Journal of Medicinal Chemistry
Article
dissolved in methanol/water (2:1, 12 mL), and the resulting solution
was treated with K2CO3 (0.26 g, 1.86 mmol) and refluxed for 3 h. After
cooling, the reaction mixture was extracted with CH2Cl2 (3 × 10 mL),
and the combined organic layers dried over Na2SO4. The solvent was
removed in vacuo, and the residue submitted to mpc. Elution by light
petroleum−ethyl acetate (80:20) afforded pure sample of 13: 36% yield;
mp 173.2−175.4 °C; 1H NMR (400 MHz) δ 0.69 (s, 3H), 2.28−2.29
(m, 2H), 2.49−2.50 (m, 1H), 2.75 (dd, 1H, J = 9.32 and 2.21 Hz), 3.52
(m, 1H), 5.35−5.36 (m, 1H); 13C NMR (100 MHz) δ 11.82, 12.45,
16.17, 19.37, 19.54, 20.17, 20.85, 21.01, 24.53, 27.93, 29.13, 31.61, 31.88
(2C), 36.48, 37.22, 38.66, 39.55, 42.24, 42.62, 48.28, 50.07, 53.42, 56.35,
62.14 (2C), 71.74, 121.54, 140.79; Anal. calcd for C29H48O2: C, 81.25%;
H, 11.29%. Found: C, 81.17%; H, 11.24%.
(22S,23S)-22,23-Epoxystigmast-5-ene-3β-ol (14). The epoxide 31
was treated as reported for compound 30 to furnish 14 in 29% yield; mp
127.8−130.2 °C; 1H NMR (400 MHz) δ 0.68 (s, 3H), 2.24−2.30 (m,
2H), 2.49−2.54 (m, 2H), 3.53 (m, 1H), 5.35−5.37 (m, 1H); 13C NMR
(100 MHz) 11.97, 12.36, 16.28, 19.36 (2C), 20.92, 21.06, 24.51, 27.07,
29.31, 29.68, 31.62, 31.87 (2C), 36.48, 37.25, 38.87, 39.67, 42.27, 42.67,
48.77, 50.17, 56.02, 56.32, 58.55, 63.13, 71.77, 121.67, 140.67; Anal.
calcd for C29H48O2: C, 81.25%; H, 11.29%. Found: C, 81.33%; H,
11.27%.
(22R,23R)-22,23-Epoxyergosta-5,7-diene-3β-ol (15). 2M KOH
solution (0.2 mL) was added to a solution of 42 (0.037 g, 0.08 mmol)
in EtOH (3.8 mL), and the resulting mixture was refluxed for 15 min.
After cooling, the reaction mixture was extracted with EtOAc (4 × 5
mL). The combined organic layers were washed with brine (8 mL),
dried over Na2SO4, and filtered, and the solvent was removed in vacuo to
give a residue, which was submitted to flash chromatography. Elution
with light petroleum−ethyl acetate (80:20) afforded 15 in 64% yield;
mp: 163.8−165.2 °C; 1H NMR (400 MHz) δ 0.61 (s, 3H), 3.61−3.65
(m, 1H), 5.39−5.41 (m, 1H), 5.57−5.58 (m, 1H); 13C NMR (100
MHz) 11.9, 13.7, 16.2, 16.3, 19.5, 20.4, 21.0, 23.2, 26.8, 31.1, 31.9, 37.0,
38.3, 39.0 (2C), 40.7, 42.3, 43.2, 46.2, 54.0, 55.6, 60.4, 64.3, 70.3, 116.5,
119.5, 139.8, 140.8; Anal. calcd for C28H44O2: C, 81.50%; H, 10.76%.
Found: C, 81.17%; H, 10.74%.
11.85, 18.89, 19.10, 19.38, 21.05, 23.27, 24.18, 27.8, 28.84, 31.58, 31.80,
31.88, 35.77, 36.43, 37.20, 39.72, 39.92, 42.04, 42.20 (2C), 50.02, 52.58,
56.61, 71.71, 71.86, 121.60, 140.73; Anal. calcd for (C29H50O2): C,
80.87%; H, 11.70%. Found: C, 80.70%; H, 11.65%.
(23R)-3β-Stigmast-5-ene-3,23-diol (19). A solution of 36 (0.03 g,
0.06 mmol) in MeOH (3 mL) was treated with 2M KOH solution (1
mL), and the resulting mixture was refluxed for 30 min. After cooling,
the reaction mixture was extracted with EtOAc (3 × 10 mL). The
combined organic layers were dried over Na2SO4 and filtered, and the
solvent was removed in vacuo. The residue, thus obtained, was purified
by flash chromatography. Elution with light petroleum−ethyl acetate
(80:20) afforded 19 in 55% yield; mp 158.1−158.6 °C; 1H NMR (400
MHz) δ 0.72 (s, 3H), 2.27−2.28 (m, 2H), 3.51 (m, 1H), 3.69−3.74 (m,
1H), 5.35 (d, 1H, J = 5.25 Hz); 13C NMR (100 MHz) 11.94, 14.48,
18.63, 18.96, 19.17, 19.36, 21.09, 21.44, 24.25, 27.80, 28.50, 31.67, 31.90
(2C), 32.78, 36.51, 37.28, 39.87, 41.11, 42.32, 42.51, 50.17, 52.49, 56.92
(2C), 70.25, 71.76, 121.60, 140.81; Anal. calcd for C29H50O2: C, 80.87%;
H, 11.70%. Found: C, 80.67%; H, 11.66%.
(22R)-3β-Stigmast-5-ene-3,22-diol (20). The derivative 37 was
treated as reported for compound 36 to furnish 20 in 72% yield; mp:
149.2−149.9 °C; 1H NMR (400 MHz) δ 0.72 (s, 3H), 3.51 (m, 1H),
3.69−3.74 (m, 1H), 5.35 (d, 1H, J = 5.25 Hz); 13C NMR (100 MHz)
11.75 (2C), 12.32, 17.74, 19.38, 20.45, 21.11, 23.60, 24.45, 27.50, 28.92,
29.65, 30.11, 31.70, 31.92, 36.54, 37.30, 39.81, 41.53, 42.33, 42.59, 42.70,
50.22, 53.07, 56.39, 71.39, 71.78, 121.59, 140.85; Anal. calcd for
C29H50O2: C, 80.87%; H, 11.70%. Found: C, 80.91%; H, 11.69%.
(23R)-3β-Ergost-5,7-diene-3,23-diol (23) and (23S)-3β-Ergost-5,7-
diene-3,23-diol (21). NaBH4 (0.13 g, 3.44 mmol) was added to a
solution of the ketone 48 (0.10 g, 0.2 mmol) in THF-2-propanol (2:1, 6
mL), and the resulting mixture was stirred at room temperature
overnight. The reaction mixture was then diluted with H2O (5 mL) and
extracted with Et2O (3 × 5 mL). The combined organic layers were
washed with brine (10 mL), dried over Na2SO4, and filtered, and the
solvent was removed in vacuo. The residue, thus obtained, was dissolved
in EtOH (10 mL) and treated with PPTS (0.012 g, 0.047 mmol). After
refluxing for 1 h, the reaction mixture was allowed to cool to room
temperature, and the solvent was removed in vacuo to give a residue,
which was submitted to mpc. Elution with light petroleum−ethyl acetate
(90:10) afforded pure samples of the desired compounds in 78% total
(22S,23S)-22,23-Epoxyergosta-5,7-diene-3β-ol (16). The derivative
43 was treated as reported for 42 to furnish 16 in 89% yield; mp: 138.3−
1
139.6 °C; H NMR (400 MHz) δ 0.60 (s, 3H), 3.60−3.66 (m, 1H),
1
5.39−5.41 (m, 1H), 5.56−5.58 (m, 1H); 13C NMR (100 MHz) 11.8,
12.5, 16.2, 17.1, 18.5, 20.2, 21.0, 23.3, 27.8, 31.0, 31.9, 37.0, 38.3, 39.0,
39.8, 40.7, 42.5, 43.2, 46.1, 53.3, 54.0, 63.1, 63.8, 70.3, 116.5, 119.4,
140.0, 140.7; Anal. calcd for C28H44O2: C, 81.50%; H, 10.76%. Found:
C, 81.32%; H, 10.77%.
yield; 21: mp 129.8−130.7 °C; H NMR (400 MHz) δ 0.65 (s, 3H),
3.62−3.69 (m, 2H), 5.40−5.42 (m, 1H), 5.59 (dd, 1H, J = 7.89, 2.35
Hz). 13C NMR (100 MHz) 10.54, 11.71, 16.27, 17.92, 20.67, 21.04,
21.75, 23.05, 27.74, 28.40, 31.92, 35.75, 36.97, 38.32, 39.09, 40.70
(2xC), 43.00, 45.32, 46.16, 54.37, 56.76, 70.44, 73.30, 116.33, 119.54,
139.80, 141.24; Anal. calcd for C28H46O2: C, 81.10%; H, 11.18%. Found:
C, 80.97%; H, 11.19%.
(23S)-3β-Stigmast-5-ene-3,23-diol (17) and (22S)-3β-Stigmast-5-
ene-3,22-diol (18). LiAlH4 (0.25 g, 6.71 mmol) was portion wise added
to the solution of the epoxide 30 (0.27 g, 0.61 mmol) in anhydrous THF
(15 mL). The resulting mixture was refluxed for 36 h under an argon
atmosphere. After cooling, first EtOAc and then water were carefully
added. The organic phase was separated, and the water phase extracted
with EtOAc (3 × 15 mL). The combined organic phases were washed
with brine (20 mL) and then dried over Na2SO4. After filtration, the
solvent was evaporated in vacuo to give a residue, which was dissolved in
glacial acetic acid (5 mL), and the resulting solution refluxed for 6 h.
After cooling, the mixture of 32 + 33, obtained by the removal of the
solvent in vacuo, was directly dissolved in methanol (16 mL) and treated
with 2 M KOH solution (8 mL). After refluxing for 3 h, the reaction
mixture was extracted with EtOAc (3 × 15 mL). The combined organic
layers were dried over Na2SO4 and filtered, and the solvent was removed
in vacuo to give a residue, which was submitted to mpc. Elution by light
petroleum−ethyl acetate (70:30) afforded pure samples of the desired
compounds in 69% total yield; 17: mp 178.2−181.4 °C; 1H NMR (400
MHz) δ 0.69 (s, 3H), 2.23−2.31 (m, 2H), 3.53 (m, 1H), 3.91 (m, 1H),
5.36 (m, 1H); 13C NMR (100 MHz) 11.79, 13.82, 18.28, 19.38 (2C),
19.85, 21.03, 21.12, 24.24, 28.45, 28.54, 31.58, 31.82 (2C), 34.16, 36.44,
37.19, 39.73, 42.22, 42.35, 42.46, 49.13, 50.01, 56.66, 56.88, 70.55, 71.73,
121.62, 140.72; Anal. calcd for C29H50O2: C, 80.87%; H, 11.70%. Found:
C, 80.63%; H, 11.72%. 18: mp 168.9−172.4 °C; 1H NMR (400 MHz) δ
0.70 (s, 3H), 2.24−2.31 (m, 2H), 3.53 (m, 1H), 3.75 (t, 1H, J = 6.77
Hz), 5.35 (d, 1H, J = 5.21 Hz); 13C NMR (100 MHz) 11.39, 11.73,
(22R)-3β-Ergost-5,7-diene-3,22-diol (24) and (22S)-3β-Ergost-5,7-
diene-3,22-diol (22). The derivative 49 was treated as reported for 48 to
furnish pure samples of the desired compounds 24 and 22 in 83% total
yield. 22: mp 117.3−121.0 °C; 1H NMR (400 MHz) δ 0.65 (s, 3H), 0.79
(d, 3H, J = 6.84 Hz), 3.63−3.66 (m, 1H), 3.78−3.81 (m, 1H), 5.40−5.42
(m, 1H), 5.59−5.61 (m, 1H); 13C NMR (100 MHz) 11.79, 12.49, 15.55,
16.02, 16.22, 21.04, 21.12, 23.15, 23.80, 27.39, 29.53, 31.92, 34.60, 35.27,
36.98, 38.30, 39.09, 40.72, 43.00, 46.14, 52.73, 54.01, 70.40, 71.67,
116.43, 119.54, 139.90, 140.96; Anal. calcd for C28H46O2: C, 81.10%; H,
11.18%. Found: C, 81.09%; H, 11.17%.
(23R)-3β-Ergost-5,7-diene-3,23-diol (23). PPTS (0.010 g, 0.039
mmol) was added to a solution of 44 (0.050 g, 0.1 mmol) in EtOH (5
mL), and the resulting mixture was refluxed for 5 h. After cooling, the
solvent was removed in vacuo, and the residue was purified by flash
chromatography. Elution with light petroleum−ethyl acetate (80:20)
furnished 23 in 70% yield; mp 167.8−169.4 °C; 1H NMR (400 MHz) δ
0.68 (s, 3H), 3.68 (m, 1H), 3.82 (m, 1H), 5.44 (s, 1H), 5.61 (s, 1H). 13C
NMR (100 MHz) 9.83, 11.88, 16.25, 18.45, 18.79, 21.05, 21.50, 22.98,
28.33, 29.55, 31.92, 33.11, 36.97, 38.33, 39.18, 40.73, 42.11, 43.01, 45.37,
46.17, 54.52, 56.47, 70.38, 70.64, 116.34, 119.53, 139.80, 141.19; Anal.
calcd for C28H46O2: C, 81.10%; H, 11.18%. Found: C, 81.26%; H,
11.16%.
(22R)-3β-Ergost-5,7-diene-3,22-diol (24). The derivative 47 was
treated as reported for 44 to furnish 24 in 73% yield; mp 197.7−201.2
K
J. Med. Chem. XXXX, XXX, XXX−XXX