase was filtered through an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor. The mixture was washed twice with a 5% acetic acid solution, which was prepared from acetic acid (2.24 kg) in purified water (42.50 kg). The solution was added at the rate of 10-20 kg/h. The organic phase was washed twice with a sodium carbonate solution, which was prepared from sodium carbonate (9.41 kg) in purified water (48.00 kg). The organic phase was washed twice with a sodium chloride solution, which was prepared from sodium chloride (16.00 kg) in purified water (44.80 kg). The organic phase was transferred into a 300 L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added into the mixture and the mixture was stirred for 2-4 h at 15-30C. The mixture was filtered with a nutsche filter, which was pre-loaded with about 1 cm thick silica gel (7.50 kg). The filter cake was soaked and washed with ethyl acetate (14.40 kg) before filtration. The filtrates were combined and the combined filtrate was added into a 72 L flask through an in-line fluid filter. The mixture was concentrated at T40C under reduced pressure (P-0.08 MPa) until 3-4 L remained. Then MTBE (4.78 kg) was added into the mixture. The mixture was cooled to 0-10C for crystallization with stirring. After 1 h, the mixture was sampled and analyzed by wt% every 1-2 h until the wt% of the mother liquor was 5% or the change of wt% between consecutive samples was 1 %. The mixture was filtered with a vacuum filter flask and the filter cake was dried in the tray drier under nitrogen at 30-40C until KF0.5%. 3.55 kg of product was recovered as an off-white solid at 100% purity.
64%
With benzotriazol-1-ol; 1-ethyl-(3-(3-dimethylamino)propyl)-carbodiimide hydrochloride; N-ethyl-N,N-diisopropylamine; In DMF (N,N-dimethyl-formamide); at 20℃;Product distribution / selectivity;
4-(5-Chloro- 2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid [1-(3- chlorophenyl) -2-hydroxyethyl] amide: To a suspension of 4-(5-chloro-2- isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid (1.93 g, 6.9 mmol, 1.0 equivalent) in DMF (5.0 mL) was added EDCI (1.45 g, 7.6 mmol, 1.1 equivalents), HOBt (0.94 g, 6.9 mmol, 1.0 equivalent) and (S)-3-chlorophenylglycynol (1.58 g, 7.6 mmol, 1.1 equivalents). Diisopropylethylamine (2.7 mL) was then added and the resulting mixture was stirred a room temperature overnight. The mixture was then poured into water and extracted with ethyl acetate. After drying over sodium sulfate, the solvent was removed and the crude was adsorbed on silica gel. Purification was effected by flash chromatography on silica, eluting with mixtures of hexanes/acetone (from 80: 20 to 60: 40) to afford the title compound as white solid (1.9 g, 64%). Rt(min) 4.981s. FIA MS: 433.1 ES+; 431.2 ES-. (at)HNMR (CD30D) 8 1.31 (d, 6H), 3.85 (in, 3H), 5.15 (t, 1H), 7.01 (s, 1H), 7.25 (m, 3H), 7.4 (s, 1H), 7.45 (s, 1H), 7.7 (s, 1H), 7.95 (s, 1H).
With benzotriazol-1-yloxyl-tris-(pyrrolidino)-phosphonium hexafluorophosphate; N-ethyl-N,N-diisopropylamine; In dichloromethane; at 20℃; for 48.0h;
(25)-(S)-2-(4-(5-chloro-2-(isopropylamino)pyridine-4-yl)-lH-pyrrole-2- carboxamido)-2-(3-chlorophenyl)ethyl-2-amino-2-methylbutanoate (11-1): To a solution of compound 1-9 (1 g, 2.3 mmol, 1.0 equivalent) in dichloromethane (50 mL) were added DIEA (1.1 mL, 6.9 mmol, 3.0 equivalent) and N-BOC-L-Valine (1.2 g, 5.52 mmol, 2.4 equivalents). PyBOP (2.9 g, 5.75 mmol, 2.5 equivalents. ) was then added slowly and the resulting mixture was stirred at room temperature for 48 hours. The mixture was then washed with water and dried over sodium sulfate. The crude solid was adsorbed on silica and then purified by flash chromatography eluting with mixtures of hexane/ethyl acetate (from 90:10 to 50: 50), yielding the Boc-protected compound as a white solid (786 mg). This intermediate (761 mg, 1.2 mmol) was dissolved in dioxane (1 mL) and treated with a solution of 4 M HCL in dioxane. The resulting mixture was stirred for 16 hours at room temperature. The solvent was then removed and the 2xHCl salt of the title compound was obtained as a white solid (571 mg). HPLC Rt: 4.56 minutes. FIA MS: 531.9 ES+; 529.8 ES-. LC/MS: Rt: 2.07 minutes; 532.0 ES+; 530.1 ES-. 'HNMR (CD30D) No. 0.9 (dd, 6H), 1.35 (d, 6H), 2.2 (m, 1 H), 3.9 (m, 2H), 4.7 (m, 2H), 5 .6 (m, 1H), 7.1 (s, 1H), 7.3 (d, 1H), 7.35 (t, 1H), 7.4 (d, 1 H), 7.5 (s, 1 H), 7.6 (s, 1 H), 7.75 (s, 1 H), 7.95 (s, 1 H).
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide hydrochloride[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
88.65%
With hydrogenchloride; In methanol; ethanol; isopropyl alcohol; at 70 - 75℃; for 2.0h;
4-(5-Chloro-2-isopropylam inopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide Form C was prepared from 4-(5-Chloro-2- isopropylam inopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2- hydroxyethyl]amide free base as follows. ASYM-1 11935 (10.4 kg) was added to a stirred mixture of anhydrous ethanol (73.9 kg), methanol (4.1 kg) and isopropanol (4.1 kg). The mixture was heated to 70-75C and stirred until all the solids dissolved. Anhydrous HCI (37 wt%, 1.leq) in a mixture of ethanol/methanol/isopropanol (90:5:5) was added and the mixture maintained at 70-75C for 2 hours after the addition was completed. The mixture was then cooled to 15-25C at a rate of 5- 15C per hour and stirred at this temperature until the desired polymorphic purity was reached. The end point of the crystallization/polymorph conversion was determined by the absence of an XRPD peak at about 10.5 2 in three successive samples.The mixture was then filtered and washed successively with a preprepared solution of anhydrous ethanol (14.8 kg), methanol (0.8 kg) and isopropanol (0.8 kg), followed by MTBE (2 x 21 kg). Avoidance of delay in the washing of the filter cake is preferable because the polymorph may be unstable in the wet filter cake in the presence of reagent alcohol and improved stability was observed after the MTBE wash has been performed. The wet filter cake was then dried in a heated filter funnel or a tray drier at 40-50C until dry. Typical yields were about 85-90%.
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylicacid[1-(3-chlorophenyl)-2-hydroxyethyl]amide hydrochloride[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
90%
With hydrogenchloride; In methanol; ethanol; isopropyl alcohol; at 70 - 75℃; for 2.0h;Large scale;
[0155] 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-ch lorophenyl)-2-hydroxyethyl]am ide Form C was prepared from 4-(5-Chloro-2- isopropylam inopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2- hydroxyethyl]amide free base as follows. ASYM-1 11935 (10.4 kg) was added to a stirred mixture of anhydrous ethanol (73.9 kg), methanol (4.1 kg) and isopropanol (4.1 kg). The mixture was heated to 70-75C and stirred until all the solids dissolved. Anhydrous HCI (37 wt%, 1.leq) in a mixture of ethanol/methanol/isopropanol (90:5:5) was added and the mixture maintained at 70-75C for 2 hours after the addition was completed. The mixture was then cooled to 15-25C at a rate of 5- 15C per hour and stirred at this temperature until the desired polymorphic purity was reached. The end point of the crystallization/polymorph conversion was determined by the absence of an XRPD peak at about 10.5 20 in three successive samples.[0156] The mixture was then filtered and washed successively with a preprepared solution of anhydrous ethanol (14.8 kg), methanol (0.8 kg) and isopropanol (0.8 kg), followed by MTBE (2 x 21 kg). Avoidance of delay in the washing of the filter cake is preferable because the polymorph may be unstable in the wet filter cake in the presence of reagent alcohol and improved stability was observed after the MTBE wash has been performed. The wet filter cake was then dried in a heated filter funnel or a tray drier at 40-50C until dry. Typical yields were about 85-90%.
4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylicacid[1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
at 20℃; for 24.0h;
[0168] 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide free base was reacted at 100 mg scale in an ethanol slurry with malonic acid (1:1 molar ratio). After 1 day at room temperature the product was purified by vacuum filtration and evaporation of the filtrate. The resulting 4-(5-Ch loro-2-isopropylam inopyridin-4-yl )-1 H-pyrrole-2- carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A formed crystalline needles.[0169] In a scale-up experiment 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 Hpyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide free base (Ig) was suspended in ethanol at ambient temperature with stirring and about 1.1 equivalents of malonic acid (268 mg) were added. The mixture was then stirred at ambient temperature for approximately four days, after which solids were collected by vacuum filtration. Out of solids isolated approximately 500 mg were vacuum-dried at 45C overnight and analyzed by XRPD. The remaining solids were analyzed without drying. Both samples were consistent with 4-(5-Chloro-2- isopropylam inopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2- hydroxyethyl]amide malonate Form A.
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carboxamido)cyclopropane carboxylate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(3-(5-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-hydroxy-4H-1,2,4-triazol-4-yl)-1H-indol-1-yl)propanamido)cyclopropanecarboxylate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-hydroxy-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carboxamido)cyclopropanecarboxylate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carbonyl)piperidine-4-carboxylate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-hydroxy-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carbonyl)piperidine-4-carboxylate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 2-(1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carbonyl)piperidin-4-yl)acetate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 2-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carboxamido)-2-methylpropanoate[ No CAS ]
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carbonyl)azetidine-3-carboxylate[ No CAS ]
(S)-(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-(1-(4-(3-(2,4-dihydroxy-5-isopropylphenyl)-5-(ethylcarbamoyl)-4H-1,2,4-triazol-4-yl)-2-fluorobenzyl)piperidine-4-carbonyl)pyrrolidine-3-carboxylate[ No CAS ]
(S)-tert-butyl 4-(2-(2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethoxy)-2-oxoethyl)piperidine-1-carboxylate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
96%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a mixture of (S)-4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-N- (i-(3- chl orophenyi)-2-hydroxyethyl)- 1 H-pyrroie-2-carboxarnide (65 mg, 0. 15 mm ol) and 2-(1 -(tertbutoxycarbonyl)piperidin-4-vl)acetic acid (40 mg, 0.165 rnmol) in dry CH2CI2 (4 mL) was added EDC (32 mg, 0.165 mmol) and DMAP (20 mg, 0.165 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous NH4CI solution. The organic layer was dried over Na2SO4 and concentrated in vacuum. The crude residue was purified by ISCO using DCMMeOH (0-10%) as eluent to afford 95 mg (96%) of product. ESMS calculated (C33H41C12N505): 658.62; found: 659.7 M+H).
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 1-((tertbutoxycarbonyl)amino)cyclopropane carboxylate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
96.9%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a suspension of(S)-4-{5--chioro--2--(isopropy1amino)pyridin-4vi)--N-(i (3 chlorophenyl)-2--hydroxyethyl) iH.-pyrrole2carboxamide (250 mg, 0.576 mmol) and 1- ((tertbutoxycarbonyi)amino)cyclopropanecarboxylic acid (145 rng, 0.634 mmol) in dry CH2CI2 (15 mL) was added EDC (121 mg, 0.634 mmol) and DMAP (77 mg, 0.634 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous NH4CI solution. The organic layer was dried over Na2SO4 and concentrated in vacuum. The crude residue was purified by ISCO using DCMJMeOH (0-40%) as eluent to afford 343 mg (966%) of product. ESMS calculated for C30H35C12N5O5: 616.54; found: 617 (M±H).
(S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl 2-((tert-butoxycarbonyl)amino)-2-methylpropanoate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
96%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a mixture of (i(3- chioropheny 1)24iydroxyethyi)- 1H-pyrroie--2-carboxami de (44 ing, 0,1 mmoi) and 2((tert.- butoxycarbonyl)amino).-2.methylpropanoic acid (23 mg, 0.11 mmol) in dry CH2C12 (4.5 mL) was added EDC (21 mg, 0.11 mmol) and DMAP (14 mg, 0.11 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous NH4CI solution. The organic layer was dried over Na2SO4 and concentrated in vacuum. The crude residue was purified by ISCO using DCM/MeOEI (0-J0%) as eluent to afford 59 mg (96%) of product. ESMS calculated (C30H37C12N505): 618.55; found: 619 (M+H).
(S)-1-tert-butyl 3-(2-(4-(5-chloro-2-(isopropylamino) pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl)azetidine-1,3-dicarboxylate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
90%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a mixture of (S)-4-(5.-chioro.-2-(isopropyiainino)pyridiri-4.-yi>-N-( I - (3- chlorophenvl)-2-hydroxyethyl).-1 H-pyrrole--2-carboxamide (44 mg, 0.1 mrnol) and 1 -(tertbutoxycarbonyi)azetidine-3-carboxylic acid (23 mg, 0.11 mmol) in dry CH2CI2 (4.5 ml.) was added EDC (21 mg, 0.11 mrnol) and DMAP (14mg, 0.11 rnmol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous NHiCi solution. The organic layer was dried over Na2SO. and concentrated in vacuum. The crude residue was purified by ISCO using DCMJIVIeOH (0-10%) as eluent to afford 55 rng (90%) of product. ESMS calculated (C30H35C12N505): 61 6.54; found: 61 7 (M+H).
(S)-1-tert-butyl 3-((S)-2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl) pyrrolidine-1,3-dicarboxylate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
99%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a mixture of (S)-4-(5-chioro-2-(isopropylamino)pyridin-4-yi)-N- (i-(clii orophenyi)-2-hydroxyethyi)- 1 H-pyrroie-2-carboxamide (44 mg, 0. 1 mmnol) and (5)-i -(tertbutoxycarbonyl)pyrrolidine-3-carboxylic acid (24 mg, 0.11 mmol) in dry CH2CI2 (4.5 mL) was added EDC (21 mg, 0.11 mnioi) and DMAP (14 mg, 0.11 nimol). The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous N}14C1 solution. The organic layer was dried over Na2SO4 and concentrated in vacuum. The crude residue was purified by ISCO using DCMIMeOH (0-10%) as eluent to afford 64 mg (99%) of product. ESMS calculated (C31H37C12N505): 630.56; found: 631(M+H).
(S)-1-tert-butyl 4-(2-(4-(5-chloro-2-(isopropylamino)pyridin-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl)piperidine-1,4-dicarboxylate[ No CAS ]
Yield
Reaction Conditions
Operation in experiment
98%
With dmap; 1,2-dichloro-ethane; In dichloromethane; at 20℃;
To a mixture of (S)-.4.-(5.-chioro.-2(isopropyiainino)pyridiri.-4.-yi).-N-.( I .-(3 chl orophenyi)2-hydroxyethyl)- 1 El-pyrrol e-2-carboxamide (65 m, 0.15 mmol) and I -(tert.butoxycarbonyl)piperidine -4-carhoxylic acid (38 mg, 0.165 mmol) in dry CH2CI2 (4 mU) was added EDC (32 mg, 0.165 mmoi) and DMAP (20 mg, 0. 165 mmol), The reaction mixture was stirred at room temperature overnight. The mixture was diluted with DCM and washed with aqueous NHiCi solution. The organic layer was dried over Na2SO. and concentrated in vacuum. The crude residue was purified by ISCO using DCMIMeOH (0.-10%) as eluent to afford 95 mg (98%) of product. ESMS calculated (C32H39C12N505): 64459; found: 645 (M+H).
(2S)-(S)-2-(4-(5-chloro-2-(isopropylamino)pyridine-4-yl)-1H-pyrrole-2-carboxamido)-2-(3-chlorophenyl)ethyl-2-amino-2-methylbutanoate dihydrochloride[ No CAS ]