The mixture was stirred at room PX-478 concentration temperature 4 h. After removing the solvent under reduced pressure, a liquid product appeared. This was recrystallized by column chromatography (n-hexane:ethyl acetate, 4:1). Yield 58 %. FT-IR (KBr, GSK3326595 mouse ν, cm−1): 3373 (OH + NH), 2980, 2974 (aliphatic CH), 1676 (4C=O), 1432 (C=N), 1232 (C=S). Elemental analysis for C33H47FN8O8S2 calculated (%) C: 51.68; H: 6.18; N: 14.61. Found (%): C: 51.47;
H: 6.00; N: 14.67. 1H-NMR (DMSO-d 6 ) δ ppm: 1.12 (t, 12H, 4CH3, J = 7.0 Hz), 1.99 (s, 3H, CH3), 2.98–3.18 (m, 12H, 6CH2), 3.82 (brs, 8H, 4CH2), 4.00 (s, 2H, CH2), 4.56 (s, 2H, CH2), 4.65 (s, 1H, CH), 5.19 (s, 1H, CH), 6.40 (brs, 2H, 2NH), 6.90 (brs, 1H, ar–H), 6.94 (brs, 2H, ar–H). 13C-NMR (DMSO-d 6 ) δ ppm: 9.33 (3CH3), 15.15 (CH3), 21.39 (CH3), 25.75 (CH2), 40.94 (CH2), 43.66 (CH2), 44.04 (CH2), 46.26 (2CH2), 48.64 (CH2), 50.95 (3CH2), 61.71 (CH2), 67.38
(CH2), 67.73 (CH), 70.89 (CH), arC: [107.63 (d, CH, J C–F = 11.8 Hz), 113.45 (CH), 115.47 (CH), 120.42 VX-809 molecular weight (d, C, J C–F = 34.7 Hz), 122.05 (C), 150.83 (d, C, J C–F = 273.3 Hz)], 130.04 (C), 134.26 (C), 155.50 (C=O), 155.65 (C=O), 162.28 (C), 175.25 (2C=O), 189.74 (C=S). ([(5R,6R)-6-([5-[(4-[4-(Ethoxycarbonyl)piperazin-1-yl]-3-fluorophenylamino)methyl]-2-thioxo-1,3,4-oxadiazol-3(2H)-yl]methylamino)-3,3-dimethyl-7-oxo-4-thia-1-aza bicyclo[3.2.0]hept-2-yl]carbonyloxy)(triethyl)ammonium (22) To the mixture of compound 20 (10 mmol), triethylamine (20 mmol), and formaldehyde (50 mmol) in tetrahydrofurane, 6-apa (10 mmol) was added. The mixture was stirred at room temperature 6 h. After removing the solvent under reduced pressure, 5-Fluoracil order a liquid product appeared. This was recrystallized by column chromatography (n-hexane:ethyl acetate, 4:1). Yield 66 %. FT-IR (KBr, ν, cm−1): 3676 (OH), 2901, 2987 (aliphatic CH), 1768 (C=O),
1683 (2 C=O), 1431 (C=N), 1231 (C=S). Elemental analysis for C31H47FN8O6S2 calculated (%): C, 52.38; H, 6.66; N, 15.76. Found (%): C, 52.18; H, 6.79; N, 15.55. 1H-NMR (DMSO-d 6 , δ ppm): 0.99–1.21 (m, 18H, 6CH3), 2.90 (q, 8H, 4CH2, J = 7.0 Hz), 3.38 (q, 8H, 4CH2, J = 7.2 Hz), 3.98–4.08 (m, 4H, 2CH2), 4.55 (s, 1H, CH), 5.26 (s, 1H, CH), 5.30 (s, 1H, CH), 5.38, 5.45 (brs, 2H, 2NH), 6.80 (brs, 1H, ar–H), 6.94 (brs, 2H, ar–H). 13C-NMR (DMSO-d 6 , δ ppm): 9.32 (3CH3), 15.25 (CH3), 27.77 (CH3), 32.62 (CH3), 44.13 (CH2), 45.67 (2CH2), 51.09 (CH2), 51.50 (CH2), 52.61 (CH2), 56.73 (C–(CH3)2), 61.52 (CH2), 62.23 (CH2), 62.99 (CH2), 63.59 (CH2), 65.39 (CH), 67.00 (CH), 73.68 (CH), arC: [107.41 (d, CH, J C–F = 9.8 Hz),113.72 (d, CH, J C–F = 33.0 Hz), 120.07 (CH), 134,64 (d, C, J C–F = 9.1 Hz), 143.12 (d, C, J C–F = 9.5 Hz), 154.47 (d, C, J C–F = 81.2 Hz)], 163.63 (C), 170.45 (C=O), 170.91 (C=O), 172.13 (C=O), 175.29 (C=S).