全 文 ： 94 Chin J Nat Med Mar. 2010 Vol. 8 No. 2 2010 年 3 月 第 8 卷 第 2 期

Chinese Journal of Natural Medicines 2010, 8(2): 0094−0096
doi: 10.3724/SP.J.1009.2010.00094
Chinese
Journal of
Natural
Medicines







Diterpenoids from Euphorbia esula
WANG Yu-Bo, JI Ping, WANG Hong-Bing, QIN Guo-Wei*
Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 201203,
China
Available online Mar. 2010
[ABSTRACT] AIM: To study the chemical constituents of the aerial parts of Euphorbia esula. METHODS: Compounds were iso-
lated by repeated column chromatography over silica gel and Sephadex LH-20. Structures were elucidated based on spectral analysis
and comparison with literatures. RESULTS: Three ingenane-type diterpenoids were isolated and identified as 16-benzoyloxy-
20-deoxyingenol 5-benzoate (1), ingenol-3, 20-dibenzoate (2) and 13, 16-dibenzoyloxy-20-deoxyingeno-3-benzoate (3), respectively.
CONCLUSION: Compound 1 is a new ingenane-type diterpenoid diester, and compound 3 was isolated from the plant for the first
time. In vitro bioassay data showed that diterpenoids 2 and 3 exhibited weak cytotoxicity against Namalwa cells.
[KEY WORDS] Euphorbia esula; Chemical constituents; Diterpenoids; Cytotoxicity
[CLC Number] 284.1 [Document code] A [Article ID] 1672-3651(2010)02-0094-03

Euphorbia is the largest genus in the family Euphor-
biaceae, comprising more than 1 000 species, among which
about 80 species grow in China. According to our literature
review about 40 Chinese Euphorbia plants have been used
medicinally. The plant Euphorbia esula L. is a perennial herb,
distributed widely in China and used as a folk medicine for
the treatments of cancer, swelling and wart [1]. The plant was
first studied by Kupchan et al in 1976, resulting in the isola-
tion of ingenol-3, 20-dibenzoate, which showed antileu- ke-
mic activities against P-388 lymphocytic leukemia in mice at
doses of 130-360 μg·kg−1 [2]. Besides ingenane-type diter-
penoids [3-4], the plant also contains jatrophane-type diter-
penoids [5-9] as well as flavonoids [10-11]. In the course of our
investigation on the plant E. esula, a novel norparaliane
diterpenoid named eslatin G has been isolated from its aerial
parts [12]. Our further study has led to the isolation of three
additional diterpenoids (1-3, Fig. 1). They were identified as
a new ingenane diterpenoid, 16-benzoyloxy-20-deoxyingenol
5-benzoate (1), and two known ingenane diterpenoids, in-
genol-3, 20-dibenzoate (2) [2] and 13, 16-dibenzoyloxy-20-
deoxyingeno-3-benzoate (3) [13]. Compounds 2 and 3 showed
weak cytotoxic activity toward Namalwa cells.

[Received on] 12-Sep-2009
[Research Funding] This project was supported by Science and
Technology Commission of Shanghai Municipality (No. 06DZ22028)
and Shanghai Institute of Materia Medica (No. SIMM0709QN-05)
[*Corresponding author] QIN Guo-Wei: Prof., Tel: 86-21-5080
5853; E-mail: gwqin@mail.shcnc.ac.cn
1 Results and Discussion
Compound 1, obtained as resinous solids with the mo-
lecular formula C34H36O7, determined by HR-EI-MS (m/z
556.246 6, [M]+ calcd., 556.246 1). In the IR spectrum, 1
exhibited carbonyl absorption band at νmax 1 720 cm−1 and a
strong hydroxyl band at νmax 3 442 cm−1. The 1H NMR data
at δ 7.45-8.12 (m, 10H) and 13C NMR data at δ 166.2, 169.9
(two ester carbonyls) and 128.4-133.5 (two phenyl carbons)
revealed the presence of two benzoyloxyl groups in com-
pound 1. Besides, the remained moiety showed 20 signals for
four methyls (two olefinic), two methylenes (one oxygena-
ted), eight methines (two oxygenated, two olefinic) and six
quaternary carbons (one carbonyl, two olefinic, two oxygena-
ted). Moreover, the 1H NMR spectrum of 1 showed signals
for two olefinic protons at δ 6.00 (s, H-1), 5.89 (q, J = 1.6 Hz,
H-7), two oxygenated methine protons at δ 3.83 (s, H-3), 5.42
(s, H-5), three methyl groups at δ 1.83 (s, H-19), 1.55 (s,
H-20), 1.01 (d, J = 7.1 Hz, H-18) and one methelene group at
δ 4.43, 4.62 (ABq, each 1H, J =12.0 Hz, 16- CH2OBz), indi-
cating that 1 was an ingenane-type diterpenoid dibenzoate.
In the ingenane-type diterpenoids, C-3, 5, 13, 16, 20 are
often found to be esterificated. Our further analysis showed
that the 1H NMR data of 1 were very similar to those of
16-benzoyloxy-20-deoxyingenol 3-benzoate [2]. The HMBC
correlations between δH 4.43, 4.62 (H2-16) and δC 27.9
(C-14), 30.9 (C-15), 24.5 (C-17) and 169.9 (carbonyl of ben-
zoate) indicated a benzoyloxy group at C-16. Besides, the
HMBC correlations between δH 5.42 (H-5) and C-7 (δ 125.0),
WANG Yu-Bo, et al. /Chinese Journal of Natural Medicines 2010, 8(2): 94−96
2010 年 3 月 第 8 卷 第 2 期 Chin J Nat Med Mar. 2010 Vol. 8 No. 2 95

HOBzO
HO
CH2OBz
H
H
O
H
2
HOHO
BzO
CH3
CH2OBz
H
H
O
H
HOBzO
HO
CH3
CH2OBz
H
BzO
O
H
1 3
20
2
4 6
810
12
14 16
17
18
19

Fig. 1 Structures of compounds 1-3

between δH 3.83 (H-3) and C-1 (δ 130.1), and δH 5.42 (H-5)
and 166.2 (carbonyl of benzoate) indicated that another ben-
zoyloxyl group was linked at C-5. Therefore, the structure of
1 was concluded to be 16-benzoyloxy-20-deoxyingenol
5-benzoate, a new ingenane-type diterpenoid diester. Regar-
ding biogenesis and spectral similarity, 1 was considered to
have the same stereochemistry as compounds 2, 3 and 16-
benzoyloxy-20-deoxyingenol 3-benzoate.
Diterpenoids 2 and 3 were identified by spectral analysis
and comparison with literatures, among which 3 was first
isolated from the plant, and 2 and 3 showed weak cyto- tox-
icity on Namalwa cells (human B-lymphoma cells) with IC50
of 13.16 and 11.46 μmol·L−1 respectively with positive con-
trol Vinblastine sulfate (IC50 0.029 μmol·L−1).
2 Experimental
2.1 General
Optical rotations were measured on a Perkin Elmer
model 341 polarimeter. UV spectra were recorded on a Shi-
maduz UV-2550 UV-visible spectrophotometer. FT-IR spec-
tra were recorded on a Nicolet Magna FT-IR spectro- pho-
tometer. NMR spectra were recorded on a Bruker AM- 400
or an AV-500 spectrometer with TMS as the internal stan-
dard. EI and HR-EI-MS were carried out on a MAT 95 mass
spectrometer. Silica gel is from Qingdao Marine Chemical
Group Co. and Sephadex LH-20 from Pharmacia, Sweden.
All solvents are in chemical grades (Shanghai Chemical
Company, Ltd).
2.2 Plant Material
The aerial parts of E. esula were collected from A’ba
Tibet area of Sichuan province, October, 2005, and identified
by Prof. LONG Xing-Jie, Southwest Herbal Medicine Store,
Chengdu, Sichuan Province. A voucher specimen (No.
20051022) was deposited in the herbarium of Shanghai In-
stitute of Materia Medica, Chinese Academy of Sciences.
2.3 Extraction and Isolation
The powdered aerial parts of E. esula (20 kg) were ex-
tracted with 95% EtOH. The ethanol extracts were suspended
in water and extracted successively with CHCl3 and EtOAc
to give each residue 360 g and 210 g, respectively. The
CHCl3 residue was subjected to silica gel column, eluted with
petroleum ether containing increasing amounts of acetone to
give four combined fractions according to TLC detection.
The third and fourth fractions were separated by column
chromatography on silica gel eluting with petroleum
ether-acetone (20∶1), followed over LH-20 to give 1 (14
mg), 2 (24 mg) and 3 (17 mg).
2.4 Identification
16-Benzoyloxy-20-deoxyingenol 5-benzoate (1) Resi-
nous solids; C34H36O7, [α]D24 -38.3º (c 0.47, MeOH); IR (KBr)
νmax 3 417, 2 937, 1 635, 1 035 cm−1; 1H NMR (500 MHz,
CDCl3) δ: 6.00 (1H, s, H-1), 3.83(1H, s, H-3), 5.42 (1H, s,
H-5), 5.89 (1H, m, H-7), 4.50 (1H, m, H-8), 2.48 (1H, m,
H-11), 1.93 (2H, m, H2-12), 4.43, 4.62 (ABq, each 1H, J
=12.0 Hz, H2-16), 1.24 (3H, s, H3-17), 1.01 (3H, d, J = 7.1
Hz, H3-18), 1.83 (3H, s, H3-19), 1.55 (3H, s, H3-20);
7.45-8.12 (10H, m, 2 × OBz); 13C NMR (125 MHz, CDCl3) δ:
130.1 (C-1); 139.5 (C-2); 80.2 (C-3); 85.3 (C-4); 77.3 (C-5);
135.5 (C-6); 125.0 (C-7); 43.7 (C-8); 202.6 (C-9); 72.9
(C-10); 39.6 (C-11); 31.0 (C-12); 24.1 (C-13); 27.9 (C-14);
30.9 (C-15); 66.3 (C-16); 24.5 (C-17); 17.0 (C-18); 15.4
(C-19); 21.4 (C-20); 166.2, 130.4, 130.1, 128.4, 132.9
(5-OBz), 169.9, 129.2, 129.6, 128.6, 133.5 (16-OBz); EI-MS
m/z 556 [M]+, 538, 510, 447, 105; HR-EI-MS m/z 556.246 6
[M]+ (calcd. for C34H36O7, 556.246 1).
Ingenol-3,20-dibenzoate[2] (2) Resinous solids; C34H36O7,
1H NMR (400 MHz, CDCl3) δ: 6.13 (1H, d, J = 0.4 Hz, H-1),
5.71 (1H, s, H-3), 3.74 (1H, s, H-5); 5.74 (1H, br s, H-7),
4.23 (1H, m, H-8), 2.57 (1H, m, H-11), 1.92 (2H, m, H2-12),
1.21 (3H, s, H3-16), 1.71 (3H, s, H3-17); 1.03 (3H, d, J = 7.8
Hz, H3-18), 1.82 (3H, d, J = 0.4 Hz, H3-19); 4.76, 4.34 (2H,
ABq, J = 16.0 Hz, H2-20); 7.39-8.03 (10H, m, 2 × OBz); 13C
NMR (100 MHz, CDCl3) δ: 132.5 (C-1), 137.9 (C-2), 83.7
(C-3), 85.3 (C-4), 77.2 (C-5), 135.9 (C-6), 123.1 (C-7), 43.1
(C-8), 206.1 (C-9), 72.0 (C-10), 39.1 (C-11), 31.0 (C-12),
24.1 (C-13), 23.8 (C-14), 27.8 (C-15), 24.5 (C-16), 30.9
(C-17), 17.0 (C-18), 15.7 (C-19), 66.2 (C-20), 167.1, 130.3
(3-OBz) 166.8, 129.5 (20-OBz), 128.3-133.4 (other data of
3,20-OBz); EI-MS m/z 556 [M]+.
13,16-Dibenzoyloxy-20-deoxyingeno-3-benzoate[13] (3)
Resinous solids; C41H40O9, [α]D20 +40º (c 0.18, CHCl3); IR
(KBr) νmax 3 461, 1 724, 1 274, 711 cm−1; 1H NMR (400
MHz, CDCl3) δ: 5.72 (1H, s, H-1), 5.73 (1H, s, H-3), 3.77
(1H, s, H-5), 6.16 (1H, s, H-7), 4.27 (1H, m, H-8), 2.71 (1H,
m, H-11), 2.60 (1H, dd, J = 4.7, 16.8 Hz, H-12a), 3.00 (1H,
dd, J = 3.1, 16.8 Hz, H-12b), 1.62 (1H, d, J = 12.6 Hz, H-14),
4.62, 4.43 (2H, ABq, J = 12.0 Hz, H2-16), 1.25 (3H, s, H3-17),
1.09 (3H, d, J = 7.2 Hz, H3-18), 1.84 (3H, s, H3-19), 1.71
WANG Yu-Bo, et al. /Chinese Journal of Natural Medicines 2010, 8(2): 94−96
96 Chin J Nat Med Mar. 2010 Vol. 8 No. 2 2010 年 3 月 第 8 卷 第 2 期

(3H, s, H3-20), 7.45-8.12 (15H, m, 3 × OBz); 13C NMR (125
MHz, CDCl3) δ: 122.2 (C-1); 138.3 (C-2); 83.6 (C-3); 85.3
(C-4); 77.3 (C-5); 136.3 (C-6); 132.1 (C-7); 42.9 (C-8); 205.1
(C-9); 72.0 (C-10); 38.6 (C-11); 35.3(C-12); 69.3 (C-13);
28.9 (C-14); 34.4 (C-15); 65.6 (C-16); 18.7 (C-17); 18.0
(C-18); 15.6 (C-19); 21.8 (C-20); 167.2, 130.1, 129.8, 128.6,
133.2 (3-OBz), 166.2, 130.2, 129.7, 128.4, 133.2 (13-OBz),
166.7, 129.5, 129.8, 128.4, 132.9 (16-OBz); EI-MS m/z 676
[M]+, 658, 584, 554, 511, 105.
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乳浆大戟的化学成分
王玉波, 季 萍, 王红兵, 秦国伟*
中国科学院上海生命科学院上海药物研究所, 上海 201203
【摘 要】 目的：研究乳浆大戟(Euphorbia esula)地上部分的化学成分。方法：利用反复硅胶和凝胶 LH-20 柱层析进行分离
和纯化, 通过理化性质和波谱数据分析鉴定化合物结构。结果：分离并鉴定出 3 个二萜化合物, 分别为 3, 20-dibenzoyloxyingenol
(1), 3, 16-dibenzoyloxy-20-deoxyingenol (2) 和 3, 13, 16-tribenzoyloxy-20-deoxyingenol (3)。结论：化合物 1 为新二萜化合物, 3 为
本植物中首次报道, 2 和 3 对 Namalwa 细胞显示弱的细胞毒活性。
【关键词】 乳浆大戟; 化学成分; 二萜; 细胞毒

【基金项目】 上海市科委基金(No. 06DZ22028)和中国科学院青年人才基金(No. SIMM0709QN-05)