全 文 ：NEW BIFLAVANONES AND BIOACTIVE COMPOUNDS
FROM STELLERA CHAMAEJASME L.
XU Zhi-hong , QIN Guo-wei , LI Xiao-yu , XU Ren-sheng＊
(Shanghai Institute of Materia Medica , Shanghai Institutes of Biological Sciences , Chinese Academy of Sciences , Shanghai 200031 , China)
ABSTRACT:AIM　To study the chemical constituents of the root of Stellera chamaejasme L.METHODS　Various column
chromatographies on silica gel and RP-18 were employed for isolation and purification.Structures of compounds were elucidated by
spectral analysis.RESULTS　Eight lignans and three biflavonoids possessing a C-3 C-3″ linkage were isolated.They are
ruixianglangdusu A(1)and B(2), 4′, 4 , 5 , 5″, 7 , 7″-hexahydroxy-3 , 3″-biflavone (3), (+)-kusunokinin (4), lirioresinol-B
(5), magnolenin C (6), (-)-pinoresinol monomethyl ether(7), (-)-pinoresinol(8), (+)-matairesinol(9), isohinokinin
(10) and (-)-eudesmin (11).CONCLUSION　Compounds 1 and 2 are new biflavanones , 1 is enantiomeric to known
chamaejasmenin C , 4 , 6 , 8 , 9 , 10 and 11 were isolated from this plant for the first time , and 7 was isolated from natural resources
for the first time. In vitro bioassays showed that 3 and 8 exhibited antibacterial activity , and 1 , 2 , 9 and 11 exhibited
immunomodulatory activity.
KEY WORDS:Stellera chamaejasme;biflavanones;in vitro bioassay
CLC number:R284.1;R284.2　　　Document code:A　　　Article ID:0513-4870(2001)09-0668-04
　　As part of a search for bioactive components from
Chinese traditional medicines “ Lang Du” , of which the
original sources are roots of three plants:Euphorbia
ebracteolata , E.fisheriana (Euphorbiaceae)and Stellera
chamaejasme (Thymelaeaceae)[ 1 ,2] , we have studied the
roots of S .chamaejasme L.In the course of our study ,
eight lignans and three biflavonoids possessing a C-3 C-3″
linkage have been separated and structurally identified ,
which include two new biflavanones named
ruixianglangdusu A(1)and B(2)and known compounds
4′, 4 , 5 , 5″, 7 , 7″-hexahydroxy-3 , 3″-biflavone (3)[ 3] ,
(+)-kusunokinin (4)[ 4] , lirioresinol-B (5)[ 5] ,
magnolenin C(6)[ 6] , (-)-pinoresinol monomethyl ether
(7)[ 7, 8] , (-)-pinoresinol (8)[ 9] , (+)-matairesinol
(9)[ 9] , isohinokinin (10)[ 9] and (-)-eudesmin
(11)[ 5] .Among these compounds , 4 , 6 , 8 , 9 , 10 and
11 were isolated for the first time from this plant , and 7
was for the first time separated from natural resource.The
roots of S.chamaejasme L.have been used for clinical
treatment of mange , stubborn skin ulcer , malignant
tumor , chronic tracheitis and tuberculosis for many years
in China.In vitro bioassays showed that the petroleum
ether and ether extracts were bioactive.The tests also
Received date:2001-03-19.
Biographies:XU Zhi-hong(1966-), female , Ph.D;
XU Ren-sheng(1931-),male ,professor , tutor for Ph.D.
＊Corresponding author　Tel Fax:001-650-9418568 ,
E-mail:rsxu@yahoo.com
showed that 1 , 2 , 9 and 11 exhibited immunomodulatory
activity (Table 1), and 3 and 8 exhibited antibacterial
activity. In this communication , the isolation and
structure identification of two new biflavanones ,
ruixianglandusu A (1)and B(2)were reported.
Table 1　Immunomodulatory activity of compounds
from S .chamaejasme
Compounds
Prolif eration rate(% of cont rol)
B Lymphocyte μmol T Lymphocyte μmol
0.1 1 10 0.1 1 10
1 90 311 148 - - -
2 - - - 89 183 322
9 - - - 94 185 161
11 262 123 107 99 204 245
　　Ruixianglangdusu A (1)　White plate crystals ,
was assigned the molecular formula C33H28O10 by HRMS
(m z 584.1696 [ M+ ]).It turned red with HCl-Mg
reagent.This indicates that compound 1 is a flavonoid.
The IR absorption at 3500 , 1640 , 1620 , 1580 , 1570 and
1510 cm
-1
suggested the presence of hydroxyl , conjugated
carbonyl and benzyl groups.The UV spectrum showed a
maximum absorption at 298 nm(lgε4.44)and a shoulder
peak at 330 nm , characteristics of A-ring oxygen-
substituted flavanone.
　　The molecular formula of 1 required twenty degrees
of unsaturation.Its 1HNMR(in DMSO-d6)showed three
methoxyl signals (δ:3.77 , 3.78 , 3.78), twelve
·668· 药学学报 Acta Pharmaceutica Sinica 2001 , 36(9):668-671
DOI :10.16438/j.0513-4870.2001.09.008
aromatic proton signals(δ:5.73-7.02), flavanone′s 2-
H (δ:5.52 , 5.57 , each 1H , s), 3-H (δ:2.92 , 2H ,
s)signals and three hydroxyl proton signals(δ:10.77 ,
11.49 , 11.50), respectively.The single peaks of 3-H
and 2-H suggested that compound 1 should be a C-3 C-3″
linkaged biflavanone. The suggestion was further
confirmed by two C-2(δ:79.42 , 79.52)and two C-3
(δ:45.99 , 45.99)signals in 13CNMR , compatible with
those of C-3 C-3″linkaged biflavonone in their chemical
shifts.The C-4 carbonyl groups showed H-bonded
conjugated absorption in IR (1640 cm-1)and 13CNMR
signals at lower field (δ:195.56 , 196.23), indicating
the existence of two 5-hydroxyl groups.Its 1HNMR
showed two groups of typical 4′-oxygenated B ring
protons:δ6.79(4H , d , J=8.4 Hz)and δ7.02(4H ,
d , J = 8.4 Hz).NOESY experiment revealed the
correlations between the two methoxyl groups (δ3.78)
and two pairs of 3′, 5′and 3 , 5 protons on B ring (δ
6.79 , 4H).The other four aromatic protons should be of
two A rings:δ5.93(1H , d , J=2 Hz)and 6.05(1H ,
d , J =2 Hz);δ5.73(1H , s)and 5.85(1H , s).
Correlations between one methoxyl group(δ3.77)and 6-
H , 8-H protons on A ring (δ:5.93 , 6.05)were also
observed on NOESY spectrum.This indicated that the
remaining one methoxyl group and one hydroxyl group
were located on either C-7 of each A ring.The
stereochemistry at the C-2 C-3 and C-2″ C-3″positions
were determined by comparison of the J-values of the
corresponding protons (0 and 0 Hz)[ 10] .Thus ,
ruixianglangdusu A has the geometry of cis-cis at C-2 C-3
and C-2″ C-3″positions as shown in Figure 1.Its plane
structure is the same as chamaejasmenin C (Figure
2)[ 10] , but not for the three-dimensional structure.The
difference between the two compounds is on their optical
activities.They all have large [ α] D values in the optical
rotation indicating that they both have no symmetrical
plane and no free rotation of the single bond between C-3
and C-3″, but the opposite direction:[ α] D -141°(c
1.0 , EtOH)for chamaejasmenin C and [α] D +175.97°
(c 0.106 , MeOH) for ruixianglangdusu A (1).
Therefore , each compound should be an antipode of the
other
[ 11] .Compound 1 should be ent-chamaejasmenin C.
However , the chirality of C-3 C-3″remains uncertain ,
like those biflavanones isolated from same plant in
previous literatures.
Figure 1　Structures of Ruixianglandusu A(1)and B(2), and pinoresinol monomethyl ether (7)
Figure 2　Structures of isochamaejasmin(a), chamaijasmenin A(b), and chamaejasmenin C(c)
　　Ruixianglangdusu B (2)　Light yellow powder ,
was assigned as C33H28 O10 by HRMS (m z 584.1677
[M +]).It turned red with HCl-Mg reagent.Analyses of
its spectral data suggested 2 was the same type of
compound as 1.They have the same kind and number of
functional groups.The difference is the stereochemistry at
the C-2 C-3 and C-2″ C-3″positions.The two groups of
proton signals at 2-H and 3-H in compound 1 were
·669·药学学报 Acta Pharmaceutica Sinica 2001 , 36(9):668-671
singlets while doublets in compound 2:δ2.84(1H , d , J
=12 Hz , 3-H)and 2.86(1H , d , J=12 Hz , 3″-H);δ
5.73(1H , d , J=12Hz , 2-H)and 5.78(1H , d , J=12
Hz , 2″-H).Thus 2 is a C-3 C-3″biflavanone having the
geometry of trans-trans at the C-2 C-3 and C-2″ C-3″
positions and the chirality at C-3 C-3″position still
remains unsettled.The reported trans-trans biflavanones
were two types of compounds as isochamaejasmin(a)and
chamaejasmenin A (b) shown in Figure 2[ 12] .The
different stereochemistry resulted the big difference of the
chemical shift of 2-H and 3-H.
EXPERIMENTAL
　　Melting points were determined with a Kofler mp
apparatus and were uncorrected;Optical rotations were
measured with a JASCOO DIP-181 polarimeter;The IR
spectra were run on a Perkin-Elmer 599B spectrometer
and UV spectra obtained on a Shimatzu UV-250
spectrometer;1H and 13CNMR were recorded on Bruker
AM-300 or 400 instrument in DMSO-d6 or CDCl3;MS
were performed with a Finnigan MAT-711 instrument.
The roots of Stellera chamaejasme L.were bought from
De-Yang Herb Corporation of Sichuan Province in
August , 1993 and identified by Dr.Dao-Feng Chen of
School of Pharmacy , Fudan University.A voucher
specimen was deposited in the Herbarium of Shanghai
Institute of Materia Medica , Chinese Academy of
Sciences.
1　Extraction and separation
The air-dried ground plant materials (20 kg)were
extracted with 95% EtOH and a portion of the extracts
after concentration were subsequently partitioned with
petrol and ether.The petrol extract (830 g)was
fractionated over silica gel , using petroleum ether-acetone
mixtures of increasing polarity as eluents.The petroleum
ether-acetone (9∶1)fractions gave 4 (56 mg), and
petroleum ether-acetone (7∶3)fractions gave 5(8 mg)
and 6(5 mg).The ether extract(1750 g)was subjected
to repeated column chromalographyies over silica gel ,
using hexane and acetone mixtures of increasing polarity
as eluents.Compound 3(14 mg)was obtained in hexane-
acetone(7∶3)fractions and 1 (15 mg)was obtained in
the fractions eluted by hexane-acetone (1∶1).The rest
hexane-acetone(1∶1)fractions were combined and were
further fractionated by RP-8 chromatography , using
ethanol-water as eluents.40%EtOH fractions gave 7(7
mg), 8(21 mg), and 9(26 mg), 60%EtOH fractions
gave 10 (13 mg)and 11 (21 mg), and 70% EtOH
fractions gave 2(26 mg).
2　Structure detemination
　　Ruixianglangdusu A(1)　White lamellar crystal ,
mp 232-236℃, [α] 19D +176°(c 0.106 , MeOH), IR
(KBr)cm-1:3500 , 1640 , 1620 , 1580 , 1570 , 1510 ,
1160;UVλMeOHmax (nm):218(lgε4.61), 298(lgε4.44);
EIHRMS m z:584.1696[ M+ ] , EIMS m z:57 , 77 ,
83 , 105 , 135(base), 149 , 165 , 192 , 219 , 239 , 299 ,
356 , 382 , 484 , 584(M +);1HNMR(300MHz , DMSO-
d6):δ2.92(2H , s , 3 ,3″-H), 3.77(3H , s , 7-OCH3),
3.78(6H , s , 4′,4 -OCH3), 5.52(1H , s , 2-H), 5.57
(1H , s , 2″-H), 5.73(1H , s , br , 8-H), 5.85(1H , s ,
br , 6-H), 5.93(1H , d , J=2 Hz , 8″-H), 6.05(1H ,
d , J=2 Hz , 6″-H), 6.79(4H , d , J=8.4 Hz , 3′,5′,
3 ,5 -H), 7.02(4H , d , J=8.4 Hz , 2′, 6′, 2 , 6 -
H), 10.77(OH , s), 11.49(OH , s), 11.50(OH , s);
13
CNMR(75 MHz , DMSO-d6):δ45.99(dx2 , C-3 , 3″),
55.08(q , OCH3×2), 55.86(q , OCH3), 79.42(d , C-
2), 79.59(d , C-2″), 93.30(d , C-8), 94.32(d , C-
8″), 94.56(d , C-6), 95.45(d , C-6″), 101.89(s , C-
10), 102.72(s , C-10″), 113.63(dx4 , C-3′, 3 , 5′,
5 ), 127.00(dx4 , C-2′, 2 , 6′, 6 ), 127.70(sx2 , C-
1′, 1 ), 158.66(sx2 , C-4′, 4 ), 162.50(s , C-9),
162.90(s , C-9″), 163.19(s , C-5), 166.20(s , C-5″),
166.33(s , C-7), 167.31(s , C-7″), 195.56(s , C-4),
196.23(s , C-4″).
　　Ruixianglangdusu B (2)　Amorphous powder ,
[α] 15D +181°(c 0.28 , MeOH), IR(KBr)cm-1:3400 ,
1640 , 1620 , 1515 , 1254 , 1155;UVλMeOHmax (nm):219
(lgε4.57), 296(lgε4.45);EIHRMS m z:584.1677
[M + ] , EI-MS m z:57 , 71 , 105 , 121 , 149 , 167 ,
178 , 266 , 285 , 299(base), 311 , 370 , 386 , 431 , 463 ,
584(M +);1HNMR(300MHz , DMSO-d6):δ2.84(1H ,
d , J=12 Hz , 3-H), 2.86(1H , d , J=12 Hz , 3″-H),
3.76(3H , s , 7-OCH3), 3.79(6H , s , 4′, 4 -OCH3),
5.73(1H , d , J=12 Hz , 2-H), 5.78(1H , d , J=12
Hz , 2″-H), 5.82(1H , s , br , 8-H), 5.90(1H , s , br ,
6-H), 6.04(1H , d , J=2.3 Hz , 8″-H), 6.09(1H , d ,
J=2.3 Hz , 6″-H), 6.89(4H , d , J=8.5 Hz , 3′, 5′,
3 ,5 -H), 6.99(4H , d , J=8.5 Hz , 2′, 6′, 2 , 6 -
H), 10.91(OH , s), 11.73(OH , s), 11.74(OH , s);
13
CNMR(75 MHz , DMSO-d6):δ49.05(dx2 , C-3 , 3″),
55.23(q , OCH3×2), 55.96(q , OCH3), 82.70(d , C-
2), 82.89(d , C-2″), 93.89(d , C-8), 95.00(dx2 , C-
8″, 6), 96.10(d , C-6″), 101.00(s , C-10), 102.80(s ,
C-10″), 114.05(dx4 , C-3′, 3 , 5′, 5 ), 129.18(dx4 ,
C-2′, 2 , 6′, 6 ), 127.91(sx2 , C-1′, 1 ), 159.91
(sx2 , C-4′, 4 ), 162.33(s , C-9), 162.90(s , C-9″),
·670· 药学学报 Acta Pharmaceutica Sinica 2001 , 36(9):668-671
163.12(s , C-5), 166.80(s , C-5″), 166.90(s , C-7),
167.65(s , C-7″), 196.00(s , C-4), 196.60(s , C-4″).
　　(-)-Pinoresinol Monomethyl ether(7)　Yellow
amorphous powder , Gibbs test (-), [ α] 30D -58°(c
0.05 , EtOH), IR(KBr)cm-1:3400(hydroxyl group),
1600 , 1510(benzyl group), 1260 , 760;UVλMeOHmax (nm):
223(lgε3.75), 279(lgε4.15);EI-MS m z:55 , 77 ,
105(base), 122 , 135 , 149 , 198 , 218 , 266 , 291 , 354
(M +);1HNMR(400MHz , CDCl3):δ3.09(2H , m , 1 ,
5-H), 3.86(3H , s , OCH3), 3.88(3H , s , OCH3),
3.89(3H , s , OCH3), 3.91(2H , m , 4a ,8a-H), 4.24
(2H , dd , J=6.8 , 9.1 Hz , 4e ,8e-H), 4.73(1H , d , J
=5.4 Hz , 6-H), 4.74(1H , d , J=4.5 Hz , 2-H),
-6.86(6H , m , aromatic protons).
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瑞香狼毒中新的双黄酮和活性化合物
徐志红 , 秦国伟 , 李晓玉 , 徐任生
(中国科学院上海生命科学研究院上海药物研究所 , 上海 200031)
摘要:目的　研究中药狼毒药材来源之一瑞香狼毒(Stellera chamaejasme L)根的化学成分 。方法　用各
种柱色谱进行分离纯化 ,用各种波谱分析方法鉴定其结构 。结果　从瑞香狼毒的根中分离并鉴定出 3种双
黄酮和 8 种木脂素:瑞香狼毒素 A(1)和 B(2), 4 , 4″, 5 , 5″, 7 , 7″-hexahydroxy-3 , 3″-biflavanone (3), (+)-
kusunokinin(4), lirioresinol-B (5), magnolenin C (6), (-)-pinoresinol monomethyl ether (7), (-)-pinoresinol
(8), (+)-matairesinol(9), isohinokinin(10)和(-)-eudesmin(11)。结论　化合物 1和 2为新化合物 , 1是已
知双黄酮 chamaejasmenin C的对映体 ,4 ,6 ,8 ,9 ,10和11是从本植物中首次分得 ,7是首次从天然界分得 ,体外
生物测试表明 3和8有抗菌活性 ,1 ,2 ,9和 11有免疫调节活性。
关键词:狼毒;双黄酮;体外生物测定
·671·药学学报 Acta Pharmaceutica Sinica 2001 , 36(9):668-671