全 文 ：中华双扇蕨中一个新的贝壳杉烷型二萜?
王 扣1 , 2 , 李明明1 , 2 , 成 晓1 , 许 刚1 ,
彭丽艳1 , 赵勤实1??
(1 中国科学院昆明植物研究所植物化学与西部植物资源持续利用国家重点实验室 ,
云南 昆明 650204; 2 中国科学院研究生院 , 北京 100049)
摘要 : 从中华双扇蕨 ( Dipteris chinensis) 中分离得到一个新的对映贝壳杉烷型二萜和反式桂皮酸的二聚体 :
16β-hydroxy -17-[ ( Z) -p-coumaroyl ]- ent-kauran-19-oic acid, 命名为 dipterinoid A ( 1)。同时首次从该植物中分离
得到其他 13 个已知化合物。
关键词 : 双扇蕨科 ; 中华双扇蕨 ; 对映贝壳杉烷型二萜 ; Dipterinoid A
中图分类号 : Q 946 文献标识码 : A 文章编号 : 0253 - 2700 (2009) 03 - 279 - 05
A New ent-Kaurane Diterpenoid from the Aerial
of Dipteris chinensis (Dipteridaceae)
WANG Kou
1 , 2
, LI Ming-Ming
1 , 2
, CHENG Xiao
1
, XU Gang
1
,
PENG Li-Yan1 , ZHAO Qin-Shi1 * *
(1 State KeyLaboratory of Phytochemistry and Plant Resources in West China, Kunming Instituteof Botany, Chinese Academy
of Sciences, Kunming 650204 , China; 2 Graduate University of ChineseAcademy of Sciences, Beijing 100049 , China)
Abstract: A new ent-kaurane diterpenoid, dipterinoid A (1 ) , was isolated from the aerial parts of Dipteris chinensis to-
gether with thirteen known compounds . Its structure was determined as 16β-hydroxy-17- [ ( Z) -p-coumaroyl ]- ent-kauran-
19-oic acid by extensive NMR and MS analysis . All of known compounds were isolated fromthis plant for thefirst time .
Key words: Dipteridaceae; Dipteris chinensis; ent-Kaurane-type diterpene; Dipterinoid A
Dipteris, theonly living genus of the family Dip-
teridaceae, includes eight species which are distributed
in the tropics of Asia . Threeof the species have a dis-
tribution in the southwest, south China and Taiwan
(Delectis FloraeReipublicaePopularis SinicaeAgendae
Academiae Sinicae Edita, 2000 ) . To the best of our
knowledge, only investigation about chemical constitu-
entsof Dipterisplants has been reported which revealed
thepresenceof ent-kaurane-type diterpenes in D. con-
jugat (Tanaka et al. , 1985 ) . Dipteris chinensis, dis-
tributed in Yunnan, Guizhou, and Guangxi provinces
of China, is used as herbal medicine to treat edema
and kidney deficiency ( Ching, 1980) , andoccasional-
ly planted for ornament because of its large showy fan-
shaped leaves . There is no phytochemical report on this
plant up to now . As apart of our research work onbio-
activemetabolites fromfernsof China (Li et al. , 2006;
2007) , phytochemical investigation on the aerial parts
of D. chinensis was studied, and a new ent-kaurane
diterpenoid, dipterinoid A ( 1) , was isolated from the
云 南 植 物 研 究 2009 , 31 (3) : 279～283
Acta Botanica Yunnanica DOI : 10 .3724?SP. J . 1143 .2009.09007
?
?? ?Author for correspondence; E-mail : qinshizhao@ mail . kib. ac. cn; Tel : + 86 - 871 - 5223058
Received date: 2009 - 01 - 12 , Accepted date: 2009 - 03 - 03
作者简介 : 王扣 ( 1983 - ) 女 , 在读博士研究生 , 主要从事蕨类植物化学与生物学的研究。 ?
Foun ?dation items: Thefoundations from the project ( P-06-04) of State KeyLaboratory of Phytochemistry and Plant Resources in West China, Kun-
ming Instituteof Botany, Chinese Academy of Sciences, and the 973 Program ( No . 2009CB522300 )
Fig . 1 The structures of compound 1 and 2
95% ethanol extract together with thirteen known com-
pounds . Herein, the isolation and the structural eluci-
dation of 1 was reported .
Results and Discussion
Compound 1 , obtained as white amorphous pow-
der, has the molecular formula C29 H38 O6 based on the
negative HR-ESI-MS ( [ M-H] - at m?z 481.2583;
calcd for C29 H37 O6 , 481 .2590) as well as NMR spec-
tral data, requiring eleven degreesof unsaturation . The
IR spectrum showed the absorption bands for the
hydroxyl ( 3423 cm- 1 ) and carbonyl groups ( 1696
cm- 1 ) . The 13 C NMR and DEPT spectra showed 29
carbon signals including twenty carbon signals for a
diterpenoid unit together withnine characteristic signals
for a p-coumaroyl unit atδC 167 .1 (s) , 159 .6 (s) ,
144 .1 ( d) , 133 .6 ( 2×d) , 127 .4 (s) , 116 .8 ( d)
and 115 .7 ( 2×d) . The 1 H NMR data also indicated
the presence of the p-coumaroyl moiety atδH 5 .81
(1H, d, J = 12 .8 Hz, H - 2′) , 6 . 88 ( 1H, d, J =
12 .8 Hz, H - 3′) , 7 . 74 (2H , d, J = 8 .8 Hz, H - 5′
and H - 9′) , and 6 .82 ( 2H, d, J = 8 .8 Hz, H - 6′
and H - 8′) . This was further supported by the 1 H-1 H
COSY ( H - 2′with H - 3′, H - 5′?H - 9′with H - 6′?H
- 8′) and HMBC correlations ( H - 2′and H - 3′with
C - 1′and C - 4′) . In addition, the double bond was
suggested as cis-double bond due to the coupling con-
stant (Macias et al. , 1997) .
The remaining diterpenoid unit was consisted of
two angle methyl groups (δH 0.98 , 1 .17 ; δC 16 .0 ,
29 .2) , ten methylenes ( includingone oxygenated car-
bon atδC 71 .6 for C - 17 ) , four methines ( including
δC 57 .4 , 57 .1 , 42 .4 for C - 5 , C - 9 , C - 13 respec-
tively, and one oxygenated carbon atδC 78 .5 for C -
16) , three quaternary carbons ( includingδC 44 .5 ,
44 .0 , 40 .4 for C - 8 , C - 4 , C - 10 respectively) , and
one carbonyl group (δC 178 .9) . Considering the spec-
tral characteristic discussed above, together with the
fact that several ent-kauranediterpenoids such as16β,
17-dihydroxy- ent-kauran-19-oic acid ( 2 ) ( Tanaka et
al. , 1985) and 9 , 16 , 17-trihydroxy-19-kauranoic ac-
id ( Richter et al. , 1977 ) isolated from D. chinensis,
this diterpenoid unit was ascribable to be an ent-kau-
rane diterpenoid . The NMR data of 1 was very similar
to those of 2 except for the nine more signals for the
( Z) -p-coumaroyl unit discussed above, which indicat-
ed that 1 was an ent-kaurane diterpenoid with p-cou-
maroyl (Table1) .The ( Z) -p-coumaroyl unit in 1 was
connected to C - 17 , as suggested by the HMBC corre-
lationsof H - 17 with C - 1′. In addition, the 1 H-1 H
COSY spectrum of 1 indicated the presence of the
structural fragments - CH2 CH2 CH2 - (C-1 - C-2 - C-3) ,
- CHCH2 CH2 - (C-5 - C-6 - C-7) , and - CHCH2 CH2 CH
- (C-9 - C-11 - C-12 - C-13) . Analysis of the HMBC
spectrum demonstrated the correlations of H - 5 (δH
1 .07 , 1H, m) with C - 4 , C - 6 , C - 7 , C - 10 , C -
18 , and C - 19 , H - 9 (δH 1 .09 , 1H, m) with C - 5 ,
C - 8 , C - 10 , C - 12 , C - 15 , and Me - 20 , and from
H - 13 (δH 2.01 , 1H, m) to C - 8 , C - 12 , and C -
16 . All the 2D NMR data discussed above confirmed
the structure of ent-kaurane diterpenoid further more .
The relative stereochemistry of 1 was suggested as
the same with that of 2 , and the hydroxyl group at C -
16 was also suggested to be β-oriented because the
082 云 南 植 物 研 究 31 卷
Table 1 1 H and 13 C NMR Dataof 1 in CD3 COCD3
No
1 ?
δC δH HMBC
2 ?
δC
1 ?41 N. 5 t 0 . 85 , 1 . 89 m C-9 , C-10 , C-11 , C-20 41 ?. 2 t
2 ?19 N. 9 t 1 . 38 , 1 . 91 m C-3 , C-4 , C-10 , C-18 19 ?. 8 t
3 ?38 N. 8 t 1 . 18 , 1 . 97 ma C-5 , C-15 , C-18 38 ?. 6 t
4 ?44 J. 0 s 43 ?. 9 s
5 ?57 F. 4 d 1 . 07 m C-4 , C-6 , C-7 , C-10 , C-18 , C-19 57 ?. 1 d
6 ?22 N. 6 t 1 . 81 m C-5 , C-9 22 ?. 5 t
7 ?42 N. 6 t 1 . 44 m C-5 , C-8 , C-9 42 ?. 5 t
8 ?44 J. 5 s 43 ?. 9 s
9 ?57 F. 1 d 1 . 09 m C-5 , C-8 , C-10 , C-12 , C-15 , C-20 56 ?. 7 d
10 ?40 J. 4 s 40 ?. 1 s
11 ?19 N. 5 t 1 . 55 m, 2 ?. 11 m C-8 , C-12 , C-13 19 .4 t
12 ?27 N. 5 t 1 . 43 m, 1 ?. 84 m C-14 , C-16 27 .6 t
13 ?42 F. 4 d 2 . 01 m C-8 , C-12 , C-16 41 ?. 6 d
14 ?38 N. 8 t 1 . 01 m, 2 ?. 11 mb C-9 , C-12 38 .7 t
15 ?53 N. 6 t 1 ?. 45 d, J = 14 .0 Hz
1 ?. 51 d, J = 14 .0 Hz C-7 , C-8 , C-9 , C-16 , C-17 53 ?. 4 t
16 ?78 J. 5 s 79 ?. 1 s
17 ?71 N. 6 t 3 ?. 97 d, J = 11 .2 Hz
4 ?. 05 d, J = 11 .2 Hz C-13 ?, C-15 , C-16 , C-1′ 70 .4 t
18 ?29 F. 2 q 1 . 17 s C-3 , C-4 , C-5 , C-19 29 ?. 4 q
19 ?178 ]. 9 s 180 #. 0 s
20 ?16 F. 0 q 0 . 98 s C-10 ?, C-19 16 .0 q
1 u′ 167 ]. 1 s
2 u′ 116 X. 8 d 5 ?. 81 d, J = 12 .8 Hz C-1′, C-3′, C-4′
3 u′ 144 X. 1 d 6 ?. 87 d, J = 12 .8 Hz C-1′, C-4′, C-5′, C-9′
4 u′ 127 ]. 4 s
5 3′, 9′ 133 ?. 6 2×d 7 . 74 dd, J = 8 .8 , 2 L. 0 Hz C-3′, C-7′
6 3′, 8′ 115 ?. 7 2×d 6 . 82 dd, J = 8 .8 , 2 L. 0 Hz C-4′, C-7′
7 u′ 159 ]. 6 s

a, b Assignments with the same superscript may be reversed, although those given here arepreferred .
chemical shift of α-oriented C - 17 CH2 OH signal
(aboutδC 70.0) (Yamasaki et al. , 1976; Satake et
al. , 1984) should bemoredownfield than that of β-ori-
ented C - 17 CH2 OH signal (aboutδC 66.0) (He et
al. , 2005; Nishimura et al. , 2007; Wang et al., 1995) .
Therefore, the structure of compound 1 was elucidated
as 16β-hydroxy-17-[ ( Z ) -p-coumaroyl ] - ent-kauran-
19-oic acid and named as dipterinoid A (Fig . 1 ) .
The known compounds were identified to be san-
daracopimaric acid ( Comte et al. , 1995) , 16β, 17-di-
hydroxy- ent-kauran-19-oic acid (Tanaka et al. , 1985) ,
9, 16, 17-trihydroxy-19-kauranoic acid ( Richter et al. ,
1977 ) , (20 R )-dammar-24-en-20-ol (Yamashita et al. ,
1998) , bergapten (Masuda et al. , 1998) , kaempferol
(Markham et al. , 1978 ) , quercitrin ( H?rhammer et
al. , 1966; Zhong et al. , 1997) , quercetin 3-robinobi-
oside ( Brasseur and Angenot, 1986 ) , nicotiflorin
(Markham et al. , 1978 ) , 20-hydroxyecdyson 20 , 22-
acetonate (Homvisasevongs et al. , 2004) , 4-hydroxy-
benzoic acid (Scott, 1972 ) , β-sitosterol (Nes et al. ,
1992) and daucosterol (Seo et al. , 1978) . However,
20-hydroxyecdyson 20 , 22-acetonatemay be an artefact
because acetone was used in the course of isolation .
The naturally occurring compound may be 20-hydroxy-
ecdyson (Suksamrarn et al. , 2002) . Their structures
were determined bycomparisonof thespectroscopic da-
ta with those reported in the literature .
Experimental
General experimental procedures Optical rotation was
measured on aHoribaSEPA-300 polarimeter . IR spectrawas ob-
tained with a Tensor 27 FT-IR spectrometer with KBr pellets .
The1 H and 13 C NMR spectra were recorded on Bruker AV-400
and DRX-500 spectrometers in CD3 COCD3 at room temperature
(δ in ppm, J in Hz) . FAB-MS was carried out on a VG Auto-
spec-3000 spectrometer . HR-ESIMS was recorded with an API
1823 期 WANG Kou et al. : A New ent-Kaurane Diterpenoid fromthe Aerial of Dipteris chinensis (Dipteridaceae)
QSTAR Pulsar i spectrometer . Silica gel F254 , Silica gel H
(Qingdao Marine Chemical Ltd . , China) and LiChroprep RP-18
silica gel (40 - 63μm, Merck, Dramstadt, Germany) were used
for column chromatography . Fractions were monitored by TLC
and spots were visualized by heating silica gel plates immersed
with 15% H2 SO4 in ethanol . Solventswere distilled prior to use .
Plant material The aerial parts of D. chinensiswere col-
lected from Dawei Mountain, Pingbian County, Yunnan Prov-
ince, PRC, in July 2007 and were identified by professor Cheng
Xiaoof Kunming Institute of Botany, Chinese Academy of Sci-
ences . A voucher specimen has been deposited in the State Key
Laboratoryof Phytochemistry and Plant ResourcesinWest China,
Kunming Institute of Botany, Chinese Academy of Sciences .
Extraction and isolation Air-dried and powdered plant
materials (2 . 3 kg) were extracted with 95 % EtOH ( 10 L×3 ,
each3 days) at room temperature . After evaporation of the sol-
vent in vacuo at 50℃ , the residue was dissolved in H2 O and
then extracted exhaustively with EtOAc ( 2 L× 4 ) . The EtOAc
extract (88 g) was dissolved in 95% ethanol then subjected to
column chromatography over MCI elutingwith 90 % ethanol . The
elution was concentrated in vacuo to give a residue ( 71 g) ,
which was subjected to column chromotography over silica gel
(100 - 200 mesh) and elutedwith petroleumether-acetone (1∶0 ,
9∶1 , 8∶2 , 7∶3 , 6∶4 , 1∶1 , 0∶1 ) to give six fractions A-F .
Fractions B-E were separated repeatedly on silica gel , and then
further purified using Sephadex LH-20 with ( CHCl3 -MeOH 0∶1
and 1∶1) and RP-18 (MeOH-H2 O gradient elution of increasing
concentration) . Fraction B gave ( 20 R )-dammar-24-en-20-ol
(65.0 mg) . FromfractionC , bergapten (4 . 5 mg) , sandaracopi-
maric acid (5 .0 mg) , 4-hydroxybenzoic acid (3 .0 mg) , andβ-
sitosterol ( 2 . 0 g) were obtained . Fraction D afforded 1 ( 4 .0
mg) , 16β, 17-dihydroxy- ent-kauran-19-oic acid (40 .0 mg) and
kaempferol (4 .5 mg) . Fraction E gave quercitrin (1 . 7 mg) , 9 ,
16 , 17-trihydroxy-19-kauranoic acid (9 . 2 mg) and daucosterol
(200 .0 mg) . Fraction F (60 g) was subjected to silica gel CC
and elutedwith amixtureof CHCl3 -MeOH (100% CHCl3 , 9∶1 ,
8∶2 , 7∶3 , 6∶4 , 100% MeOH ) to afford five subfractions F1-
F5 . 20-Hydroxyecdyson 20 , 22-acetonate ( 55.0 mg) was ob-
tained fromF2 by silica gel CC repeatedly and then followed by
preparation HPLC . Quercetin 3-robinobioside ( 12 .0 mg) and
nicotiflorin (120 mg) were yielded fromF3 by silica gel CC and
Sephadex LH-20 .
dipterinoid A ( 1 , 16β-hydroxy-17- [ ( Z) -p-coumaroyl ]-
ent-kauran-19-oic acid) : white amorphous solid; [α] 25. 8D = -
58 .06 ( c= 0 .155 , MeOH) ; UVλMeOHmax ( logε) : 311 (4 .07) ;
IRνKBrmax cm- 1 : 3423 , 2941 , 2871 , 2852 , 1696 , 1605 , 1514 ,
1164; negative FAB-MS m?z ( % ) : 481 [M-H] - ( 100) ; HR-
ESIMS m?z: 481 .2583 [ M-H ] - ( calcd for C29 H37 O6 ,
481 .2590) ; NMR data see Table 1 .
Acknowledgements : The authors aregrateful to the members of
the Analytical Group in State Key Laboratory of Phytochemistry
and Plant Resources in West China, Kunming Institute of Bota-
ny, for the spectral measurements .
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