AZD5363

Capivasertib (AZD5363) is a potent pan-AKT kinase inhibitor with IC50 of 3, 7 and 7 nM for Akt1,Akt2 and Akt3, respectively.

alternative route 1: (S)-4-amino-N-Q-(4-chlorophenyl)-3-hvdroxypropyl)- l-(7H-pyrrolo [2, 3-dl pyrimidin-4-yl)piperidine-4-carboxamideN-Ethyldiisopropylamine (1.676 ml, 9.62 mmol) was added to (S)-4-amino-N-(l-(4- chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide (Intermediate 49) (Ig, 3.21 mmol) and 4-chloro-7H-pyrrolo[2, 3-d]pyrimidine (0.493 g, 3.21 mmol) in butan-1-ol (15 ml). The resulting solution was stirred at 60(S)-4-amino-N-Q-(4-chlorophenyl)-3-hvdroxypropyl)-l-(7H-pyrrolo[2., 3- dl pyrimidin-4-yl)piperidine-4-carboxamide (E9)HCl (4M in Dioxane) (3.00 mL, 12.00 mmol) was added to (S)-tert-butyl 4-(l-(4- chlorophenyl)-3-hydroxypropylcarbamoyl)- 1 -(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidin- 4-ylcarbamate (Intermediate 22) (1.27 g, 2.40 mmol) in dichloromethane (20 mL). The resulting suspension was stirred at 20 To a stirred suspension of 8-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)-2-oxa-4, 8-diazaspiro [4.5]- decane-l, 3-dione free base (2, 3.0 g, 10.45 mmole) in acetonitrile (10.0 rel.vols, 30.0 mL) was added potassium bicarbonate (2.5 mol.eq, 2.64 g, 26.36 mmole), (3S)-3-amino-3-(4- chlorophenyl)propan-l-ol (1.2 mol.eq, 2.61 g, 12.67 mmole), water (1.5 rel.vols, 4.5 mL) and acetonitrile (3.5 rel.vol, 10.5 mL). The reaction mixture stirred at 25°C for lOhrs. Water (5.0 rel.vols, 15.0 mL) was charged to the reaction mixture and the reaction mass distilled at 50- 55°C under reduced pressure, until the residual volume reaches to 5-6 rel.vols. Isopropyl alcohol (4.0 rel.vols, 12.0 mL) was added to the concentrated reaction mass and the pH adjusted to pH -12.0-12.5 using 10percentw/v sodium hydroxide solution (0.6 rel.vols, 1.8 mL) at 25°C. The reaction mixture was heated to 55-60 °C and the pH re-adjusted to pH ~ 12.0-12.5 using 10percent w/v sodium hydroxide solution (0.6 rel.vol, 1.8 mL) at 55-60°C. The mixture was stirred for 90 mins and water (10.0 rel.vols, 30.0 mL) was added at 55-60°C. The reaction mixture was cooled slowly to 22-25°C over a period of 60-90mins. Water (10.0 rel.vols, 30.0 mL) was added and stirred for 18-20 hrs at 22-25°C. The mixture was filtered and the precipitated solid isolated and then dried at 60°C under vacuum to give desired 4-amino-N- [(l S)-l-(4-chlorophenyl)-3-hydroxy-propyl]-l-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidine- 4-carboxamide 3 as an off white solid, 3.8 g (80.0percent), Purity (99.0 percent by HPLC area).Chromatographic conditions: Chromatographic separation was achieved on Agilent LC systems equipped with PDA detectors using Atlantis T3 column and a mixture of Water: ACN: TFA as an eluent. (0148) 1H NMR-(400.13 MHz, DMSO-d6) delta: 11.68 (1H, s), 8.48 (1H, d), 8.13 (lH, s), 7.37-7.31 (4H, m), 7.16-7.15 (1H, m), 6.57 (1H, m), 4.88 (1H, d), 4.53 (1H, t), 4.41-4.34 (2H, m), 3.59-3.50 (2H, m), 3.40-3.35 (2H, m), 2.17 (2H, s), 2.02-1.80 (4H, m), 1.47-1.39 (2H, m).(S)-4-amino-N-Q-(4-chlorophenyl)-3-hvdroxypropyl)-l-(7H-pyrrolo[2., 3- dl pyrimidin-4-yl)piperidine-4-carboxamide (E9)HCl (4M in Dioxane) (3.00 mL, 12.00 mmol) was added to (S)-tert-butyl 4-(l-(4- chlorophenyl)-3-hydroxypropylcarbamoyl)- 1 -(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidin- 4-ylcarbamate (Intermediate 22) (1.27 g, 2.40 mmol) in dichloromethane (20 mL). The resulting suspension was stirred at 20 0C for 16 hours. The reaction mixture was filtered through a PTFE filtercup and the crude solid was purified by preparative HPLC (Waters XTerra Cl 8 column, 5mum silica, 19 mm diameter, 100 mm length), using decreasingly polar mixtures of water (containing 1percent TFA) and MeCN as eluents. Fractions containing the desired compound were purified by ion exchange chromatography, using an SCX column. The desired product was eluted from the column using 7M NH3ZMeOH and pure fractions were evaporated to dryness to afford (S)-4-amino-N-(l-(4-chlorophenyl)-3- hydroxypropyl)-l-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidine-4-carboxamide (0.200 g, 19.4 percent) as a white solid. IH NMR (399.9 MHz, DMSO-d6) delta 1.45 (2H, d), 1.86 (IH, d), 1.90 - 1.93 (IH, m), 2.19 (2H, s), 3.38 (2H, q), 3.51 - 3.58 (2H, m), 4.35 - 4.38 (2H, m), 4.53 (IH, t), 4.88 (IH, d), 6.58 (IH, t), 7.16 (IH, t), 7.32 - 7.38 (4H, m), 8.12 (IH, s), 8.43 (IH, d), 11.63 (IH, s), m/z (ESI+) (M+H)+ = 429; HPLC tR = 1.46 min.alternative route 2: (S)-4-amino-N-Q-(4-chlorophenyl)-3-hvdroxypropyl)- l-(7H-pyrrolo [2, 3-dl pyrimidin-4-yl)piperidine-4-carboxamide(S)-3-Amino-3-(4-chlorophenyl)propan-l-ol (Intermediate 47) (2.055 g, 11.07 mmol) was added in one portion to 4-(tert-butoxycarbonylamino)-l-(7H-pyrrolo[2, 3-d]pyrimidin-4- yl)piperidine-4-carboxylic acid (Intermediate 1) (4g, 11.07 mmol) and DIPEA (5.80 ml, 33.20 mmol) in DMA (40 ml). HATU (4.63 g, 12.18 mmol) was added and the resulting solution was stirred at 200C for 24 hours. The reaction mixture was evaporated to dryness then diluted with EtOAc (300 mL), and washed sequentially with water (50 mL) and saturated brine (50 mL). The organic layer was dried over MgSO4, filtered and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 2 to 6percent MeOH with ammonia in DCM. Pure fractions were evaporated to dryness and triturated with dioxane (40ml) to afford (S)-tert-butyl 4-(l-(4-chlorophenyl)-3- hydroxypropylcarbamoyl)-l-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate (Intermediate 22) (4.82 g, 82 percent) as a white solid. (S)-tert-butyl 4-(l-(4-chlorophenyl)-3- hydroxypropylcarbamoyl)-l-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidin-4-ylcarbamate (Intermediate 22) (4.82 g, 82 percent) was suspended in dioxane (40.0 ml) and 4M hydrogen chloride in dioxane (7.69 ml, 221.36 mmol) added. The reaction was stirred at ambient temperatre for 2 hours. The crude product was purified by ion exchange chromatography, using an SCX column. The desired product was eluted from the column using 3.5MNH3ZMeOH and pure fractions were evaporated to dryness. The crude product was purified by preparative HPLC (Waters XBridge Prep Cl 8 OBD column, 5mum silica, 19 mm diameter, 100 mm length), using decreasingly polar mixtures of water (containing 1percent NH3) and MeCN as eluents. Fractions containing the desired compound were evaporated to dryness to afford (S)-4-amino-N-(l-(4-chlorophenyl)-3-hydroxypropyl)-l-(7H-pyrrolo[2, 3- d]pyrimidin-4-yl)piperidine-4-carboxamide (1.200 g, 25.3 percent) as a white solid. alternative route 1: (S)-4-amino-N-Q-(4-chlorophenyl)-3-hvdroxypropyl)- l-(7H-pyrrolo [2, 3-dl pyrimidin-4-yl)piperidine-4-carboxamideN-Ethyldiisopropylamine (1.676 ml, 9.62 mmol) was added to (S)-4-amino-N-(l-(4- chlorophenyl)-3-hydroxypropyl)piperidine-4-carboxamide (Intermediate 49) (Ig, 3.21 mmol) and 4-chloro-7H-pyrrolo[2, 3-d]pyrimidine (0.493 g, 3.21 mmol) in butan-1-ol (15 ml). The resulting solution was stirred at 600C for 18 hours. The reaction mixture was diluted with EtOAc (50 mL), and washed sequentially with water (25 mL) and saturated brine (25 mL). The organic layer was dried over MgSO4, filtered and evaporated to afford crude product. The crude product was purified by flash silica chromatography, elution gradient 0 to 6percent MeOH with ammonia in DCM. Pure fractions were evaporated to dryness to afford (S)-4-amino-N-(l-(4-chlorophenyl)-3-hydroxypropyl)-l-(7H-pyrrolo[2, 3- d]pyrimidin-4-yl)piperidine-4-carboxamide (842mg) as a white foam. (S)-4-amino-N-(l- (4-chlorophenyl)-3-hydroxypropyl)-l-(7H-pyrrolo[2, 3-d]pyrimidin-4-yl)piperidine-4- carboxamide was stirred in ethyl acetate (7 mL) for 18 hours. The solid was collected by filtration, washed with a small amount of ethyl acetate and vacuum oven dried at 55°C for 18 hours to afford (S)-4-amino-N-(l-(4-chlorophenyl)-3-hydroxypropyl)-l-(7H- pyrrolo[2, 3-d]pyrimidin-4-yl)piperidine-4-carboxamide (0.585 g, 42.5 percent) as a white solid, m/z (ES+) (M+H)+ = 429; HPLC tR= 1.60 min. IH NMR (400.13 MHz, DMSO-d6) delta 1.39 - 1.47 (2H, m), 1.80 - 2.02 (4H, m), 2.17 (2H, s), 3.35 - 3.40 (2H, m), 3.50 - 3.59 (2H, m), 4.34 - 4.41 (2H, m), 4.53 (IH, t), 4.88 (IH, d), 6.57 (IH, m), 7.14 - 7.16 (IH, m), 7.31 - 7.37 (4H, m), 8.12 (IH, s), 8.42 (IH, d), 11.62 (IH, s)