Journal of Medicinal Chemistry
ARTICLE
J = 12.1, 5.3, 3.6 Hz, 3R-H). 13CNMR (75 MHz, CDCl3) δppm 12.1, 15.4,
18.6, 20.3 (CH2), 21.4, 22.2 (CH2), 22.5, 22.8, 23.8 (CH2), 24.1 (CH2),
28.0, 28.2 (CH2), 30.3, 31.9 (CH2), 34.3 (CH2), 35.8, 36.1 (CH2), 38.1
(C), 39.5 (CH2), 39.8 (CH2), 42.7 (C), 46.7, 56.0, 56.2, 72.2, 73.4 (C5),
77.5, 77.9, 170.2 (CH3COO). MS m/z (%): 479.9 (33) [M + H]+, 441.6
(53), 419.1 (19), 102.6 (100), 100.6 (38). HRMS (ESI), positive mode,
m/z [M + Na]+ calcd for C29H50O5Na, 501.3556; found, 501.3586.
5r,6β,7β-Trihydroxycholestan-3β-yl Acetate (41). To a
solution of cholestane-3β,5R,6β,7β-tetrol (34, 40 mg) in toluene
(8 mL) and vinyl acetate (300 μL), lipase AY (400 mg) was added.
The mixture was shaken at 200 rpm at 45 °C. After 24 h, the enzyme was
filtered and the solvent evaporated. FCC (petroleum ether, ethyl acetate
1:2 to 1:3) afforded the 5R,6β,7β-trihydroxycholestan-3β-yl acetate
(41, 40.3 mg, 92%). Mp 178ꢀ179 °C (EtOH); lit.,61 179ꢀ180 °C. IR
(film) 3442, 2927, 2855, 1733, 1460, 1379, 1272, 1123, 1070, 1030, 891,
740 cmꢀ1. 1H NMR (300 MHz, CDCl3) δ ppm 0.68 (3H, s, 18-CH3),
0.86 and 0.86 (each 3H, 2d, J = 6.6 Hz, 26-CH3 and 27-CH3), 0.91 (3H,
d, J = 6.5 Hz, 21-CH3), 1.16 (3H, s, 19-CH3), 2.03 (3H, s, 3β-
CH3COO), 2.22 (1H, dd, J = 13.0, 11.6 Hz), 3.46 (1H, d, J = 3.7 Hz,
6R-H), 3.83 (1H, dd, J = 9.4, 3.7 Hz, 7R-H), 5.13 (1H, m, 3R-H). 13C
NMR (101 MHz, DMSO-d6) ppm 12.0, 16.6, 18.6, 20.7 (CH2), 21.1,
22.3, 22.6, 23.2 (CH2), 26.6 (CH2), 26.7 (CH2), 27.3, 28.3 (CH2), 31.6
(CH2), 35.2, 35.6 (CH2), 37.0 (CH2), 37.1 (C), 37.8, 38.9 (CH2), 39.7
(CH2), 43.0 (C), 43.2, 54.9, 55.3, 70.6, 71.2, 75.0 (C), 77.1, 169.8 (C).
MS m/z (%): 477.9 (21) [M ꢀ H]+, 407.9 (32), 378.9 (28), 339.4 (58),
337.5 (22), 325.3 (35), 311.3 (46), 165.0 (65), 149.0 (100).
(CH2), 21.4, 21.4, 22.5, 22.8, 23.9 (CH2), 24.1 (CH2), 26.6 (CH2), 28.0,
28.2 (CH2), 30.7, 31.4 (CH2), 31.8 (CH2), 35.8, 36.1 (CH2), 36.8
(CH2), 38.4 (C), 39.5 (CH2), 39.8 (CH2), 42.7 (C), 45.0, 55.7, 56.2,
70.7, 74.9 (C5), 76.2, 170.2 (CH3COO), 170.7 (CH3COO). MS m/z
(%): 505.3 (55) [M + H]+, 447.6 (79), 429.0 (100), 413.3 (84), 382.2
(73), 322.0 (90), 180.5 (91).
3β,5r-Dihydroxycholestan-6β-yl Acetate (45). To a solution
of 5R-hydroxycholestane-3β,6β-diyl diacetate (44, 85 mg) in DIPE
(aqueous, saturated, 20 mL), lipase AY (3600 mg) was added. The
mixture was shaken at 200 rpm at 45 °C. After 10 days the enzyme was
filtered and the solvent evaporated. FCC (petroleum ether, ethyl acetate
1:1) afforded the 3β,5R-dihydroxycholestan-6β-yl acetate (45, 48.3 mg,
62%). Mp 145ꢀ146 °C (EtOH); lit.,65 143 ꢀ 144 °C. IR (film) 3474,
2935, 2863, 1716, 1457, 1464, 1372, 1266, 1248, 1160, 1038, 928 cmꢀ1
.
1H NMR (400 MHz, CDCl3) δ ppm 0.68 (3H, s, 18-CH3), 0.86 and
0.86 (each 3H, 2d, J = 6.6 Hz, 26-CH3 and 27-CH3), 0.90 (3H, d, J = 6.5
Hz, 21-CH3), 1.14 (3H, s, 19-CH3), 2.06 (3H, s, CH3COO), 4.08 (1H,
tt, J = 10.4, 5.1 Hz, 3R-H), 4.71 (1H, m, 6R-H); 13C NMR (101 MHz,
CDCl3) δ ppm 12.2, 16.5, 18.6, 21.1 (CH2), 21.4, 22.5, 22.8, 23.8
(CH2), 24.1 (CH2), 28.0, 28.2 (CH2), 30.6 (CH2), 30.7, 31.4 (CH2),
32.0 (CH2), 35.8, 36.1 (CH2), 38.4 (C), 39.5 (CH2), 39.9 (CH2), 40.5
(CH2), 42.7 (C), 45.4, 55.8, 56.2, 67.3, 75.3 (C5), 76.1, 170.3
(CH3COO). MS m/z (%): 463.5 (51) [M + H]+, 413.1 (44), 384.9
(50), 358.0 (73), 328.5 (41), 259.9 (100), 224.5 (50), 109.3 (46), 85.2
(59), 60.8 (46).
3β,5r,6β-Trihydroxycholestan-7β-yl Acetate (46). To a
solution of 5R,6β-dihydroxycholestane-3β,7β-diyl diacetate (33, 130
mg) in DIPE (aqueous, saturated, 30 mL), lipase AY (2730 mg) was
added . The mixture was shaken at 200 rpm at 45 °C. After 30 days the
enzyme was filtered and the solvent evaporated. FCC (petroleum ether,
ethyl acetate 1:1 to 1:5) afforded the 3β,5R,6β-trihydroxycholestan-7β-
yl acetate (46, 88.4 mg, 74%). Mp 218ꢀ219 °C (EtOH). IR (film) 3478,
2951, 2872, 1714, 1468, 1374, 1266, 1142, 1086, 1031, 973, 888, 757,
3β-Hydroxycholest-5-en-7β-yl Acetate (42). To a solution of
cholest-5-ene-3β,7β-diyl diacetate (30, 120 mg) in DIPE (aqueous
saturated, 10 mL), lipase AY (2500 mg) was added. The mixture was
shaken at 200 rpm at 45 °C. After 10 days, the enzyme was filtered and
the solvent evaporated. FCC (petroleum ether, ethyl acetate 8:1 to 1:1)
afforded the 3β-hydroxycholest-5-en-7β-yl acetate (42, 70 mg, 64%).
Mp 132ꢀ133 °C (MeOH); lit.,63 83ꢀ84 °C. IR (film) 3402, 2948,
2867, 1733, 1466, 1372, 1239, 1175, 1141, 1057, 1018, 949, 757 cmꢀ1
.
695 cmꢀ1. H NMR (300 MHz, DMSO-d6) δ ppm 0.63 (3H, s, 18-
1
1H NMR (300 MHz, CDCl3) δ ppm 0.67 (3H, s, 18-CH3), 0.83 and
0.84 (each 3H, 2d, J = 6.6 Hz, 26-CH3 and 27-CH3), 0.88 (3H, d, J = 6.5
Hz, 21-CH3), 1.04 (3H, s, 19-CH3), 2.00 (3H, s, 7β-CH3COO), 3.51
(1H, tt, J = 11.4, 4.6 Hz, 3R-H), 4.99 (1H, td, J = 8.7, 1.6 Hz, 7R-H), 5.17
(1H, t, J = 1.6 Hz, 6-H); 13C NMR (75 MHz, CDCl3) δ ppm 11.7, 18.7,
19.0, 21.0 (CH2), 21.6, 22.5, 22.8, 23.7 (CH2), 25.1 (CH2), 27.9, 28.3
(CH2), 31.3 (CH2), 35.6, 36.0 (CH2), 36.3 (C), 36.4, 36.7 (CH2), 39.3
(CH2), 39.4 (CH2), 41.5 (CH2), 42.7 (C), 48.1, 55.3, 55.5, 71.0, 75.7,
121.2 (C6), 145.3 (C5), 171.3 (CH3COO). MS m/z (%): 443.9 (85)
[M ꢀ H]+, 431.1 (63), 369.7 (37), 353.8 (64), 339.7 (100), 310.4 (99),
136.6 (63), 116.9 (45). HRMS (ESI), positive mode, m/z [M + Na]+
calcd for C29H48O3Na, 467.3501; found, 467.3526.
5r-Hydroxycholestane-3β,6β-diyl Diacetate (44). To a so-
lution of 5R,6β-dihydroxycholestan-3β-yl acetate (44, 120 mg, 0.259
mmol) and DMAP (15.8 mg, 0.130 mmol) in THF (2 mL), acetic
anhydride 98.5% (99.4 μL, 1.036 mmol) was added. The reaction
mixture was stirred overnight at room temperature, then stirred with
MeOH for 30 min, concentrated under vacuum, and the residue was
dissolved in diethyl ether. The resulting organic phase was stirred with
NH4Cl (saturated aqueous solution) for 1 h and then washed sequen-
tially with NaHCO3 (saturated aqueous solution) and water, dried with
anhydrous Na2SO4, filtered, and evaporated. FCC (petroleum ether,
ethyl acetate 4:1) yielded the 5R-hydroxycholestane-3β,6β-diyl diace-
tate (44, 113.9 mg, 87%). Mp 168ꢀ169 °C (EtOH); lit.,64 165ꢀ167 °C.
IR (film) 3446, 2937, 2867, 1734, 1716, 1458, 1375, 1242, 1164, 1031,
960 cmꢀ1. 1H NMR (400 MHz, CDCl3) δ ppm 0.67 (3H, s, 18-CH3),
0.85 and 0.86 (each 3H, 2d, J = 6.6 Hz, 26-CH3 and 27-CH3), 0.90 (3H,
d, J = 6.5 Hz, 21-CH3), 1.14 (3H, s, 19-CH3), 2.01 (3H, s, CH3COO),
2.06 (3H, s, CH3COO), 4.69 (1H, m, 6R-H), 5.14 (1H, tt, J = 10.8, 5.3
Hz, 3R-H). 13C NMR (101 MHz, CDCl3) δ ppm 12.2, 16.3, 18.6, 21.0
CH3), 0.83 and 0.84 (each 3H, 2d, J = 6.6 Hz, 26-CH3 and 27-CH3),
0.88 (3H, d, J = 6.5 Hz, 21-CH3), 1.01 (3H, s, 19-CH3), 1.93 (3H, s,
CH3COO), 3.41 (1H, dd, J = 5.0, 3.5 Hz, 6R-H), 3.76 (1H, m, 3R-H),
4.13 (1H, s, 5-OH), 4.21 (1H, dd, J = 5.7 Hz, 3-OH), 4.63 (1H, dd, J =
5.0 Hz, 6-OH), 4.90 (1H, dd, J = 10.7, 3.5 Hz, 7R-H); 13C NMR (75
MHz, DMSO-d6) δ ppm 11.8, 16.8, 18.6, 20.8, 21.7 (CH2), 22.3, 22.6, 23.2
(CH2), 25.5 (CH2), 27.3, 28.2 (CH2), 31.0 (CH2), 32.0 (CH2), 34.6, 35.0,
35.6 (CH2), 37.2 (C), 38.9 (CH2), 39.6 (CH2), 40.6 (CH2), 43.1 (C),
43.7, 54.5, 54.7, 65.2, 74.3, 75.1 (C5), 75.8, 169.9 (CH3COO). MS m/z
(%): 477.8 (100) [M ꢀ H]+, 468.3 (83), 384.9 (81), 326.7 (69), 316.7
(51), 265.1 (56), 263.3 (75), 237.9 (64), 236.6 (81). HRMS (ESI), positive
mode, m/z [M + Na]+ calcd for C29H50O5Na, 501.3556; found, 501.3540.
5r-Hydroxycholestane-3β,6β,7β-triyl Triacetate (47). Acetyla-
tion of 5R,6β-dihydroxycholestane-3β,7β-diyl diacetate (33, 150 mg, 0.288
mmol), as described for the preparation of compound 44, using acetic
anhydride (114.6 μL, 1.152 mmol) afforded, after FCC (chloroform,
ethanol 100:0 to 95:5), the 5R-hydroxycholestane-3β,6β,7β-triyl triacetate
(47, 145.9 mg, 90%). Mp 144.5ꢀ146 °C (MeOH); lit.,61 100ꢀ103 °C. IR
(film) 3396, 2934, 2867, 1742, 1716, 1457, 1364, 1241, 1135, 1070, 1028,
879 cmꢀ1. 1H NMR (300 MHz, CDCl3) δ ppm 0.70 (3H, s, 18-CH3), 0.86
and 0.86 (each 3H, 2d, J= 6.6 Hz, 26-CH3 and 27-CH3), 0.90 (3H, d, J=6.5
Hz, 21-CH3), 1.16 (3H, s, 19-CH3), 1.91 (3H, s, CH3COO), 2.03 (3H, s,
CH3COO), 2.10 (3H, s, CH3COO), 5.02 (1H, d, J = 4.0 Hz, 6R-H), 5.19
(1H, m, 3R-H), 5.20 (1H, dd, J = 10.8, 4.0 Hz, 7R-H). 13C NMR (75 MHz,
CDCl3) δppm 12.2, 17.1, 18.7, 21.1, 21.2 (CH2), 21.3, 21.4, 22.5, 22.8, 23.8
(CH2), 25.7 (CH2), 26.6 (CH2), 28.0, 28.5 (CH2), 31.9 (CH2), 35.5, 35.7,
36.1 (CH2), 36.6 (CH2), 37.9 (C), 39.4 (CH2), 39.7 (CH2), 43.5 (C),
44.0, 54.4, 55.2, 70.4, 73.0, 74.7, 75.3 (C5), 170.3 (CH3COO), 170.7
(CH3COO), 171.0 (CH3COO). MS m/z (%): 561.6 (23) [M ꢀ H]+,
515.1 (26), 487.1 (100), 361.1 (25), 326.2 (40).
6390
dx.doi.org/10.1021/jm200803d |J. Med. Chem. 2011, 54, 6375–6393