1,4-dioxane-2,5-diol

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

Procedure: 2-(6-Chloro-benzothiazol-2-ylamino)-1-(2-methoxy-ethyl)-1H-benzoimidazole-5-carboxylic acid [1-(2-hydroxy-ethyl)piperidin-4-ylmethyl]-amide (45 mg) was prepared by following General Procedure P starting from 2-(6-chloro-benzothiazol-2-ylamino)-1-(2-methoxy-ethyl)-1H-benzoimidazole-5-carboxylic acid (piperidin-4-ylmethyl)-amide dihydrochloride (136 mg), [1,4]dioxane-2,5-diol (54 mg), and Na(OAc)3BH (95 mg) in DCM (2 mL). LC/MS: m/z 542.7.

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

Procedure: 2-(6-Chloro-benzothiazol-2-ylamino)-1-(2-methoxy-ethyl)-1H-benzoimidazole-5-carboxylic acid [1-(2-hydroxy-ethyl)-piperidin-4-yl]-amide (52 mg) was prepared by following General Procedure P starting from 2-(6-chloro-benzothiazol-2-ylamino)-1-(2-methoxy-ethyl)-1H-benzoimidazole-5-carboxylic acid piperidin-4-ylamide dihydrochloride (145 mg), [1,4]dioxane-2,5-diol (55 mg), and Na(OAc)3BH (95 mg) in DCM (2 mL). LC/MS: m/z 528.6.

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

Procedure: A solution of methyl-{4-[4-(3-pyrrolidin-1-ylpropoxy)phenyl]-tetrahydropyran-4-ylmethyl}amine (350 mg, 1.05 mmol), glycolaldehyde dimer (130 mg, 1.05 mmol) and AcOH (0.15 ml, 2.1 mmol) in DCM (5 ml) was stirred at room temperature for 18 hours. Saturated aqueous sodium carbonate (10 ml) and DCM (10 ml) were added and the mixture was shaken and partitioned. The aqueous phase was extracted with DCM (2×10 ml) and the combined organics dried (K2CO3), filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel eluting with dichloromethane:methanol:ammonia (92:8:1) to provide the title compound as a clear colourless oil (223 mg, 0.59 mmol, 56%). 1H NMR (400 MHz, CDCl3) δ 1.70-1.90 (m, 6H), 1.96 (s, 3H), 2.00 (quintet, 2H), 2.14 (m, 2H), 2.37 (t, 2H), 2.40-2.60 (m, 6H), 2.65 (t, 2H), 3.35 (t, 2H), 3.50 (m, 2H), 3.76 (m, 2H), 4.01 (t, 2H), 6.89 (d, 2H), 7.19 (d, 2H). HRMS ESI+ m/z 377.2796 [MH]+. partitioned The aqueous phase was extracted with DCM (2×10 ml) the combined organics dried (K2CO3) filtered concentrated in vacuo The crude product was purified by flash chromatography on silica gel eluting with dichloromethane:methanol:ammonia (92:8:1)

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

Procedure: A solution of methyl-{4-[4-(3-pyrrolidin-1-ylpropoxy)phenyl]-tetrahydropyran-4-ylmethyl}amine (350 mg, 1.05 mmol), glycolaldehyde dimer (130 mg, 1.05 mmol) and AcOH (0.15 ml, 2.1 mmol) in DCM (5 ml) was stirred at room temperature for 18 hours. Saturated aqueous sodium carbonate (10 ml) and DCM (10 ml) were added and the mixture was shaken and partitioned. The aqueous phase was extracted with DCM (2×10 ml) and the combined organics dried (K2CO3), filtered and concentrated in vacuo. The crude product was purified by flash chromatography on silica gel eluting with dichloromethane:methanol:ammonia (92:8:1) to provide the title compound as a clear colourless oil (223 mg, 0.59 mmol, 56%). 1H NMR (400 MHz, CDCl3) δ 1.70-1.90 (m, 6H), 1.96 (s, 3H), 2.00 (quintet, 2H), 2.14 (m, 2H), 2.37 (t, 2H), 2.40-2.60 (m, 6H), 2.65 (t, 2H), 3.35 (t, 2H), 3.50 (m, 2H), 3.76 (m, 2H), 4.01 (t, 2H), 6.89 (d, 2H), 7.19 (d, 2H). HRMS ESI+ m/z 377.2796 [MH]+. partitioned The aqueous phase was extracted with DCM (2×10 ml) the combined organics dried (K2CO3) filtered concentrated in vacuo The crude product was purified by flash chromatography on silica gel eluting with dichloromethane:methanol:ammonia (92:8:1)

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

Procedure: 120 mg (1.0 mmol) of glycolaldehyde dimer and 2 ml of acetic acid were added to 10 ml of a methanol solution containing 473 mg (1.55 mmol) of the 7-amino-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one obtained in Example 38. Thereafter, 754 mg (12.0 mmol) of sodium cyanoborohydride was further added thereto, and the obtained mixture was then stirred at a room temperature for 3 hours. Thereafter, 0.82 ml (10 mmol) of a 37% formalin aqueous solution was added to the reaction solution, and the obtained mixture was further stirred for 2.5 hours. Thereafter, the reaction solution was concentrated, and a saturated sodium bicarbonate aqueous solution and methylene chloride were then added to the concentrate. An organic layer was separated, and was then dried over anhydrous sodium sulfate. Thereafter, the solvent was distilled away under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate methanol=15:30:1), so as to obtain 344 mg (61%) of 7-[(2-hydroxyethyl)methylamino]-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one in the form of a yellow solid. the obtained mixture was further stirred for 2.5 hours Thereafter, the reaction solution was concentrated a saturated sodium bicarbonate aqueous solution and methylene chloride were then added to the concentrate An organic layer was separated was then dried over anhydrous sodium sulfate Thereafter, the solvent was distilled away under reduced pressure The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate methanol=15:30:1)

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

Procedure: 120 mg (1.0 mmol) of glycolaldehyde dimer and 2 ml of acetic acid were added to 10 ml of a methanol solution containing 473 mg (1.55 mmol) of the 7-amino-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one obtained in Example 38. Thereafter, 754 mg (12.0 mmol) of sodium cyanoborohydride was further added thereto, and the obtained mixture was then stirred at a room temperature for 3 hours. Thereafter, 0.82 ml (10 mmol) of a 37% formalin aqueous solution was added to the reaction solution, and the obtained mixture was further stirred for 2.5 hours. Thereafter, the reaction solution was concentrated, and a saturated sodium bicarbonate aqueous solution and methylene chloride were then added to the concentrate. An organic layer was separated, and was then dried over anhydrous sodium sulfate. Thereafter, the solvent was distilled away under reduced pressure. The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate methanol=15:30:1), so as to obtain 344 mg (61%) of 7-[(2-hydroxyethyl)methylamino]-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one in the form of a yellow solid. the obtained mixture was further stirred for 2.5 hours Thereafter, the reaction solution was concentrated a saturated sodium bicarbonate aqueous solution and methylene chloride were then added to the concentrate An organic layer was separated was then dried over anhydrous sodium sulfate Thereafter, the solvent was distilled away under reduced pressure The obtained residue was purified by silica gel chromatography (hexane:ethyl acetate methanol=15:30:1)

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

Procedure: 350 mg (1.15 mmol) of the 7-amino-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one obtained in Example 38 was dissolved in 11.5 ml of methanol, and thereafter, 1.15 ml of acetic acid and 552.5 mg (4.60 mmol) of glycolaldehyde dimer were added thereto. Thereafter, 722 mg (11.5 mmol) of sodium cyanoborohydride was added to the mixture under cooling on ice. The temperature of the mixture was increased to a room temperature, and the mixture was then stirred for 8 hours. Thereafter, a saturated sodium bicarbonate aqueous solution was added to the reaction solution, and the mixture was then extracted with ethyl acetate. The extract was dried over anhydrous sodium sulfate and was then concentrated. The obtained residue was purified by silica gel column chromatography (methylene chloride:methanol=20:1 to 12:1), so as to obtain 61.8 mg (15%) of 7-(2-hydroxyethylamino)-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one in the form of a yellow amorphous substance, as well as 369.3 mg (82%) of 7-[bis(2-hydroxyethyl)amino]-3-(2-trifluoromethylphenyl)-2H-isoquinolin-1-one in the form of a yellow foaming substance. under cooling on ice the mixture was then extracted with ethyl acetate The extract was dried over anhydrous sodium sulfate was then concentrated The obtained residue was purified by silica gel column chromatography (methylene chloride:methanol=20:1 to 12:1)

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

Procedure: Na(OAc)3BH (14.2 mg, 67.0 μmol, Eq: 1.4) was added to a solution of (S)-2-amino-N-((S)-5-((1-(2-cyanophenyl)-1H-indazol-3-yl)methyl)-4-oxo-2,3,4,5-tetrahydrobenzo[b][1,4]oxazepin-3-yl)propanamide (23 mg, 47.9 μmol, Eq: 1.00) and glycolaldehyde dimer (3.16 mg, 26.3 μmol, Eq: 0.55) in DCM (1.5 mL). After 72 h. at RT, the mixture was diluted with NaHCO3 and extracted with DCMThecombined extracts were concentrated. The residue was purified by preparative TLC to give the title compound (16 mg, 95%) as a solid. MS m/z 525 (MH+) extracted with DCMThecombined extracts were concentrated The residue was purified by preparative TLC

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

Procedure: Na(OAc)3BH (14.2 mg, 67.0 μmol, Eq: 1.4) was added to a solution of (S)-2-amino-N-((S)-5-((1-(2-cyanophenyl)-1H-indazol-3-yl)methyl)-4-oxo-2,3,4,5-tetrahydrobenzo[b][1,4]oxazepin-3-yl)propanamide (23 mg, 47.9 μmol, Eq: 1.00) and glycolaldehyde dimer (3.16 mg, 26.3 μmol, Eq: 0.55) in DCM (1.5 mL). After 72 h. at RT, the mixture was diluted with NaHCO3 and extracted with DCMThecombined extracts were concentrated. The residue was purified by preparative TLC to give the title compound (16 mg, 95%) as a solid. MS m/z 525 (MH+) extracted with DCMThecombined extracts were concentrated The residue was purified by preparative TLC

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

Procedure: Sodium cyanoborohydride (5.14 mg, 81.8 μmol, Eq: 1.50) was added to a solution of (S)-N-((S)-5-((6-bromo-2-methoxynaphthalen-1-yl)methyl)-4-oxo-2′,3′,4,5,5′,6′-hexahydro-3H-spiro[benzo[b][1,4]oxazepine-2,4′-pyran]-3-yl)-2-amino-propanamide (31 mg, 54.5 μmol, Eq: 1.00), glycolaldehyde dimer (3.6 mg, 30.0 μmol, Eq: 0.55) and acetic acid (3.27 mg, 3.15 μl, 54.5 μmol, Eq: 1.00) in MeOH (1 mL) and the mixture was stirred at RT overnight. The mixture was diluted with 1 N HCl and H2O/1 N NaOH was added to adjust the pH to ˜8-9. The mixture was extracted with EtOAc, the combined extracts were washed with brine, dried over Na2SO4 and concentrated. The resulting material was purified by flash chromatography to afford the title compound (30 mg, 90% yield). MS m/z 613.8 (MH+). was added The mixture was extracted with EtOAc the combined extracts were washed with brine dried over Na2SO4 concentrated The resulting material was purified by flash chromatography

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

Procedure: Sodium cyanoborohydride (5.14 mg, 81.8 μmol, Eq: 1.50) was added to a solution of (S)-N-((S)-5-((6-bromo-2-methoxynaphthalen-1-yl)methyl)-4-oxo-2′,3′,4,5,5′,6′-hexahydro-3H-spiro[benzo[b][1,4]oxazepine-2,4′-pyran]-3-yl)-2-amino-propanamide (31 mg, 54.5 μmol, Eq: 1.00), glycolaldehyde dimer (3.6 mg, 30.0 μmol, Eq: 0.55) and acetic acid (3.27 mg, 3.15 μl, 54.5 μmol, Eq: 1.00) in MeOH (1 mL) and the mixture was stirred at RT overnight. The mixture was diluted with 1 N HCl and H2O/1 N NaOH was added to adjust the pH to ˜8-9. The mixture was extracted with EtOAc, the combined extracts were washed with brine, dried over Na2SO4 and concentrated. The resulting material was purified by flash chromatography to afford the title compound (30 mg, 90% yield). MS m/z 613.8 (MH+). was added The mixture was extracted with EtOAc the combined extracts were washed with brine dried over Na2SO4 concentrated The resulting material was purified by flash chromatography

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

Procedure: 1 g (2.2 mmol) of (7S)-N-[(3,4-dimethoxybicyclo[4.2.0]octa-1,3,5-trien-7-yl)methyl]-3-(7,8-dimethoxy-1,2,4,5-tetrahydro-3H-3-benzazepin-3-yl)-3-oxopropan-1-amine (obtained by returning the compound of Example 4 to the base), 0.4 g (3.3 mmol) of 1,4-dioxane-2,5-diol, 0.7 g (3.3 mmol) of sodium triacetoxyborohydride and 8 mL of methylene chloride are mixed together. After 45 minutes at ambient temperature, the reaction mixture is rendered basic with 10 mL of 1N sodium hydroxide solution and stirring is carried out for half an hour. The phases are separated, the aqueous phase is extracted twice with ethyl acetate, the organic phases are combined, washed with water and then with saturated NaCl solution and dried over MgSO4. After evaporating off the solvent, a residue is isolated which is chromatographed over silica (eluant: CH2Cl2/EtOH/NH4OH 95/5/0.5) to yield the expected compound, which is converted into its hydrochloride using 2N ethereal HCl solution. are mixed together is carried out for half an hour The phases are separated the aqueous phase is extracted twice with ethyl acetate washed with water with saturated NaCl solution and dried over MgSO4 After evaporating off the solvent a residue is isolated which is chromatographed over silica (eluant: CH2Cl2/EtOH/NH4OH 95/5/0.5)