5096 J ournal of Medicinal Chemistry, 1996, Vol. 39, No. 26
Goldstein
1
3H, C-7, C-22, C-23), 3.88 (m, 1H, C-3), 2.90 (d, 1H, C-4, J )
1.0 Hz), 2.68-2.65 (dd, 1H, C-6), 1.03 (s, 3H, C-19), 0.95 (d,
3H, C-21, J ) 6.9 Hz), 0.85 (d, 3H, C-28, J ) 6.9 Hz), 0.76 (t,
6H, C-26, C-27, J ) 7.1 Hz), 0.59 (s, 3H, C-18); MS (EI) m/z
413 (M, 11), 91 (M - 322, 100). Anal. (C28H44O2) C, H.
7,22(E)-Er gosta d ien e-3â,5r-d iol (6). To 4R,5-epoxy-7,22-
(E)-ergostadien-3-one (3) (0.066 g, 0.16 mmol) in 25 mL of
anhydrous Et2O was added LiAlH4 (0.019 g, 0.50 mmol). The
reaction mixture was refluxed for 3 h under Ar and then cooled
to 0 °C. EtOAc (3 mL) and 3 drops of saturated Na2SO4(aq)
were added. The resultant solution was dried with anhydrous
MgSO4 and filtered, and the solvent was evaporated in vacuo.
The residue was purified by flash chromatography (3:1-0:1
hexane:EtOAc) to yield 6 as a white solid (0.015 g, 22%): mp
237-238 °C (lit.25 mp 227-234 °C); Rf 0.55 (1:1 hexane:EtOAc);
1H NMR δ 5.24-5.14 (dd, 2H, C-22, C-23), 5.08-5.05 (m, 1H,
C-7), 4.04 (m, 1H, C-3), 2.23 (d, 1H, J ) 16.8 Hz), 1.03 (d, 3H,
C-21, J ) 6.6 Hz), 0.92 (s, 3H, C-19), 0.91 (d, 3H, C-28, J )
5.1 Hz), 0.83 (t, 6H, C-26, C-27, J ) 7.3 Hz), 0.57 (s, 3H, C-18);
IR 3590, 3423 cm-1; MS (CI) m/z 415 (M + 1, 3), 397 (M + 1
- H2O, 20), 379 (M + 1 - 2 × H2O, 2), 125 (C9H17, 100). Anal.
(C28H46O2) C, H.
10: mp 120-123 °C; Rf 0.15 (3:1 hexane:EtOAc); H NMR
δ 5.78 (s, 1H, C-7), 5.18-5.07 (m, 2H, C-22, C-23), 2.94 (s, 1H,
C-4 axial), 2.35 (d, 1H, C-4), 1.19 (s, 3H, C-19), 0.95 (d, 3H,
C-21, J ) 6.5 Hz), 0.84 (d, 3H, C-28, J ) 6.5 Hz), 0.76 (t, 6H,
C-26, 27 J ) 7.5 Hz), 0.68 (s, 3H, C-18); IR 2240, 1670 cm-1
;
MS (CI) m/z 422 (M + 1, 100), 396 (M + 1 - CN, 8), 296 (M +
1 - C9H17, 5), 125 (C9H17, 27). Anal. (C29H43NO) H, N; C:
calcd, 82.60; found, 82.01.
5r-Cya n o-7,22(E)-er gosta d ien -3â-ol (11) a n d 5r-Cya n o-
7,22(E)-er gosta d ien -3r-ol (12). To 5R-cyano-7,22(E)-ergo-
stadien-3-one (9) (2.7 g, 6.4 mmol) were added 25 mL of CH2-
Cl2, 175 mL of 95% EtOH, and NaBH4 (0.7 g, 19.2 mmol). The
solution was stirred for 12 h. The solvent was evaporated in
vacuo and the residue purified by flash chromatography (8:
1-2:1 hexane:EtOAc) to yield in order of elution 11, as a white
solid which was recrystallized from Et2O/hexane to form white
needles (1.0 g, 37%), and 12, as a white solid (1.7 g, 63%).
1
11: mp 170-174 °C; Rf 0.48 (3:1 hexane:EtOAc); H NMR
δ 5.24-5.14 (m, 3H, C-7, C-22, C-23), 4.06 (m, 1H, C-3), 1.02
(d, 3H, C-21, J ) 7 Hz), 0.94 (d, 3H, C-28, J ) 8.5 Hz), 0.92 (s,
3H, C-19), 0.84 (d, 3H, C-26, J ) 7 Hz), 0.82 (d, 3H, C-27, J )
7 Hz), 0.55 (s, 3H, C-18); IR 3443, 2223 cm-1; MS (CI) m/z 424
(M + 1, 100), 397 (M + 1 - HCN, 20), 378 (M + 1 - HCN -
H2O, 18), 271 (M + 1 - HCN - C9H17, 20), 125 (C9H17, 80).
Anal. (C29H45NO) C, H, N.
5-(S-Acetylth io)-7,22(E)-er gosta d ien -3-on e (7). After
4,7,22(E)-ergostatrien-3-one (2) (0.503 g, 1.27 mmol) was
stirred in 25 mL of thiolacetic acid with benzoyl peroxide (0.077
g, 0.3 mmol) for 5 days, the solvent was evaporated in vacuo.
The resultant residue was dissolved in Et2O and washed to
neutrality with 5% NaOH(aq). The organic layer was evapo-
rated in vacuo. The residue was then recrystallized from
MeOH to give 7 as white needles (0.025 g, 4%): mp 120-122
°C; Rf 0.50 (1:1 hexane:EtOAc); 1H NMR δ 5.25-5.14 (m, 3H,
C-7, C-22, C-23), 2.26 (s, 3H, SAc), 1.08 (s, 3H, C-19), 1.02 (d,
3H, C-21, J ) 6.6 Hz), 0.92 (d, 3H, C-28, J ) 6.9 Hz), 0.83 (t,
1
12: mp 174-176 °C; Rf 0.40 (3:1 hexane:EtOAc); H NMR
δ 5.24-5.14 (m, 3H, C-7, C-22, C-23), 4.14 (m, 1H, C-3), 1.03
(d, 3H, C-21, J ) 6.6 Hz), 0.92 (d, 3H, C-28, J ) 7.1 Hz), 0.89
(s, 3H, C-19), 0.83 (t, 6H, C-26, C-27, J ) 7.8 Hz), 0.56 (s, 3H,
C-18); IR 3462, 2211 cm-1; MS (CI) m/z 424 (M + 1, 100), 406
(M + 1 - H2O, 91), 380 (M + 1 - H2O - HCN, 5), 125 (C9H17
40). Anal. (C29H45NO) H, N; C: calcd, 82.21; found, 81.37.
,
5r-F or m yl-7,22(E)-er gosta d ien -3â-ol (13). 5R-Cyano-7,
22(E)-ergostadien-3â-ol (11) (0.7 g, 3.1 mmol) was dissolved
in 100 mL of anhydrous toluene. The solution was cooled to
-10 °C, and 1.5 M DIBAL in toluene (12.3 mL, 18.4 mmol)
was added. The reaction mixture was stirred for 40 min under
argon at -10 °C and then refluxed with 20 mL of 1.0 M H2-
SO4(aq) for 40 min. The organic layer was washed with 40
mL of H2O and 40 mL of saturated NaCl(aq). The aqueous
layer was back-extracted with 2 × 30 mL of CH2Cl2. All
organic layers were evaporated in vacuo. The residue was
purified by flash chromatography (6:1-2:1 hexane:EtOAc) to
yield 13 as a white solid (0.6 g, 86%): mp 149-152 °C; Rf 0.29
6H, C-26, J ) 7.1 Hz), 0.59 (s, 3H, C-18); IR 1700, 1678 cm-1
MS (CI) m/z 471 (M + 1, 8), 395 (M + 1 - SAc, 100).
;
5-Mer capto-7,22(E)-er gostadien -3â-ol (8). A flask charged
with 5-(S-acetylthio)-7,22(E)-ergostadien-3-one (7) (0.265 g,
0.56 mmol), 25 mL of anhydrous Et2O, and LiAlH4 (0.064 g,
1.7 mmol) was refluxed for 5 h and then allowed to cool to
room temperature. EtOAc (5 mL) was added followed by 3
drops of 10% Na2SO3(aq). The precipitate was removed by
vacuum filtration and the liquid evaporated in vacuo. The
residue was purified by flash chromatography (12:1 toluene:
EtOAc) to afford 8 as a white solid (0.111 g, 46%). Recrystal-
lization from CH3CN gave fine white needles: mp 139-140
°C; Rf 0.28 (3:1 hexane:EtOAc); 1H NMR δ 5.24-5.14 (m, 2H,
C-22, C-23), 5.06 (s, 1H, C-7), 4.17 (m, 1H, C-3), 1.01 (d, 3H,
C-21, J ) 6.6 Hz), 0.99 (s, 3H, C-19), 0.91 (d, 3H, C-28, J )
6.9 Hz), 0.84 (d, 3H, C-26, J ) 7.3 Hz), 0.82 (d, 3H, C-27, J )
7.3 Hz), 0.56 (s, 3H, C-18); IR 3440, 1057 cm-1; MS (CI) m/z
431 (M + 1, 6), 413 (M + 1 - H2O, 31), 397 (M + 1 - H2S, 63),
379 (M + 1 - H2O - H2S, 13), 288 (M + 1 - H2O - C9H17, 3),
273 (M + 1 - H2S - C9H17, 18), 255 (M + 1 - H2O - H2S -
C9H17, 9), 125 (C9H17, 100). Anal. (C28H46OS) C, H, S.
5r-Cya n o-7,22(E)-er gosta d ien -3-on e (9) a n d 5â-Cya n o-
7,22(E)-er gosta d ien -3-on e (10). To a solution of 4,7,22(E)-
ergostatrien-3-one (2) (1.5 g, 3.8 mmol) in 300 mL of anhydrous
THF was added 1.0 M Et2AlCN (11.4 mL, 11.4 mmol) in
toluene. Upon addition of the cyanide reagent the solution
turned dark red. After stiring for 3.5 h the mixture was
washed with 2 × 40 mL of ice cold 10% NaOH(aq). (Caution!
When the reaction is performed on a large scale, add the
aqueous components dropwise to avoid a violent exotherm.)
The aqueous layer was exhaustively extracted with CH2Cl2,
the combined organic layers were dried with anhydrous MgSO4
and filtered, and the solvent was evaporated in vacuo. The
residue was purified by flash chromatography (8:1-0:1 hexane:
EtOAc) to provide 9, which was recrystallized from EtOH
giving white plates (1.3 g, 82%), and 10, as a white solid (0.2
g, 13%).
1
(3:1 hexane:EtOAc); H NMR δ 9.59 (s, 1H, CHO), 5.24-5.13
(m, 3H, C-7, C-22, C-23), 3.78 (m, 1H, C-3), 1.02 (s, 3H, C-19),
0.91 (d, 3H, C-28, J ) 6.8 Hz), 0.84 (d, 3H, C-26, J ) 6.8 Hz),
0.82 (d, 3H, C-27, J ) 6.8 Hz), 0.56 (s, 3H, C-18); IR 3419,
1708 cm-1; MS (CI) m/z 427 (M + 1, 43), 409 (M + 1 - H2O,
100), 381 (M + 1 - CHO, 6), 125 (C9H17, 77); HRMS m/z 425,
3417 (M + 1), calcd for C29H45O2 425.3419. Anal. (C29H45O2)
C: calcd, 79.44; found, 81.41. H: calcd, 10.87; found, 10.24.
3â-Hyd r oxy-7,22(E)-er gost a d ien e-5r-for m a m id e (14).
5R-Formyl-7,22(E)-ergostadien-3â-ol (13) (0.9 g, 2.1 mmol), 100
mL of CH2Cl2, pyridine (0.19 mL, 2.3 mmol), and acetyl
chloride (0.16 mL, 2.3 mmol) were combined and stirred
overnight. The solution was washed with 3 × 10 mL of 5%
HCl(aq), the organic layer was rotary evaporated, and the
residue was purified by flash chromatography (20:1-8:1
hexane:EtOAc) to give 3â-acetoxy-7,22(E)-ergostadiene-5R-
formaldehyde as a white solid (0.9 g, 90%): mp 149-152 °C;
Rf 0.66 (3:1 hexane:EtOAc); 1H NMR δ 9.57 (s, 1H, CHO),
5.23-5.13 (m, 3H, C-7, C-22, C-23), 4.84 (m, 1H, C-3), 2.00 (s,
3H, OAc), 1.03 (s, 3H, C-19), 1.01 (d, 3H, C-21, J ) 6.4 Hz),
0.91 (d, 3H, C-28, J ) 7.1 Hz), 0.83 (t, 6H, C-26, C-27, J ) 7.0
Hz), 0.56 (s, 3H, C-18); IR 1720 cm-1; MS (CI) m/z 469 (M +
1, 15), 409 (M + 1 - OAc, 100), 125 (C9H17, 18). Anal.
(C31H48O3) C: calcd, 79.44; found, 81.41. H: calcd, 10.32;
found, 10.85.
To 3â-acetoxy-7,22(E)-ergostadiene-5R-formaldehyde (0.300
g, 0.64 mmol) in 80 mL of t-BuOH, 20 mL of H2O, and 20 mL
of 2-methyl-2-butene were added NaH2PO4 (0.691 g, 5.76
mmol) and NaClO2 (0.695 g, 7.68 mmol). After stirring for 24
h, the solvents were evaporated in vacuo. The residue was
dissolved in 100 mL of a 1:1 mixture of CH2Cl2 and H2O. The
aqueous layer was back-extracted with 3 × 20 mL of CH2Cl2.
9: mp 280-282 °C; Rf 0.30 (3:1 hexane:EtOAc); 1H NMR δ
5.24-5.14 (m, 3H, C-7, C-22, C-23), 2.58 (bs, 2H, C-4), 1.13 (s,
3H, C-19), 1.03 (d, 3H, C-21, J ) 6.5 Hz), 0.92 (d, 3H, C-28, J
) 7 Hz), 0.84 (d, 3H, C-26, J ) 6.8 Hz), 0.83 (d, 3H, C-27, J )
6.8 Hz), 0.59 (s, 3H, C-18); IR 2221, 1717 cm-1; MS (EI) m/z
422 (M, 5), 109 (C8H13, 100). Anal. (C29H43NO) C, H, N.