全 文 ：The chem ical constituents ofLigularia pleurocaulis
ZHANG M ian
* , ZHANG Chao-feng, WANG Zheng-tao
(D epartm ent of Pharm acognosy, China Pharm aceutica lUn iversity, Nan jing 210038, China)
Abstract:Ami 　To study the chem ical constituents o f the underground part ofLigularia pleurocau lis
(Franch. )Hand-M azz. . M ethods　The dried roo ts and rhizomes o f L. pleurocau lis were extracted w ith
me thano .l Iso lation and purification we re performed by silica ge l column chromatog raphy and
recry sta llization etc. S tructures of the pure compounds w ere estab lished on basis of spectra l analysis.
Results　 Tw elve compounds w ere obtained from L. pleurocaulis, they we re 6-ange loy loxy-furanoligu-
larenone (1), 2-oxo-3-hydroxy-eremoph ila-1(10), 3 , 7(11), 8-tetraen-8, 12-o lide (2), tig lic acid (3),
o leano lic acid (4), lupeol (5), β-sitostero l(6), daucoste ro l(7), caffeic acid (8), emodin (9), 7-
me thoxy-coumarin ( 10), ferulic acid (11 ) and 4-hydroxy-2, 5-dimethoxy-benzaldehyde (12).
Conc lusion　Compound 1 is a new eremophilane and compound 2 is a new natural compound. A ll above
compounds we re obta ined for the first time from L. pleurocaulis.
Key words:Ligu laria pleurocau lis;6-angeloy loxy-furano ligula renone;2-oxo-3-hydroxy-eremoph ila-1
(10), 3, 7(11), 8-tetraen-8, 12-olide
CLC number:R284. 1;R284. 2　　　Docum ent code:A　　　A rtic le ID:0513 -4870(2005)06 - 0529 -04
Received date:2004-09-23.
Foundat ion item:N ational Natu ral S cience Foundation of Ch ina
(30270157).
* Correspond ing au thor　Tel:86 -25 - 85391246,
Fax:86 - 25 -85309639,
E-m ail:m ianzhang@h otm ai.l com
侧茎橐吾化学成分的研究
张　勉* , 张朝凤 , 王峥涛
(中国药科大学 生药学研究室 , 江苏 南京 210038)
摘要:目的　侧茎橐吾(L igularia pleurocaulis)为菊科橐吾属植物 , 其根和根茎入药 , 有止咳化痰 、活血化瘀等功
效 , 为中药 “紫菀” (中国药典 2000年版收载为 Aster tataricus L. .f的根及根茎)的代用品之一 , 称为 “山紫菀 ”。有关
侧茎橐吾的化学成分研究尚未见报道 ,因此 , 本文对侧茎橐吾的根及根茎的化学成分进行了研究。方法　侧茎橐吾
的根和根茎用甲醇提取 ,硅胶柱色谱和重结晶等方法进行分离纯化。 根据化合物的理化性质和光谱数据进行结构
鉴定。结果　分离得到了 12个化合物 ,即 6-ange loy loxy-furano ligularenone (1), 2-oxo-3-hyd roxy-erem ophila-1(10), 3, 7
(11), 8-te traen-8, 12-o lide (2), 顺芷酸(3),齐墩果酸 (4),羽扇豆醇(5), β-谷甾醇 (6), 胡萝卜苷(7), 咖啡酸(8),大
黄素(9), 7-甲氧基香豆素 (10), 阿魏酸 (11)和 4-羟基-2, 5-二甲氧基-苯甲醛 (12)。结论 　化合物 1为新的
e remoph ilane型倍半萜 , 化合物 2为新的天然产物。所有化合物均为首次从本植物中得到。
关键词:侧茎橐吾;6-angeloy loxy-furanoligula renone;2-oxo-3-hydroxy-erem ophila-1(10), 3, 7(11), 8-tetraen-8, 12-o lide
Introduction
Ligularia Cass. is a big genus be long ing to fam ily
Compositae. The roo ts and rh izomes o fmany Ligularia
plants, called “ Shanziw an ” in traditiona l Chinese
medicine w ith the action o f e lim inating ph legm and
reliev ing cough, aremain ly used to treat acute or ch ro-
nic cough, asthma and hemoptysis. A s a sy stematic
study of Ligularia species, we investigated the chem i-
ca l constituents o f L. pleurocau lis (Franch. ) Hand-
Mazz. and 7 known eremoph ilane compounds have
been iso lated and charac te rized prev iously
[ 1]
. Th is
pape r reports the iso la tion and structure e lucidation o f a
new eremophilane sesquiterpene, 6-angeloy loxy-furano-
ligu larenone (1), and a new na tu ra l compound, 2-
oxo-3-hydroxy-eremoph ila-1(10), 3, 7(11), 8-te traen-
8, 12-o lide (2). Ten known compounds, tig lic ac id
529 药学学报 Ac ta Pharm aceu tica S in ica 2005, 40(6):529 - 532
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(3), oleano lic acid (4) lupeol (5), β-sito sterol
(6), daucesterol (7), caffeic acid (8), emodin
(9), 7-methoxy-couma rin (10), feru lic acid (11)
and 4-hydroxy-2, 5-d imethoxy-benzaldehyde (12)were
also ob ta ined fo r the first time from this plant(Figure 1).
F igu re 1　Structures of compounds 1 and 2
Results and discussion
Compound 1 was obtained as co lo rless need les.
Itsmo lecu lar formula w as estab lished as C20H24O 4 by
HR-E IM S a tm /z 328.164 1 (calcd. 328.167 5).
The
1
H NMR and
13
C NMR spec tra show ed signals due
to an angeloy loxy group atδH 6.20(1H , qq, J =7.3,
1.4 H z, H-3′), 2.05 (3H , dq, J =7.3, 1.4 Hz,
H-4′), 1.91(3H , dq, J =1.4, 1.4 H z, H-5′) and δC
167.2(s, C-1′), 127.3(s, C-2′), 139.9(d, C-3′),
15.7(q, C-4′), 20.7(q, C-5′);three methy l g roups
atδH 1.79(3H , d, J =1.1 H z, H-13), 1.10(3H , d,
J =6.7 Hz, H-15), 0.92(3H , s, H-14) and δC 8.7
(C-13), 7.8(C-15), 6.8 (C-14); four me theny l
groups a tδH 7.07(1H , br s), 6.55(1H , dd, J =10.0,
2.0H z), 6.28(1H , q, J =1.1 H z) and 6.12(1H ,
dd, J =10.0, 3.8 H z). By a HMQC expe riment,
above fou r me theny l groups w ere assigned to H-12,
H-1 , H-6 and H-2 respective ly, which separate ly
co rrela ted w ith them etheny l carbon signals atδ138.6
(d, C-12), 148.0(d , C-1), 72.4(d, C-6) and 128.7
(d, C-2). The IR spec trum show ed absorption bands
of este r carbony l group at 1 712 cm
-1
and α, β-
unsa turated ke tone a t 1 697 , 1 681 and 1 642 cm -1 ,
which w ere confirmed by the carbon signa ls atδ167.2
(s, C-1′) and 199.9(s, C-3) in the 13C NMR spec-
trum. Hence the structu re of compound 1was suggested
to be a fu ranoeremophilane w ith α, β-unsa tura ted
ketone and ange loy loxy groups. Comparison o f NMR
spectra l parame ters w ith furano ligularenone
[ 2] , com-
pound 1 re tains the same enone system from the ABX
sy stem d isp layed by the hydrogen a tom s H-1(δ6.55,
1H , dd, J =10.0 , 2.0H z), H-2(δ6.12, 1H , dd, J =
10.0, 3.8 H z) and H-10(δ2.63, 1H , m ), and the
alipha tic methy l signa ls of M e-14 and M e-15 a lso
appea red nearly at the same field, bu t the signals of
angeloy loxy g roup was absen t in furanoligu larenone. In
the HMBC spectrum of compound 1 (Figure 2) the
corre lations o fH-6 to C-1′, C-2′, C-7, C-8 and C-14
established loca tion of the angeloy loxy group at C-6.
Ste reochem ically, Me-14 and M e-15 are biogen tically
in β o rientions[ 3] . The re lative configuration o f
angeloy loxy l g roup at C-6 w as de term ined to be β from
the NOESY spectrum , in w hich c ross peak as obse rved
be tw een H-6α and H-4α. On the basis of the above
evidence, the structure of compound 1 was de term ined
to be 6β-ange loy loxy-furano ligularenone.
C ompound 2 was iso lated as yellow powde r. The
UV spectrum gavemaximum absorption at 344 and 277
nm. The IR spectrum gave a strong absorption at 3 363
cm
-1
indicating the presence of a hydroxy l group, and
the absorptions at 1 774, 1 759, 1 613 cm - 1 for a t
least tw o carbony l groups w ere con firmed by the carbon
signals atδ178.8(s, C-2) and 169.3(s, C-12) in the
13
C NMR spec trum. Its molecular formula w as estab-
lished as C15 H14O 4 by HREIMS at m /z 258.090 9
(ca lcd. 258.089 2). The 1H NMR spectra show ed
signals due to a hydroxy l g roup atδ6.57(1H , br s);
two ole finic g roups atδ6.41(1H , s) and 6.25(1H ,
s);ame thy lene group atδ3.09(1H , d, J =16.6H z)
and 2.48(1H , dd, J =16.6, 0.7 Hz); three methy l
g roups atδ2.06(3H , s), 2.02(3H , br s) and 1.35
(3H , s). In the HMQC spectrum , corre lations
be tw een δH 6.41(H-1) and δC 124.6(d, C-1), δH
6.28(H-9) andδC 105.9(d, C-9), δH 3.09(H-6β),
2.46(H-6α) and δC 31.4( t, C-6), δH 2.06(H-15)
andδC 10.7(q, C-15), δH 2.02(H-13) and δC 8.6
(q, C-13), δH 1.35(H-14) and δC 28.7(q, C-14)
were obse rved. The HMBC corre lations o f δH 6.57
(1H , br s) w ith C-3, C-2 , C-4 , C-1 and C-15
established the position o f the hydroxy l g roup at C-3.
Extensive analysis of HMBC spectrum determ ined the
structure of compound 2 as 2-oxo-3-hydroxy-e remoph ila-
1(10), 3, 7(11), 8-te traen-8 , 12-o lide (Figure 2),
wh ich is a new natural compound and has been
obtained by chem ical synthesis
[ 4]
.
F igure 2　M ain corre lations inHMBC (H→C) spectra
o f compounds 1 and 2
530 药学学报 Ac ta Pharm aceu tica S in ica 2005, 40(6):529 - 532
Table 1　NMR spectral data of compounds 1 and 2 (500MHz, CDCl3)
No.
1*
δH δC
2
δH δC
1 6. 55(1H , dd, 10. 0, 2. 0) 148. 0(d) 6.41(1H , s) 124. 6(d)
2 6. 12(1H , dd, 10. 0, 3. 8) 128. 7(d) 178. 8(s)
3 199. 9(s) 144. 4(s)
4 2. 63(1H , m) 52. 8(d) 130. 5(s)
5 44. 7(s) 42. 1(s)
6 6. 28(1H , q, 1. 1) 72. 4(d) 2.48(α, 1H , dd, 16. 6, 0. 7) 31. 4( t)
3.09(β , 1H , d, 16. 6)
7 117. 2(s) 144. 5(s)
8 149. 2(s) 152. 3(s)
9 2. 65(α, 1H , dd, 15.8, 5. 2) 25. 4( t) 6.25(1H , s) 105. 9(d)
2. 63(β, 1H , m)
10 2. 96(1H , m) 43. 0(d) 160. 2(s)
11 119. 5(s) 125. 5(s)
12 7. 07(1H , br s) 138. 6(d) 169. 3(s)
13 1. 08(3H , d, 1. 1) 8. 7(q) 2.02(3H , br s) 8. 6(q)
14 0. 92(3H , s) 6. 8(q) 1.35(3H , s) 28. 7(q)
15 1. 10(3H , d, 6. 7) 7. 8(q) 2.06(3H , s) 10. 7(q)
OH 6.57(1H , br s)
*OAng:δH 6.20(1H , qq, 7. 3, 1.4, H-3′), 2.05(3H , dq, 7. 3, 1.4, H-4′), 1. 91(3H , dq, 1. 4, 1. 4,
H-5′) and δC 167.2( s, C-1′), 127.3( s, C-2′), 139.9(d, C-3′), 15. 7(q, C-4′), 20.7(q, C-5′)
Expermi ental
M elting po ints we re measu red w ith X-4 m icro-
me lting po in t appara tus and uncorrected. Optical
ro tations w ere determ ined w ith Perk in-E lmer 241
po larime ter. UV spectra performed on Shimadzu UV-
2501 spectrometer and IR spec tra w ere recorded on
N ico le t impact 410 spectrom eter.
1
H and
13
C NMR
spectra w ere obta ined on B ruke r ACF-500. HREIMS
and ESI M S measurements we re pe rformed on
HP5989A spectrome ter andA gilen t1100 series LC M/ S
Trap appa ra tus, respectively. TLC and co lumn
chromatography w ere ca rried out on p la tes precoa ted
w ith silica gel F254 and silica ge l (200 - 300 mesh)
(Q ingdao M arine Chem ical L td. , China ),
respective ly.
The roots and rhizomes of Ligularia pleurocaulis
(F ranch. )H and-Mazz. w ere co llected in Kangding,
S ichuan prov ince of China in August 2001. The p lant
w as identified by assoc iate professo r ZHANG M ian and
the vouche r specimen w as deposited in the herbarium
of China Pharmaceu tical Unive rsity.
The dried roots and rhizomes (4.0 kg) of
L. pleurocaulis were ground and perco lated w ith
me thano l at room tempe ra tu re fo r three times, each fo r
seven days. A fter removal o f the methanol under
reduced pressure, 520 g crude ex tract w as obtained
and suspended in wa ter, ex tracted w ith ethy l aceta te
and n-bu tano l to g ive ethy l ace tate so lub le fraction 380
g and bu tano l soluble fraction 60 g. The ethy l ace tate
frac tion (300 g) was subjected to silica ge l co lumn
chrom atog raphy using a g radien t solvent sy stem o f
pe tro leum-E tOAc (80∶1 to 0∶1) to afford five ma jor
frac tions (Fr. A to F r. E). F r. B (16.5 g) was
chrom atog raphied ove r silica ge l co lumn e lu ted w ith
pe tro leum-ac tone (9∶1 to 8∶2), and then Sephadex
LH-20 e lu ted w ith CHC l3-MeOH (1∶1) to y ie ld
compounds 1 (21 mg) and 2 (12 mg). F r. D (32.1
g) was extensive ly chrom atog raphied over silica ge l
co lumn w ith CHC l3-MeOH (98∶2 to 9∶1) and
Sephadex LH-20 e lu ted w ith CHC l3-MeOH (1∶1) to
y ield compounds 3 (5 mg), 4 (21 mg) and 5 (3
mg). The bu tano l fraction (55 g)was a lso separated
by silica ge l co lumn chrom atog raphy w ith a CHC l3-
MeOH gradient, yie ld ing compounds 8 (9 mg), 9 (2
mg), 10 (23 mg), 11 (35 mg) and 12 (48 mg).
Identification
6β-Angeloyloxy-furano ligularenone ( 1 )
C20H24O4 , co lo rless need les (pe tro leum-E tOAc), mp
178 - 179 ℃, [ α] 21D +41.8°(c 2.50, CHC l3 ).
HREIM Sm /z:328.164 1(calcd. 328.167 5). IR
υmax cm - 1:1 712, 1 697, 1 681, 1 642. 1H and 13C
NMR data listed in Table 1.
2-Oxo-3-hydroxy-eremophila-1(10), 3, 7(11),
8-tetraen-8, 12-olide (2)　C15H14O4 , ye llow powder.
HREIM Sm /z:258.090 9(calcd. 258.089 2). UV
531 ZHANG M ian, et a l:The chem ical constituents o f Ligu laria pleurocaulis
λm ax (MeOH ) nm:344, 277. IR υmax cm - 1:3 363
(OH), 1 774, 1 759 , 1 613. 1H and 13C NMR da ta
listed in Tab le 1.
T iglic acid (3)　C5 H8 O 2 , co lorless plate let
cry sta ls (pe tro leum), mp 63 - 65℃. 1H NMR (300
MH z, CDC l3) δ:12.54(1H , s, COOH), 6.25(1H ,
m , H-3), 2.04(3H , s, H-4) and 1.92(3H , t, H-5).
13
C NMR (75MHz, CDC l3)δ:174.1(s, C-1), 127.2
(s, C-2), 141.2(d, C-3), 20.32(q, C-4) and 16.0
(q, C-5). Th is w as iden tified by co-TLC chroma to-
gram w ith an au thentic sample.
O leanolic acid (4)　C30H48O 3 , wh ite pow der.
1
H NMR (500 Hz, CDC l3 ) δ:5.28(1H , t, 3.6,
H-12), 3.22(1H , dd, 4.3, 11.2, H-3), 1.13(3H , s,
CH 3), 0.99(3H , s, CH3), 0.93(3H , s, CH3), 0.91
(3H , s, CH3), 0.90(3H , s, CH 3), 0.77(3H , s,
CH 3) and 0.75(3H , s, CH3). The spectra data w ere
sim ilar to those recorded in re ference
[ 5]
.
Lupeol (5 ) 　 C30 H 50 O , colo rless need les
(E tOAc-H2O), mp 215 - 216℃. 1H NMR (500Hz,
CDC l3) δ:4.68(1H , dd, J =11.1, 4.8 H z, H-28),
4.56(1H , d, J =1.8 Hz, H-29), 1.68(3H , s, H-30),
1.03(3H , s, CH3), 0.96(3H , s, CH3), 0.94(3H , s,
CH 3), 0.83(3H , s, CH3 ), 0.79(3H , s, CH3 ) and
0.76(3H , s, CH3). The spectra data w ere sim ilar to
those recorded in refe rence
[ 5]
.
β-Sitosterol (6)　C29H 50 O , co lorless need les
(pe tro leum-E tOAc), mp 137 - 138 ℃. Identified by
co-TLC chromatog ram and co-me lting po int w ith an
authentic sample.
Daucosterol(7)　C35H60O 6 , colo rless pow der.
1
H NMR (500 H z, C5D5N) δ:5.36(1H , t, 2.4, H-
6), 5.05(1H , d, 7.7, H-1′G lu), 0.67(s, CH3),
0.88(d , 2.8, CH3 ), 0.89( t, CH 3), 0.90(d , 3.5,
CH 3), 0.95(s, CH3 ), 1.00(d , 6.5, CH3 ). The
spectra da ta w ere sim ilar to tho se o f the standard
substance.
Caffe ic ac id (8)　C9H8O 4 , pa le-ye llow need les
(MeOH-H2O), mp 223 -225℃. 1H NMR (500Hz,
acetone-d6) δ:7.53(1H , d, J =15.9H z, H-8), 7.32
(1H , d, J =1.1H z, H-2), 7.02(1H , dd, J =1.1, 8.2
H z, H-5), 6.87(1H , d, J =8.2 H z, H-6) and 6.24
(1H , d , J =15.9 H z, H-7). The spec tra data w ere
sim ilar to those recorded in re ference
[ 6]
.
Emodin (9)　C15H10O 5 , orange needles(MeOH),
mp 263 - 265℃. 1H NMR (500H z, acetone-d6) δ:
12.10(1H , br s, α-OH), 12.08(1H , br s, β-OH),
7.65(1H , br s, H-4), 7.31(1H , d, J =2.4H z, H-2),
7.11(1H , br s, H-5), 6.68(1H , J =2.4 Hz, H-7)
and 2.43(3H , s, Ar-CH3). The spec tra data were sim ilar
to tho se recorded in re ference
[ 5]
.
7-Methoxy-coumarin (10)　C10H8O3 , co lo rless
need les (MeOH ). 1H NMR (500 H z, CDC l3 ) δ:
7.64(1H , d, J =9.4 Hz, H-4), 7.38(1H , d, J =8.5
Hz, H-5), 6.84(1H , dd, J =2.2, 8.5H z, H-6), 6.82
(1H , d , J =2.5 Hz, H-8), 6.25(1H , d, J =9.4 H z,
H-3) and 3.82(3H , s, OCH3). The spectra data w ere
sim ilar to those reco rded in refe rence
[ 5]
.
Ferulic acid (11)　C10H10O4 , co lorless needles
(M eOH), mp 173 - 174 ℃. 1H NMR (500 H z,
ace tone-d6)δ:7.60(1H , d, J =15.9 Hz, H-8), 7.32
(1H , d, J =2.0 Hz, H-2), 7.13(1H , dd, J =2.0 , 8.1
Hz, H-5), 6.87(1H , d , J =8.1 Hz, H-6) and 6.37
(1H , d, J =15.9 Hz, H-7). The spectra data w ere
sim ilar to those reco rded in refe rence
[ 7]
.
4-Hydroxy-2, 5-dmi ethoxy-benzaldehyde (12)
C9H10O4 , colo rless need les (M eOH-H 2O), mp 105 -
106 ℃. N ative ES IM Sm /z:181[M - 1] -. 1H NMR
(500 H z, ace tone-d6 ) δ:11.37(1H , OH ), 9.79
(1H , CHO), 7.23(1H , H-3), 6.54 (1H , H-6),
3.92(3H , OM e) and 3.81(3H , OM e). The spectra
data w ere sim ilar to those recorded in re ference
[ 8]
.
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