Biologically active phenols from Saussurea medusa

Article abstract of DOI:10.1016/S0968-0896(02)00470-4

Sixteen phenolic compounds were isolated from the polar fraction of Saussurea medusa and were structurally elucidated by chemical evidences and spectral methods. These compounds include two new lignan glucosides, namely medusasides A (1) and B (2), and fourteen known phenolic compounds (3-16). One major compound, apigenin 7-O-[α-L-rhamnopyranosyl-(1→6)-β-D-glucopyranoside] (6) showed remarkable activity to attenuate the scopolamine induced memory deficit of mice. Compound 6 and another major one, quercetin (8) also exhibited moderate cell protecting activities against hydrogen peroxide (H₂O₂) induced PC12 cell damage.

Full text of DOI:10.1016/S0968-0896(02)00470-4

Bioorganic & Medicinal Chemistry 11 (2003) 703–708
Biologically Active Phenols from Saussurea medusa
Cheng-Qi Fan and Jian-Min Yue*
State Key Laboratory of Drug Research, Institute of Materia Medica, Shanghai Institutes for Biological Sciences,
Chinese Academy of Sciences, 294Taiyuan Road, Shanghai, 200031, PR China
Received 16 August 2002; accepted 28 September 2002
Abstract—Sixteen phenolic compounds were isolated from the polar fraction of Saussurea medusa and were structurally elucidated by
chemical evidences and spectral methods. These compounds include two new lignan glucosides, namely medusasides A (1) and B (2), and
fourteen known phenolic compounds (3–16). One major compound, apigenin 7-O-[a-l-rhamnopyranosyl-(1!6)-b-d-glucopyranoside]
(6) showed remarkable activity to attenuate the scopolamine induced memory deficit of mice. Compound 6 and another major one,
quercetin (8) also exhibited moderate cell protecting activities against hydrogen peroxide (H2O2) induced PC12 cell damage.
# 2002 Elsevier Science Ltd. All rights reserved.
Introduction
C26H34O12 from its quasi-molecular ions at m/z 561.3
[M+Na]+ in positive ion mode ESI-MS and at m/z
537.2 [M–H]À in negative ion mode ESI-MS, and the
13C NMR (with DEPT) spectral data (Table 1). The
molecular composition of C26H34O12 was finally con-
firmed by HREIMS at m/z 376.1525 (C20H24O7, calcu-
lated: 376.1522) [M-glc (162)]+. Hydroxyl groups,
carbonyl group and aromatic ring(s) could be existed in
the structure of compound 1 judged from the absorp-
tion bands at 3406 (OH), 1655, 1591 and 1516 cmÀ1 in
its IR spectrum. In the 13C NMR, 12 carbon signals
were appeared at the down field, four of which at dC
146.2, 149.2, 149.3 and 153.6 were oxygenated, suggest-
ing the presence of two benzene rings. One carbon sig-
nal atdC 203.5 showed the possible presence of a
conjugated ketone group. Two methoxyl groups and a
glucopyranosyl moiety were also distinguished in the
structure of 1 on the basis of 1H and 13C NMR spectra,
and the remaining carbon signals were observed as three
methylenes (two of them were oxygenated) and two
methines. The above-mentioned data proposed that
compound 1 was most likely a lignan glucoside. The key
carbon signals for two methines at dC 43.1 (C-8) and
50.1 (C-80) and the coupling patterns of key proton sig-
nals of H-8 and H-80 indicated that the aglycone of
compound 1 was diphenylbutane type lignan7 and con-
firmed by HMBC spectrum. In the HMBC spectrum,
the correlations were observed among H-8 at dH 2.20
with C-7 (dC 37.0), C-9 (dC 70.5) and C-80 (dC 51.0), and
among H-80 at dH 2.18 with C-70 (dC 203.5), C-90 (dC
62.0) and C-8 (43.1). In the 1H NMR spectrum of 1, the
coupling patterns of the aromatic proton signals at dH
6.62 (d, J=1.8 Hz), 6.55 (d, J=8.1 Hz) and 6.51 (dd,
J=1.8, 8.1 Hz), and aromatic proton signals at dH 7.26
The whole plants of the Saussurea genus (Asteraceae) were
used in the both of traditional Chinese medicine and Tibet
folklore medicine for the cure of rheumatic arthritis, dys-
menorrhea and gynopathy. A series of compounds with a
variety of biological activities have been reported from the
genus of Saussurea.1À3 From the plant S. medusa Maxim,
nine flavonoids and flavonoid glycosides have been repor-
ted,4,5 and one flavonoid glycoside A1 was tested for effects
on CNS of mouse.6 Here, we present the isolation and
identification of two new lignan glucosides, namely medu-
sasides A (1) and B (2), together with 14 known phenolic
compounds from the polar fraction of S. medusa. One
major compound, apigenin 7-O-[a-L- rhamnopyranosyl-
(1!6)-b-d-glucopyranoside] (6) showed remarkable
activity to attenuate the scopolamine induced memory
deficit of mice by the animal model of mouse behavioral
tests in the water maze. The compounds 4, 6–9 and 14 with
relatively rich quantity were tested for cell protecting
activity against hydrogen peroxide (H2O2) induced PC12
cell damage, only compound 6 and another major one,
quercetin (8) exhibited moderate cell protecting activity.
Results and Discussion
Structure elucidation of the new compounds 1 and 2
Compound 1 was obtained as an amorphous powder.
The possible molecular formula of 1 was inferred to be
*Corresponding author. Tel.: +86-21-6431-1833; fax: +86-21-6437-
0269; e-mail: jmyue@mail.shcnc.ac.cn
0968-0896/03/$ - see front matter # 2002 Elsevier Science Ltd. All rights reserved.
PII: S0968-0896(02)00470-4

704
Cheng-Qi Fan, Jian-Min Yue / Bioorg. Med. Chem. 11 (2003) 703–708
Table 1. 1H (400 MHz) and 13C (100 MHz) NMR of 1 (CD3OD, d in
ppm)
configuration on the basis of the coupling constant of
the anomeric proton at dH 4.04 (d, J=7.7 Hz) was
attached to the C-9 judged from the correlation signals
between C-9 and the anomeric proton in HMBC spec-
trum, and the NOE correlation between H-9 and the
anomeric proton in NOESY spectrum. The structure of
compound 1 was thus elucidated to be 4-O-(b-d-gluco-
pyranosyl)-2-hydroxymethyl-3-[(4-hydroxy-3-methox-
yphenyl)methyl]-1-(4-hydroxy-3-methoxyphenyl)butan-
1-one (Fig. 1). The complete assignments of the proton
and the carbon signals of compound 1 were achieved by
1D NMR and 2D NMR (1H-1H COSY, HMQC,
HMBC and NOESY) spectra (Table 1).
1H
13C
1H
13C
1
2
3
4
5
6
7
131.3
114.5
149.2
146.2
116.3
40
50
60
70
80
90
153.6
116.0
6.62,d,1.8
6.61,d,8.4
7.18,dd,1.8,8.4 125.4
203.5
6.55,d,8.1
2.18,m
3.86,dd,7.7,11.0
3.72,dd,3.8,11.0
3.67, s
4.04, d, 7.7
3.06,m
3.15,m
3.15,m
3.06,m
3.66,m
3.50,dd,5.5,11.7
50.1
62.0
6.51,dd,1.8,8.1 123.5
2.89,dd,7.7,13.6
2.73,dd,6.8,13.6
2.20,m
3.30,dd,5.6,10.2
3.76,dd,4.8,10.2
3.62,s
37.0
30-OMe
Glc-1
Glc-2
Glc-3
Glc-4
Glc-5
Glc-6
56.7
105.0
75.5
78.4
71.9
78.2
63.0
8
9
43.1
70.5
3-OMe
56.7
133.3
112.7
149.3
10
20
30
Compound 2 was obtained as an amorphous powder.
Its negative ion mode ESI-MS showed a quasi-mole-
cular ion at m/z 537.3 [M–H]À and a significant frag-
ment at m/z 374.9 [M–H-162]À. A quasi-molecular ion
at m/z 561.3 [M+Na]+ and a remarkable fragment at
m/z 399.1 [M+Na-162]+ were also observed in its
positive ion mode ESI-MS. The molecular formula of 2
was then deduced to be C26H34O12 on the basis of a
combination of ESI-MS and 13C NMR (DEPT) (Table
2), and confirmed by HREIMS at m/z 376.1530
(C20H24O7, calcd: 376.1522) [M-glc (162)]+. Further
7.26,d,1.8
(d, J=1.8 Hz), 6.61 (d, J=8.4 Hz) and 7.18 (dd, J=1.8,
8.4 Hz), suggested the presence of two 1,3,4-trisub-
stituted benzene rings. In the NOESY spectrum, The
NOE correlations between 3-OMe at dH 3.62 and H-2 at
dH 6.62 (d, J=1.8 Hz), and between 30-OMe at dH 3.67
and H-20 at dH 7.26 (d, J=1.8 Hz) strongly indicated the
two methoxyl groups were connected to the C-3 and
C-30, respectively. The glucose moiety assigned as b-
1
analysis of its H and 13C NMR indicated the presence
Figure 1. Structures of compounds 1–16.

Cheng-Qi Fan, Jian-Min Yue / Bioorg. Med. Chem. 11 (2003) 703–708
Table 2. 1H (400 MHz) and 13C (100 MHz) NMR of 2 (CD3OD, d in ppm)
705
1H
13C
1H
13C
1
2
3
4
5
6
7
8
9
134.9
113.9
150.7
147.8
117.8
123.5
85.7
50
60
70
6.92,d,7.9
6.85,dd,1.8,7.9
3.30,dd,3.2,12.9
2.70,dd,12.6,12.9
2.80,m
4.30,dd,6.6,8.4
3.90,overlapped
4.05,s
5.07, d, 7.3
3.68,m
3.58,m
3.58,m
3.65,m
116.5
122.6
35.2
7.23,d,1.9
7.33,d,8.4
7.07,dd,1.8,8.4
5.07,s
80
90
52.3
62.0
83.6
64.8
30-OMe
Glc-1
Glc-2
Glc-3
Glc-4
Glc-5
Glc-6
56.7
103.3
75.3
78.5
71.7
78.2
62.9
4.00,d,11.4
3.83,d,11.4
4.10,s
3-OMe
57.0
133.6
113.8
149.4
146.2
10
20
30
40
7.00,d,1.8
3.90,m
4.12,m
of two 1,3,4-trisubstituted benzene rings, a glucopyr-
anosyl moiety, two methoxyl groups, three methylenes
(two oxygenated), two methines (including one oxyge-
nated) and one oxygenated quaternary carbon in the
determined to be (À)-berchemol 4-O-b-d-glucopyrano-
side (Fig. 1), and its NMR data were assigned as in
Table 2.
1
structure of 2. The H NMR and 13C NMR patterns of
Structural identification of the known compounds 3–16
aglycone moiety of 2 showed great similarity with those
of (À)-berchemol (4), which was also isolated from this
plant. The oxygenated methine at dC 85.7 and a singlet
proton dH 5.07 were respectively assigned to C-7 and H-
7. An oxygenated methylene at dC 64.8, bearing two
AB-type-coupling protons at dH 4.00, 3.83 (each 1H, d,
J=11.4 Hz, H-9) correlated with the quaternary carbon
C-8 at dC 83.6 in the HMBC spectrum, was attributable
to C-9. The carbon signals at d 35.2, 52.3 and 72.4 were
respectively assigned to C-70, C-80 and C-90 according to
the chemical shift and DEPT spectrum. The proton sig-
nals of H-70, H-80 and H-90 were then respectively dis-
tinguished by the correlations with C-70, C-80 and C-90
in the HMQC. A 7, 90- epoxy linkage was indicated by
the strong correlations among H-7 with C-80 and C-90 in
the HMBC spectrum. A NOESY spectrum was per-
formed to reveal the relative configuration of compound
2. The key NOE correlations (Fig. 2) revealed that the
aglycone of 2 had the same planar structure and relative
configuration as that of (À)-berchemol.
Among those of the known compounds, apigenin (5),
luteolin (7), quercetin (8), quercetin 3-O-b-d-glucopyr-
anoside (9) and vanillic acid (16) were identified by
spectral data (1H NMR and MS) and comparison with
authentic
drodiconiferyl alcohol 90-O-b-d-glucopyranoside (3),9
(À)-berchemol 4-(2-hydroxyethyl)-2-methox-
(4),8
yphenyl b-d-glucopyranoside (10),10 2-(4- hydroxy-3-
methoxyphenyl)propan-1,3-diol (2S)-3-(4-
(12),11
samples
(co-TLC).
Dihydrodehy-
hydroxy-3-methoxy phenyl)propan-1,2-diol (13),12 syr-
ingin (14)13 and glucovanillyl alcohol (15)14 were iden-
tified by comparison of their spectral data (1H, 13C
NMR, MS and [a]D) with those reported. 2-(4-
Hydroxy-3-methoxyphenyl)ethyl b-d-glucopyranoside
11 was identified by spectral data for the first time as a
natural product, which was synthesized before.15 Com-
pound 6 was identified as apigenin 7-O-[a-l-rhamno-
pyranosyl-(1!6)-b-d-glucopyranoside] by comparison
1
of H and 13C NMR spectral data with those of its iso-
mer apigenin 7-O-[a-l-rhamnopyranosyl-(1!2)-b-d-
glucopyranoside].5 The glc-6 carbon signal in com-
pound 6 was down-field shifted to 66.0 ppm in the 13C
NMR caused by the glucosylation, while that of the
latter was appeared at 61.0 ppm (both in DMSO-d6).
The spectral data of compounds 6 and 11 were pre-
sented in the experimental part. Except for compounds
5, 7 and 9, all other known compounds were isolated
from this plant for the first time.
The glucopyranosyl moiety was linked to C-4 with b-
configuration was ascertained according to the 1H
NMR, HMBC and NOESY spectra. Acidic hydrolysis
of compound 2 afforded d-glucose and (À)-berchemol
(4), [a]2D0À9.3ꢀ (c 0.081, MeOH) (reported value À7.9ꢀ,
c=0.30, in MeOH).8 The structure of compound 2 was
Evaluation of biological activities
Effects on scopolamine-induced memory deficits of mice.
In the course of our search for biologically active
components from natural resources, the polar fraction
of the EtOH extract of S. medusa showed a moderate
activity to attenuate the scopolamine induced memory
deficit by mouse behavioral tests in the water maze.
After purification, one major compound 6 obtained
from the polar fraction presented remarkable activity
Figure 2. Main NOE correlations of compound 2.

706
Cheng-Qi Fan, Jian-Min Yue / Bioorg. Med. Chem. 11 (2003) 703–708
to attenuate the scopolamine induced memory deficit in
the same animal model. Oral administration of com-
pound 6 at the dosages of 10, 15 and 20 mg/kg and
then injection of scopolamine at the dose of 4.5 mg/kg
for each group of mice 30 min later, the mice taking
compound 6 in the tested groups obviously reduced the
errors to enter non-exit and shortened the time to reach
the platform than those injected only 4.5 mg/kg of sco-
polamine (Table 3).
Cell protective activities of major phenolics against
H2O2-iduced PC12 cell damage
The major phenolic compounds 4, 6–9 and 14 were tes-
ted for protecting activity against H2O2-induced
damage in rat pheochromocytoma line PC12 cells. Two
of them, apigenin 7-O-[a-l-rhamnopyranosyl-(1!6)-b-
d-glucopyranoside] (6) and quercetin (8), showed mod-
erate protective activities against H2O2-induced cell
damage (Fig. 3).
Experimental
General
Optical rotation [a]D values were measured with a Per-
kin–Elmer polarimeter 341. UV spectra were recorded
on a Shimadizu UV-210A spectrometer. IR spectra were
performed on a Nicolet Magna 750 spectrometer with
KBr discus. NMR spectra were obtained on a Brucker
AM-400 MHz spectrometer. General 1H NMR data
were run at 400 MHz, and 13C NMR were measured at
100.6 MHz. Chemical shifts are expressed in ppm rela-
tive to TMS. ESI-MS were recorded with a Finnigan
LCQDECA mass spectrometer.
Figure 3. Effects of compounds 6 and 8 on cell viability. Cells were
incubated with 200 mM H2O2 for 30 min and the cultures were further
developed for another 6 h. Samples were added to the cultures 2 h
prior to H2O2 addition. Cell viability was obtained by measuring the
MTT reduction. Two independent experiments were carried out in
triplicate. All data were expressed as percentage of control value. Sta-
tistical comparison was made by using one-way ANOVA, and fol-
lowed by Duncan’s test. The data were expressed as meansÆSEM;
*P<0.05, **P<0.01 vs H2O2 control group.
Plant material
Extaction and isolation
The whole plant of S. medusa Maxim was collected in
September 2000 in the Tibet Autonomous Region of
China, and was identified by professor Zeng-Tao Wang
of Shanghai Chinese Traditional Medical Unversity. A
voucher specimen (No. Sm-2000-1Y) was deposited in
Shanghai Institute of Materia Medica, Shanghai Insti-
tutes for Biological Sciences, Chinese Academy of Sci-
ences, PR China.
The dried whole plant of S. medusa (2.5 kg) was pow-
dered and extracted by reflux in 95% EtOH for three
times (2LÂ3), and the combined EtOH solution was
evaporated to dryness under vacuum to give an extract
(230 g). The extract was suspended in 1.0 L water and
then extracted successively with dichloromethane and n-
butanol to give fractions DM (60 g) and Bu (45 g),
Table 3. Effects on scopolamine-induced mice memory deficits of compound 6
Sample combinations
Dose (mg/kg+mg/kg) po+ip
N
Time to find the platform
Number of entering non-exit
Saline+saline
/+4.5
/+4.5
10+4.5
15+4.5
20+4.5
12
16
10
10
10
15.8Æ2.8
51.4Æ3.4a
44.1Æ5.6
36.7Æ6.5
43.8Æ8.3
0.9Æ0.3
7.5Æ0.6a
4.8Æ1.0b
3.8Æ1.0c
5.9Æ1.4
Saline+scopolamine
Compound 6
+scopolamine
N stands for the numbers of mice in the tested groups; data were expressed as meansÆSEM. All mice were trained to follow a criterion of finding the
platform within 20 min and the errors of entering non-exit were less than 2 (<2). The samples were orally administered 30 min prior to injection (ip)
of scopolamine. The time to reach the platform and the numbers of entering non-exit were measured after 20 min.16
aP<0.05 vs saline+saline group.
bP<0.05 vs saline+scopolamine group.
cP<0.01 vs saline+scopolamine group.

Cheng-Qi Fan, Jian-Min Yue / Bioorg. Med. Chem. 11 (2003) 703–708
707
respectively, and the aqueous part was condensed to
give water-soluble fraction W (122 g).
(37); EI-MS m/z (%): 376 [M–glc]+(25), 153 (36), 137
(100) and 124 (34); 1H and 13C NMR data: see Table 2.
The fraction Bu (45 g) was subjected to column chro-
matography (CC) on MCI GEL CHP 20P eluted with
H2O, 20% and 50% methanol in water, MeOH and
acetone to give fractions 1–5. Fraction 5 was passed
through a column of Sephadex LH-20 eluted with
EtOH to give daucosterol. Fraction 4 was re-chromato-
graphed on MCI GEL CHP 20P eluted with 70%
methanol in water to give a major fraction 4a contain-
ing flavonoids and four other fractions 4b–4e. Fraction
4d was then subjected to CC on silica gel eluted with
CHCl3–MeOH (10:1) to give compound 4 (18 mg).
Fraction 4a was subjected to CC on silica gel eluted
with petroleum ether–EtOAc–formic acid (2:1:0.02) to
give three parts and then passed through columns of
Sephadex LH-20 eluted with EtOH to give compounds
5 (75 mg), 7 (134 mg) and 8 (55 mg). Fraction 4c was
chromatographed on a reversed phase C18 silica gel
eluted with 80% methanol in water to give compound
16 (83 mg). Part of fraction 3 was passed through a
column of Sephadex LH-20 eluted with 70% ethanol in
water to give two flavonoid glycosides, compound 6
(784 mg) and 9 (51 mg). Fraction 2 was re-chromato-
graphed on a column of MCI GEL CHP 20P eluted
with 10–50% methanol in water to obtain six fractions
2a–2f. Compounds 12 (13.5 mg) and 13 (6.5 mg) were
obtained from fraction 2a by extensive column chro-
matography on reversed phase C18 silica gel eluted with
30–40% methanol in water. Fraction 2bwas passed
through a column of Sephadex LH-20 eluted with 70%
ethanol in water and then subjected to CC on silica gel
eluted with CHCl3–MeOH (6:1) to give compound 15
(16 mg). Fraction 2c was subjected to CC on silica gel
eluted with CHCl3–MeOH (6:1) to give compound 14
(68 mg). Fraction 2d was passed through a column of
Sephadex LH-20 eluted with 70% ethanol in water and
then re-chromatographed on reversed phase C18 silica gel
eluted with 40% methanol in water to give compounds 1
(15 mg) and 2 (12.5 mg). Fraction 2e was chromato-
graphed on reversed phase C18 silica gel eluted with 40%
methanol in water to give compound 3 (13 mg).
Acidic hydrolysis of 2. Compound 2 (6 mg) was added
to 4 mL 5% H2SO4 in MeOH and refluxed for 4 h. The
reaction mixture was neutralized with 5% NaHCO3 and
then worked up to give yellow residue. The residue was
passed through a Sephadex LH-20 column eluted with
EtOH to obtain glucose and (À)-berchemol (3.2 mg)
which was identified by comparison of co-TLC and
optical rotation [a]D20 À9.3ꢀ (c 0.081, MeOH) with those
of authentic sample.
Apigenin 7-O-[ꢀ-L-rhamnopyranosyl-(1!6)-ꢁ-D-gluco-
pyranoside] (6). Yellow amorphous power. Positive
ESI-MS m/z (%): 601.3 [M+Na]+ (100), 579.3
[M+H]+ (80), 433.3 [M+H-146]+ (47), 271.5 [M+H-
146-162]+ (66); 1H NMR data (400 MHz, DMSO-d6) d:
6.87 (1H, s, H-3), 6.45 (1H, s, H-6), 6.78 (1H, s, H-8),
6.98 (2H, d, J=8.8 Hz, H-30, 50), 7.98 (2H, d, J=8.8 Hz,
H-20, 60), 5.08 (1H, d, J=7.3 Hz, glc-1), 4.54 (1H, s,
rhm-1), 1.08 (1H, s, rhm-6); 13C NMR data (100 MHz,
DMSO-d6) d: 164.4 (C-2), 103.1 (C-3), 182.0 (C-4),
105.4 (C-4a), 161.2 (C-5), 99.9 (C-6), 162.9 (C-7), 94.8
(C-8), 157.0 (C-8a), 121.1 (C-10), 128.7 (C-20, 60), 161.2
(C-40), 116.1 (C-30, 50), 99.5 (glc-1), 72.0 (glc-2), 76.2
(glc-3), 70.7 (glc-4), 75.6 (glc-5), 66.0 (glc-6), 100.5 (rhm-
1), 70.3 (rhm-2), 69.5 (rhm-3), 73.1 (rhm-4), 68.3 (rhm-
5) and 17.8 (rhm-6).
2-(4-Hydroxy-3-methoxyphenyl)ethyl ꢁ-D-glucopyrano-
side (11). White amorphous powder. Positive ESI-MS
m/z (%): 353.3.2 [M+Na]+ (100); negative ESI-MS m/z
(%): 329.3 [M–H]À (100), 167.1 [M–H-162]À (30), 113.0
1
(95); H NMR data (400 MHz, CD3OD) d: 3.55, 3.90
(2H, m, H-1), 2.67 (2H, m, H-2), 6.70 (1H, d, J=1.8 Hz,
H-20), 6.53 (1H, d, J=8.0 Hz, H-50), 6.49 (1H, dd,
J=1.8, 8.0 Hz, H-60), 3.66 (3H, s, H-OMe), 4.13 (1H, d,
J=7.7 Hz, glc-1), 3.00-3.18 (4H, m, glc-2, 3, 4, 5), 3.50,
3.70 (2H, m, glc-6); 13C NMR data (100 MHz, CD3OD)
d: 72.0 (C-1), 36.7 (C-2), 131.6 (C-10), 113.8 (C-20), 148.8
(C-30), 145.9 (C-40), 116.1 (C-50), 122.4 (C-60), 56.4 (C-
OMe), 104.3 (glc-1), 75.1 (glc-2), 78.1 (glc-3), 71.7 (glc-
4), 77.9 (glc-5), 62.8(glc-6).
Medusaside
A
(1). White amorphous powder.
[a]2D0 À 60:9ꢀ (c 0.755, MeOH); UV lmax (MeOH) (log
"): 229 (4.08), 279 (3.85), 306 (3.68) nm; IR (KBr disc)
ꢀ=3406 (OH), 2933 (C–H), 1655, 1591, 1516 (Ar–
C¼O), 1450, 1427 (C–H), 1275, 1161, 1076, 1031 (C–O)
cmÀ1; positive ESI-MS m/z (%): 561.3 [M+Na]+ (100);
negative ESI-MS m/z (%): 537.2 [M–H]À (100); EI–MS
m/z (%): 376 [M–glc]+(5), 358 [M–glc–H2O]+(8), 340
[M–glc–2H2O]+(12), 191 (20), 151 (48), 137 (100) and
Biology
Effects on scopolamine-induced memory deficits of mice.
Effects on scopolamine-induced memory deficits of mice
were carried out according to the procedure described in
the literature.16
Tests of cell protective activities. Evaluation of cell pro-
tective activities of major phenols against H2O2-iduced
PC12 cell damage was made according to the protocol
reported in the literatures.17
1
98 (88); H and 13C NMR data: see Table 1.
Medusaside B (2). White amorphous powder. [a]D20
À34.9ꢀ (c 0.53, MeOH); UV lmax (MeOH) (log "): 227
(3.89), 279 (3.43) nm; IR (KBr disc) ꢀ=3415 (OH), 2924
(C–H), 1601, 1514 (Ar), 1452, 1425 (C–H), 1269, 1070,
1032 (C–O) cmÀ1; Positive ion mode ESI-MS m/z (%):
561.2 [M+Na]+ (48), 399.1 [M+Na-162]+ (100);
Negative ion mode ESI-MS m/z (%): 537.3 [M–H]À (23),
489.0 (25), 374.9 [M–H-162]À (100), 345.0 (20) and 327.0
Acknowledgements
Financial support of the National Scientific Foundation
(for J. M. Yue, 30025044) and the Shanghai Municipal
Scientific Foundation for Fundamental Research (for J.

708
Cheng-Qi Fan, Jian-Min Yue / Bioorg. Med. Chem. 11 (2003) 703–708
M. Yue, 00JC14053) is gratefully acknowledged. We
thank professor Xi-Can Tang of Shanghai Institute of
Materia Medica for biological assays. We thank Pro-
fessor Zeng-Tao Wang of Shanghai Chinese Traditional
Medical University for identification of plant resource.
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