全 文 ：书天然产物研究与开发 Nat Prod Ｒes Dev 2014，26:983-986
文章编号:1001-6880(2014)7-0983-04
Ｒeceived:April 3，2014 Accepted:May 16，2014
Foundation 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:snowcpx@126． com
石岩枫中两个新的木脂素类化合物
杨宁线1，2，梁光义1，曹佩雪1*
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
摘 要:从石岩枫 Mallotus repandus (willd)muell． -arg．全草中分离得到两个新的木脂素类化合物，根据理化性
质和波谱鉴定其化学结构，分别为:repandusin(1)、repanduthylin(2)。化合物 1 对 H2O2 诱导的 PC12 细胞损伤
有一定的保护作用，化合物 2 对 H2O2 诱导的 PC12 细胞损伤有微弱的保护作用。
关键词:石岩枫;木脂素类;H2O2 诱导;PC12 细胞
中图分类号:Ｒ284 文献标识码:A
Two New Coumarinolignoids from Mallotus repandus(Willd)Muell． Arg．
YANG Ning-xian1，2，LIANG Guang-yi1，CAO Pei-xue1*
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
Abstract:Two new coumarinolignoids were isolated from the Ｒhizome of Mallotus repandus (willd)Muell． -Arg． Silica
gel and Sephadex LH-20 column chromatographies were used to isolate the chemical constituents，and spectroscopic
techniques were employed to elucidate their structures． Their structures were identified as repandusin (1)and repan-
duthylin (2)． Compound 1 and 2 had medium and feeble protective effects to scathing PC12 cells which was induced by
H2O2 respectively．
Key words:Mallotus repandus;coumarinolignoids;H2O2-induction;PC12 cells
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 previously［2-4］． In this
study，we investigated the constituents of chloroform ex-
tract of M． repandus，and two new coumarinolignoids
were isolated．
Materials and Methods
Instruments
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，America)．
Plant materials
M． repandus was collected from Luodian county of
Guizhou，China，in Oct 2007． It was identified by Prof．
WEI Sheng-hua (Guiyang College of Traditional Chi-
nese Medicine，Guiyang 550002，China)．
Extraction and isolation
Air-dried powder of M． repandus 9． 0 kg was extracted
by 82% ethanol for three times and 60% ethanol for
once． The extracts were combined and concentrated un-
der vacuum． The residue obtained was suspended in
water，extracted respectively with petroleum ether，
chloroform and ethyl acetate to yield the petroleum e-
ther extract，chloroform extract and ethyl acetate ex-
DOI:10.16333/j.1001-6880.2014.07.006
tract． The chloroform layer (33 g)was concentrated
and then subjected to silica gel column chromatography
by eluting with petrol / EtOAc (80∶ 1-2∶ 1)and EtOAc
/ CH3OH (20 ∶ 1-1 ∶ 1)． Ten fractions were obtained，
and fraction 8 (2 g)were purified over silica gel col-
umn and Sephadex LH-20 to obtain compound 1 (38
mg) ;Fractions 9 (4 g)were purified over silica gel
column and recrystallization to afford compound 2(49
mg)．
Bioassay
PC12 cells were preactioned with certain concentration
of samples for 24 hours，and the culture of the samples
were discarded． Then join incubation media with H2O2
for some time to injury PC12 cells，and join H2O2 en-
zyme to terminate reaction and culture for 24 hours，test
cells surviving and growing by MTT method．
Compounds 1 and 2 were analyzed with three concen-
trations in a range between 2 μg /mL and 200 μg /mL，
respectively by MTT method． Compound 1 showed me-
dium protective effects to scathing PC12 cells which
was induced by H2O2，and compound 2 showed feeble
protective effects to that cells (Table 1)．
Table 1 Protective effects of compound 1 and 2 against in-
jury PC12 cells induced by H2O2
Sample
Ultimate
concentration
(μg /mL)
Ｒestraining
rate to
normal
PC12 cells (%)
Ｒestraining
rate to
PC12 cells
injured by
H2O2(%)
Compound 1 200 7． 11 34． 18 ± 9． 41
20 0 43． 40 ± 1． 26
2 0 9． 880 ± 3． 15
Compound 2 200 12． 05 13． 22 ± 9． 18
20 10． 24 27． 84 ± 9． 51
2 3． 73 6． 995 ± 10． 8
α-Tocopherol 4． 3 0 22． 91 ± 2． 45
Note:1． α-Tocopherol is positive control;2． the diffluence of compounds
will influence the result．
Ｒesults and Discussion
Structural elucidation
Compound 1 was obtained as buff powder，found to be
possess a molecular formula of C21H20O9，as evidenced
by positive mode HＲ-FAB-MS (m/z 417． 1209，［M +
H］+，calc 417． 1186)． The IＲ spectra of compound 1
showed absorption bands for hydroxyl (3525 cm-1) ，
carbonyl group (1699 cm-1)． The 1H NMＲ，13 C NMＲ
and DEPT spectra displayed signals for 21 carbons，
three methyl groups (δC 55． 9，56． 2，56． 4) ，all of them
were linked to oxygen;one methylene (δC 60． 8) ;sev-
en methines (δC 113． 9，144． 6，101． 1，116． 4，112． 3，
77． 6，80． 0) ;ten quaternary carbons (δC 160． 9，
106. 4，133． 1，139． 4，112． 0，106． 4，149． 4，148． 8，
127． 6，146． 5) ;δC 160． 9 were matched closely to the
carbonyl group of IＲ spectra． In the 1H NMＲ，the cou-
pling constant of δH 6． 45 and 7． 75 was 9． 6 Hz，which
is representative coumarin that H-3 and H-4 have no
substituent;H-5 is double apices，so H-6 has no sub-
stituent． In the 13C NMＲ，the chemical shift of δC 77． 6
and 80． 0 suggested their linking to an oxygen atom re-
spectively，δC 160． 9 confirm that compound 1 has a
carbonyl group［5］．
Detailed analysis of the 2D-NMＲ data，including 1H-1H
COSY，HMQC and HMBC，confirmed the above deduc-
tion． In the 1H-1H COSY，H-7and H-8，H-3 and H-4，
H-5 and H-6 were connected． In the HMBC spectrum，
the C-2，C-5 and C-10 showed correlations to H-4，
meanwhile，the C-7 and C-8 showed correlations to
H-9． The detailed1 H NMＲ，13 C NMＲ and 2D-NMＲ
data were shown in Table 2． The key 1H-1H COSY and
HMBC spectra were expressed in Fig． 1．
Compound 2，a white solid，had the molecular formula
C21 H24 O8 based on the positive-mode HＲ-FAB-MS
(m/z 404． 1477，［M + H］+，calc 405． 1533)． The IＲ
spectrum of compound 2 showed absorption bands for
hydroxyl (3490 cm-1) ，1469、1090 cm-1 and 750 cm-1
indicated the presence of benzene ring，but there is no
carbonyl group comparing to compound 1． The 1H
NMＲ，13 C NMＲ and DEPT spectrum displayed signals
for 21 carbons，three methyl groups (δC 49． 7，56． 3，
56． 4) ，all of them were linked to oxygen;three methyl-
enes (δC 61． 4，62． 9，62． 9) ;seven methines
(δC129. 6，100． 7，145． 4，105． 6，105． 6，77． 4，79． 9) ;
eight quaternary carbons δC 130． 4，130． 0，134． 0，
108. 6，149． 6，138． 2，149． 6) ;In the 1H NMＲ，two
methylenes in 6． 82 and 6． 94，which is representative
coumarin that H-3 and H-4 changed to unsaturated
bonds and had no substituent． In the 1H-1H COSY
489 Nat Prod Ｒes Dev Vol. 26
spectrum，the link between H-7 /H-8，H-8 /H-9，H-
3 /H-4 and H-2 /H-3 were observed clearly． In the HM-
BC spectrum，the C-5，C-9 and C-10 showed correla-
tions to H-4;the C-1 and C-2 showed correlations to
H-7． Meanwhile，C-7 and C-8 showed correlations to
H-9． The detailed data of compound 2 were shown in
Table 3，and the key correlations of 1H-1H COSY and
HMBC spectrum were indicated in Fig． 1［6］． Ｒepan-
dusin (1)and repanduthylin (2)were separated from
M． Ｒepandus for the first time，and were related to cou-
marin． According to Naming Organic Compounds of
IUPAC，compound 1 relating to coumarinic anhydride
and hence was named as repandusin and compound 2
was similar to lignan and hence was named as repan-
duthylin．
Fig. 1 Key HMBC (arrow， )and 1H-1H COSY (bold
line， )correlations of compounds 1 and 2
Table 2 The 1H NMＲ (400 MHz) ，13C NMＲ (100 MHz)and 2D-NMＲ data of compound 1 (δ in ppm，J in Hz)
No． δH δC HMBC 1H-1H COSY
2 160． 9
3 6． 45 (1H，d，9． 6) 113． 9 C-2，C-10 H-4
4 7． 75 (1H，d，9． 6) 144． 6 C-2，C-5，C-8，C-9，C-10 H-3
5 6． 74 (1H，d，6． 0) 101． 1 C-3，C-4，C-10 H-6
6 7． 28 (1H，d，8． 0) 116． 4 H-5
7 106． 4
8 133． 1
9 139． 5
10 112． 0
1 106． 4
2 149． 4
3 148． 8
4 127． 7
5 7． 41 (1H，s) 112． 3 C-1，C-3，C-4，C-10
6 146． 5
7 5． 58 (1H，m，8． 0) 77． 7 C-1，C-6 H-8
8 4． 50 (1H，m，6． 8) 80． 0 C-7 H-7
9 4． 30 (1H，t) ，3． 93 (1H，t) 60． 8 C-7，C-8
2-CH3 3． 59 (3H，s) 56． 2 C-3
3-CH3 3． 69 (3H，s) 55． 9 C-2
6-CH3 3． 76 (3H，s) 50． 0
Table 3 The 1H NMＲ (400 MHz) ，13C NMＲ (100 MHz)and 2D-NMＲ data of compound 2 (δ in ppm，J inHz)
No． δH δC HMBC 1H-1H COSY
2 4． 60 (1H，d，4． 8) 129． 6 C-2 H-3
3 6． 82 (2H，m，11． 6) 62． 9 C-2，C-3 H-2，H-4
4 6． 94 (2H，t，9． 6) 62． 9 C-2，C-4，C-5，C-9，C-10 H-3
5 6． 92 (1H，d，6． 4) 100． 8 C-3，C-5，C-9，C-10
589Vol. 26 YANG Ning-xian． et al:Two New Coumarinolignoids from Mallotus repandus (willd)muell． arg．
6 7． 09 (1H，d，1． 6 Hz) 145． 4 C-5，C-6，C-8，C-9
7 130． 4
8 130． 1
9 134． 0
10 108． 6
2-CH3 3． 62 (3H，s) 49． 7
3-CH3 3． 86 (3H，s) 56． 4 C-3
5-CH3 3． 82 (3H，s) 56． 4 C-5
1 127． 5
2 7． 16 (1H，s) 105． 6 C-1，C-3，C-4，C-2，C-7
3 149． 6
4 138． 2
5 149． 6
6 7． 16 (1H，s) 105． 6 C-1，C-3，C-4，C-6，C-7
7 5． 50 (1H，d，8． 0) 77． 4 C-1，C-2 ，C-6，C-8 H-8
8 4． 36 (1H，d，8． 0) 79． 9 C-7 H-7，H-9
9 3． 94 (1H，t) ，3． 91 (1H，t) 61． 4 C-7，C-8 H-8
Acknowledgments
This work was supported by the Characteristic Key Dis-
ciplines of Guizhou Province． We are grateful to Prof．
Sheng-Hua Wei for identifying the plant material． We
also thank Jian-Xin Zhang for NMＲ measures and Dao-
Ping Wang for mass spectra and infrared spectrum．
Meanwhile，we acknowledged for the Drug Screening
Lab of the key Laboratory of Chemistry for Natural
Products of Guizhou Province and Chinese Academy of
Science．
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