Reaction Steps
4.00 mmol of cesium carbonate was added to the solution of 2.00 mmol of carboline aldehyde in 20 ml DMF. After stirring 1 hour at room temperature 3.00 mmol of N-(chloroacetyl)morpholine was added to the reaction mixture. The reaction mixture was heated to 65° C. for 24 hours and then poured to ice-water. The precipitate was filtered off, and washed with water and dried. The product was pure enough to use for the next step (75% yield). ESI+ MS: m/z (rel intensity) 324.10 (100, [M+H]+). 1H-NMR (400 MHz, CDCl3): δ 10.21 (s, 1H), 8.66 (d, 1H, J=4.8 Hz), 8.20 (d, 1H, J=4.4 Hz), 8.17 (d, 1H, 6.8 Hz), 7.64 (dt, 1H, J=1.2, 7.2 Hz), 7.38 (q, 2H, J=7.2 Hz), 5.81 (s, 2H), 3.92 (t, 2H, J=4.4 Hz), 3.74 (t, 4H, J=4.8 Hz), 3.61 (t, 2H, J=4.8 Hz). Step b: Preparation of (S)-1-morpholino-2-(1-((5,6,7,8-tetrahydroquinolin-8-ylamino)methyl)-9H-pyrido[3,4-b]indol-9-yl)ethanone. 1.5 mmol of 9-(2-morpholino-2-oxoethyl)-9H-pyrido[3,4-b]indole-1-carbaldehyde, 1.5 mmol of (S)-5,6,7,8-tetrahydroquinolin-8-amine, and 3.00 mmol of sodium triacetoxyborohydride in 20 ml dichloromethane were stirred for 2 hours at room temperature, then reaction was quenched with saturated sodium bicarbonate solution. Organic layer was dried over magnesium sulfate, filtered off, and evaporated. Desired product was purified with column chromatography using dichloromethane:methanol:NH4OH (9:1:0.1) solvent system (85% yield). ESI+ MS: m/z (rel intensity) 456.2 (100, [M+H]+). 1H-NMR (400 MHz, CDCl3): δ 8.38 (d, 1H, J=4.8 Hz), 8.36 (d, 1H, J=1.6 Hz), 8.12 (d, 1H, J=7.6 Hz), 7.92 (d, 1H, J=5.6 Hz), 7.55 (dt, 1H, J=1.2, 8.0 Hz), 7.37 (d, 1H, J=7.6 Hz), 7.29 (q, 2H, J=4.8 Hz), 7.07 (q, 1H, 4.8 Hz), 6.11 (d, 1H, J=18.0 Hz), 5.74 (d, 1H, J=17.6 Hz), 4.46 (d, 1H, J=12.0 Hz), 4.33 (d, 1H, J=12.0 Hz), 4.00 (t, 1H, J=5.6 Hz), 3.82-3.76 (m, 1H), 3.73-3.60 (m, 6H), 3.55-3.50 (m, 1H), 2.78 (t, 2H, J=6.4 Hz), 2.18-2.08 (m, 1H), 1.96-1.66 (m, 4H). Step c: Preparation of (S)-4-(((9-(2-morpholino-2-oxoethyl)-9H-pyrido[3,4-b]indol-1-yl)methyl)(5,6,7,8-tetrahydroquinolin-8-yl)-(4-(Bis-N-tertbutoxycarbonyl)amino) butylcarbamate. 1.00 mmol of (S)-1-morpholino-2-(1-((5,6,7,8-tetrahydroquinolin-8-ylamino)methyl)9H-pyrido[3,4-b]indol-9-yl)ethanone, 1.00 mmol of Bis-1-(N-tertbutoxycarbonyl)-4-oxobutylcarbamate, and 2.00 mmol of sodium triacetoxyborohydride in 20 ml 1,2-dichloroethane were stirred at room temperature for 2 hours. The reaction was quenched with saturated sodium bicarbonate solution. Organic layer was dried over magnesium sulfate, filtered off, and evaporated. The desired product was purified with column chromatography using dichloromethane:methanol:NH4OH (9:1:0.1) in 50% dichloromethane (95% yield). ESI+ MS: m/z (rel intensity) 727.3 (100, [M+H]+). 1H-NMR (400 MHz, CDCl3): δ 8.29 (d, 1H, J=5.2 Hz), 8.21 (d, 1H, J=4.4 Hz), 8.09 (d, 1H, J=7.6 Hz), 7.86 (d, 1H, J=4.8 Hz), 7.56 (dt, 1H, J=1.2, 8.0 Hz), 7.30-7.23 (m, 3H), 7.17 (d, 1H, 18.4 Hz), 6.88 (dt, 1H, J=3.2, 4.4 Hz), 5.80 (d, 1H, J=17.2 Hz), 4.36 (s, 2H), 4.13 (q, 1H, J=6.8 Hz), 3.97 (d, 1H, J=12.8 Hz), 3.89 (d, 1H, J=14.4 Hz), 3.81-3.77 (m, 1H), 3.67-3.56 (m, 4H), 3.46-3.33 (m, 2H), 3.11-3.01 (m, 2H), 2.79-2.65 (m, 1H), 2.64-2.60 (m, 2H), 2.48-2.31 (m, 3H), 2.10-2.04 (m, 2H), 1.93-1.86 (m, 1H), 1.68-1.55 (m, 2H), 1.50 (s, 9H), 1.37 (s, 9H). Step d: Preparation of (S)-2-(1-(((4-aminobutyl)(5,6,7,8-tetrahydroquinolin-8-yl)amino)methyl)-9H-pyrido[3,4-b]indol-9-yl)-1-morpholinoethanone. 0.95 mmol of (S)-4-(((9-(2-morpholino-2-oxoethyl)-9H-pyrido[3,4-b]indol-1-yl)methyl)(5,6,7,8-tetrahydroquinolin-8-yl)-(4-(Bis-N-tertbutoxycarbonyl)amino)butylcarbamate was dissolved in 2 ml of dichloromethane and treated with 2 ml of trifluoroacetic acid. After the reaction was stirred at room temperature for 2 hours it was cooled to 0° C. with ice bath and then neutralized with 1M NaOH solution carefully. The mixture was warmed to ambient temperature. The organic layer was separated, dried over magnesium sulfate, filtered off, and evaporated. The desired product was purified with column chromatography using dichloromethane:methanol:NH4OH (9:1:0.1). ESI+ MS: m/z (rel intensity) 527.2 (100, [M+H]+). 1H-NMR (400 MHz, CDCl3): δ 8.31 (d, 1H, J=5.2 Hz), 8.25 (d, 1H, J=4.0 Hz), 8.11 (d, 1H, J=8.0 Hz), 7.89 (d, 1H, J=4.8 Hz), 7.57 (dt, 1H, J=1.2, 8.0 Hz), 7.31-7.25 (m, 3H), 7.18 (d, 1H, 19.6 Hz), 6.90 (dt, 1H, J=2.8, 4.8 Hz), 5.76 (d, 1H, J=17.2 Hz), 4.36 (s, 2H), 4.12 (t, 1H, J=9.6 Hz), 3.97 (d, 1H, J=12.8 Hz), 3.89 (d, 1H, J=13.6 Hz), 3.79 (td, 1H, J=3.6, 12.0 Hz), 3.70-3.55 (m, 3H), 3.45-3.40 (m, 1H), 3.35-3.30 (m, 1H), 2.82-2.76 (m, 1H), 2.68-2.59 (m, 2H), 2.42-2.32 (m, 2H), 2.21-2.17 (m, 2H), 2.10-2.04 (m, 2H), 1.63-1.59 (m, 1H), 1.25 (s, 2H), 0.97-0.76 (m, 4H). Elemental Analysis (C/H/N): C31H38N6O2×1.25 mol H2O; Calculated: 67.80/7.43/15.30; Found: 67.65/7.12/15.21
