(1R,2S)-2-[(2-methylpropan-2-yl)oxycarbonylamino]cyclohexane-1-carboxylic acid

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

Procedure: A solution of cis-2-(tert-butoxycarbonylamino)-cyclohexane carboxylic acid (255 mg, 1.05 mmol) in pyridine (10 ml) was stirred at RT. 6-chloro-9H-β-carbolin-8-ylamine (218 mg, 1.00 mmol) was added, followed by EDCI (315 mg, 1.64 mmol) and the slightly turbid pale orange solution was stirred at RT for 16 hr. The solution was diluted with H2O (20 ml) and poured into a separatory funnel containing H2O (50 ml) and EtOAc (50 ml). The mixture was shaken and the layers were separated. The aqueous layer was extracted with EtOAc (50 ml) and the combined organic layers were washed with brine. The organic layer was dried, filtered and concentrated to yield a yellow oil which was placed under high vacuum for 4 hr. The resulting yellow-brown glass was slurried in CH2Cl2 (10 ml) at RT. Trifluoroacetic acid (5 ml) was added and the slurry instantly dissolved to form a clear orange solution. The solution was stirred at RT for 45 min, then concentrated to a brown residue. The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid. A solution of dilute aqueous Na2CO3 was prepared by adding a small volume of 10% aqueous Na2CO3 to H2O (50 ml) until the aqueous solution reached a pH of 10. The yellow solid was dissolved in minimal MeOH and was added drop-wise to the aqueous solution with stirring. The precipitate which formed was filtered, washed with H2O and placed under high vacuum to yield 2-amino-cyclohexanecarboxylic acid (6-chloro-9H-β-carbolin-8-yl)-amide as pale yellow solid (147 mg). 1H-NMR (300 MHz, DMSO-d6): δ 9.00 (s, 1); 8.37-8.34 (m, 1); 8.16-8.13 (m, 2); 7.83 (m, 1); 5.66-5.00 (br s, 2); 3.42-3.40 (m, 1); 2.70-2.62 (m, 1); 2.02-1.90 (m, 1); 1.70-1.54 (m, 5); 1.42-1.29 (m, 2). NH4OAc standard conditions. DAD Rf=1.46 min. M+H=343. poured into a separatory funnel The mixture was shaken the layers were separated The aqueous layer was extracted with EtOAc (50 ml) the combined organic layers were washed with brine The organic layer was dried filtered concentrated to yield a yellow oil which was placed under high vacuum for 4 hr the slurry instantly dissolved to form a clear orange solution The solution was stirred at RT for 45 min concentrated to a brown residue The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid was added drop-wise to the aqueous solution with stirring The precipitate which formed was filtered washed with H2O

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

Procedure: To a mixture of cis-2-tertbutoxycarbonylamino-cyclohexanecarboxylic acid (Acros, 2 g, 8.22 mmol), 2,2,2-trifluoroethylamine (0.969 mL, 12.33 mmol), HOBT (1.888 g, 12.33 mmol) and DIPEA (4.31 mL, 24.66 mmol) in DMF (10 mL) was added EDC (2.364 g, 12.33 mmol) and the mixture allowed to stir at RT for 1 hr. The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL). Organic washed with aq. NaHCO3 (50 mL) and then brine (50 mL). Aqueous extracts backwashed with ethyl acetate (50 mL) and organic extracts combined, dried over sodium sulfate, filtered and concentrated to give 2.55 gm of a white solid, I-5a. 1H NMR (600 MHz, CDCl3) δ 6.18 (m, 1H), 5.07 (m, 1H), 3.75-4.0 (m 3H), 2.62 (m, 1H), 1.3-2.0 (br m, 8H), 1.40 (s, 9H). The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL) Organic washed with aq. NaHCO3 (50 mL) dried over sodium sulfate filtered concentrated

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

Procedure: To a mixture of cis-2-tertbutoxycarbonylamino-cyclohexanecarboxylic acid (Acros, 2 g, 8.22 mmol), 2,2,2-trifluoroethylamine (0.969 mL, 12.33 mmol), HOBT (1.888 g, 12.33 mmol) and DIPEA (4.31 mL, 24.66 mmol) in DMF (10 mL) was added EDC (2.364 g, 12.33 mmol) and the mixture allowed to stir at RT for 1 hr. The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL). Organic washed with aq. NaHCO3 (50 mL) and then brine (50 mL). Aqueous extracts backwashed with ethyl acetate (50 mL) and organic extracts combined, dried over sodium sulfate, filtered and concentrated to give 2.55 gm of a white solid, I-5a. 1H NMR (600 MHz, CDCl3) δ 6.18 (m, 1H), 5.07 (m, 1H), 3.75-4.0 (m 3H), 2.62 (m, 1H), 1.3-2.0 (br m, 8H), 1.40 (s, 9H). The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL) Organic washed with aq. NaHCO3 (50 mL) dried over sodium sulfate filtered concentrated

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

Procedure: A solution of cis-2-(tert-butoxycarbonylamino)-cyclohexane carboxylic acid (255 mg, 1.05 mmol) in pyridine (10 ml) was stirred at RT. 6-chloro-9H-β-carbolin-8-ylamine (218 mg, 1.00 mmol) was added, followed by EDCI (315 mg, 1.64 mmol) and the slightly turbid pale orange solution was stirred at RT for 16 hr. The solution was diluted with H2O (20 ml) and poured into a separatory funnel containing H2O (50 ml) and EtOAc (50 ml). The mixture was shaken and the layers were separated. The aqueous layer was extracted with EtOAc (50 ml) and the combined organic layers were washed with brine. The organic layer was dried, filtered and concentrated to yield a yellow oil which was placed under high vacuum for 4 hr. The resulting yellow-brown glass was slurried in CH2Cl2 (10 ml) at RT. Trifluoroacetic acid (5 ml) was added and the slurry instantly dissolved to form a clear orange solution. The solution was stirred at RT for 45 min, then concentrated to a brown residue. The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid. A solution of dilute aqueous Na2CO3 was prepared by adding a small volume of 10% aqueous Na2CO3 to H2O (50 ml) until the aqueous solution reached a pH of 10. The yellow solid was dissolved in minimal MeOH and was added drop-wise to the aqueous solution with stirring. The precipitate which formed was filtered, washed with H2O and placed under high vacuum to yield 2-amino-cyclohexanecarboxylic acid (6-chloro-9H-β-carbolin-8-yl)-amide as pale yellow solid (147 mg). poured into a separatory funnel The mixture was shaken the layers were separated The aqueous layer was extracted with EtOAc (50 ml) the combined organic layers were washed with brine The organic layer was dried filtered concentrated to yield a yellow oil which was placed under high vacuum for 4 hr the slurry instantly dissolved to form a clear orange solution The solution was stirred at RT for 45 min concentrated to a brown residue The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid was added drop-wise to the aqueous solution with stirring The precipitate which formed was filtered washed with H2O

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

Procedure: A solution of cis-2-(tert-butoxycarbonylamino)-cyclohexane carboxylic acid (255 mg, 1.05 mmol) in pyridine (10 ml) was stirred at RT. 6-chloro-9H-β-carbolin-8-ylamine (218 mg, 1.00 mmol) was added, followed by EDCI (315 mg, 1.64 mmol) and the slightly turbid pale orange solution was stirred at RT for 16 hr. The solution was diluted with H2O (20 ml) and poured into a separatory funnel containing H2O (50 ml) and EtOAc (50 ml). The mixture was shaken and the layers were separated. The aqueous layer was extracted with EtOAc (50 ml) and the combined organic layers were washed with brine. The organic layer was dried, filtered and concentrated to yield a yellow oil which was placed under high vacuum for 4 hr. The resulting yellow-brown glass was slurried in CH2Cl2 (10 ml) at RT. Trifluoroacetic acid (5 ml) was added and the slurry instantly dissolved to form a clear orange solution. The solution was stirred at RT for 45 min, then concentrated to a brown residue. The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid. A solution of dilute aqueous Na2CO3 was prepared by adding a small volume of 10% aqueous Na2CO3 to H2O (50 ml) until the aqueous solution reached a pH of 10. The yellow solid was dissolved in minimal MeOH and was added drop-wise to the aqueous solution with stirring. The precipitate which formed was filtered, washed with H2O and placed under high vacuum to yield 2-amino-cyclohexanecarboxylic acid (6-chloro-9H-β-carbolin-8-yl)-amide as pale yellow solid (147 mg). poured into a separatory funnel The mixture was shaken the layers were separated The aqueous layer was extracted with EtOAc (50 ml) the combined organic layers were washed with brine The organic layer was dried filtered concentrated to yield a yellow oil which was placed under high vacuum for 4 hr the slurry instantly dissolved to form a clear orange solution The solution was stirred at RT for 45 min concentrated to a brown residue The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid was added drop-wise to the aqueous solution with stirring The precipitate which formed was filtered washed with H2O

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

Procedure: To 5.05 g (20.75 mmol) of the acid of step 2 dissolved in 40 mL DMF was added 1.92 g (20.75 mmol) aminoacetonitrile hydrochloride, 3.98 g (20.75 mmol) of EDCI, 2.80 g (20.75 mmol) of HOBT and 7.98 mL (72.63 mmol) of N-methylmorpholine. The reaction mixture was stirred at room temperature overnight, partitioned between ethyl acetate and water, dried over magnesium sulfate and concentrated to obtain 5.83 g of the crude tert-butyl (1S,2R)-2-{[(cyanomethyl)amino]carbonyl}cyclohexylcarbamate. partitioned between ethyl acetate and water dried over magnesium sulfate concentrated

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

Procedure: To 6.05 grams (22.3 mmol) of the ester of step 1 dissolved in 25 mL methanol was added 2.6 g (55.74 mmol) lithium hydroxide monohydrate dissolved in 10 mL water. The mixture was stirred for 48 hours, partitioned between ethyl acetate and 1 N HCl, dried over magnesium sulfate and concentrated to obtain 5.05 g of the crude (1R,2S)-2-[(tert-butoxycarbonyl)amino]cyclohexanecarboxylic acid. partitioned between ethyl acetate and 1 N HCl dried over magnesium sulfate concentrated

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

Procedure: N-Cyclopropyl-N-piperidin-4-yl-3-trifluoromethyl-benzenesulfonamide (0.348 g, 1.00 mmol), HOBt (0.135 g, 1.0 mmol), EDCI (0.230 g, 1.2 mmol) and rac-cis-2-(tert-butoxycarbonylamino)-cyclohexanecarboxylic acid (0.276 g, 1.1 mmol) were dissolved in 5 ml of dry CH2Cl2. The mixture was allowed to stir at room temperature for 12 hours. The reaction mixture was diluted with 30 ml of CHCl3 and washed with 10 ml of H2O. The organic layer was dried over MgSO4 and concentrated to dryness. The crude product was purified through a column of silica gel with a gradient of 30% to 70% EtOAc in hexane to afford the rac-((1R,2S)-2-{4-[cyclopropyl-(3-trifluoromethyl-benzenesulfonyl)-amino]-piperidine-1-carbonyl}-cyclohexyl)-carbamic acid tert-butyl ester (573 mg, 100%). The product (0.120 g, 0.21 mmol) was taken up in 1 ml of EtOAc, and cooled in ice bath. 4N HCl-EtOAc (1.0 ml) was added slowly, and the mixture was stirred at room temperature for 5 hours. The solvent was removed and remaining material was triturated with ethyl ether to give the desired product 4 (colorless amorphous, 90 mg, 84%) as hydrochloride salt. LC: 100%. MS: m/z=473. washed with 10 ml of H2O The organic layer was dried over MgSO4 concentrated to dryness The crude product was purified through a column of silica gel with a gradient of 30% to 70% EtOAc in hexane

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

Procedure: A solution of cis-2-(tert-butoxycarbonylamino)-cyclohexane carboxylic acid (255 mg, 1.05 mmol) in pyridine (10 ml) was stirred at RT. 6-chloro-9H-β-carbolin-8-ylamine (218 mg, 1.00 mmol) was added, followed by EDCI (315 mg, 1.64 mmol) and the slightly turbid pale orange solution was stirred at RT for 16 hr. The solution was diluted with H2O (20 ml) and poured into a separatory funnel containing H2O (50 ml) and EtOAc (50 ml). The mixture was shaken and the layers were separated. The aqueous layer was extracted with EtOAc (50 ml) and the combined organic layers were washed with brine. The organic layer was dried, filtered and concentrated to yield a yellow oil which was placed under high vacuum for 4 hr. The resulting yellow-brown glass was slurried in CH2Cl2 (10 ml) at RT. Trifluoroacetic acid (5 ml) was added and the slurry instantly dissolved to form a clear orange solution. The solution was stirred at RT for 45 min, then concentrated to a brown residue. The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid. A solution of dilute aqueous Na2CO3 was prepared by adding a small volume of 10% aqueous Na2CO3 to H2O (50 ml) until the aqueous solution reached a pH of 10. The yellow solid was dissolved in minimal MeOH and was added drop-wise to the aqueous solution with stirring. The precipitate which formed was filtered, washed with H2O and placed under high vacuum to yield 2-amino-cyclohexanecarboxylic acid (6-chloro-9H-β-carbolin-8-yl)-amide as pale yellow solid (147 mg). 1H-NMR (300 MHz, DMSO-d6): δ 9.00 (s, 1); 8.37-8.34 (m, 1); 8.16-8.13 (m, 2); 7.83 (m, 1); 5.66-5.00 (br s, 2); 3.42-3.40 (m, 1); 2.70-2.62 (m, 1); 2.02-1.90 (m, 1); 1.70-1.54 (m, 5); 1.42-1.29 (m, 2). NH4OAc standard conditions. DAD Rf=1.46 min. M+H=343. poured into a separatory funnel The mixture was shaken the layers were separated The aqueous layer was extracted with EtOAc (50 ml) the combined organic layers were washed with brine The organic layer was dried filtered concentrated to yield a yellow oil which was placed under high vacuum for 4 hr the slurry instantly dissolved to form a clear orange solution The solution was stirred at RT for 45 min concentrated to a brown residue The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid was added drop-wise to the aqueous solution with stirring The precipitate which formed was filtered washed with H2O

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

Procedure: To a mixture of cis-2-tertbutoxycarbonylamino-cyclohexanecarboxylic acid (Acros, 2 g, 8.22 mmol), 2,2,2-trifluoroethylamine (0.969 mL, 12.33 mmol), HOBT (1.888 g, 12.33 mmol) and DIPEA (4.31 mL, 24.66 mmol) in DMF (10 mL) was added EDC (2.364 g, 12.33 mmol) and the mixture allowed to stir at RT for 1 hr. The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL). Organic washed with aq. NaHCO3 (50 mL) and then brine (50 mL). Aqueous extracts backwashed with ethyl acetate (50 mL) and organic extracts combined, dried over sodium sulfate, filtered and concentrated to give 2.55 gm of a white solid, I-5a. 1H NMR (600 MHz, CDCl3) δ 6.18 (m, 1H), 5.07 (m, 1H), 3.75-4.0 (m 3H), 2.62 (m, 1H), 1.3-2.0 (br m, 8H), 1.40 (s, 9H). The reaction was quenched into 10% aq. KHSO4 (100 mL) and product extracted with ethyl acetate (100 mL) Organic washed with aq. NaHCO3 (50 mL) dried over sodium sulfate filtered concentrated

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

Procedure: A solution of cis-2-(tert-butoxycarbonylamino)-cyclohexane carboxylic acid (255 mg, 1.05 mmol) in pyridine (10 ml) was stirred at RT. 6-chloro-9H-β-carbolin-8-ylamine (218 mg, 1.00 mmol) was added, followed by EDCI (315 mg, 1.64 mmol) and the slightly turbid pale orange solution was stirred at RT for 16 hr. The solution was diluted with H2O (20 ml) and poured into a separatory funnel containing H2O (50 ml) and EtOAc (50 ml). The mixture was shaken and the layers were separated. The aqueous layer was extracted with EtOAc (50 ml) and the combined organic layers were washed with brine. The organic layer was dried, filtered and concentrated to yield a yellow oil which was placed under high vacuum for 4 hr. The resulting yellow-brown glass was slurried in CH2Cl2 (10 ml) at RT. Trifluoroacetic acid (5 ml) was added and the slurry instantly dissolved to form a clear orange solution. The solution was stirred at RT for 45 min, then concentrated to a brown residue. The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid. A solution of dilute aqueous Na2CO3 was prepared by adding a small volume of 10% aqueous Na2CO3 to H2O (50 ml) until the aqueous solution reached a pH of 10. The yellow solid was dissolved in minimal MeOH and was added drop-wise to the aqueous solution with stirring. The precipitate which formed was filtered, washed with H2O and placed under high vacuum to yield 2-amino-cyclohexanecarboxylic acid (6-chloro-9H-β-carbolin-8-yl)-amide as pale yellow solid (147 mg). poured into a separatory funnel The mixture was shaken the layers were separated The aqueous layer was extracted with EtOAc (50 ml) the combined organic layers were washed with brine The organic layer was dried filtered concentrated to yield a yellow oil which was placed under high vacuum for 4 hr the slurry instantly dissolved to form a clear orange solution The solution was stirred at RT for 45 min concentrated to a brown residue The residue was azeotroped from toluene (3×10 ml) to yield a yellow solid was added drop-wise to the aqueous solution with stirring The precipitate which formed was filtered washed with H2O