J. Woo Bok et al. / Phytochemistry 51 (1999) 891±898
897
solution turned pink, then quickly returned to a clear
colourless solution. After 10 min the solution was
allowed to warm slowly to room temperature. The
reaction was quenched with the addition of Et3N
(60 ml) after 40 min. The clear solution was diluted
with CH2Cl2, ®ltered to remove the molecular sieves
and washed twice with H2O. The organic layer was
concentrated under reduced pressure. Column chroma-
tography eluted with 5% MeOH in CHCl3 aorded 5
(0.0466, mmol) in 16 % yield (Rf=0.78, 15 % MeOH
in CHCl3) followed by unreacted starting material
(0.1192 g, 300 mmol) (Rf=0.66, 15 % MeOH in
Water was circulated into the dewar to maintain room
temperature for the reaction. After 3 h the solution
was concentrated under reduced pressure, with the
resulting residue applied to a silica gel column what
we eluted with 15% MeOH in CHCl3 to aord 1a
(0.0138 g, 0.023 mmol) in 62% yield as a white solid.
IR,1H,13C and LRMS were identical to spectra of 1a
isolated from the mycelia of C. sinensis.
3.5.4. Synthesis of 5a,8a-epidioxy-24(R )-methyl-
cholesta-6,22-dien-3b-ol (1b) (ergosterol peroxide)
from ergosterol (6)
1
CHCl3). H NMR (400 MHz, CDCl3) d: 0.59 (3H, s,
Following the same procedure as used for the prep-
aration of 1a from 3, ergosterol (6) (0.1040 g ,
0.26 mmol) was oxidized, then puri®ed by silica gel
column eluted with 15% EtOAc in CH2Cl2 to aord
1b (0.0932 g, 0.22 mmol) in 85% yield as a white
solid. 1H NMR (500 MHz, CDCl3) 0.61(3H, s, Me-
18), 0.67 (3H, s, Me-19), 0.907 (3H, d, J=6.67 Hz,
Me-26 or 27), 0.913(3H, d, J=6.71 Hz, Me-26 or -27),
0.99 (3H, d, J=6.78 Hz, Me-28), 1.00 (3H, d, J=6.49,
Me-21), 1.1±2.0 (20H, m ), 3.92 (m, 1H, H-3), 5.14
(1H, m, H-23), 5.25 (1H, m, H-22), 5.95 (1H, d,
J=8.37 Hz, H-6), 6.29 (1H, d, J=8.47 Hz, H-7); 13C
NMR (50 MHz, CDCl3): Table 2.
Me-18), 0.79 (3H, d, J=6.7, Me-26 or 27), 0.80 (3H, d,
J=6.7, Me-26 or 27), 0.86 (3H, d, J=6.9, Me-28),
0.88 (3H, s, Me-19), 1.00 (3H, d, J=6.5, Me-21), 1.7±
2.0 (18H, m ) 1.98 (3H, s ), 1.99 (3H, s), 2.01 (3H, s),
2.03 (3H, s), 2.21 (1H, m ), 2.40(1H, m ), 3.56 (1H, m,
H-3), 3.66 (1H, m ), 4.10 (1H, m ), 4.24 (1H, m ), 4.60
(1H, d, J=7.7, H-1'), 4.94 (1H, m ), 5.04 (1H, m ), 5.18
(3H, m, H-22, H-23), 5.36 (1H, m, H-7), 5.55 (1H, m,
H-6); 13C NMR (50 MHz, CDCl3) 12.0, 16.1, 17.6,
19.2, 19.6, 19.6, 19.9, 20.5, 20.6, 20.7, 21.0, 22.9, 28.2,
29.7, 33.0, 33.0, 37.1, 37.4, 38.2, 39.0, 40.3, 42.8, 46.1,
54.5, 55.7, 62.0, 68.5, 71.4, 71.7, 72.8, 78.6, 99.5, 116.1,
119.9, 131.9, 135.4, 139.0, 141.6, 169.2, 169.3, 170.3,
170.6.
Acknowledgements
3.5.2. Ergosteryl-3-O-b-D-glucopyranoside (3)
To a stirring solution of 5 (0.0564 g, 0.078 mmol) in
20 ml MeOH was added 3 ml Et3N and 0.5 ml H2O.
After stirring for 48 h the solvent was removed under
reduced pressure. The residue was applied to a silica
gel column, and eluted with 15% MeOH in CHCl3
(Rf=20.6) to aord a white solid. Recrystallization
from MeOH provided 3 (0.0351 g, 0.063 mmol) in
81% yield as small white needles. IR (cm 1,CHCl3):
3400, 2950, 2875, 1681, 1458, 1371; 1H NMR
(400 MHz, DMSO) d: 0.58 (3H, s, Me-18), 0.79 (3H,
d, J=6.7, Me-26 or -27), 0.80 (3H, d, J=6.7, Me-26 or
-27), 0.86 (3H, s, Me-19), 0.88 (3H, d, J=6.9, Me-28),
1.00 (3H, d, J=6.5, Me-21), 1.24 (2H, m ) 1.45 (1H,
m ), 1.6 (1H, m ) 1.68 (1H, m ), 1.85 (2H, m ), 2.0 (1H,
m ), 2.18 (1H, m ), 2.54 (1H, m ), 2.98 (1H, m ), 3.13
(1H, m ), 3.14 (1H, m ), 3.3 (1H, bs), 3.46 (2H, m ),
3.61 (1H, m ), 4.24 (1H, d, J=7.7 Hz, H-1'), 5.20(2H,
m, H-22, H-23), 5.34 (1H, m, H-7), 5.52 (1H, m, H-6);
13C NMR (50 MHz, CDCl3,): Table 2.
This work was supported by North American Reishi
Inc., British Columbia. We also thank Dr S. G.
Withers for providing 2,3,4,6-tetra-O-acetyl-a-D-gluco-
syltrichloroacetimidate.
References
Ahmed, W., Ahmed, Z., & Malik, A. (1992). Phytochemistry, 31,
4038.
Aiello, A., Fattorusso, E., Magno, S., Mayol, L., & Menna, M.
(1990). J. Nat. Prod., 53, 487.
Cerny, I., Pouzar, V., Drasar, P., Budesinsky, M., & Havel, M.
(1984). Collct. Czech. Chem. Commun., 49, 881.
Cheng, K. P., Nagano, H., Bang, L., Ourrison, G., & Beck, J. P.
(1977). Chem. Res., Sup., 9, 217.
Cunningham K.G., Herchinson S.A., Manson W., Spring F.S. 1951
J. Chem. Soc. 2299.
Furuya, T., Hirotani, M., & Matsuzawa, M. (1983). Phytochemistry,
22, 2509.
Gonzalez, A. G., Barrera, J. D., Perez, E. M. R., & Padron, C. E.
H. (1992). Planta Med., 58, 214.
Gunatilaka, A. A. L., Gopichand, Y., Schmitz, F. J., & Djerassi, C.
(1981). J. Org. Chem., 46, 3860.
3.5.3. 5a,8a-Epidioxy-24(R )-methylcholesta-6,22-dien-
3b-D-glucopyranoside (1a)
To a solution of 3 (0.0211 g, 0.038 mmol) in 20 ml
dry EtOH was added 2 drops of a 10% solution of
eosin in EtOH. Oxygen was bubbled through the sol-
ution. The reaction vessel was then placed in a silver
dewa and irradiated with a 500 W tungsten lamp.
Guyot, M., & Durgeat, M. (1981). Tetrahedron Lett., 22, 1391.
Hirayama, T., Fujikawa, F., Yosioka, I., & Kitagawa, I. (1975).
Chem. Pharm. Bull. Jpn, 23, 693.
Isaacs, S., Berman, R., & Kashman, Y. (1991). J. Nat. Prod., 54, 83.
Jones, K. (1993). A brief pharmacologic review of Cordyceps sinensis
and C. ophiolglossoides. Armana Research.