全 文 ： 2011 年 3 月 第 9 卷 第 2 期 Chin J Nat Med Mar. 2011 Vol. 9 No. 2 105

Chinese Journal of Natural Medicines 2011, 9(2): 0105−0107
doi: 10.3724/SP.J.1009.2011.00105
Chinese
Journal of
Natural
Medicines







New Cerebrosides from Acanthopanax gracilistylus
ZHANG Jing-Yan1, 2, PU She-Ban1*, QIAN Shi-Hui2*, LIU Dan1, 2
1China Pharmaceutical University, Nanjing 211198, China;
2Jiangsu Institute of Traditional Chinese Medicine, Nanjing 210028, China
Available online Mar. 2011
[ABSTRACT] AIM: To study the chemical constituents from the fruits of Acanthopanax gracilistylus. METHODS: Compounds
were isolated and purified by various chromatography techniques and their structures were elucidated by physicochemical properties
and spectroscopic data. RESULTS: A mixture of cerebrosides, called acanthopanax cerebrosides were isolated and characterized as
1-O-β-D-glucopyranosides of phytosphingosine type ceramides comprised of a common long chain base (2S, 3S, 4R, 8E)-2-[(2′R)-2′-
hydroxydocosanosylamino]-8(E)-alkene-1, 3, 4-triol. The alkene chain is eicosene(1), nonadecene(2), octadecene (3, momor-cere-
broside), heptadecene (4) and hexadecene (5).CONCLUSION: Compounds 1, 2 and 5 were new ones named as acanthopanx cere-
broside A (1), acanthopanax cerebroside B (2), and acanthopanax cerebroside C (5), and compound 3, 4 were isolated from this plant
for the first time.
[KEY WORDS] Acanthopanax gracilistylus; Chemical constituent; Acanthopanax cerebroside A; Acanthopanax cerebroside B
[CLC Number] R284.1 [Document code] A [Article ID] 1672-3651(2011)02-0105-03

1 Introduction
There are more than 26 species of Acanthopanax (Ar-
aliaceae) widely distributed in China, and the dried root barks
of Acanthopanax gracilistylus W.W. Smith are listed offi-
cially in Chinese Pharmacopoeia as Cortex Acanthopanacis
(Wujiapi), which has been used as medicines for the
treatment of paralysis, arthritis, rheumatism, lameness, and
liver diseases [1]. The leaves and fruits of Acanthopanax
gracilistylus as folk medicines are similar to Wujiapi in po-
tency, efficacy and clinical use. In our previous papers [2-3],
some lignans, phytosterols and lupine-triterpene glycosides
from the leaves and stem barks of Acanthopanax gracilistylus
have been isolated and identified.
In this paper as a continuing study on Acanthopanax ge-
nus, we reported the isolation and structure characterization
of a mixture of cerebrosides from the fruits of Acanthopanax
gracilistylus. These compounds were identified as acan-
thopanax cerebroside A (1), acanthopanax cerebroside B (2),

[Received on] 13-Dec.-2010
[Research Funding] This project was supported by the Public Re-
search Project in Jiangsu Province of Jiangsu Science and Technol-
ogy Departmeng (No. BM2006104).
[*Corresponding author] PU She-Ban: Associate Prof., Tel: 86-
25-86185139, E-mail: pusheban@126.com; Qian Shi-Hui: Prof., Tel:
86-25-85639644, E-mail: njqsh2005@126.com
These authors have no any conflict of interest to declare.
momor-cerebroside (3), 1-O-β-D-glucopyranose-(2S, 3S, 4R,
8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-heptade-
cene-1, 3, 4-triol (4), and acanthopanax cerebroside C (5).
Among these, compounds 3, 4 were found from the genus for
the first time, and compounds 1, 2 and 5 were new ones.
2 Apparatus and Reagents
1H NMR (500 MHz) and 13C NMR (125 MHz) spectra
were recorded with Bruker ACF-500 NMR spectrometer.
Mass spectra were obtained on MS Agilent 1100 Series
LC/MSD Trap Mass spectrometer (ESI-MS). All solvents
used were of analytical grade. Column chromatography was
performed with: macroporous resin D101 (Tianjin Hongyuan
Chemical Co., Ltd., Tianjin, China), silica gel (100-200, 200-
300 mesh ), and GF254 silica gel for TLC (Qingdao Marine
Chemical Co., Ltd., Qingdao, China), Sephadex LH-20
(Pharmacia Fine Chemical Co., Ltd.), and ODS-C18 (Merck).
The fruits of Acanthopanax gracilistylus were offered by
Xuyi Medicine Plant and identified by Qian Shi-Hui, one of
the authors. The voucher specimen was deposited in the De-
partment of Pharmacognosy, Jiangsu Institute of Traditional
Chinese Medicine.
3 Extraction and Isolation
The air-dried fruits of Acanthopanax gracilistylus (4 kg)
were refluxed with 80% ethanol and concentrated. The ex-
tract (0.5 kg) was dissolved in H2O and filtrated. Then the
ZHANG Jing-Yan, et al. /Chinese Journal of Natural Medicines 2011, 9(2): 105−107
106 Chin J Nat Med Mar. 2011 Vol. 9 No. 2 2011 年 3 月 第 9 卷 第 2 期

insoluble part was partitioned by EtOAc and n-BuOH suc-
cessively. n-BuOH fraction (100 g) was chromatographed on
macroporous resin D101 by elution with H2O, 30%, 50% and
95% aqueous EtOH successively. A saponin eluted with 95%
EtOH was subsequently chromatographed on silica gel using
CHCl3-MeOH-H2O (100 : 2 : 0.1-7 : 3 : 0.5) to give five
fractions (1-5). Fr. 5 was chromatographed on silica gel using
CHCl3-MeOH-H2O (100 : 10 : 0.5) to give six frs. A-F. Fr. B
was chromatographed on Sephadex LH-20 using CHCl3-
MeOH (1 : 1) to give a mixture of cerebrosides compounds
1-5 (10 mg).
4 Structure identification
Compounds 1-5 Amorphous white powder (MeOH).
ESI-MS m/z 872, 858, 844, 830 and 816 [M + H] +.
In the ESI-MS spectra of 1, (CH2)n functionalities and a
series of [M + H]+ ion peaks were observed. The NMR data
of 1 indicated the presence of a sugar (δH 4.94, 1H, d, J = 8.0
Hz, anomeric H; δC 105.6), an amide linkage (δH 8.52, 1H, d,
J = 8.5 Hz, N-H; δC 175.6), two long chain aliphatic moieties
(δH 1.26-1.32) and a triplet proton ( δH 0.87, 6H, t, J = 7.0 Hz)
which were essentially identical to those of momor-cerebro-
side [4-5], suggesting a glycosphingolipid nature [4-7]. The 1H
and 13C NMR spectra were typical of a sphingosine-type
cerebroside possessing 2-hydroxy fatty acid and β-gluco-
pyranose moieties (Table 1).
The positive ESI-MS spectra of 1-5 showed a series of
molecular ion [M + H]+ peaks at m/z 872, 858, 844, 830, 816,
and fragment ion at m/z 718 (Fig. 1) [glucosyl-long chain
base + H]+.
The presence of a fragment ion at m/z 718 in the ESI-MS
spectrum indicated that 1-5 comprise a common long-chain
base linked to varying chain lengths of fatty acid residue [8].
The presence of a 1, 3, 4-trihydroxy unsaturated C18 long
chain base was deduced from the 1H-1H correlation spectros-
copy (COSY) and MS data.The signal at δH 8.52 gave a cross
peak with the signal at δH 5.27 (H-2) in the 1H-1H COSY
spectrum of 1, which in turn showed cross peaks with
methylene protons (H-1) at δH 4.56 and 4.68 and δH 4.27
(H-3). The latter correlated with the signal at δH 4.17 (H-4).
The chemical shift of the H-2 signal and the 13C chemical
shifts of C-1-C-4, C-1 and C-2 of glucosphin- golipids are
especially suitable for the determination of the absolute
stereochemistry of the phytosphingosine moiety [6]. The
chemical shift of H-2 (δH 5.27) and the carbon chemical
shifts at δC 70.4 (C-1), 51.7 (C-2), 75.9 (C-3), 72.4 (C-4),
Table 1 1H and 13 C NMR data of 1 (C5D5N)
No. δC δH COSY HMBC
1 70. 4 4.68 dd (11.0, 6.5) 4.56 dd (10.5, 4.0) 2-H 1″- H,3-H
2 51.7 5. 27 m 1-H 1-H, 3-H
3 75.9 4.27m 4-H
4 72.4 4.17 m 3-H, 5-H 3-H
5 33.8 2.24 m, 1. 91 m 4-H, 6-H
6 26. 6 2. 18 m 5-H, 7-H
7 33.3 2.00 m 8-H, 6-H
8 130.8 5.47 m 7-H, 9-H 7-H, 10-H
9 130.6 5.52 m 8-H,10-H 6-H, 7-H, 10-H
10 32.9 2.00 m (11-17)-H, 9-H
11-17 29. 51 - 30. 1 1.26 - 1.32 brs
18 32.1 1.26 - 1.32 brs 20-H
19 22.9 1.26 - 1.32 brs (11-17)-H, 20-H
20 14.3 0.87 t (7. 0) (11-17)-H
1′ 175.6
2′ 72.4 4. 47 m 3′-H
3′ 35.5 2.18 m, 1.98 m 2′-H, 4′-H
4′ 25.8 1.70 m 3′-H, (5′-21′)-H
5′-21′ 29. 51 - 30. 1 1.26 - 1.32 brs
22′ 32.1 1.26 - 1.32 brs
23′ 22.9 1.26 - 1.32 brs
24′ 14.3 0. 87 t (7. 0) (5′-23′)-H
1″ 105. 6 4.94 d (8. 0) 2″- H 2″- H
2″ 75. 1 3.99 t (7.5) 1″- H, 3″- H
3″ 78. 5 4.18 m 2″- H 2″- H, 4″- H
4″ 71.5 4.18 m 5″- H 3″- H, 5″- H
5″ 78. 4 3.86 m 4″- H, 6″- H 4″- H, 6″- H
6″ 62. 6 α 4. 50 dd (10.5, 4.5) β 4. 32 dd (11.5, 5.0) 5″- H
NH 8. 52 d (8.5) 2- H

ZHANG Jing-Yan, et al. /Chinese Journal of Natural Medicines 2011, 9(2): 105−107
2011 年 3 月 第 9 卷 第 2 期 Chin J Nat Med Mar. 2011 Vol. 9 No. 2 107


Fig. 1 ESI-MS pattern of 1

Fig. 2 Key 1H-1H COSY( ) and HMBC(→) correlations
of 1
175.6 (C-1) and 72.4 (C-2) in 1 were essentially identical to
those of the momor-cerebroside [4-5] and the data reported for
other (2S, 3S, 4R)-phytosphingosine moieties[6, 9]. These re-
sults clearly indicate that the 1, 3, 4-trihydroxy phytos-
phingosine moiety in 1 possesses the 2S, 3S, 4R configuration.
The position of the double bond in the long chain base
was determined at C-8 by HMBC (Fig. 2) analysis as indi-
cated in momor-cerebroside, and 3 was determined as momor-
cerebroside.
The trans (E) configuration of the double bond was evi-
denced by the chemical shifts of the carbons next to the dou-
ble bond at δC 32.9 (C-7) and 33.3 (C-10) in 1. The geometry
is also verified from the fact that the olefinic protons ap-
peared at δH 5.47-5.52 as a m-like signal in 1 [6]. In the light
of the above evidence, the structures of these cerebrosides
were deduced to be 1-O-β-D-glucopyranose-(2S, 3S, 4R,
8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-eicosene-1,
3, 4-triol (1), 1-O-β-D-glucopyranose-(2S, 3S, 4R, 8E)-2-
[(2′R)-2′-hydroxydocosanosylamino]-8(E)-nonadecene-1, 3,
4-triol (2), 1-O-β-D-glucopyranose-(2S, 3S, 4R, 8E)-2-
[(2′R)-2′-hydroxydocosanosylamino]-8(E)-octadecene-1, 3, 4-
triol (momor-cerebroside) (3), 1-O-β-D-glucopyranose-(2S,
3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-
hepta- decene-1, 3, 4-triol (4), 1-O-β-D-glucopyranose-(2S,
3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-
hexadecene-1, 3, 4-triol (5).
Compounds 3 and 4 have been found for the first time in
Acanthopanax gracilistylus. Compounds 1, 2 and 5 were new
cerebrosides and named as acanthopanax cerebroside A,
acanthopanax cerebroside B, and acanthopanax cerebroside
C .
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细柱五加中新的脑苷酯类成分
张静岩 1, 2 , 濮社班 1*, 钱士辉 2*, 刘 丹 1, 2
1中国药科大学, 南京 211198;
2江苏省中医药研究院, 南京 210028
【摘 要】 目的：研究细柱五加 Acanthopanax gracilistylus 果实的化学成分。为阐明其有效成分提供科学依据。方法：采
用硅胶、Sephadex LH-20 及 Rp-18 等柱层析色谱方法进行分离纯化, 并根据理化性质和光谱数据鉴定其结构。结果：分离得到
5 个脑苷酯类同系物, 分别鉴定为 1-O-β-D-glucopyranose-(2S, 3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-eicosene-1, 3,
4-triol(1), 1-O-β-D-glucopyranose-(2S, 3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-nonadecene-1, 3, 4-triol (2)，1-O-β-D-
glucopyranose-(2S, 3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-octadecene-1, 3, 4-triol (momor-cerebroside) (3)，1-O-β-D-
glucopyranose-(2S, 3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-heptadecene-1, 3, 4-triol (4)，1-O-β-D-glucopyranose-(2S,
3S, 4R, 8E)-2-[(2′R)-2′-hydroxydocosanosylamino]-8(E)-hexadecene-1, 3, 4-triol (5)。结论：化合物 3 和 4 苦瓜脑苷为首次从该属植
物中分离得到, 化合物 1，2, 5 均为新化合物, 分别命名为：细柱五加脑苷 A (1)、细柱五加脑苷 B (2)、细柱五加脑苷 C (5)。
【关键词】 细柱五加; 化学成分；细柱五加脑苷 A；细柱五加脑苷 B

【基金项目】江苏省公益研究项目(No. BM2006104)