全 文 ：　　　　 天然产物研究与开发　 　　　　　
2003　Vo1.15　No.2 NATU RAL PRODUCT RESEARCH AND DEVELOPMENT　 　　　　　　　　　　
　
　
　
　
　　收稿日期:2002-11-22　　　修回日期:2002-12-06
　　＊Corresponding author:Tel.021-64311833×349;Emai l:yw guo
@mail.shcnc.ac.cn
　　This research is supported by Nat ional Science Foundation for Out-
standing You th(30125044)and the “Foundation for S cholars Come Back
f rom Abroad” provided by Minist ry of Education , Minist ry of Personnel ,
and Chinese Academy of S ciences.
ECDYSTEROIDS FROM THE STEMSOF
DIPLOCLISIA GLAUCESCENS
HUANG Xiao-chun1 　GUO Yue-wei1＊　ZHOU Wen-liang1
ZUO Jian-ping1 　WANG Zheng-tao2
(1.State Key Laboratory of Drug Research , Institute of Materia Medica , Shanghai Institutes for
Biological Sciences , Chinese Academy of Sciences , Shanghai　200031 , China;
2.I nstitute of Chinese Material Medica , Shanghai University of Traditional Chinese
Medicine , Shanghai　200032 , China)
Abstract　Five ecdysteroids w ere isolated from the EtOH ex tract o f the stems of Diploclisia glaucescens.Their
structures were identified as paristerone 20 , 22-monoacetonide(1), paristerone(2), β-ecdy sterone(3), makis-
terone C(4)and capitasterone(5), respectively , on the basis of chemical and phy sical evidence.This is the first
report of paristerone 20 , 22-monoacetonide isolated from natural source.Compounds 2 , 4 , and 5 were also iso-
lated for the first time from D .glaucescens.
Key words　Menispermaceae;Diploclisia glaucescens;ecdysteroid;paristerone 20 , 22-monoacetonide;paris-
terone;ecdy sterone;makisterone C;capitasterone
　Introduction
Ecdysteroids are an important g roup of natural prod-
ucts w hich have been found in many plants especially
in ferns , gymnosperms and angiosperms , as w ell as in
marine o rganisms.More than 150 ecdy steroids have
been repo rted to date.These compounds exhibi t inter-
esting pharmacological effects on mammals , including
the stimulation of protein synthesis , the reduction of
blood g lucose and cholesterol levels , antiulcer , an-
tirheumatic , insulin regulation and diuretic or tonic ef-
fects[ 1-3] .
Diploclisia glaucescens is g row ing widely in the re-
gion of south and southw est China.The leaves and
stems of the plant have been used in China as a folk
medicine fo r the therapy of rheumatism and venereal
diseases[ 4] .Previous chemical investig at ions of this
plant collected f rom India and Pakistan proved that
D .glaucescen was a rich source of ecdy steroids[ 5-7] .
However , no any phytochemical investigation had
been done on the ti tle species f rom China.On our re-
cent research fo r plant-derived medicinal agents , the
stems of D .glaucescens collected f rom Guangxi
Province w ere chemically studied.Five ecdysteroids
(1-5), namely paristerone 20 , 22-monoacetonide[ 8] ,
paristerone[ 8] , β-ecdy sterone[ 9] , makisterone C[ 10] and
capitasterone
[ 11] , respectively , were isolated.In the
present paper we describe the isolation , structure elu-
cidation and the complete 1H-and 13C-NMR assign-
ments of ecdysteroid 1 which is reported now fo r the
fi rst time as a natural product .The biological act ivit ies
of 1-5 are also presented.
1　Results and Discussion
Ecdysteroids 1-5 were isolated as described in the Ex-
perimental section.The known compounds 2-5 were i-
dentified by MS , 1H-NMR and 13C-NMR spect ra evi-
dence and by comparison w ith the spect ral data re-
ported in the literatures[ 8-11] .Structure determina-
tion of paristerone 20 , 22-monoacetonide (1) was
based on the following.
93
Fig.1　Compounds 1-7 isolated from Diploclisia glaucescens
Compound 1 , colorless needles ,mp.108-110 ℃, [α] 25D
=48.7°(c 0.21 , MeOH), possesses the composition
C30H48 O7{ESIMS [ M +H ] + at m /z 521 , [ M +
Na] + at m/ z 543}.The UV spectrum of 1 showed
absorption at 241 nm(log ε=4.0)and an intense IR
band at 1651 cm-1 corresponding to an enone sys-
tem.1H NMR spectrum displayed a broad sing let at δ
5.78 which could be assigned to a proton α-to the
enone carbonyl.These data together with 13C-NMR
chemical shift at δ206.6 st rongly suggested the pres-
ence of a β , β-substituted-α, β-unsaturated carbonyl
moiety in the molecule[ 8] .Furthermore a st rong IR
band at 3410 cm
-1
indicated this compound to be an
ecdy steroid[ 8] .The 1H-NMR spect rum of this com-
pound w as very similar to that of paristerone (2)[ 8] .
The spect ral features and relative positions of H-2 ,H-
3 ,H-7 ,H-9 and H-17 , as w ell as those of 18-Me and
19-Me indicated that those two ecdysteroids possessed
the same nucleus.Careful comparison 13C-NMR data
of compounds 1 and 2 revealed that 1 deffers from 2
only in the side chain , where 13 C-NMR chemical
shift s at C-20 and C-22 were shif ted downf ield f rom
δ78.4 to 86.3 and from δ78.9 to 83.8 , respective-
ly , except that three mo re peaks corresponding to the
ketal group were observed(δ108.5 ,29.9 and 29.7).
This fact indicated that the presence of C-20 , 22-ace-
tonide at the side-chain of 1.Comparison 13C-NMR
data of 1 w ith those of model compound 6[ 12] (see
Table 1)further conf irmed above assignment accord-
ing to the structure 1.In o rder to ensure the st ructure
of 1 , compound 2 was t reated w ith anhydrous acetone
affording two expected products 1 and 7 , respectively.
Based on the above information the structure of paris-
terone C-20 , 22-monoacetonide w as ambiguously elu-
cidated.The complete 1H- and 13C-NMR assignments
of 1(see Table 1)were based on the detailed analy sis
of i ts
13
C-NMR , DEPT , 1H-1H COSY , HMQC , and
HMBC spectra.
Table 1　1H- and 13C-NMRdata of compound 1a and compari-
son with 13C-NMR data of compounds 2a and 612
Posi tion
δ1H(mult.) δ13C
1 1 2 612
1ax 28.1 28.1 38.0
1 eq 1.69(1 H , m)
2 eq 3.95(1 H , m , ω1/2=20) 69.4 69.4 68.1
3 eq 3.72(1 H , d , J =2.5 Hz) 74.2 74.2 68.1
4ax 1.60(1 H , m) 26.1 26.1 31.7
4 eq 1.54(1 H , m)
5 2.28(1 H , m) 52.6 52.6 51.4
6 - 206.6 206.6 203.5
7 5.78(1 H , d , J =2.1 Hz) 122.5 122.5 121.8
8 - 167.3 167.6 165.4
9ax 3.13(1 H , m) 36.3 36.2 34.5
10 - 44.1 44.1 38.6
11ax 1.72(1 H , m) 22.4 22.4 21.1
11 eq 1.69(1 H , m)
12ax 2.08(1 H , m) 32.8 32.8 32.4
12 eq 1.84(1 H , m)
13 - 48.9 48.8 47.9
14 - 85.6 85.6 84.2
15a 1.95(1 H , m) 32.2 32.4 31.7
15b 1.61(1 H , m)
16a 2.03(1 H , m) 22.8 21.9 22.1
16b 1.81(1 H , m)
17 2.31(1 H , m) 51.0 51.1 50.0
18 0.83(3 H , s) 18.2 18.5 17.3
19 1.03(3 H , s) 20.7 20.7 24.4
20 - 86.3 78.4 82.1
21 1.17(3 H , s) 23.1 21.5 22.4
22 3.67(1 H , m , ω1/2=13) 83.8 78.9 85.6
23a 1.51(1 H , m) 25.2 27.8 24.4
23b 1.54(1 H , m)
24a 1.46(1 H , m) 42.7 42.9 42.2
24b 1.70(1 H , m)
25 - 71.6 71.8 69.2
26 1.19(3 H , s) 29.5 29.5 30.1
27 1.20(3 H , s) 30.0 30.2 29.9
28 - 108.5 - 106.9
29b 1.31(3 H , s) 27.7 - 27.2
30b 1.38(3 H , s) 29.9 - 29.4
aBruker DR×400 spectrometer;δvalue are reported in ppm referenced
to TMS as internal standard.Assignments deduced from the analysi s of
mononuclear and heteronuclear spect ra and comparison wi th know n 3-epi
ecdysterone(2).
94　　　　　　　　　 天然产物研究与开发 2003　Vo1.15　No.2
Compound 1 was first reported as a synthetic product
during the st ructural determination of paristerone[ 8] .
But the complete NMR data of 1 were not given
then.This is the first report of paristerone 20 , 22-
monoacetonide as a natural product .To our knowl-
edge , 1 is the second ecdy steroid C 20 , 22-monoace-
tonide isolated f rom a natural source.
2　Experimental
2.1　General
The IR spectra were recorded on a Nicolet M agna
FT-IR750 spect rometer , and UV spect ra were ob-
tained on a Varian CARY 300 BIO spectropho-
tometers.1H- and 13C-NMR spect ra were measured
on a Bruker DR×400 spect rometer.Chemical shif ts
w ere given in ppm referenced to TMS as internal
standard.1H- and 13 C-NMR assignments were sup-
po rted by 1H-1H COSY , HMQC and HMBC experi-
ments.EIMS , ESIMS and HRESIMS spectra were
obtained on a Finnigan-MA T-95 mass inst rument.
Optical rotation w as measured on a Perkin-Elmer 241
MC Polarimeter in M eOH .Reversed-phase HPLC pu-
rifications were performed on an Agilent 1100 series
liquid chromatography using a VWD G1314A detec-
tor at 254 nm.One semi-preparat ive ODS-HG-5[ 5
μm , 10 mm(i.d.)×25 cm] column w as employed fo r
the purification.
2.2　Plant material
Plant material was collected in Guangxi Province ,
China , and identified by Prof.Lai M-X of China Phar-
maceutical Universi ty.Voucher specimen is deposited
in SIBS for inspection.
2.3　Extraction and isolation
Air dried stems of D .glaucescens(4.3 kg)was
g round and ex tracted 3 times w ith refluxing 95%E-
tOH.The combined EtOH solution w as concentrated
under reduced pressure to yield the residue as a dark
brow nish gum(324 g).The residue w as suspended in
w ater and ex tracted wi th pet rolume ether , CHCl3 ,E-
tOAc and n-BuOH successively .The CHCl3 soluble
portion(15.9 g)was subjected to silica gel(500 g)col-
umn chromatog raphy eluting w ith pet roleum ether-
CHCl3 ,CHCl3 , CHCl3-MeOH in g radient to give sev-
en fractions(f rs.1-7).Fraction 6(1.56 g)was rechro-
matorgraphed on silica gel(50 g)column eluting w ith
CHCl3-MeOH in g radient to give six f ractions(f rs.6-
1—6-6).Fraction 6-3(59.6 mg)was further separat-
ed by a silica gel column(2.5 g)eluting w ith CHCl3-
MeOH system to yield capi tasterone(5 ,38.4 mg).
The EtOAc ex tract(18.7 g)was subjected to silica gel
(550 g)column chromatog raphy using pet roleum
ether-acetone in gradient to give f ractions 1′-13′.
Fraction 10′(1.4 g)was chromatog raphed repeatedly
on a silica gel column eluting w ith CHCl3-MeOH sys-
tem affo rding makisterone C(4 , 16.4 mg).Fract ion
11′(1.5 g)was subjected to column chromatog raphy
over silica gel(30 g)eluting wi th petroleum ether-ace-
tone system.Three fractions(11′-1—11′-3)were ob-
tained.Fraction 11′-1(50.3 mg)was further purified
on a silica gel(3.0 g)column eluting with CHCl3-
MeOH to affo rd paristerone C-20 , 22-monoacetonide
(1 ,16.9 mg).
The residue of n-BuOH extraction(32.6 g)was chro-
matographed on silica gel(1 kg)column developing
w ith CHCl3-MeOH , and w as further purified by semi-
preparative ODS-HG-5[ 5μm ,10 mm(i.d.)×25 cm]
column using M eOH-H2O(1∶1)as eluent to afford
ecdysterone(3 ,24.2 mg)and paristerone(2 ,7.1 mg),
respect ively .
2.4　Identi fication
Paristerone C-20 , 22-monoacetonide 1　Colorless nee-
dles, mp.108-110 ℃[ α] 25D = +48.7°(c 0.21 ,
MeOH).ESIMS m/ z 521 [ M +H] + , HRESIMS
m /z :543.3298 , [ M +Na] + for C30 H48 O7(calcd
543.3313).IRυKBrmax cm-1:3490 , 2923 , 1650 , 1463 ,
1382 , 1107 , 1070.UV(MeOH)λmax 241 nm(log ε
4.0).1H- and 13C-NMR:see Table 1.
Paristerone 2　Colorless needles , posi tive ion ESIMS
m /z :503[ M +Na] + ,983[ 2M +Na] + , negat ive ion
ESIM S m/ z 479[ M-H] -.1H-NMR:δ0.89(3 H , s ,
H3-18),1.03(3 H , s ,H3-19), 1.18(3 H , s , H3-21),
1.18(3 H , s , H3-26), 1.19(3 H , s , H3-27), 2.30(1
H ,dd , J =5.3 , 11.1 Hz ,H-5), 2.38(1 H , t , J =8.6
Hz ,H-17), 3.14(1 H , m , H-9), 3.32(1 H , brd , J =
5.2 Hz ,H-22), 3.72(1 H ,d , J =2.4 Hz ,H-2), 3.95
952003　Vo1.15　No.2 黄孝春等:苍白秤钩风中脱皮甾酮类化学成分的研究 　　　　　　　　　
(1 H , m , H-3), 5.78(1 H , d , J =2.2 Hz ,H-7).13C-
NMR:see Table 1.
Preparation of acetonide of paristerone　A suspension
of 2(1.7 mg)in anhydrous acetone(1 ml)was stirred
w ith anhydrous CuSO4 in water bath at the tempera-
ture of 30℃ fo r 24 hours and the react ion w as moni-
tored by TLC.After completion of the reaction , the
reaction mix ture w as concentrated under reduced
pressure to yield the residue(tw o spots exhibited on
TLC).The residue w as subjected to silica gel CC , e-
luting with CHCl3-MeOH 95∶5 to yield paristerone
C-20 , 22-monoacetonide(1 , 0.4 mg)and paristerone
C-2 ,3 , 20 ,22-diacetonide(7 ,1.1 mg), respect ively .
Paristerone C-2 , 3 , 20 , 22-diacedonide 7 　Colo rless
needles , 1H-NMR:δ0.66(3 H , s ,H3-18), 0.89(3 H ,
s , H3-19), 1.00(3 H , s , H3-21), 1.02(3 H , s , H3-
31),1.03(3 H , s , H3-32),1.14(3 H , s ,H3-26),1.15
(3 H , s , H3-27), 1.21(3 H , s , H3-29), 1.31(3 H , s ,
H3-30), 2.10(1 H , m , H-5), 2.14(1 H , m , H-17),
2.80(1 H , m ,H-9),3.51(1 H ,m ,H-22), 3.98(1 H ,
d , J =4.4 Hz ,H-2), 4.07(1 H , m ,H-3),5.63(1 H ,
d , J =2.2 Hz ,H-7).
Ecdysterone 3　Colorless needles , EIM S m /z :363 ,
345 ,327 , 301 , 149(100%).1H-NMR:δ0.72(3 H ,
s , H3-18), 0.80(3 H , s , H3-19), 1.02(3 H , s , H3-
21),1.03(3 H , s , H3-26),1.03(3 H , s ,H3-27),2.09
(1 H ,m , H-5), 2.22(1 H , m , H-17), 2.98(1 H , m ,
H-9), 3.18(1 H , brs , H-22), 3.67(1 H , m , H-3),
3.79(1 H , d , J =2.6 Hz ,H-2),5.64(1 H , d , J =2.3
Hz ,H-7).13C-NMR:δ18.5(C-18), 21.6(C-21),
22.0(C-11 , 16), 24.9(C-19), 27.8(C-23), 29.5(C-
26), 30.2(C-27)32.3(C-12),33.0(C-15), 33.3(C-
4), 35.6(C-9), 37.8(C-1), 39.8(C-10), 42.9(C-
24), 51.0(C-17), 52.3(C-5), 69.0(C-2), 69.2(C-
3),71.8(C-25),78.4(C-20),78.9(C-22), 85.7(C-
14), 122.6(C-7),168.5(C-8),207.0(C-6).
Makisterone C 4　Colo rless needles ,positive ion ES-
IM S m/ z :531[ M +Na] + , negative ion ESIM S m/
z :507[ M-H] -.1H-NMR:δ0.73(3 H , s , H3-18),
0.80(3 H , s , H3-19), 0.87(3 H , t , J =7.3 Hz , H3-
29),0.96(3 H , s , H3-21),1.05(3 H , s ,H3-26),1.08
(3 H , s , H3-27), 2.20(1 H , brd , J =4.8 Hz , H-5),
2.24(1 H , m ,H-17),3.00(1 H ,m ,H-9), 3.24(1 H ,
brd , J =10.6 Hz ,H-22),3.66(1 H ,m ,H-3), 3.78(1
H ,brs ,H-2),5.65(1 H ,brs ,H-7).13C-NMR:δ14.9
(C-29),18.6(C-18), 21.5(C-21), 22.1(C-11 , 16),
24.9(C-19),26.1(C-26),26.5(C-28), 29.6(C-27),
32.3(C-12),33.0(C-15), 33.4(C-4), 33.5(C-23),
35.6(C-9), 37.9(C-1), 39.8(C-10), 50.8(C-24),
50.9(C-17), 52.3(C-5), 69.0(C-2), 69.2(C-3),
74.6(C-25),77.7(C-22),78.5(C-20), 85.8(C-14),
122.6(C-7),168.5(C-8), 207.0(C-6).
Capitasterone 5　Colo rless needles ,EIMS m / z:363 ,
345 , 327 , 149.Negative ion ESIMS m /z :503[ M-
H] -.1H-NMR:δ0.71(3 H , s ,H3-18),0.76(3 H , t ,
J =7.5 Hz ,H3-29),0.82(3 H , s ,H3-19),0.97(3 H ,
d , J =7.0 Hz , H3-27), 1.12(3 H , s ,H3-21),2.22(1
H ,dd , J =12.6 , 4.6 Hz ,H-5), 2.30(1 H , t , J =8.8
Hz ,H-17),3.01(1 H , m , H-9), 3.67(1 H ,m ,H-3),
3.78(1 H , d , J =2.2 Hz ,H-2), 4.12(1 H , dd , J =
11.9 ,3.5 Hz ,H-22),5.65(1 H , d , J =2.2 Hz ,H-7)
.13C-NMR:δ11.0(C-29), 11.8(C-27), 17.5(C-
18),20.4(C-11 ,16),20.7(C-21),23.9(C-19), 24.4
(C-28), 26.7(C-23), 31.3(C-4 , 12), 33.7(C-9),
36.0(C-24), 36.9(C-1), 37.3(C-25), 38.1(C-9),
47.4(C-13), 49.0(C-17), 50.2(C-5), 67.2(C-3),
67.4(C-2), 75.6(C-20), 83.9(C-22), 84.1(C-14),
121.5(C-7),164.7(C-8), 203.7(C-6).
Acknowledgement:This research is supported by Na-
tional Science Foundation for Outstanding Youth
(30125044)and the “Foundation for Scholars Come
Back from Abroad” provided by M inistry of Educa-
tion , Minist ry of Personnel , and Chinese Academy of
Sciences.The authors are g rateful to Prof.Lai M-X
fo r identification of the plant .
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苍白秤钩风中脱皮甾酮类化学成分的研究
黄孝春1　郭跃伟1＊　周文亮2　左建平2　王峥涛2
(1.中国科学院上海生命科学院药物研究所新药研究国家重点实验室　上海　200031;
2.上海中医药大学中药研究所　上海　200032)
摘　要　从中国广西药用植物苍白秤钩风(Diploclisia glaucescens)藤茎的 95%乙醇提取物中分离得到 5
个脱皮甾酮类化合物(1-5)。通过 NMR、MS 以及化学沟通等方法分别将其结构鉴定为 paristerone C-
20 , 22-monoacetonide(1), paristerone(2), ecdysterone(3), makisterone C(4)和 capisterone(5)。化合物 1为
首次从自然界分得的天然产物 , 2 , 4 , 5亦为首次从该植物中分得。
关键词　防己科;苍白秤钩风;ecdy steroid;paristerone 20 , 22-monoacetonide;paristerone;ecdy sterone;makis-
terone C;capitasterone
《天然产物研究与开发》杂志入选
1992年美国《化学文摘》千名表
美国《化学文摘》 ,简称 CA , 1907年创刊 ,是国际上化学化工及相关学科最负盛名的权威性的检索刊物
和数据库 。
美国《化学文摘》收录了 150多个国家近一万四千多种期刊等文献 ,文种达 56种 ,其中所收录的化学化
工文献占全世界该类文献量的 98%。据《美国化学文摘资料来源索引》统计表明 ,1992年中国科技期刊(含
台湾)仅有 45种入选《CA》千名表 。本刊在《CA》千名表中排名第 755位。在我国(包括台湾)入围千名表的
45种期刊中 ,列第 29名。
《天然产物研究与开发》是 1989年 6月新创刊的季刊 ,公开发行后 ,迅速被《CA》收录 ,仅两年就跨入千
名表。其后 ,收录率提高很快 , 1998 年 、1999年及 2000 年 ,收录率均高达 90%左右 ,如 2000年全年该刊共
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