1H-indole-2-carboxamide

Conditions: temp:23.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portion to a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF (0.15 mL) at 0° C. under an argon atmosphere and the solution was stirred for 5 min. Indole-2-carboxylic acid (0.40 mg, 2.5 μmol, 1.3 equiv), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.48 mg, 2.5 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.31 mg, 2.3 μmol, 1.2 equiv) were then added separately, each in one portion, to the above solution at 0° C. The reaction mixture was warmed to 23° C. over 15 h 30 min, then was quenched with saturated aqueous ammonium chloride solution (3 mL). The mixture was diluted with ethyl acetate (10 mL) and the layers were separated. The aqueous layer was extracted with ethyl acetate (10 mL) and the combined organic layer was dried over sodium sulfate. Concentration in vacuo left a white solid, which was purified by flash column chromatography (60% ethyl acetate-hexanes) to give the indole-2-carboxylic acid amide derivative (1.1 mg, 87%) as a white solid. at 0° C was quenched with saturated aqueous ammonium chloride solution (3 mL) The mixture was diluted with ethyl acetate (10 mL) the layers were separated The aqueous layer was extracted with ethyl acetate (10 mL) the combined organic layer was dried over sodium sulfate Concentration in vacuo left a white solid, which was purified by flash column chromatography (60% ethyl acetate-hexanes)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The precipitate formed was filtered under nitrogen

Conditions: temp:-5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The dry ice/acetone bath was removed the reaction mixture stirred for 18 h under nitrogen atmosphere was stirred for an additional 2 h was stirred for 30 min Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added The layers were partitioned the aqueous layer was extracted with dichloromethane The combined organic layers were washed with water and brine dried over magnesium sulfate filtered

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The precipitate formed was filtered under nitrogen

Conditions: temp:-5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The dry ice/acetone bath was removed the reaction mixture stirred for 18 h under nitrogen atmosphere was stirred for an additional 2 h was stirred for 30 min Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added The layers were partitioned the aqueous layer was extracted with dichloromethane The combined organic layers were washed with water and brine dried over magnesium sulfate filtered

Conditions: temp:-5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The dry ice/acetone bath was removed the reaction mixture stirred for 18 h under nitrogen atmosphere was stirred for an additional 2 h was stirred for 30 min Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added The layers were partitioned the aqueous layer was extracted with dichloromethane The combined organic layers were washed with water and brine dried over magnesium sulfate filtered

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 ml/min 100-100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mil/min) Rt 2.67 min (100%). The precipitate formed was filtered under nitrogen

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 m/min 100—100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mi/min) Rt 2.67 min (100%). The precipitate formed was filtered under nitrogen

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 m/min 100—100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mi/min) Rt 2.67 min (100%). The precipitate formed was filtered under nitrogen

Conditions: temp:-5.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: A solution of 3-(4-amino-3-methoxyphenyl)-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (0.500 g, 1.15 mmol) in pyridine (8 mL) at −5° C. was treated with a solution of 1H-2-indolecarbonyl chloride (0.413 g, 2.3 mmol) in dichloromethane (1 mL). The reaction mixture stirred for 20 min at −5° C. The dry ice/acetone bath was removed and the reaction mixture stirred for 18 h under nitrogen atmosphere. 1H-2-indolecarbonyl chloride (0.207 g, 1.15 mmol) was added and was stirred for an additional 2 h. Sodium hydroxide (1 N) solution (10 mL) was added and was stirred for 30 min. Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added. The layers were partitioned, and the aqueous layer was extracted with dichloromethane. The combined organic layers were washed with water and brine, dried over magnesium sulfate, filtered, and reduced under pressure to give crude N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide was purified by flash chromatography on silica gel using 15% (methanol with 2% ammonium hydroxide) in dichloromethane to still give a crude product. A second column using a gradient of 10% (methanol with 2% ammonium hydroxide) to 50% (methanol with 2% ammonium hydroxide) gave 0.139 g (21%) pure N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide. N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide (0.139 g, 0.24 mmol) in warmed ethyl acetate was treated with a warmed solution of maleic acid (0.083 g, 0.719 mmol) in ethyl acetate. The precipitate formed was filtered under nitrogen to give 0.166 g of N1-(4-{4-amino-1-[1-(1-methylpiperidin-4-yl)piperidin-4-yl]-1H-pyrazolo[3,4-d]pyrimidin-3-yl}-2-methoxyphenyl))-1H-2-indolecarboxamide di-maleate. 1H NMR (DMSO-d6, 400 MHz) δ 11.83 (s, 1H), 9.442 (s, 1H), 8.283 (s, 1H), 8.154-8.134 (d, 1H, J=8.12 Hz), 7.694-7.674 (d, 1H, J=8.04 Hz), 7.498-7.477 (d, 1H, J=8.20 Hz), 7.407-7.402 (m, 1H), 7.352-7.325 (m, 2H), 7.267-7.229 (m, 1H), 7.112-7.074 (m, 1H), 6.078 (s, 4H), 5.10-5.00 (m, 1H), 3.974 (s, 3H), 3.525 (m, 4H), 3.178 (m, 2H), 2.975 (m, 2H), 2.771 (s, 3H), 2.457 (s, 3H), 2.208 (m, 4H), 1.909 (m, 2H); HPLC Perkin Elmer Pecosphere C18, 3 μM, 33×4.6, 3.5 m/min 100—100% 50 mM ammonium acetate to acetonitrile in 4.5 minutes, C36H44N6O3 (581.2), 95%. LCMS (Perkin Elmer, Pecosphere C18 column, 3 um particle size, 33×4.6 mm; 100% 50 mM ammonium Acetate in Water to 100% Acetonitrile over 5 min, 3.0 to 3.5 mi/min) Rt 2.67 min (100%). The dry ice/acetone bath was removed the reaction mixture stirred for 18 h under nitrogen atmosphere was stirred for an additional 2 h was stirred for 30 min Organic solvent was removed under reduced pressure, and dichloromethane (25 mL) was added The layers were partitioned the aqueous layer was extracted with dichloromethane The combined organic layers were washed with water and brine dried over magnesium sulfate filtered

Conditions: temp:23.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: Diethylaniline (0.33 μL, 2.1 μmol, 1.1 equiv) was added in one portion to a stirred solution of the amine (1.0 mg, 1.9 μmol, 1 equiv) in THF (0.15 mL) at 0° C. under an argon atmosphere and the solution was stirred for 5 min. Indole-2-carboxylic acid (0.40 mg, 2.5 μmol, 1.3 equiv), 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (0.48 mg, 2.5 μmol, 1.3 equiv) and 1-hydroxybenzotriazole (0.31 mg, 2.3 μmol, 1.2 equiv) were then added separately, each in one portion, to the above solution at 0° C. The reaction mixture was warmed to 23° C. over 15 h 30 min, then was quenched with saturated aqueous ammonium chloride solution (3 mL). The mixture was diluted with ethyl acetate (10 mL) and the layers were separated. The aqueous layer was extracted with ethyl acetate (10 mL) and the combined organic layer was dried over sodium sulfate. Concentration in vacuo left a white solid, which was purified by flash column chromatography (60% ethyl acetate-hexanes) to give the indole-2-carboxylic acid amide derivative (1.1 mg, 87%) as a white solid. at 0° C was quenched with saturated aqueous ammonium chloride solution (3 mL) The mixture was diluted with ethyl acetate (10 mL) the layers were separated The aqueous layer was extracted with ethyl acetate (10 mL) the combined organic layer was dried over sodium sulfate Concentration in vacuo left a white solid, which was purified by flash column chromatography (60% ethyl acetate-hexanes)

Conditions: temp:0.00; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: To a solution of indole-2-carboxylic acid (1e: 1 mmol) in DMF (4 mL) was added DIPEA (2 eq.) followed by HATU (1.2 eq.). The mixture was stirred for 15 minutes before the desired amine (R6—NH2) was added. The mixture was left to stir at room temperature for 5-16 hrs before saturated NH4Cl solution was added. The mixture was extracted with ethyl acetate and the combined organic extracts were washed with water and brine, dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The crude material was purified on CombiFlash® purification system (Teledyne Isco) using appropriate mixtures of ethyl acetate and cyclohexanes to obtain the desired product indole-2-carboxamide (I). was added The mixture was left was added The mixture was extracted with ethyl acetate the combined organic extracts were washed with water and brine dried over anhydrous Na2SO4 filtered concentrated in vacuo The crude material was purified on CombiFlash® purification system (Teledyne Isco)