全 文 ：天然产物研究与开发 Nat Prod Res Dev 2013，25:733-735
文章编号:1001-6880(2013)6-0733-03
Received:March 25，2013 Accepted:May 20，2013
Fundation item:National Basic Research Program of China (973 pro-
gram，No． 2012CB722601) ;Key Subjects of the Chemistry of Chinese
materia of State Administration of TCM of P． R． China(GZYYRJF，No．
2012，32)
* Corresponding author Tel:86-851-3807713;E-mail:bixue_xu@ 126．
com
石岩枫中一个新的三萜类环内酯
杨宁线1，2，梁光义1，3，曹佩雪2，徐必学2*
1贵州省中国科学院天然产物化学重点实验室，贵阳 550002;
2贵阳护理职业学院，贵阳 550081;3 贵阳中医学院，贵阳 550002
摘 要:从石岩枫全草中分离得到三个三萜类化合物，根据理化性质和波谱鉴定其化学结构，分别为:repandula-
sin(1)、齐墩果-12-烯-3β，11α-二醇(2)及齐墩果酸(3)。化合物 1 为新化合物，化合物 2 为首次从该植物中提
出，其中化合物 1 对肿瘤细胞有微弱的抑制作用。
关键词:石岩枫;肿瘤细胞;抑制作用
中图分类号:R284 文献标识码:A
A New Triterpenoid Lactone from Mallotus repandus (willd)Müell． Arg
YANG Ning-xian1，2，LIANG Guang-yi1，3，CAO Pei-xue2，XU Bi-xue2*
1The Key Laboratory of Chemistry for Natural Products of Guizhou Province and Chinese Academy of Sciences，Guiyang 550002，China;
2Guiyang Nursing Vocational College，Guiyang 550081，China;3Guiyang College of Traditional Chinese Medicine，Guiyang 550002，China
Abstract:A new triterpenoid lactone repandulasin (1) ，together with two known triterpenoids:3β，11α-di-hydroxyolean-
12-oleanene (2)and oleanolic acid (3) ，were isolated from the whole plant of Mallotus repandus (willd)Muell． -Arg．
Their structures were elucidated on the basis of spectroscopic data and chemical methods． Compound 1 showed feeble re-
straining effect to K562 tumor cells．
Key words:Mallotus repandus;riterpenoid;repandulasin;restraining effect
Introduction
Mallotus repandus (willd)Muell． -Arg． (Euphorbiace-
ae)is a Chinese ethnic drug native to the south of
Guangdong province，Guizhou province and Taiwan． It
is widely used as a traditional medicine for rheumatic
arthrositis，snake-bite，hepatitis and liver cirrhosis［1］．
Two diterpene lactones，mallorepine and bergenin have
been isolated from this plant［2］． In this paper，we report
the isolation and structural elucidation of a new triter-
penoid repandulasin (1) ，along with two known triter-
penoids:3β，11α-di-hydroxy -olean-12-oleanene (2)
and oleanolic acid (3) (Fig． 1)．
Fig. 1 Structures of 1，2 and 3
Materials and Methods
General experimental procedures
IR spectra was obtained using a Vector 22 (KBr)
spectrometer． 1H NMR (400 MHz) ，13 C NMR (100
MHz) ，and 2D NMR spectra were recorded with an
INOVA-400 spectrometer using TMS as internal stand-
ard and values were given in ppm． Column chromatog-
raphy was performed on silica gel (Qingdao Haiyang
Chemical Co． Ltd．，China) ，Sephadex LH-20 (Sigma-
Aldrich，USA)．
Plant material
M． repandus (willd)Müell． Arg． was collected from
Luodian county of GuiZhou province，China，in Oct
DOI:10.16333/j.1001-6880.2013.06.001
2007． It was identified by Prof WEI Sheng-hua (Guiy-
ang College of Traditional Chinese Medicine，Guiyang
550002，China)．
Extraction and isolation
Air-dried powder of M． repandus (willd)Müell． Arg
(9． 0 kg)was extracted by 82% ethanol for three
times and 60% ethanol for once． The extracts were
combined and concentrated under vacuum． The residue
obtained was suspended in water，extracted respectively
with petroleum ether，chloroform and ethyl acetate to
yield the petroleum ether extract，chloroform extract
and ethyl acetate extract (37 g)． The ethyl acetate ex-
tract was chromatographed on a silica gel column elu-
ting with petroleum ether /EtOAc (1 ∶ 1 to 0 ∶ 1)and
EtOAc /MeOH (1 ∶ 0 to 1 ∶ 5)to yield twelve fractions
(F1-F12)． The fraction 5 was chromatographed on a
silica gel column eluting with CHCl3 /MeOH，followed
by a Sephadex LH-20 column eluted with CHCl3 /
MeOH，to yield a new compound 1，the fraction 6-8 was
subjected to a Silica gel column with CHCl3-MeOH as
the eluent to obtain two known triterpenoids 2 and 3．
Results and Discussion
Structural elucidation
Compound 1 was obtained as white powder． Its molecu-
lar formula，C37 H52 O4，was established using HR-ESI-
MS with m/z 583． 3780［M +Na］+(calc． 583． 3763)． The
IR spectrum exhibited no absorption band beyond 3000
cm-1，which indicated the disappearance of hydroxyl
group． There were carbonyl groups for absorption band
at 1702 cm-1，and benzene substitute absorption band at
1467，1099 and 688 cm-1 ．
The 1H NMR spectrum of compound 1 displayed sig-
nals of benzoyl part at 7． 44 (2H，t，J = 7． 2 Hz) ，
7. 57 (1H，t，J = 8． 0 Hz)and 8． 04 (2H，t，J = 6． 8
Hz)． In the up-field region，signals of two methenyl
groups at 3． 99 (1H，s，H-19)and 4． 89 (1H，m，H-
3) ，including seven methyl groups at 0． 81 (3H，d，J
= 6． 8 Hz ) ，0． 87 (3H，s) ，0． 88 (3H，s) ，0． 93
(3H，s) ，0． 98 (3H，s) ，1． 19 (3H，s)and 1． 20 (3H，
s)． The 13C NMR and DEPT spectral data of compound
1 exhibited 37 carbon signals (Table 1)including sev-
en methyls，ten methylenes，eleven methenyls and nine
quarternary carbons atoms． Among them，two carbonyl
groups were observed at δ 166． 4 and δ 177． 0;Two
methenyl signals at δ 75． 6 and δ 83． 5 were deduced to
be oxygenated，benzene carbon signals were detected at
δ 129． 5 (two carbons) ，128． 3 (two carbons) ，130． 8
and 132． 7 (Table 1)．
In the HMBC spectrum，the H-2 and H-6 (δH 8． 04)
showed correlations to C-7，and C-2，C-6，mean-
while，the correlations among H-3 /C-1，H-5 /C-1
and H-3 /C-7 were clearly observed． The correlations
of methyls were as follows:correlations between H3-23 /
C-3，C-4 and C-5 illustrated that the methyl (C-23)
was bonded to C-4，correlations among H3-25 /C-8，C-9
and C-10 indicated the position of the methyl (C-25)
bond at C-10，and correlations among H3-24 /C-4，C-5
and C-6 showed methyl (C-24)was linked at C-5． In
addition，C-29 and C-30 were also confirmed to link at
C-20． Finally，C-26 and C-27 were exhibited to link at
C-8 and C-14 respectively (Fig． 2)．
Fig. 2 Key HMBC，1H-1H COSY and ROESY correla-
tions of 1
Based on the above evidence，the structure of 1 was de-
termined as showed in Fig． 2 and it was named as
repandulasin． Assignment for compound 1 was based on
COSY，HMQC，HMBC experiments．
The structures of two known compounds 2 and 3 were i-
dentified as:3β-11α-di-hydroxy-olean-12-ene［3］ and
oleanolic acid［4］ respectively，by comparison of their
spectral data with those reported in the literature．
Biological results
The anti-tumor activity of compound 1 against K562
tumor cells and A549 tumor cells were investigated by
MTT assay and SRB method respectively． Three con-
centration levels (0． 2 mg /mL，0． 02 mg /mL and 0. 002
mg /mL)of compound 1 were analyzed． The MTT assay
results of compound 1 and adriamycin against K562
tumor cells were shown in Table 2．
437 Nat Prod Res Dev Vol. 25
Table 1 1H NMR (400 MHz)and 13C NMR (100 MHz)data of 1 in CDCl3
No． 13 C(δ，ppm) 1H (multi，J in Hz) No． 13C(δ，ppm) 1H (multi，J in Hz)
1 39． 40 (t) 1． 77 (1H，m) ，1． 82 (1H，m) 20 37． 99 (s)
2 21． 53 (t) 1． 87 (2H，m) 21 29． 99 (t) 0． 86 (1H，m) ，0． 92 (1H，m)
3 75． 57 (d) 4． 89 (1H，m) 22 35． 92 (t) 1． 20 (2H，m)
4 49． 77 (d) 0． 98 (1H，m) 23 10． 05 (q) 0． 81 (3H，d，J = 6． 8)
5 38． 47 (s) 24 14． 32 (q) 0． 88 (3H，s)
6 40． 46 (t) 0． 97 (1H，m) ，1． 06 (1H，m) 25 17． 97 (q) 0． 87 (3H，s)
7 38． 47 (t) 1． 96 (2H，m) 26 30． 51 (q) 0． 98 (3H，s)
8 36． 44 (s) 27 34． 59 (q) 0． 93 (3H，s)
9 58． 72 (d) 1． 02 (1H，m) 28 177． 04 (s)
10 37． 18 (s) 29 20． 35 (q) 1． 19 (3H，s)
11 19． 31 (t) 0． 98 (2H，m) 30 23． 27 (q) 1． 20 (3H，s)
12 18． 47 (t) 1． 46 (2H，m) 1 130． 80 (s)
13 38． 98 (d) 1． 86 (1H，m) 2 129． 52 (d) 8． 04 (1H，d，J = 6． 8)
14 27． 99 (s) 3 128． 27 (d) 7． 44 (1H，t，J = 8． 0)
15 31． 42 (t) 1． 42 (2H，m) 4 132． 67 (d) 7． 57 (1H，t，J = 6． 8)
16 32． 38 (t) 2． 18 (2H，m) 5 128． 27 (d) 7． 44 (1H，t，J = 8． 0)
17 51． 36 (s) 6 129． 52 (d) 8． 04 (1H，d，J = 6． 8)
18 57． 42 (d) 0． 85 (1H，m) 7 166． 38 (s)
19 83． 45 (d) 3． 99 (1H，s)
Table 2 MTT assay results of compound 1 and adriamycin
against K562 tumor cells
Sample Concentration Restraining rate (%)
Compound 1 0． 2 mg /mL 97． 26 ± 0． 16
0． 02 mg /mL 66． 96 ± 0． 41
0． 002 mg /mL 9． 42 ± 0． 12
Adriamycina 1． 00 × 10-6mol /L 64． 94 ± 2． 27
5． 00 × 10-7mol /L 63． 70 ± 2． 81
2． 50 × 10-7mol /L 62． 64 ± 2． 08
1． 25 × 10-7mol /L 54． 80 ± 2． 57
6． 25 × 10-8mol /L 45． 32 ± 7． 11
a adriamycin is positive control．
These results indicated that compound 1 had feeble re-
straining effect to K562 tumor cells． However，com-
pound 1 were found to be inactive to A549 tumor cells，
other relevant biological results were being investiga-
ted．
Acknowledgments We are grateful to Prof． Sheng-
Hua WEI for identifying the plant material． We also
thank Mr． Jian-Xin ZHANG for conducting NMR exper-
iments and Dao-Ping WANG for MS and IR analysis．
The bio-activity results were detected by Drug Screen-
ing Lab，the Key Laboratory of Natural Products of
Guizhou province and Chinese Academy of Science．
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537Vol. 25 YANG Ning-xian，et al:A New Triterpenoid Lactone from Mallotus repandus (willd)Müell． Arg