Chemical constituents from Eupatorium chinense

Article abstract of DOI:10.1080/10286020.2011.598149

Two new monoterpenoid glycosides, α-(E)-acaridiol 8-O-β-d-glucopyranoside (1) and α-(E)-acaridiol 9-O-β-d- glucopyranoside (2), as well as a new thiophen, 2-acetyl-3-hydroxy-5-(prop-1- ynyl)thiophen 3-O-β-d-glucopyranoside (3), were isolated from the roots of Eupatorium chinense. Their structures were determined on the basis of extensive spectroscopic and chemical analyses.

Full text of DOI:10.1080/10286020.2011.598149

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Chemical constituents from Eupatorium
chinense
Wen-Jing Wang a b , Ying Wang a b , Qin-Wen Zhang c , Xiao-Qi
Zhang a b , Ting-Ting Yang a b , Yi Dai a b , Lei Wang a b , Lei-Hong
Zhang d & Wen-Cai Ye a b
a Institute of Traditional Chinese Medicine and Natural Products,
College of Pharmacy, Jinan University , Guangzhou, 510632, China
b Guangdong Province Key Laboratory of Pharmacodynamic
Constituents of TCM and New Drugs Research , Jinan University ,
Guangzhou, 510632, China
c Institute of Chinese Medical Sciences, University of Macau ,
Taipa, Macau, China
d Guangdong Province Institute of Traditional Chinese Medicine ,
Guangzhou, 510520, China
Published online: 10 Aug 2011.
To cite this article: Wen-Jing Wang , Ying Wang , Qin-Wen Zhang , Xiao-Qi Zhang , Ting-
Ting Yang , Yi Dai , Lei Wang , Lei-Hong Zhang & Wen-Cai Ye (2011) Chemical constituents
from Eupatorium chinense , Journal of Asian Natural Products Research, 13:9, 845-850, DOI:
10.1080/10286020.2011.598149
To link to this article: http://dx.doi.org/10.1080/10286020.2011.598149
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Journal of Asian Natural Products Research
Vol. 13, No. 9, September 2011, 845–850
Chemical constituents from Eupatorium chinense
Wen-Jing Wangab1, Ying Wangab1, Qin-Wen Zhangc, Xiao-Qi Zhangab, Ting-Ting Yangab,
Yi Daiab, Lei Wangab, Lei-Hong Zhangd and Wen-Cai Yeab*
aInstitute of Traditional Chinese Medicine and Natural Products, College of Pharmacy, Jinan
University, Guangzhou 510632, China; Guangdong Province Key Laboratory of
b
Pharmacodynamic Constituents of TCM and New Drugs Research, Jinan University, Guangzhou
510632, China; cInstitute of Chinese Medical Sciences, University of Macau, Taipa, Macau, China;
dGuangdong Province Institute of Traditional Chinese Medicine, Guangzhou 510520, China
(Received 18 April 2011; final version received 14 June 2011)
Two new monoterpenoid glycosides, a-(E)-acaridiol 8-O-b-D-glucopyranoside (1) and
a-(E)-acaridiol 9-O-b-D-glucopyranoside (2), as well as a new thiophen, 2-acetyl-3-
hydroxy-5-(prop-1-ynyl)thiophen 3-O-b-D-glucopyranoside (3), were isolated from
the roots of Eupatorium chinense. Their structures were determined on the basis of
extensive spectroscopic and chemical analyses.
Keywords: Eupatorium chinense; monoterpenoid glycoside; thiophen
1. Introduction
(4–8) (Figure 1). In this paper, the
isolation and structural elucidation of the
two new isomeric monoterpenoid glyco-
sides and the new thiophen are described.
The plants of genus Eupatorium (Compo-
sitae) are mainly spread in the temperate
areas of USA and East Asia. Previous
phytochemical investigation revealed that
the main constituents in the plants of this
genus were sesquiterpene lactones [1–5],
triterpenoids [6,7], flavonoids [5,6], ben-
zofurans [7,8], and lignans [7]. The dried
root of Eupatorium chinense had been
widely used as a Chinese folk medicine for
the treatment of multifarious throat dis-
orders for a long time. The potential
medicinal importance of E. chinense
prompted us to initiate a phytochemical
study on the roots of this plant, which led
to the isolation of two new monoterpenoid
glycosides, a-(E)-acaridiol 8-O-b-D-glu-
copyranoside (1) and, a-(E)-acaridiol 9-O-
b-D-glucopyranoside (2), and a new
thiophen, 2-acetyl-3-hydroxy-5-(prop-1-
ynyl)thiophen 3-O-b-D-glucopyranoside
(3), along with five known compounds
2. Results and discussion
Compound 1 was obtained as a colorless
gum. The molecular formula of 1 was
established as C16H28O7 by HR-ESI-MS at
1
m/z 355.1730 [M þ Na]þ. The H NMR
spectrum of 1 showed signals for two vinyl
protons [dH 5.63 and 5.12 (each 1H, m)],
two oxo-methylenes [dH 4.40 (1H, m),
4.29 (1H, m) and 4.00 (2H, br s)], two
methylenes [dH 2.14 (2H, m) and 2.10 (2H,
m)], two methyls [dH 1.67 and 1.60 (each
3H, s)], and an anomeric proton [d 4.28
(1H, d, J ¼ 7.8 Hz)]. The 13C NMR and
DEPT spectra of 1 exhibited 16 carbon
signals including 4 olefinic carbons, 4
methylenes, 2 methyls and a sugar unit.
With the aid of 1H–1H COSY, HSQC,
HMBC, and NOESY experiments, the
*Corresponding author. Email: chywc@yahoo.com.cn
ISSN 1028-6020 print/ISSN 1477-2213 online
q 2011 Taylor & Francis
DOI: 10.1080/10286020.2011.598149
http://www.informaworld.com

846
W.-J. Wang et al.
OH
5′
OR1
6′
HO 4′
HO
8
O
1
1′
3
5
O
7
2′ OH 10
3
4
3′
2
6
4
9
6
9
10
2
5
S
OR2
8
1
7
O
1 R1 = Glc, R2 = H
3
2 R1 = H, R2 = Glc
OH
OH
O
OCH3
OH
O
O
HO
HO
HO
HO
O
O
O
HO
HO
O
OH
OH
OH
H3CO
R
HO
4
5 R = OH
8
6 R = COOH
7 R = CH2OH
Figure 1. Chemical structures of compounds 1–8.
NMR signals of 1 were assigned as shown
in Table 1.
The above 1H and 13C NMR signals of
the aglycone and sugar unit of 1 were
similar to those of the known compound a-
(E)-acaridiol [9] and glucopyranosyl [10].
The HMBC correlations between the
proton at dH 5.12 (H-3) and the carbons
at dC 25.9 (C-1)/17.8 (C-10), and between
the proton at dH 5.63 (H-7) and the carbons
at dC 29.2 (C-5)/66.3 (C-9), confirmed the
planar structure of the aglycone of 1
Table 1. 1H and 13C NMR spectral data of compounds 1 and 2 (CD3OD, d in ppm, J in Hz)a.
1
2
No.
dC
dH
dC
dH
1
2
3
4
5
6
7
8
25.9 q
133.0 s
125.0 d
28.2 t
1.67 s
–
5.12 m
2.10 m
2.14 m
–
5.63 m
4.40 m
4.29
25.9 q
132.8 s
125.1 d
28.0 t
1.67 s
–
5.12 m
2.10 m
2.14 m
29.2 t
29.1 t
144.4 s
122.5 d
66.2 t
139.3 s
128.7 d
59.0 t
–
5.68 t (6.6)
4.13 dd (6.6, 2.8)
9
66.3 t
4.00 br s
73.2 t
4.30 d (12.3)
4.08 d (12.3)
1.61 s
4.27 d (7.8)
3.19
3.25
10
10
20
30
40
50
60
17.8 q
103.1 d
75.1 d
77.9 d
71.7 d
78.2 d
62.7 t
1.60 s
4.28 d (7.8)
3.19
3.25
3.28
3.33
17.8 q
103.1 d
75.1 d
77.9 d
71.8 d
78.2 d
62.8 t
3.28
3.33
3.85 dd (12.0, 2.2)
3.66 dd (12.0, 5.5)
3.86 dd (12.0, 2.2)
3.65 dd (12.0, 5.4)
Note: a Overlapped signals were reported without designating multiplicity.

Journal of Asian Natural Products Research
847
HO
OH
HO
OH
OH
OH
H
OH
H
HO
O
HO
O
O
H2C
CH
O
O
2
H
OH
H3C
O
H
HO
HO
S
OH
CH3
OH
O
1
2
3
Figure 2. Key HMBC ( ! ) and NOESY ( $ ) correlations of 1–3.
(Figure 2). The NOESY correlation
between H-5 at dH 2.14 and H-8a at dH
4.40 suggested that the geometry of the
double bond was E form. Furthermore, the
HMBC correlation between H-8a at dH
4.40 and C-10 at dC 103.1 indicated that the
glucopyranosyl unit was located at C-8.
Thus, the structure of 1 was elucidated as
a-(E)-acaridiol 8-O-b-D-glucopyranoside.
Compound 2 was obtained as a color-
less gum. Compound 2 showed the same
molecular formula as 1 by its HR-ESI-MS.
The 1H and 13C NMR spectral data
(Table 1) of 2 were also very similar to
those of 1, except that the C-9 resonance in
2 exhibited a comparative downfield shift,
whereas the C-8 resonance displayed
somewhat upfield shift. Analysis of the
1H–1H COSY, HSQC, HMBC, and
NOESY spectral data of 1 and 2 led to
the conclusion that the two compounds
were isomers due to the different location
of the sugar unit. The HMBC correlation
between H-9b at dH 4.08 and C-10 at dC
103.1 suggested that the monosaccharide
moiety was located at C-9. The NOESY
correlation between H-5 at dH 2.14 and H-
8 at dH 4.13 confirmed that the geometry of
the double bond was also E form. Thus, the
structure of 2 was elucidated as a-(E)-
acaridiol 9-O-b-D-glucopyranoside.
ESI-MS indicated that the molecular
formula of 3 was C15H18O7S. The IR
spectrum of 3 suggested the presence of
hydroxyl group (3299 cm21), acetylenic
bond (2234 cm21), and carbonyl group
(1624 cm21). The UV spectrum showed
absorption maxima at 226 and 316 nm.
The above data suggested that 3 may be a
1
thiophen. The H NMR spectral data of 3
showed the presence of an olefinic proton
[dH 7.08 (1H, s)], two methyls [dH 2.56
and 2.07 (each 3H, s)], and an anomeric
proton [dH 5.01 (1H, d, J ¼ 7.4 Hz)]. The
13C NMR and DEPT spectra of 3 showed
15 carbon signals including a carbonyl, 4
olefinic carbons, 2 methyls, and a sugar
unit. Acid hydrolysis of 3 suggested the
presence of D-glucopyranosyl residue. The
HMBC correlations between H-10 at dH
2.56 and C-2 at dC 125.1, between H-10 at
dH 5.01 and C-3 at dC 158.4, as well as
between H-8 at dH 2.07 and C-5 at dC
131.9 indicated that the acetyl, glucopyr-
anosyl unit and prop-1-ynyl group were
connected to C-2, C-3, and C-5 positions,
respectively. Thus, the structure of 3 was
characterized as 2-acetyl-3-hydroxy-5-
(prop-1-ynyl)thiophen 3-O-b-D-glucopyr-
anoside.
Other known compounds were ident-
ified as pungenin (4) [10], 2,6-dimethoxy-
4-hydroxyphenol-O-b-D-glucoside (5)
[11], syringic acid 4-O-b-D-glucopyrano-
side (6) [12], di-O-methylcrenatin (7) [11],
Compound 3 was obtained as an
amorphous powder. The quasi-molecular
ion at m/z 365.0665 [M þ Na]þ in its HR-

848
W.-J. Wang et al.
and benzyl-1-O-b-D-glucopyranoside (8)
[13], by comparison of their physical and
spectroscopic data with those reported
previously.
China, in October of 2006, and authenti-
cated by Prof. Shu-Yuan Li (Guangdong
Pharmaceutical University). A voucher
specimen (No. 2006101601) is deposited
in the Institute of Traditional Chinese
Medicine & Natural Products, Jinan
University, Guangzhou, China.
3. Experimental
3.1 General experimental procedures
Melting points were measured on an X-5
micro melting point apparatus (Beijing
Tech Instrument, Inc., Beijing, China). UV
spectra were obtained on a Jasco V-550
UV/Vis spectrometer. IR spectra were
obtained on a Jasco FT/IR-480 plus
infrared spectrometer with KBr pellets.
Optical rotations were determined on a
Jasco P-1020 polarimeter. 1D and 2D
NMR spectra were measured using a
Bruker AV-400 spectrometer. HR-ESI-
MS data were detected on an Agilent 6210
ESI/TOF mass spectrometer. Column
chromatography (CC) was carried out on
silica gel (200–300 mesh; Qingdao
Marine Chemical, Inc., Qingdao, China),
ODS (YMC, Kyoto, Japan), and Sephadex
LH-20 (Pharmacia, NJ, USA), respect-
ively. Preparative high-performance liquid
chromatography (HPLC) was carried out
on a Varian chromatograph equipped
with a Prostar 215 pump, a Prostar
325 UV-vis detector, and a Cosmosil
5C18-MS-II reversed-phase column
(20 mm £ 250 mm, 5.0 mm, Nacalai Tes-
que, Kyoto, Japan). Analytical HPLC
was carried out on a Dionex chromato-
graph equipped with a P680 pump, a PDA-
100 photodiode array detector, and a
Cosmosil 5C18-MS-II reversed-phase col-
umn (4.6 mm £ 250 mm, 5.0 mm, Nacalai
Tesque). Chiral separation was carried out
on an Agilent 1200 system equipped with a
G1315B DAD detector and a chiral CD-Ph
column (4.6 mm £ 250 mm, 5.0 mm, Shi-
seido Fine Chemicals Ltd, Tokyo, Japan).
3.3 Extraction and isolation
The dried and powdered roots of E.
chinense (10 kg) were extracted with 70%
(v/v) EtOH–H2O (3 £ 30 L, each 2 h)
under reflux. The concentrated EtOH
extract was suspended in H2O (4 L) and
then successively extracted with petroleum
ether (4 £ 4 L), EtOAc (4 £ 4 L), and n-
BuOH (4 £ 4 L). The n-BuOH solution was
concentrated to give a residue (180 g),
which was subjected to silica gel
column eluting with CHCl3 –MeOH
(100:0 ! 0:100) to afford seven fractions
(Fr. A–G). Fr. C (3 g) was further
purified using ODS column eluting with
MeOH–H2O (50:50 ! 90:10) to give five
fractions (Fr. C1ZC5). Fr. C1 (183 mg)
was subjected to preparative HPLC
(MeOH–H2O, 10:90; flow rate, 8 ml/min;
wavelength, 210 nm) to yield 4 (4 mg), 5
(17 mg), and 7 (16 mg). Fr. C2 (50 mg) was
subjected to preparative HPLC (MeOH–
H2O, 25:75; flow rate, 8 ml/min; wave-
length, 210 nm) to obtain 8 (4 mg). Fr. C3
(246 mg) was separated by preparative
HPLC (MeOH–H2O, 40:60; flow rate,
8 ml/min; wavelength, 210 nm) to yield 3
(12 mg), 6 (7 mg), and a mixture of 1 and 2
(10 mg). The mixture was further separated
into isomers 1 (0.5 mg, tR 23.9 min) and 2
(0.3 mg, tR 26.5 min) by chiral column
chromatography (MeOH–H2O, 20:80; col-
umn temperature, 308C; flow rate,
0.8 ml/min; wavelength, 210 nm).
3.3.1 a-(E)-Acaridiol 8-O-b-D-
glucopyranoside (1)
3.2 Plant material
The roots of E. chinense were collected in
Conghua city, Guangdong Province of
A colorless gum, IR nKmBaxr: 3412, 3020,
;
2961, 1673, 1102, 1060, 810 cm21 1H

Journal of Asian Natural Products Research
849
NMR (CD3OD, 400 MHz) and 13C NMR
(CD3OD, 100 MHz) spectral data, see
Table 1; HR-ESI-MS m/z 355.1730
[M þ Na]þ (calcd for C16H28O7Na,
355.1727).
aqueous layer was concentrated and dried
by N2, and treated with dry pyridine (1 ml)
and L-cysteine methyl ester hydrochloride
(2 mg), followed by heating at 608C for
2 h, and then concentrated to dryness with
N2. The residue was added to N-
(trimethylsilyl)imidazole (0.2 ml) and
kept at 608C for 1 h. Then, the solution
was diluted with H2O (1 ml) and extracted
with cyclohexane (3 £ 1 ml). The super-
natant was concentrated to 1 ml, then
subjected to GC analysis [column: AT-SE-
30 (0.5 mm £ 0.32 mm £ 30 m), detector:
FID, column temperature: 2208C, detector
temperature: 2708C, injector temperature:
2708C, and carried gas: N2]. A peak of the
derivative of 3 was observed at tR
28.78 min (D-Glc), whereas the peaks of
the standard monosaccharide derivatives
were recorded at 28.76 (D-Glc) and 30.22
(L-Glc), respectively.
3.3.2 a-(E)-Acaridiol 9-O-b-D-
glucopyranoside (2)
A colorless gum, IR nKmBaxr: 3413, 3019,
2961, 1673, 1107, 1062, 811 cm21 1H
;
NMR (CD3OD, 400 MHz) and 13C NMR
(CD3OD, 100 MHz) spectral data, see
Table 1; HR-ESI-MS m/z 355.1731
[M þ Na]þ (calcd for C16H28O7Na,
355.1727).
3.3.3 2-Acetyl-3-hydroxy-5-(prop-1-
ynyl)thiophen 3-O-b-D-glucopyranoside
(3)
An amorphous powder, mp 118–1198C;
½aꢀ2D5 237.8 (c 0.7, CH3OH); UV (MeOH)
lmax (log 1) 226 (4.11), 316 (4.50) nm; IR
Acknowledgements
(KBr) nmax 3299, 2234, 1624, 1454, 1100,
1080, 1041 cm21 1H NMR (CD3OD,
This work was financially supported by grants
from the Program for Changjiang Scholars and
Innovative Research Team in University
(IRT0965), the Natural Science Foundation of
Guangdong Province (No. 8351063201000003;
No. 9451052005001975), the Science and
Technology Development Fund of Macau
(No. 013/2008/A1), and Administration of
Traditional Chinese Medicine of Guangdong
Province (No. 2010235).
;
400 MHz) d: 7.08 (1H, s, H-4), 5.01 (1H,
d, J ¼ 7.4 Hz, H-10), 3.88 (1H, dd,
J ¼ 12.1, 2.0 Hz, H-60a), 3.69 (1H, dd,
J ¼ 12.1, 5.6 Hz, H-60b), 3.47 (1H, m, H-
20), 3.46 (1H, m, H-30), 3.43 (1H, m, H-50),
3.39 (1H, m, H-40), 2.56 (3H, s, H-10),
2.07 (3H, s, H-8); 13C NMR (CD3OD,
100 MHz) d: 192.6 (C-9), 158.4 (C-3),
131.9 (C-5), 125.1 (C-2), 123.7 (C-4), 96.0
(C-7), 73.7 (C-6), 29.8 (C-10), 4.2 (C-8),
103.5 (C-10), 78.5 (C-50), 78.1 (C-30), 74.8
(C-20), 71.1 (C-40), 62.3 (C-60); HR-ESI-
MS m/z 365.0665 [M þ Na]þ (calcd for
C15H18O7SNa, 365.0665).
Note
1. The authors contributed equally to this
work.
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