全 文 ：Journal of Chinese Pharmaceutical Sciences208
Chemical constituents from Pithecellobium clypearia and their
effects on T lymphocytes proliferation
Xiao-Yu Guo1, 3, Nai-Li Wang 2, 3, Li Bo2, 3, Yi-Hua Li4, Qiang Xu4 and Xin-Sheng Yao2, 3*
1. Department of Natural Medicines, School of Pharmaceutical Sciences, Peking University, Beijing 100083, China;
2. College of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China;
3. Traditional Chinese Medicines and Natural Products Research Center Shenzhen, Shenzhen 518057, China;
4. State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210093, China
Abstract: Aim To investigate the chemical constituents from the twigs and leaves of Pithecellobium clypearia Benth and their
immunomodulatory effects. Methods The constituents were separated and purified by various chromatographic methods and their
structures were identified on the basis of spectral analysis. The immunomodulatory effects of all the compounds were examined
by a Con A-induced T lymphocytes proliferation assay. Results Eight compounds were isolated and identified as (-)-
epigallocatechin (1), (-)-5, 7, 3, 4, 5-pentahydroxyflavan (2), (-)-epigallocatechin-7-gallate (3), (-)-5, 3, 4, 5-tetrahydroxyflavan-
7-gallate (4), quercitin-3-O-α-L-rhamnpyranoside (5), myricitin-3-O-α-L-rhamnpyranoside (6), gallic acid (7), and ethyl gallate (8),
respectively. Conclusion Compounds 3 and 8 were isolated from this genus for the first time, and compound 1 was isolated
from this species for the first time. Compound 3 exhibited a strong inhibition on the T lymphocytes proliferation induced by Con
A with an IC50 of 4.4 μmol·L–1.
Keywords: Pithecellobium clypearia; Flavonoids; Flavane; Lymphocytes proliferation
CLC number: R284.2 Document code: A Article ID: 1003–1057(2007)3–208–06
Introduction
Pithecellobium clypearia Benth, a member of
Mimosoaceaeis family, is an aiphyllium and distrib-
uted widely in the south of China[1]. Its twigs and
leaves are used as a herbal medicine in the treatment
of empyrosis and rheumatism[2]. The pharmaceutical
preparations made of the aqueous extract of the
twigs and leaves of P. clypearia have been used in
China to treat upper respiratory infection, acute
laryngopharyngitis, acute tonsillitis, and chordapsus.
Phenolic compounds, such as (7-O-galloyltricetiflavan
and 7, 4-O-di-galloyltricetiflavan) have been isolated
from this plant[3, 4].
In the current study, eight compounds were iso-
lated from the 60% ethanol extract of the twigs and
leaves of P. clypearia and their structures were
identified as (-)-epigallocatechin (1), (-)-5, 7, 3,
4, 5-pentahydroxyflavan (2), (-)-epigallocatechin-7-
gallate (3), (-)-5, 3, 4, 5-tetrahydroxyflavan-7-
gallate (4), quercitin-3-O-α?-L-rhamnpyranoside (5),
myricitin-3-O-α-L-rhamnpyranoside (6), gallic acid
(7), and ethyl gallate (8) by spectral analysis and
comparing the spectral data with those of the
literatures. The immunomodulatory effects of eight
compounds were examined by a Con A-induced T
lymphocytes proliferation assay.
Results and discussion
Compound 1, 29D[ ]a = –71.8° (MeOH, c. 1.0), responded
positively to 3% FeCl3-ethanol solution. The IR spec-
trum showed the absorption bands at 3244, 1624, 1524,
and 1458 cm 1¯ which were ascribable to the hydroxyl
and aromatic moieties. The ESI-MS spectrum showed
the ions of m/z 307 [M + H]+ and 305 [M – H] ,¯ suggesting
that the molecular weight of 1 was 306. The molecular
formula of 1 was deduced as C15H14O7 in combination
with the NMR data. The 1H and 13C NMR data of 1
were in agreement with those of (-)-epigallocatechin[5].
Thus, 1 was identified as (-)-epigallocatechin.
Compound 2, 28D[ ]a = –2.3° (MeOH, c. 1.0), showed
a positive response to spraying 3% FeCl3-ethanol solution,
and the IR spectrum of 2 was similar to that of 1.
The molecular formula of 2 was deduced as C15H14O6
according to the ions of m/z 313 [M + Na]+ and 289
[M - H] ¯in the ESI-MS spectrum and the NMR data. In
the 13C NMR spectrum, the presence of twelve aro-
matic carbon signals at ä 157.7–95.8, one oxygenated
methenyl carbon signal at ä 78.0, and two methylene
carbon signals at ä 30.4, and 19.8, suggested that 2 was a
flavan. The 1H NMR and HMQC spectra showed a pair
of meta-couple aromatic proton signals from A-ring at ä
6.00 (1H, d, J = 2.4 Hz, H-6) and 5.88 (1H, d, J = 2.4
Hz, H-8) , two aromatic proton signals from B-ring at ä
6.48 (2H, s, H-2, 6) , one oxygenated methenyl proton
Received date: 2007-01-22.
*Corresponding author. Tel.: 86-20-85225849;
fax: 86-20-85221559; e-mail: yaoxinsheng@vip.tom.com
X. Y. Guo et al. / Journal of Chinese Pharmaceutical Sciences 2007 (16) 208–213 209
signal at ä 4.78 (1H, dd, J = 9.7, 2.4 Hz, H-2) and
two methylene proton signals at ä 2.62 (2H, m, H-
4), 2.10 (1H, m, H-3b), and 1.88 (1H, m, H-3a)
from C-ring. So the structure of 2 was elucidated as
(-)-5, 7, 3, 4, 5-pentahydroxyflavan[4].
Compound 3, 28D[ ]a = –37.4° (MeOH, c. 1.0),
showed absorption bands in the IR spectrum at
3244, 1697, 1620, 1512, and 1454 cm 1¯ which were
ascribable to hydroxyl, carbonyl, and aromatic
moieties. The molecular formula of 3 was deduced
as C22H18O11 from the ions of m/z 481 [M + Na]+,
459 [M + H]+, and 457 [M - H] ¯ in the ESI-MS
spectrum and the NMR data. The 13C NMR and
DEPT spectra showed the signals of one carbonyl
carbon (ä 165.3), eighteen aromatic carbons (ä 157.2 –
101.7), two oxygenated methenyls (ä 79.5, 66.5),
and one methylene (ä 29.0), suggesting the presence
of three aromatic groups and one ester group in the
structure of 3. The 1H NMR and HMQC spectra ex-
hibited similar signal profile to that of 1, except for
the aromatic proton signal at ä 7.25 (2H, s, H-2,
6). In the HMBC spectrum, the correlations
between H-2 (ä 4.92) and C-9 (ä 156.8), C-1 (ä
131.1), C-2, and C-6 (ä 106.8), H-3 (ä 4.28) and
C-10 (ä 106.0), H-4 (ä 2.96, 2.88) and C-9 (ä 156.8),
and C-10 (ä 106.0) confirmed the presence of the
same flavanol skeleton in the structure of 3 with
that of 1. The HMBC correlation between H-2, 6
(ä 7.25) and the carbonyl carbon at ä 165.3 indi-
cated the presence of one galloyl group in the
structure. Comparing the 13C NMR data of 3 with
those of 1, the signals of C-7 was found to have an
upfield shift of 6.4 ppm, with C-6, C-8 and C-10
having downfield shiftes of 5.5, 6.5, and 6.2 ppm,
respectively. The above NMR data indicated that the
position of the galloyl ester group was located at C-7
of epigallocatechin. Thus, the structure of 3 was
elucidated as (-)-epigallocatechin-7-gallate[6, 7].
Compound 4, 28D[ ]a = –2.6° (MeOH, c. 1.0), had
the similar IR spectrum to that of 3. The HR-TOF-
ESI-MS spectrum showed the ion at m/z 443.0948
[M + H]+ to give the molecular formula C22H18O10.
The 13C NMR data of 4 were similar to those of 3,
and the 1H NMR showed the signals similar to those
of 2, except for the carbon signal at ä 30.0 (C-3)
and the aromatic proton signal at ä 7.24 (2H, s, H-
2, 6), indicating that 4 was a flavan galloyl ester.
The HMBC correlations between H-2 (ä 4.87) and
C-9 (ä 157.4), C-1 (ä 133.9), C-2 , and C-6
(ä 106.1), H-3b (ä 2.17) and C-10 (ä 108.1), H-4
(ä 2.73) and C-10 (ä 108.1), and C-9 (ä 157.4)
confirmed the presence of the same flavan skeleton
in the structure of 4 with that of 2, and the HMBC
correlation between H-2, 6 (ä 6.51) and the car-
bonyl group at ä 165.3 indicated the presence of one
galloyl group. Comparing the 13C NMR data of 4
with those of 2, the signal of C-7 showed an upfield
shift of 6.4 ppm, and the signals of C-6, C-8, and
C-1 shifted downfield for 5.7, 6.6, and 6.2 ppm,
suggesting that the position of the galloyl ester was
at C-7 of the flavan. Consequently, the structure of
4 was characterized as (-)-5, 3, 4, 5-tetrahydroxy-
flavan-7-gallate[3]. The structures of compounds 1 –
8 were showed in Figure 1.
Figure 1. Structures of compounds 1 – 8 from Pithecellobium
clypearia Benth.
1 R = OH
2 R = H
3 R = OH
4 R = H
5 R = H
6 R = OH
7 R = H
8 R = CH2CH3
X. Y. Guo et al. / Journal of Chinese Pharmaceutical Sciences 2007 (16) 208–213210
(-)-Epigallocatechin-7-gallate (3) and ethyl gallate
(8) were isolated from this genus for the first time,
and (-)-epigallocatechin (1) was isolated from this
species for the first time. In the present study, we
first examined the immunomodulatory effects of the
phenolic compoumds from P. clypearia by a Con A-
induced T lymphocytes proliferation assay. Com-
pounds 1 – 8 all inhibited the T lymphocytes prolif-
eration induced by Con A, and compound 3 showed
the strongest inhibitory effects with an IC50 value of
4.4 μmol·L–1 (see Table 1). The results suggested
that the phenolic compounds from P. clypearia may
inhibit the immunological function of mice cells by
inhibiting the T lymphocytes proliferation.
Experimental
General procedures
Melting points were determined on a YANAGIMOTO
micromelting point apparatus and uncorrected. UV
spectra were measured in MeOH using a SHIMADZU
UV2401PC spectrophotometer. IR spectra were run
in KBr disks with a SHIMADZU FTIR-8400
spectrophotometer. HR-TOF-ESI-MS and ESI-MS
spectra were recorded on a Micromass Q-TOF
mass spectrometer and a Bruker Esquire 2000 mass
spectrometer, respectively. NMR spectra were recorded
on a Bruker AVANCE-400 spectrometer with TMS as
an internal standard. Diaion HP-20 and MCI GEL
CHP20P (Mitsubisghi Chemical Corporation, Japan),
ODS-A 120-S150 (YMC Co., Ltd., Japan),
SephadexTM LH-20 (Amersham Biosciences AB,
Sweden), and Toyopearl HW-40F (Tosoh Corporation,
Japan) were used for the column chromatography.
Plant material
The twigs and leaves of Pithecellobium clypearia
were collected in October 2004, from Conghua in
Guangdong Province, China, and identified by Bai-
Ying Liu (Associate chief pharmacist, Guangdong
Institute of Drug Control). A voucher specimen
(YGXYPC-2004) was deposited in Traditional Chinese
Medicines and Natural Products Research Center
Shenzhen, Shenzhen, China.
Extract and isolation
The twigs and leaves of P. clypearia (7.6 kg) were
extracted two times with 60% ethanol for 2 h/each.
The combined extract was concentrated under re-
duced pressure to yield a dark-brown residue (1.5
kg, 19.7 %). The residue (1.2 kg) was suspended in
water, and partitioned successively with CHCl3,
EtOAc, and n-BuOH, respectively. The EtOAc extract
(100.0 g) was subjected to a Diaion HP-20 column
chromatography (5.2 cm × 46.5 cm) eluted with
MeOH-H2O gradiently to give nine fractions (Fr. 1 – 9).
Fr. 2 (10.0 g) was subjected to a Sephadex LH-20
column chromatography eluted by MeOH-H2O
gradiently and was recrystallized in MeOH-H2O (10 : 90,
V/V) to afford 7 (4100.0 mg). Fr. 3 (28.0 g) was
separated by a MCI GEI CHP 20P column chroma-
tography eluted by MeOH-H2O gradiently to give
four fractions (Fr. 3.1 – 3.4). Fr. 3.1 was subjected
to a column chromatography on Sephadex LH-20
eluted by MeOH-H2O gradiently and was recrystal-
lized in MeOH-H2O (20 : 80, V/V) to obtain 1 (30.0
mg) and 2 (1000.0 mg). Fr. 3.2 was separated by a
Sephadex LH-20 column chromatography eluted by
MeOH-H2O gradiently to afford 3 (900.0mg), 4
(5500.0 mg), and 8 (2400.0 mg). Fr. 4 (30.0 g)
was subjected to a medium pressure liquid chroma-
tography on ODS eluted by MeOH-H2O gradiently
and was recrystallized in MeOH-H2O (20 : 80, V/V) to
give 5 (842.0 mg) and 6 (117.2 mg).
Identification
(-)-Epigallocatechin (1). White amorphous powder;
29
D[ ]a = –71.8° (MeOH, c. 1.0); UV(MeOH) ëmax
(log å) nm = 212 (4.95), 270 (3.69), 392 (3.56);
IR (KBr) ímax (cm
1¯): 3244 (-OH), 1624, 1524, 1458
(-Ar); ESI-MS: m/z 307 [M + H]+, 305 [M - H] ;¯ 1H
and 13C NMR data, see Table 2.
Table 1. Inhibitory effects of compounds 1 – 8 on T lymphocytes
proliferation
Compound IC50 (μmol·L 1¯)
1 > 50.0
2 21.9
3 4.4
4 25.0
5 > 50.0
6 30.0
7 25.1
8 27.1
211X. Y. Guo et al. / Journal of Chinese Pharmaceutical Sciences 2007 (16) 208–213
(-)-5, 7, 3, 4, 5-Pentahydroxyflavan (2). White
amorphous powder; 28D[ ]a = –2.3° (MeOH, c. 1.0);
UV (MeOH) ëmax (log å) nm = 214 (4.96), 270 (3.69);
IR (KBr) ímax (cm
1¯): 3294 (-OH), 1624, 1520, 1474
(-Ar); ESI-MS: m/z 313 [M + Na]+, 289 [M - H] ;¯
1H and 13C NMR data, see Table 2.
(-)-Epigallocatechin-7-gallate (3). White amor-
phous powder; 28D[ ]a = –37.4° (MeOH, c. 1.0); UV
(MeOH) ëmax (log å) nm = 211 (5.09), 280 (4.44);
IR (KBr) ímax (cm
1¯): 3244 (-OH), 1697 (C=O),
1620, 1512, 1454 (-Ar); ESI-MS: m/z 481 [M + Na]+,
459 [M + H]+, 457 [M - H] ;¯ 1H and 13C NMR data,
see Table 2.
(-)-5, 3, 4, 5-Tetrahydroxyflavan-7-gallate (4).
White amorphous powder; 28D[ ]a = –2.6° (MeOH, c.
1.0); UV (MeOH) ëmax (log å) nm = 210 (4.97), 280
(4.31), 446 (3.45), 473 (3.45); IR (KBr) ímax (cm
1¯):
3244 (-OH), 1732 (C=O), 1620, 1535, 1443 (-Ar);
ESI-MS: m/z 465 [M + Na]+, 443 [M + H]+, 441
[M - H] ;¯ 1H and 13C NMR data, see Table 2.
Quercitin-3-O-á-L-rhamnpyranoside (5). Yellow
amorphous powder; 29D[ ]a = –205.4° (MeOH, c. 1.0);
UV (MeOH) ëmax (log å) nm = 250 (4.80), 256 (4.53),
351 (4.41); IR (KBr) ímax (cm
1¯): 3260 (-OH), 1655
Table 2. 1H NMR (400 MHz) and 13C NMR (100 MHz) data of compounds 1 – 4 (in acetone-d6)
(C=O), 1605, 1497, 1454 (-Ar); ESI-MS: m/z 471
[M + Na]+, 325 [M + Na - 146]+, 447 [M - H] ,¯
301 [M - H - 146] ;¯ 1H NMR (400 MHz, acetone-
d6): ä 6.27 (1H, d, J = 2.1 Hz, H-6), 6.47 (1H, d,
J = 2.1 Hz, H-8), 7.50 (1H, d, J = 2.2 Hz, H-2), 7.00
(1H, d, J = 8.3 Hz, H-5), 7.40 (1H, dd, J = 8.3, 2.2
Hz, H-6), 5.52 (1H, d, J = 1.4 Hz, H-1), 4.33 (1H,
dd, J = 3.5, 1.4 Hz, H-2), 3.73 (1H, dd, J = 9.8, 3.5
Hz, H-3), 3.36 (1H, t, J = 9.0 Hz,H-4), 3.38 (1H,
m, H-5), 0.92 (3H, d, J = 5.9 Hz, H-6), 12.73
(1H, s, 5-OH); 13C NMR (100 MHz, acetone-d6): ä
158.4 (C-2), 135.8 ( C-3), 179.4 (C-4), 163.2 (C-5),
99.5 (C-6), 165.0 (C-7), 94.5 (C-8), 158.0 (C-9),
105.8 (C-10), 122.9 (C-1), 116.8 (C-2), 145.9 (C-3),
149.0 (C-4), 116.2 (C-5), 122.6 (C-6), 102.8 (C-1),
71.5 (C-2), 72.2 (C-3), 73.1 (C-4), 71.3 (C-5),
17.8 (C-6). The UV, IR, and NMR data were similar
to those of quercitin-3-O-á-L-rhamnpyranoside[4, 8].
Myricitin-3-O-á-L-rhamnpyranoside (6). Yellow
amorphous powder; 28D[ ]a = –156.9° (MeOH, c. 1.0);
UV (MeOH) ëmax (log å) nm = 209 (4.89), 257 (4.53),
354 (4.44); IR (KBr) ímax (cm 1¯): 3256 (-OH), 1655
(C=O), 1605, 1500, 1454 (-Ar); ESI-MS: m/z 487
[M + Na]+, 463 [M - H] ,¯ 317 [M - H - 146] ;¯ 1H NMR
(400 MHz, acetone-d6): ä 6.26 (1H, d, J = 2.2 Hz,
1 2 3 4
äH äC äH äC äH äC äH äC
2 4.82 (1H, br s) 79.4 4.78 (1H, dd, J = 9.7, 2.4) 78.0 4.92 (1H, br s) 79.5 4.87 (1H, dd, J = 9.9, 2.2) 78.3
4.20 (1H, br s) 67.0 2.10 (1H, m) 30.4 4.28 (1H, br s) 66.5 2.17 (1H, m) 30.0
1.88 (1H, m) 1.95 (1H, m)
2.85 (1H, dd, J = 16.6, 4.6) 28.8 2.62 (2H, m) 19.8 2.96 (1H, dd, J = 17.0, 4.3) 29.0 2.73 (2H, m) 20.1
2.73 (1H, dd, J = 16.6, 3.4) 2.88 (1H, dd, J = 17.0, 2.7)
5 157.6 157.0 157.2 156.8
6 6.02 (1H, d, J = 2.2) 96.2 6.00 (1H, d, J = 2.4) 95.8 6.34 (1H, d, J = 2.2) 101.7 6.31 (1H, d, J = 2.2) 101.5
7 157.6 157.7 151.2 151.3
8 5.92 (1H, d, J = 2.2) 95.7 5.88 (1H, d, J = 2.4) 95.9 6.28 (1H, d, J = 2.2) 102.2 6.23 (1H, d, J = 2.2) 102.5
9 157.1 157.5 156.8 157.4
10 99.9 101.9 106.0 108.1
1 131.5 134.4 131.1 133.9
2, 6 6.58 (1H each, s) 106.9 6.48 (1H each, s) 106.0 6.62 (1H each, s) 106.8 6.51 (1H each, s) 106.1
3, 5 146.1 146.4 146.0 146.5
4 132.9 133.0 132.9 133.1
1 121.0 121.1
2, 6 7.25 (1H each, s) 110.2 7.24 (1H each, s) 110.3
3, 5 146.1 146.2
4 139.4 139.4
C=O 165.3 165.3
b
a3
b
a4
Position
212 X. Y. Guo et al. / Journal of Chinese Pharmaceutical Sciences 2007 (16) 208–213
H-6), 6.46 (1H, d, J = 2.2 Hz, H-8), 7.10 (2H, s, H-
2, 6), 5.49 (1H, d, J = 1.4 Hz, H-1), 4.22 (1H,
dd, J = 4.2, 1.4 Hz, H-2), 3.75 (1H, dd, J = 9.4,
4.2 Hz, H-3), 3.37 (1H, t, J = 9.4 Hz, H-4), 3.52
(1H, m, H-5), 0.94 (3H, d, J = 6.2 Hz, H-6), 12.74
(1H, s, 5-OH); 13C NMR (100 MHz, acetone-d6): δ
158.5 (C-2), 135.9 ( C-3), 179.4 (C-4), 163.2 (C-
5), 99.5 (C-6), 165.0 (C-7), 94.5 (C-8), 158.0 (C-
9), 105.8 (C-10), 121.9 (C-1), 109.3 (C-2), 146.4
(C-3, 5), 137.0 (C-4), 109.3 (C-6), 102.8 (C-1),
71.5 (C-2), 72.2 (C-3), 73.2 (C-4), 71.3 (C-5),
17.8 (C-6). The UV, IR, and NMR data were similar
to those of myricitin-3-O-α-L-rhamnpyranoside[4, 9].
Gallic acid (7). White needles; mp 250 – 251 °C;
UV (MeOH) λmax (log ε) nm = 216 (4.67), 265 (4.16);
IR (KBr) υmax (cm
1¯) = 1701 (C=O), 1616, 1539,
1450 (-Ar); ESI-MS: m/z 171 [M + H]+, 169 [M - H] ;¯
1H NMR (400 MHz, DMSO): δ 6.94 (2H, s, H-2, 6);
13C NMR (100 MHz, DMSO): δ 167.5 (C=O), 120.5
(C-1), 108.7 (C-2, 6), 145.4 (C-3, 5), 138.0 (C-4).
The 1H NMR data was in agreement with those of
gallic acid[10].
Ethyl gallate (8). White needles; mp 156 – 158 °C;
UV (MeOH) λmax (log ε) nm = 217 (4.74), 275 (4.34);
IR (KBr) υmax (cm
1¯): 3294 (-OH), 1705(C=O),
1620, 1535, 1470 (-Ar); ESI-MS: m/z 221 [M + Na]+,
199 [M + H]+, 197 [M - H] ;¯ 1H NMR (400 MHz,
DMSO): δ 6.95 (2H, s, H-2, 6), 4.20 (2H, q, J = 7.1
Hz, -O-CH2-), 1.27 (3H, t, J = 7.1 Hz, -CH3); 13C
NMR (100 MHz, DMSO): δ 165.8 (C=O), 119.6
(C-1), 108.5 (C-2, 6), 145.5 (C-3, 5), 138.3 (C-4),
60.0 (-O-CH2-), 14.2 (-CH3). The 1H and 13C NMR
data were in agreement with those of ethyl gallate [11].
T lymphocytes proliferation assay[12]
Spleens were aseptically taken from the Kunming
mice, crushed gently and homogenized in 10 mL
RPMI-1640 medium (GIBCO BRL). The cell sus-
pension was filtrated with the gauze of 400 mesh
and centrifuged at 1000 r·min 1¯ for 5 min at 4 °C.
The cell pellet was suspended in 10 mL 0.17 mol·L–1
Tris (hydroxymethyl aminomethane) –0.75% NH4Cl
(pH 7.5), and centrifuged at 1000 r·min 1¯ for 5 min at
4 °C to remove erythrocytes. After washing twice
with RPMI-1640 medium, the cells were suspended
and cultured in fresh RPMI-1640 medium. The spleen
cells were seeded in 96-well plates at a density of
5 × 105 cells/well with RPMI-1640 medium, and
treated with 5 μg·mL–1 Con A and sample solution for
72 h at 37 °C in 5% CO2. Then, the effects on the T
lymphocytes proliferation were evaluated with the
modified MTT assay[13]. In brief, 20 μL of 5
mg·mL 1¯ MTT/RPMI-1640 solution were added for a
4-h incubation. The supernatant was removed after
centrifugation, and 200 μL DMSO were added to
dissolve the formazan crystals. The absorbance was
read on an ELISA reader (Sunrise Remote/Touch
Screen, TEACAN, Austria) at 540 nm.
Acknowledgements
The authors thank associate chief pharmacist Bai-
Ying Liu, Guangdong Institute of Drug Control, for
the identification of the plant Pithecellobium
clypearia Benth. Thanks are also extended to Xue
Zhang and San-Lin Jin for NMR experiments, and
Hao Gao and Xin-Luan Wang for ESI-MS experiments,
Traditional Chinese Medicines and Natural Products
Research Center.
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猴耳环的化学成分及其对T淋巴细胞增殖的影响
郭晓宇1, 3, 王乃利2, 3, 宝丽2, 3, 李轶华4, 徐强4, 姚新生2, 3
(1.北京大学药学院 天然药物学系, 北京 100083; 2.沈阳药科大学 中药学院, 辽宁 沈阳 110016;
3.深圳中药及天然药物研究中心, 广东 深圳 518057;
4.南京大学生命科学学院 医药生物技术国家重点实验室, 江苏 南京 210093)
摘要: 目的 研究猴耳环Pithecellobium clypearia Benth枝叶的化学成分及其免疫活性。方法 采用多种色谱学方法进行
分离纯化, 利用波谱学方法进行结构鉴定; 并通过Con A诱导的T淋巴细胞增殖试验考察各化合物的免疫活性。结果 从中
分离并鉴定了 8个化合物, 分别为(-)-表没食子儿茶素(1), (-)-5, 7, 3, 4, 5-五羟基黄烷(2), (-)-表没食子儿茶素 -7-没食子酸酯
(3), (-)-5, 3, 4, 5-四羟基黄烷 -7-没食子酸酯 (4), 槲皮素 -3-O-α-L-吡喃鼠李糖苷(5), 杨梅树皮素 -3-O-α-L-吡喃鼠李糖苷(6),
没食子酸 (7), 没食子酸乙酯 (8)。结论 化合物 3和 8 为首次从该属植物中分离得到, 化合物 1为首次从该种植物中分离得
到; 化合物 3能显著抑制 Con A诱导的 T淋巴细胞增殖, 其 IC50 为 4.4 μmol·L 1¯。
关键词: 猴耳环; 黄酮类化合物; 黄烷; 淋巴细胞增殖