3-aminopropanamide;hydrochloride

Conditions: temp:80.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N-(4-fluoro-3-nitro-phenyl)-N-methyl-benzamide (1.47 g, 5.37 mmol) was taken up in dry acetonitrile under argon, DIEA (2.08 g, 16.11 mmol, 2.80 ml) and β-alanine amide hydrochloride (1.00 g, 8.06 mmol) were added. The reaction mixture was heated to 80° C. for 48 h. The reaction was poured into 1N HCl and the aqueous layer was extracted 3× with ethyl acetate. The combined organic layers were washed with water, brine and dried over magnesium sulfate. The solution was filtered and the solvent removed under reduced pressure. The residue was triturated twice with cold hexane and dried to give N-[4-(2-carbamoyl-ethylamino)-3-nitro-phenyl]-N-methyl-benzamide. (1.75 g, 95%) the aqueous layer was extracted 3× with ethyl acetate The combined organic layers were washed with water, brine dried over magnesium sulfate The solution was filtered the solvent removed under reduced pressure The residue was triturated twice with cold hexane dried

Conditions: temp:80.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N-(4-fluoro-3-nitro-phenyl)-N-methyl-benzamide (1.47 g, 5.37 mmol) was taken up in dry acetonitrile under argon, DIEA (2.08 g, 16.11 mmol, 2.80 ml) and β-alanine amide hydrochloride (1.00 g, 8.06 mmol) were added. The reaction mixture was heated to 80° C. for 48 h. The reaction was poured into 1N HCl and the aqueous layer was extracted 3× with ethyl acetate. The combined organic layers were washed with water, brine and dried over magnesium sulfate. The solution was filtered and the solvent removed under reduced pressure. The residue was triturated twice with cold hexane and dried to give N-[4-(2-carbamoyl-ethylamino)-3-nitro-phenyl]-N-methyl-benzamide. (1.75 g, 95%) the aqueous layer was extracted 3× with ethyl acetate The combined organic layers were washed with water, brine dried over magnesium sulfate The solution was filtered the solvent removed under reduced pressure The residue was triturated twice with cold hexane dried

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

Procedure: 360 mg (1.88 mmol) of EDC were added to a mixture of 400 mg (1.25 mmol) of [(tert-butoxycarbonyl)amino][3-(trifluoromethyl)phenyl]acetic acid and 254 mg (1.88 mmol) of HOBt in 12 ml of DMF, and the mixture was stirred at RT for 30 min 234 mg (1.88 mmol) of beta-alaninamide hydrochloride and 436 μl (2.50 mmol) of N,N′-diisopropylethylamine were added, and the mixture was stirred for another 1 h. The mixture was diluted with ethyl acetate and washed four times with water. The organic phase was dried over sodium sulphate and freed from the solvent on a rotary evaporator. The residue was dried under high vacuum. This gave 446 mg (91% of theory) of the title compound. were added washed four times with water The organic phase was dried over sodium sulphate freed from the solvent on a rotary evaporator The residue was dried under high vacuum

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

Procedure: 360 mg (1.88 mmol) of EDC were added to a mixture of 400 mg (1.25 mmol) of [(tert-butoxycarbonyl)amino][3-(trifluoromethyl)phenyl]acetic acid and 254 mg (1.88 mmol) of HOBt in 12 ml of DMF, and the mixture was stirred at RT for 30 min 234 mg (1.88 mmol) of beta-alaninamide hydrochloride and 436 μl (2.50 mmol) of N,N′-diisopropylethylamine were added, and the mixture was stirred for another 1 h. The mixture was diluted with ethyl acetate and washed four times with water. The organic phase was dried over sodium sulphate and freed from the solvent on a rotary evaporator. The residue was dried under high vacuum. This gave 446 mg (91% of theory) of the title compound. were added washed four times with water The organic phase was dried over sodium sulphate freed from the solvent on a rotary evaporator The residue was dried under high vacuum

Conditions: temp:50.01; pressure:0.00; stir:

Procedure: Palladium (II) acetate (0.011 g, 0.05 mmol) was added to(R)-(-)-1-[(S)-2-(dicyclohexylphosphino)ferrocenyl]ethyldi-t-butylphosphine (0.027 g, 0.05 mmol) in THF (1 mL) in a reaction vessel evacuated and purged with nitrogen. The resulting catalyst mixture was stirred and under nitrogen at 50 °C for 30 minutes. 04-Dec-2013 10:52:28 AM -0500 LiHMDS (1.194 mL, 1.19 mmol), 3-aminopropanamide, HCl (0.077 g, 0.62 mmol) and 4-bromobenzoic acid (0.1 g, 0.50 mmol) was added to a second reaction vessel and purged with nitrogen. were added at 20 °C. The catalyst solution was added to the reaction mixture and the resulting mixture was stirred at 120 °C for 16 hours. LCMS showed starting material only. Desired product not isolated from this reaction.

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

Procedure: A mixture of β-alanine amide hydrochloride (3.11 g, 25 mmol), 5-(phenylethynyl)thiophene-2-carboxaldehyde (5.31 g, 25 mmol), mercaptosuccinic acid (11.26 g, 75 mmol) and diisopropylethylamine (3.6 g, 27.5 mmol) was stirred in acetonitrile (300 mL) at reflux for forty eight hours. The solvent was evaporated and the residue was shaken with 2 N HCl (500 mL) and ethyl acetate (3×200 mL) The ethyl acetate solution was washed with water then brine and dried (MgSO4) and evaporated. The product was purified on a 600 mL. silica dry column by elution with dichloromethane:acetic acid (99:1) followed by dichloromethane:methanol:acetic acid (95:5:1). The appropriate fractions were evaporated. The residue was washed with ether and dried to provide the title compound as a white solid: (2.44 g, 23.5%): m.p. 175-178. 1H-NMR (DMSO-d6) δ 12.6 (broad, 1H), 7.56-7.57 (d, 1H, J=2.4 Hz), 7.54-7.55 (d, 1H, J=3.6 Hz), 7.4-7.46 (m, 4H), 7.28-7.32 (m, 2H), 6.90(s, 1H), 6.15 (s, 0.5 H), 6.12-6.13 (d, 0.5H, J=1.6 Hz), 4.24-4.28 (m, 1H), 4.15-4.19 (dd, 0.5H, J=3.6 Hz), 2.97-3.09 (m, 1H), 2.64-2.74 (m, 1H), 2.36-2.46 (m, 1H), 2.08-2.21 (m, 1H); MS(ESI) 413[M-H], 415[M+H]. Analytical HPLC; Capcell Pak C18, 30% acetonitrile: 70% 0.1% aqueous TFA.51 trans: 49 cis. FTIR (ATR) 1650, 1720 cm−1. Anal. (C20H18N2O4S2) calc. C, 57.95; H, 4.38; N, 6.76. obs. C, 57.59; H, 4.34; N, 6.56. at reflux for forty eight hours The solvent was evaporated was washed with water brine and dried (MgSO4) evaporated The product was purified on a 600 mL silica dry column by elution with dichloromethane:acetic acid (99:1) The appropriate fractions were evaporated The residue was washed with ether dried

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

Procedure: A mixture of β-alanine amide hydrochloride (3.11 g, 25 mmol), 5-(phenylethynyl)thiophene-2-carboxaldehyde (5.31 g, 25 mmol), mercaptosuccinic acid (11.26 g, 75 mmol) and diisopropylethylamine (3.6 g, 27.5 mmol) was stirred in acetonitrile (300 mL) at reflux for forty eight hours. The solvent was evaporated and the residue was shaken with 2 N HCl (500 mL) and ethyl acetate (3×200 mL) The ethyl acetate solution was washed with water then brine and dried (MgSO4) and evaporated. The product was purified on a 600 mL. silica dry column by elution with dichloromethane:acetic acid (99:1) followed by dichloromethane:methanol:acetic acid (95:5:1). The appropriate fractions were evaporated. The residue was washed with ether and dried to provide the title compound as a white solid: (2.44 g, 23.5%): m.p. 175-178. 1H-NMR (DMSO-d6) δ 12.6 (broad, 1H), 7.56-7.57 (d, 1H, J=2.4 Hz), 7.54-7.55 (d, 1H, J=3.6 Hz), 7.4-7.46 (m, 4H), 7.28-7.32 (m, 2H), 6.90(s, 1H), 6.15 (s, 0.5 H), 6.12-6.13 (d, 0.5H, J=1.6 Hz), 4.24-4.28 (m, 1H), 4.15-4.19 (dd, 0.5H, J=3.6 Hz), 2.97-3.09 (m, 1H), 2.64-2.74 (m, 1H), 2.36-2.46 (m, 1H), 2.08-2.21 (m, 1H); MS(ESI) 413[M-H], 415[M+H]. Analytical HPLC; Capcell Pak C18, 30% acetonitrile: 70% 0.1% aqueous TFA.51 trans: 49 cis. FTIR (ATR) 1650, 1720 cm−1. Anal. (C20H18N2O4S2) calc. C, 57.95; H, 4.38; N, 6.76. obs. C, 57.59; H, 4.34; N, 6.56. at reflux for forty eight hours The solvent was evaporated was washed with water brine and dried (MgSO4) evaporated The product was purified on a 600 mL silica dry column by elution with dichloromethane:acetic acid (99:1) The appropriate fractions were evaporated The residue was washed with ether dried

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

Procedure: A mixture of β-alanine amide hydrochloride (3.11 g, 25 mmol), 5-(phenylethynyl)thiophene-2-carboxaldehyde (5.31 g, 25 mmol), mercaptosuccinic acid (11.26 g, 75 mmol) and diisopropylethylamine (3.6 g, 27.5 mmol) was stirred in acetonitrile (300 mL) at reflux for forty eight hours. The solvent was evaporated and the residue was shaken with 2 N HCl (500 mL) and ethyl acetate (3×200 mL) The ethyl acetate solution was washed with water then brine and dried (MgSO4) and evaporated. The product was purified on a 600 mL. silica dry column by elution with dichloromethane:acetic acid (99:1) followed by dichloromethane:methanol:acetic acid (95:5:1). The appropriate fractions were evaporated. The residue was washed with ether and dried to provide the title compound as a white solid: (2.44 g, 23.5%): m.p. 175-178. 1H-NMR (DMSO-d6) δ 12.6 (broad, 1H), 7.56-7.57 (d, 1H, J=2.4 Hz), 7.54-7.55 (d, 1H, J=3.6 Hz), 7.4-7.46 (m, 4H), 7.28-7.32 (m, 2H), 6.90(s, 1H), 6.15 (s, 0.5 H), 6.12-6.13 (d, 0.5H, J=1.6 Hz), 4.24-4.28 (m, 1H), 4.15-4.19 (dd, 0.5H, J=3.6 Hz), 2.97-3.09 (m, 1H), 2.64-2.74 (m, 1H), 2.36-2.46 (m, 1H), 2.08-2.21 (m, 1H); MS(ESI) 413[M-H], 415[M+H]. Analytical HPLC; Capcell Pak C18, 30% acetonitrile: 70% 0.1% aqueous TFA.51 trans: 49 cis. FTIR (ATR) 1650, 1720 cm−1. Anal. (C20H18N2O4S2) calc. C, 57.95; H, 4.38; N, 6.76. obs. C, 57.59; H, 4.34; N, 6.56. at reflux for forty eight hours The solvent was evaporated was washed with water brine and dried (MgSO4) evaporated The product was purified on a 600 mL silica dry column by elution with dichloromethane:acetic acid (99:1) The appropriate fractions were evaporated The residue was washed with ether dried

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

Procedure: The process is carried out as in stage 3 of Example 3, but using 300 mg of 2-fluoro-4-[4-(6-fluoro-1H-benzimidazol-2-yl)carbazol-9-yl]benzonitrile, obtained according to stage 2 of Example 3, 296 mg of potassium carbonate, 1.779 g of beta-alanineamide hydrochloride and 1.445 g of triethylamine in 3 ml of dimethyl sulphoxide. 1.357 ml of a 1M aqueous solution of sodium hydroxide, 1.313 ml of a 30% aqueous solution of hydrogen peroxide and 7 ml of ethanol are then added to the reaction medium. After treatment as in stage 3 of Example 3, and then purification by flash chromatography on silica gel, elution being carried out with mixtures of dichloromethane and ethanol (from 95/5 to 90/10 by volume), followed by crystallization from 10 ml of diisopropyl ether, 200 mg of 2-(2-carbamoylethylamino)-4-[4-(6-fluoro-1H-benzimidazol-2-yl)-9H-carbazol-9-yl]benzamide are thus obtained in the form of a beige solid, the characteristics of which are the following: obtained After treatment as in stage 3 of Example 3, and then purification by flash chromatography on silica gel, elution followed by crystallization from 10 ml of diisopropyl ether, 200 mg of 2-(2-carbamoylethylamino)-4-[4-(6-fluoro-1H-benzimidazol-2-yl)-9H-carbazol-9-yl]benzamide are thus obtained in the form of a beige solid

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

Procedure: A solution of the compound of Example 1 Step 4 (2.0 g, 5.5 mmol) in EtOH (50 mL) was treated with 3-amino-propionamide hydrochloride (1.72 g, 13.8 mmol) and DIEA (0.53 g, 27.7 mmol). The resulting mixture was stirred at reflux for 6 h, then cooled to room temperature and concentrated under vacuum. The crude product was dissolved in CH3OH and purified by preparative reverse-phase HPLC (CH3OH/H2O) on a YMC S10 ODS 50×500 mm column to afford the desired product as a yellow glass (0.96 g, 49%); 1H NMR (CD3OD): δ 7.37 (m, 5H), 5.65 (d, J=16.5 Hz, 1H), 5.30 (d, J=16.5 Hz, 1H), 4.70 (t, J=6.0 Hz, 1H), 3.15 (m, 1H), 2.94 (m, 1H), 2.59 (s, 3H), 2.49 (m, 2H), 1.93 (m, 2H), 0.89 (t, J=7.5 Hz, 3H); LC/MS (ESI): 370 (M+H)+. at reflux for 6 h concentrated under vacuum The crude product was dissolved in CH3OH purified by preparative reverse-phase HPLC (CH3OH/H2O) on a YMC S10 ODS 50×500 mm column

Conditions: temp:0.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: In 10 ml of DMF, 1.0 g (4.9 mmol) of 2,4-dichloro-7-methylthieno[3,2-d]pyrimidine was dissolved, and then 668 mg (5.37 mmol) of β-alanineamide hydrochloride and 1.09 g (10.7 mmol) of triethylamine were added thereto under ice cooling. The reaction solution was stirred at 0° C. for one hour and then allowed to resume room temperature, followed by stirring for further 1 hour. After completion of the reaction, ice water was added to the reaction mixture, followed by extraction with ethyl acetate (50 ml×3). After the organic layer was washed successively with 1N hydrochloric acid, water and brine and dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by silica gel chromatography (eluent: ethyl acetate-hexane=1/8) to give 1.05 g (yield: 79.8%) of the title compound. to resume room temperature by stirring for further 1 hour was added to the reaction mixture followed by extraction with ethyl acetate (50 ml×3) After the organic layer was washed successively with 1N hydrochloric acid, water and brine dried over anhydrous sodium sulfate the solvent was distilled off under reduced pressure The residue was purified by silica gel chromatography (eluent: ethyl acetate-hexane=1/8)

Conditions: temp:80.01; pressure:0.00; stir:; See reaction.notes.procedure_details.

Procedure: N-(4-fluoro-3-nitro-phenyl)-N-methyl-benzamide (1.47 g, 5.37 mmol) was taken up in dry acetonitrile under argon, DIEA (2.08 g, 16.11 mmol, 2.80 ml) and β-alanine amide hydrochloride (1.00 g, 8.06 mmol) were added. The reaction mixture was heated to 80° C. for 48 h. The reaction was poured into 1N HCl and the aqueous layer was extracted 3× with ethyl acetate. The combined organic layers were washed with water, brine and dried over magnesium sulfate. The solution was filtered and the solvent removed under reduced pressure. The residue was triturated twice with cold hexane and dried to give N-[4-(2-carbamoyl-ethylamino)-3-nitro-phenyl]-N-methyl-benzamide. (1.75 g, 95%) the aqueous layer was extracted 3× with ethyl acetate The combined organic layers were washed with water, brine dried over magnesium sulfate The solution was filtered the solvent removed under reduced pressure The residue was triturated twice with cold hexane dried