3-Iodo-1H-pyrazolo[3,4-d]pyrimidin-4-ylamine

Synthesis of 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-amine (BA19); A solution of 3H-pyrazolo[3, 4-d]pyrimidin-4-amine (10 g, 0.074 mol) and n-iodo-succinamide (25 g, 0.111 mol) in DMF (80 mL) was heated to 80° C. overnight under an argon atmosphere. The resulting solid was filtered and rinsed with cold EtOH. Product was dried in vacuo overnight to yield BA19 (24 g, 100percent yield). ESI-MS (M+H)Step 1: 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-ylamine A mixture of 1H-pyrazolo[3, 4-d]pyrimidin-4-ylamine (5.0 g, 37 mmol) and NIS (10.7 g, 45 mmol) in DMF (100 mL) was stirred at 70° C. overnight. Then the mixture was cooled to room temperature and filtered to give 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-ylamine (4.1 g) as a white solid. The filtrate was concentrated and the residue was treated with 10percent NaNIS(1.1eq, 4.95 g, 22 mmol) was slowly added into the mixture of 9(1eq, 2.70 g, 20 mmol) and DMF (50 mL) at r.t. The reaction mixturewas continued stirred at r.t. overnight. The solvent was removed invacuo and water was added. The mixture was subjected to sonicationfor 10 min. The precipitate was collected by filtration, rinsedwith water, and dried. The solid 10 was used for the next stepwithout further purification (5.01 g, 96percent yield).NIS (250 g, 1.11 mol, 1.5 eq.) was added to a solution of 1H-pyrazolo[3, 4-d]pyrimidin-4-amine (100 g, 0.74 mol, 1.0 eq.) in DMF (800 mL). To a mixture of lff-pyrazole[3, 4-r/]pyrfniidite-4-ani.iie (5 g, 37.03 nimol) in N.N dimethylformamide (40 ml.) was added ;V-iodo nccinimide (12.5 g, 55.6 tnmol) and the reaction mixture was heated at SO 1H-Pyrazolo[3, 4-d]pyrimidin-4-amine (4, 0.10 g, 0.74 mmol) and N-iodosuccinimide (0.25 g, 1.1 mmol) in DMF (2 ml) were heated in a microwave at 90 °C for 10 min. The resulting precipitate was filtered and washed with water to afford 5 as a brown solid in 92percent yield (0.17 g, 0.65 mmol). Scheme 1 depicts the synthesis of 2-( 4-amino-lH-pyrazolo[3, 4-d]pyrimidin-3-yl) iodide (Cpd. 1-3), an intermediate in the synthesis of the compounds of the invention and its further reactions to obtain final inhibitor analogs. Cyano substituted aminopyrazole 1-1 is heated with formamide at 160Scheme 1 depicts the synthesis of 2-(4-amino-1H-pyrazolo[3, 4-d]pyrimidin-3-yl) iodide. Cyano substituted aminopyrazole 1-1 is heated with formamide at 160° C. for 5 hours to yield 2-(4-amino-1H-pyrazolo[3, 4-d]pyrimidine (compound 1-2) in 90percent yield. This intermediate is reacted with N-iodosuccinimide in dimethylformamide at 80° C. for 16 hours, to produce 2-(4-amino-1H-pyrazolo[3, 4-d]pyrimidin-3-yl) iodide (Cpd. 1-3) in 90percent yield.To a solution of 4-aminopyrazolo[3, 4-d]pyrimidine (5.00 g, 37.0 mmol) in DMF (130 mL) was added N-iodosuccinimide (10.83 g, 48.14 mmol). The reaction mixture was heated overnight at 50 °C. Additional N-iodosuccinimide (1.67 g, 7.41 mmol) was added and heating was continued at 50 °C for a second night. The mixture was then allowed to cool to rt and concentrated to one half volume under reduced pressure. Water was added and the resulting precipitate was collected by suction filtration. The solids were washed with water and ethanol, then dried in vacuo at 40 °C to afford the product, 4-amino-3-iodo-lH-pyrazolo[3, 4- d]pyrimidine (Intermediate 14, 8.66 g, 90percent). LCMS (AA): m/z 262 (M+H); 1H MR (400 MHz, -DMSO) δ 13.80 (s, 1H), 8.16 (s, 1H).A mixture of lH-Pyrazolo[3, 4-d]pyrimidin-4-ylamine (11.75 g, 0.09 mol) (Step A) and N-iodosuccinimide (25.45 g, 0.11 mol) in dimethylformamide (300 ml) was stirred at 50° C for 24 hr. A second batch of N-iodosuccinimide (3.92 g, 0.02 mol) was added and the solution stirred for additional 24 hr. Upon standing at room temperature, a precipitate was formed which was separated by filtration and washed with dimethylformamide and ethanol to afford 10.05 g of the title compound. The filtrate was concentrated in vacuo to about one half of the original volume and 500 ml of water was added. The precipitated product was separated by filtration and washed with ethanol to afford a second batch of the product (10.53 g, combined yield 20.58 g, 90.6 percent); LC/MS, API-ES, Pos, (M+H)Compound C (2.367 g, 17.5 mmol) and N-iodosuccinimide (4.810 g, 21.4 mmol) were added to dimethylformamide (60 mL) and stirred at 50°C for 24 hours. Another batch of N-iodosuccinimide (0.871 g, 3.8 mmol) was added to the reaction mixture and was allowed to stir for an additional 24 hours. The reaction mixture was cooled to room temperature and water (100 mL) was added, forming a precipitate that was collected by filtration to yield D (4.1 g, 89percent). To a four-port adding DMF flask (800 ml), by adding N-chloro succinimide (197.62g, 1 . 48mol), by a hot water bath or ice water bath temperature control, temperature control in the 30 °C, add NaI (221.84g, 1 . 48mol), stirring after adding 7.0h, then batch by adding 4-amino-pyrazolo [3, 4-d] pyrimidine (100g, 0.74mol), suck, raising the temperature to 80 °C. HPLC central control, reaction is ended. After treatment, the reaction liquid by adding water, cooling to 0-5°C, filtering, drying, to obtain the product. Yield: 89percent, purity: 99.3percent (HPLC).1H-pyrazolo [3, 4-d] pyrmidin-4-amine (20 g, 148.0 mmol, 1.0 eq) was placed in a three-necked flask. To the flask was added 150 mL of DMF (N, N-dimethylformamide), and the mixture was stirred and replaced with nitrogen 3 times. NIS (N-iodosuccinimide) (50 g, 222.0 mmol, 1.5 eq) was added and the solution was heated at 80° C. The reaction monitored by TLC was completed after 22 h, stopped and the DMF was concentrated to the remaining half of the solution. The mixture was stirred at 150 mL of saturated aqueous NaThe 1H-pyrazolo [3, 4-d] pyrimidin-4-ylamine (953 mg, 7 . 055 mm, 1 . 0equiv), N-iodo succinimide (2.381g, 10 . 58 mm, 1 . 5equiv) suspended in N, N-dimethylformamide (15 ml) in, and in the nitrogen in 80 °C lower heating sleepovers. The reaction mixture is cooled to the room temperature, and make-up water (60 ml). Collecting solid, aqueous solution of saturated sodium sulfite are used, various washing water 2 times, and dried in air, to obtain white solid compound 2 (1.58g, yield 86percent).Example 1. Synthesis of (i?)-l-(1H-pyrazolo [3, 4-d] pyrimidin-4-amine (12) (13.5 g, 0.1 mol) and N-iodosuccinimide (NIS) (25.8 g, 0.15 mol) In a three-necked flask, DMF (100 mL) was added, the temperature was raised to 50 ° C, and the reaction was carried out at 50 ° C for 18 hours(TLC test to confirm the disappearance of raw materials). Cooling to 0 ° C crystallization, filtration and drying to give intermediate (13) (yellow solid, 20.9 g, yield 80percent).:1H-[3, 4-d]pyrazolopyrimidin-4-amine(5.3g, 39mmol) andN-Iodosuccinimide (13.3 g, 59 mmol) was suspended in N, N-dimethylformamide, The reaction was heated to 80°C under a nitrogen atmosphere for 12 hours.After the reaction is completed, it is cooled to room temperature, water is added, and a large amount of solids are precipitated and filtered with a Buchner funnel.Solids in turn with saturated sodium thiosulfate solution, Water and hot ethanol are washed twice each.The oil pump was drained to give 7.8 g of the target compound in a yield of 77percent.IH-pyrazolo[3 4-dJpynmidn4arnine (1 5g 1111 mmol) was suspended in 15 nil of DMF and Nodosuccnmde (1 2 eq 3 0 g 13 34 mmol) added The mixture was heated at 180 0C in the microwave for an hour. EtOH (80 m) was added to the reacUonand a precipitate began to fo rn, hich was aided by sonicaton The preuipitate wasfiltered and Washed with EtOH (x3, 20 ml) and aUowed to dry in an oven at 40 °Coveimghtto give a sand cooured soid (2115 g, 8105 mmol 730percent) ‘H NMR (500MHz, DMS06 13.80 (s, IN), 8.i(s, 1H) 7, 796:.44 (m 2H);13C NMR (126 MHz, DM80) 6 157 60 (C) 156 08 CH) 155 04 (C), 102 50 (C), 89 82 (C) MS (ES +ve)[M+H)’: 283.9 (+Na), (ES -ye) [M-Hf: 259.9. 287.8 (+Na).To a suspension of 1H-pyrazolo[3, 4-d]pyrimidin-4-amine (1) (2.8 g, 20.7 mmol) in DMF (12 mL) was added N-iodosuccinimide (5.59 g, 24.8mrnol) at ambient temperature. The reaction mixture was heated to 80 °C and stirred for 14hr. The resulting solid was collected by filtration, rinsed with ice-cold ethanol (20 mL), and concentrated in vacuo to provide 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-amine (3.9 g, 72percent) as an off- white solid. MS m/z: 261.92 (M+1 ).87C : Compound 87B (700 mg, 5.18 mmol) and N-iodo-succinimide (1.75 g, 7.77 mmol) were dissolved in 12 mL of Ν, Ν-dimethylformamide and stirred at 80°C overnight under nitrogen atmosphere. The mixture was cooled and 30 mL of water were added. The resulting solid was filtered and rinsed with cold ethanol. The product was dried in vacuo. Compound 87C was obtained as a brown powder (935 mg, 69percent yield). NMR (ppm, DMSO): 8.17 (s, 1H). LCMS (+esi): 262.0 (M+HStep 1To a solution of lH-pyrazolo[3, 4-d]pyrimidin-4-ylamine (3, 0 g, 22.20 mmol, 1.0 eq) in DMF (30 mL), NIS (6.7g , 24.42 mmol, 1.1 eq) was added at room temperature. The reaction mixture was stirred overnight at 60 °C. The reaction mixture was cooled to room temperature and 10 percent aq. NaHCOs (150 mL) was added to the reaction mixture. The solid was filtered and re-crystallization from DMF solvent to give 3-iodo- lH~pyrazo3o[3, 4- d]pyrimidin-4-amme (4.0 g, 69 percent in yield).To a solution of 1H-pyrazoio[3, 4-d]pyrimidin4’ylamine (3.0 g, 22.20 mrnol, 1.0 eq) in I)MF (30 mE), NIS (6.7g , 24.42 mmol, 1.1 eq) was added at room temperature. The reaction mixture was stirred overnight at 60 °C. The reaction mixture was cooled to room temperature and 10 percent aq. NaHCO3 (150 added to the reaction mixture. The solid was filtered and recrystallization from DMF solvent to give 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin”amine (4.0 g, 69 percent in yield).Step 1. To a solution of 1H-pyrazolo[3, 4-d]pyrimidin-4-ylamine (3.0 g, 22.20 mmol, 1.0 eq) in DMF (30 mL), NIS (6.7g, 24.42 mmol, 1.1 eq) was added at room temperature. The reaction mixture was stirred overnight at 60 °C. The reaction mixture was cooled to roomtemperature and 10 percent aq. NaHCO3 (150 mL) was added to the reaction mixture. The solid was filtered and re-crystallization from DMF solvent to give 3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-ammne (4.0 g, 69 percent in yield).Step 1 A solution of commercially available 4-aminopyrazolo-(3, 4-d)pyrimidine (3.36 g, 24.9 mmol) in 70 mL of DMF was treated with N-iodosuccinamide (8.39 g, 37.3 mmol) and stirred at 80°C for 20 h. The reaction mixture was cooled to room temperature and added into 70 mL ofice cold water. A brown precipitate formed, which was filtered and washed with ice cold ethanol. The resulting solid was dried under reduced pressure to give 3 -iodo- 1H-pyrazolo [3, 4- d]pyrimidin-4-amine (4.50 g, 69percent), which was used directly in the next reaction.1 H-Pyrazolo[3, 4-d]pyrimidin-4-amine (20. Og, 148.01 mmol) and /V-iodosuccinimide (60.61 g, 222.01 mmol) were combined in DMF (200mL) and the mixture heated to 80°C, under nitrogen, overnight. The bulk of the solvent was removed under reduced pressure and the residue was allowed to cool and stand for a couple of hours. The resultant material was stirred in warm water (250 ml), filtered off and washed with methanol (3 x 150 ml) to give 3-iodo-1 /-/-pyrazolo[3, 4-c ]pyrimidin-4-amine (20.92g, 80.147mmol, 54.1 percent yield) LCMS (ESStep 2A mixture of lH-pyrazolo[3, 4-d]pyrimidm-4-amine (150 g, 1.1 1 mo., 1.00 equiv) and N-iodo-succinimide (375 g, 1.67 mol, 1.58 equiv) in N, N-dimethylformamide (2.5 L) was stirred at 80°C for 5 h. The reaction mixture was cooled to room temperature and then diluted with 10 L of water. The solid was collected by filtration, washed with 2 1 L of saturated aqueous sodium sulfite and dried under vacuum to give 50 g (52percent) of 3-iodo-lH- pyrazolo[3, 4-d]pyrimidin-4-amine as a yellow solid.Step 2A mixture of l H-pyrazolo[3, 4-d]pyrimidin-4-amine (150 g, 1.1 1 mol, 1.00 equiv) and N-iodo-succinimide (375 g, 1.67 mol, 1.58 equiv) in N, N-dimethylformamide (2.5 L) was stirred at 80°C for 5 h. The reaction mixture was cooled to room temperature and then diluted with 10 L of water. The solid was collected by filtration, washed with 2x1 L of saturated aqueous sodium sulfite and dried under vacuum to give 150 g (52percent) of 3-iodo-.H- pyrazolo[3, 4~d]pyrimidin-4~amine as a yellow solid.Step 2. [0121] To a solution of R-1 (50.0 g, 0.37 mol) in DMF(350.0 mL) NIS (83.3 g, 0.37 mol) is added. The reactionmixture is heated to 75° C. for 24 h. The mixture is thencooled to room temperature and poured into water. The mixtureis filtered and the percipitate is washed with water, driedunder reduced pressure to afford compound I -1 ( 45.0 g, 48percent ).A mixture of lH-pyrazolo[3, 4-d]pyrimidin-4-amine (150 g, 1.1 1 mol, 1.00 equiv) and N-iodo-succinimide (375 g, 1.67 mol, 1.58 equiv) in N, N-dimethylformamide (2.5 L) was stirred at 80°C for 5 h. The reaction mixture was cooled to room temperature and then diluted with 10 L of water. The solid was collected by filtration, washed with 2x1 L of saturated aqueous sodium sulfite, and dried under vacuum to give 150 g of 3-iodo-lH-pyrazolo[3, 4- d]pyrimidin-4-amine as a yellow solid.A solution lH-pyrazolo[3, 4-d]pyrimidin-4-amine (5.0 g, 37 mmol, 1.00 eq) in DMF (50 mL) was cooled to 0 °C. To the solution was added NIS (6.1g, 38.9 mmol, 1.05 eq) portion wise. The reaction was heated at 60 °C for 24hours, then saturated sodium bicarbonate solution was added. The reaction was stirred for another 30 minutes , filtered and concentrated to dryness under vacuum to give 3-iodo-lH-pyrazolo[3, 4-d]pyrimidin-4-amine as a yellow solid 5.8 g.To a suspension of 1H-pyrazolo[3, 4-d]pyrimidin-4-amine (7.5 g, 55.50 mmol) in anhydrous DMF (50 mL) was added N-iodosuccinimide (49 g, 222.0 mmol) portionwise under nitrogen atmosphere. The reaction mixture was heated at 80 °C for 2 h. The reaction was monitored by TLC. After completion of reaction, the mixture was cooled to RT and diluted with EtOAc (400 mL). Saturated aq. sodium thiosulfate solution (100 mL) was added, and a precipitate formed. The precipitate was collected by filtration, washed with additional amount of water, diethyl ether and then dried to obtain 14 g of 3-iodo- 1H-pyrazolo[3, 4- d]pyrimidin-4-amine as an off-white solid.1H-Pyrazolo[3, 4-d]pyrimidin-4-amine (2 g, 14.8 mol) was charged in DMF (12 mL) and to this was added N-iodosuccinimide (6.3 g, 28.6 mmol). The reaction mixture was heated at 100 °C for 4 h. The solid obtained was filtered off and washed with cold EtOH (15 mL). The solid obtained was dried under vacuum to get (4 g) 3-iodo-1H-pyrazolo [3, 4-d]pyrimidin-4-amine.A mixture of 1H-pyrazolo[3, 4-djpyrimidin-4-amine (Formula II, 20 g), Niodosuccinimide (41.6 g) and dimethylformamide (300 mL) was stirred at 75°C to 80°C for 16 hours. Water (1 L) was added to the reaction mixture, and then the mixture wasstirred at 15°C for 4 hours. The solid obtained was filtered, then washed with water (100mL), and then washed with cold ethanol (60 mL). The resulting solid was dried at 45°C under vacuum for 16 hours to obtain the title compound.Yield: 26.8 g: Preparation of 3-iodo-lH-pyrazolor3.4-dlpyrimidin-4-amine (Formula III, when X is iodine) A mixture of lH-pyrazolo[3, 4-d]pyrimidin-4-amine (Formula II, 20 g), N- iodosuccinimide (41.6 g), and dimethylformamide (300 mL) was stirred at 75°C to 80°C for 16 hours. Water (1 L) was added to the reaction mixture, and then the mixture was stirred at 15°C for 4 hours. The solid obtained was filtered, then washed with water (100 mL), and then washed with cold ethanol (60 mL). The resulting solid was dried at 45°C under vacuum for 16 hours to obtain the title compound. Yield: 26.8 gCompound g (10, 1eq), NIS (25g, 2.5eq) and N, N-dimethylformamide (DMF) (100mL) were mixed and stirredovernight under nitrogen at 80 °C, and then cooled and poured into water (500mL). Solids were filtered out and washedsuccessively with saturated sodium bisulfite (50mL) and water (100mL), and then dried under vacuum for 10h at 50 °C, giving 16g of solid.3-iodo-1H-pyrazolo[3, 4-d]pyrimidin-4-ylamine (Intermediate LA) Pyrazolo [3, 4-d] pyrimidin-4-amine (15.0 g, 111.1 mmol) was dissolved in DMF (150 mL) N-iodosuccinimide (37.5 g, 166. 6 mmol) was slowly added to the reaction mixture, and the reaction was stirred at 80 ° C for 12 h. stopWater (40 mL) was added to the reaction mixture, and the mixture was suction filtered. The solid was washed with water (80 mL), ethanol (60 mL) and dried to give to a yellow solid (24. 9 g, 86percent).

Molecular Weight:261.02
XLogP3:0.5
Hydrogen Bond Donor Count:2
Hydrogen Bond Acceptor Count:4
Exact Mass:260.95114
Monoisotopic Mass:260.95114
Topological Polar Surface Area:80.5
Heavy Atom Count:11
Complexity:154
Covalently-Bonded Unit Count:1
Compound Is Canonicalized:Yes