1404
Y. Shiono et al. / Phytochemistry 72 (2011) 1400–1405
afford 1 (35.2 mg) as colorless crystals. The filtrate was further sep-
arated by semipreparative ODS HPLC (MeOH–H2O, 80:20) to give
compounds 2 (15.4 mg), 3 (10.5 mg), 4 (5.2 mg). Fractions 1–13
were subjected to ODS CC by eluting with H2O and an increasing
ratio of MeOH (100:0–0:100) and three fractions obtained were
purified by semipreparative ODS HPLC (MeOH–H2O, 75:25) to give
compounds 5 (3.4 mg) and 6 (4.2 mg).
graphite monochromated Mo Ka radiation (k = 0.71070 Å). The
structure was solved by direct methods (SIR2004) (Burla et al.,
2005) and expanded by using Fourier techniques (DIRDIF99)
(Beurskens et al., 1999). A total of 16,248 reflections were mea-
sured and 2870 were unique (Rint = 0.044). Crystal data and refine-
ment statistics are as follows: C26H42O7, MW = 466.61, monoclinic,
space group P21 (#4); a = 7.407(3), b = 9.809(4), c = 16.628(7) Å,
b = 95.142(3)°, V = 1203.4(8) Å3; Z = 2, Dcalc = 1.288 g/cm3, F000
=
4.4. 19-(
a
-D
-glucopyranosyloxy)isopimara-7,15-dien-3b-ol (1)
508.00, (Mo K (I)),
l
a
) = 0.917 cmꢀ1, final R1 = 0.0380 (I > 2.00
r
R = 0.0397 (all data), wR2 = 0.1111 (all data), GOF = 1.009;
Complete crystallographic data, as a CIF file, have been deposited
with Cambridge Crystallographic Data Center (CCDC No. 764212).
Copies can be obtained free of charge from: The Director, CCDC,
12 Union Road, Cambridge CB2 1EZ, UK (e-mail: deposit@ccdc.
cam.ac.uk).
Colorless crystals; mp 220–221ꢀC; ½a D20
þ 49:8 (c 0.31, MeOH);
ꢂ
IR (KBr) mmax 3375, 1630, 1050 cmꢀ1; for 1H (400 MHz) and 13C
NMR (100 MHz) spectroscopic data, see Table 1; negative FABMS
m/z: 465 [M-H]ꢀ; negative HRFABMS m/z: 465.2860 [M-H]ꢀ, (calcd
for C26H41O7, 465.2852).
4.5. 19-(2-acetamido-2-deoxy-a-D-glucopyranosyloxy)isopimara-
7,15-dien-3b-ol (2)
4.11. Acid hydrolysis of 1 and 2, and identification of sugars
A solution of 1 (7.0 mg) in 1 N CF3COOH (10 mL) was heated at
100ꢀC for 3 h. After neutralizing with 1 N NaOH/H2O, the solution
was extracted with Et2O (10 mL ꢁ 3) and the combined organic
phase was evaporated to dryness under reduced pressure to yield
aglycone 1a (2.7 mg). The aqueous layer was evaporated to dryness
under reduced pressure and subjected to silica gel CC to afford
White amorphous powder; ½a D20
þ 88:6 (c 0.22, MeOH); IR (KBr)
ꢂ
m
max 3321, 1653, 1545, 1087, 1035 cmꢀ1; for 1H (400 MHz) and 13
C
NMR (100 MHz) spectroscopic data, see Table 1; negative FABMS
m/z: 506 [M-H]ꢀ; negative HRFABMS m/z: 506.3122 [M-H]ꢀ, (calcd
for C28H44NO7, 506.3118).
D
-glucose (2.5 mg). The optical rotation value was as follows:
a 2D0
ꢂ
þ 50:6 (c 0.9, H2O, measured 24 h after dissolving in
4.6. 19-(a-D-glucopyranosyloxy)isopimara-7,15-dien-3-one (3)
½
H2O)(lit. + 53.3 (Zhang et al., 2001)).
White amorphous powder; ½a D20
þ 27:7 (c 0.45, MeOH); IR (KBr)
ꢂ
A mixture of 2 (4.0 mg) and 3 N HCl (10 mL) was heated at 100
ꢀC for 3 h. This reaction mixture was treated in the same way as
described for 1 to afford aglycone 1a and a sugar fraction. From
m
max 3398, 1707, 1637, 1030 cmꢀ1; for 1H (400 MHz) and 13C NMR
(100 MHz) spectroscopic data, see Table 2; negative FABMS m/z:
463 [M-H]ꢀ; negative HRFABMS m/z: 463.2699 [M-H]ꢀ, (calcd for
the sugar fraction,
described for 1. The optical rotation value was as follows:
25 + 70.0 (c 0.2, H2O, measured 24 h after dissolving in H2O)
D-2-amino-2-deoxy-D-glucose was isolated as
C
26H39O7, 463.2696).
[a]
D
4.7. 19-(2-acetamido-2-deoxy-a-D-glucopyranosyloxy)isopimara-
7,15-dien-3-one (4)
(lit. + 72 (Armarego and Chai, 2009).
White amorphous powder; ½a D20
þ 52:1 (c 0.20, MeOH); IR (KBr)
ꢂ
Acknowledgment
mmax 3366, 1707, 1640, 1552, 1122, 1037 cmꢀ1; for 1H (400 MHz)
and 13C NMR (100 MHz) spectroscopic data, see Table 2; negative
FABMS m/z: 504 [M-H]ꢀ; negative HRFABMS m/z: 504.2969 [M-
H]ꢀ, (calcd for C28H42NO7, 504.2961).
We thank Ms. Teiko Yamada of the Faculty of Agriculture at
Tohoku University for HRMS measurements.
Appendix A. Supplementary data
4.8. 19-(a-D-glucopyranosyloxy)-3b-hydroxyisopimara-8,15-dien-7-
Supplementary data (The 1H, 13C NMR and 2D NMR spectra of
compounds 1–6, synthetic experiment and spectroscopic data,
and methods in biological test) associated with this article can
one. (5)
White amorphous powder; [
(MeOH) kmax (log
a
]
20 + 148.0 (c 0.20, MeOH); UV
D
e
) 249 (3.7); IR (KBr) mmax 3339, 1643, 1373,
1032 cmꢀ1; for 1H (400 MHz) and 13C NMR (100 MHz) spectro-
scopic data, see Table 3; negative FABMS m/z: 479 [M-H]ꢀ; nega-
tive HRFABMS m/z: 479.2643 [M-H]ꢀ, (calcd for C26H39O8,
479.2645).
References
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4.9. 19-(2-acetamido-2-deoxy-
hydroxyisopimara-8,15-dien-7-one. (6)
a-D-glucopyranosyloxy)-3b-
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White amorphous powder; [
a
]
20 + 113.9 (c 0.45, MeOH); UV
D
(MeOH) kmax (log
e
) 249 (3.7); IR (KBr) mmax 3398, 1707, 1637,
1030 cmꢀ1; for 1H (400 MHz) and 13C NMR (100 MHz) spectro-
scopic data, see Table 3; FABMS m/z: 520 [M-H]ꢀ; negative
HRFABMS m/z: 520.2915 [M-H]ꢀ, (calcd for C28H42NO8, 520.2910).
4.10. X-ray Crystallography of 1
Single crystals suitable for X-ray structure analysis were ob-
tained by recrystallization from MeOH. X-ray diffraction data were
collected at 173 K on a Rigaku Saturn CCD diffractometer with
Armarego, W.L.F., Chai, C., 2009. Purification of laboratory chemicals, sixth ed.
Butterworth-Heinemann, Oxford, p. 649.