(2S)-2-aminopropanenitrile

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

Procedure: 2-Hydroxy-2-(4-(5-(3-phenyl-4-(trifluoromethyl)isoxazol-5-yl)-1,2,4-oxadiazol-3-yl)phenyl)acetic acid (Int-Vb, 25 mg, 0.058 mmol), (S)-2-aminopropanenitrile (Int-X, 6.09 mg, 0.087 mmol), HATU (28.7 mg, 0.075 mmol), and 4-methylmorpholine (23.45 mg, 0.232 mmol) were dissolved in DMF (1 mL). The reaction mixture was stirred for 1 h, and then purified by prep HPLC (PHENOMENEX® Luna 5u 21.2×250 mm gradient elution with Method 2—CH3CN/water containing 0.1% trifluoroacetic acid as defined above, 0% B to 100% B over 30 min, 20 mL/min, 220 nM, product retention=25.5 min) to provide single enantiomer N—((S)-1-cyanoethyl)-2-hydroxy-2-(4-(5-(3-phenyl-4-(trifluoromethyl)isoxazol-5-yl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide (10 mg, 0.019 mmol, 32.1% yield): LCMS=484.0 [M+H]+; 1H NMR (400 MHz, methanol-d4) δ ppm 8.12-8.26 (2 H, m), 7.50-7.78 (7 H, m), 5.17 (1 H, s), 4.85-4.94 (1 H, m), 1.56 (3 H, d, J=7.26 Hz); HPLC peak RT=9.9 min (Method A). purified by prep HPLC (PHENOMENEX® Luna 5u 21.2×250 mm gradient elution with Method 2—CH3CN/water containing 0.1% trifluoroacetic acid as defined above, 0% B to 100% B over 30 min, 20 mL/min, 220 nM, product retention=25.5 min)

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

Procedure: 2-Hydroxy-2-(4-(5-(3-phenyl-4-(trifluoromethyl)isoxazol-5-yl)-1,2,4-oxadiazol-3-yl)phenyl)acetic acid (Int-Va, 37 mg, 0.086 mmol), (S)-2-aminopropanenitrile (Int-XI, 9.0 mg, 0.13 mmol) [see also, McLaughlin, M. et al., J. Org. Chem., 68:50-54 (2003)], 4-methylmorpholine (34.7 mg, 0.343 mmol), and HATU (42.4 mg, 0.112 mmol) were dissolved in DMF (1 mL). After stirring 1 h, the reaction mixture was purified by prep HPLC (PHENOMENEX® 21×250 mm, gradient elution with Method 1—MeOH/water containing 0.1% trifluoroacetic acid as defined above, 0% B to 100% B over 38 min, 25 mL/min, 220 nM, product retention=29.3 min) to provide single enantiomer N—((S)-1-cyanoethyl)-2-hydroxy-2-(4-(5-(3-phenyl-4-(trifluoromethyl)isoxazol-5-yl)-1,2,4-oxadiazol-3-yl)phenyl)acetamide (12 mg, 0.023 mmol, 27.3% yield): [M+H]+=484.2; 1H NMR (400 MHz, methanol-d4) δ ppm 8.13-8.25 (2 H, m), 7.54-7.77 (7 H, m), 5.19 (1 H, s), 4.87 (1 H, q), 1.54 (3 H, d, J=7.04 Hz); HPLC peak RT=11.1 min (Method A). the reaction mixture was purified by prep HPLC (PHENOMENEX® 21×250 mm, gradient elution with Method 1—MeOH/water containing 0.1% trifluoroacetic acid as defined above, 0% B to 100% B over 38 min, 25 mL/min, 220 nM, product retention=29.3 min)