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Journal of Medicinal Plants Research Vol. 6(11), pp.

2200-2205, 23 March, 2012


Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR11.1776
ISSN 1996-0875 2012 Academic Journals

Full Length Research Paper

The isolation and structure identification of four


Lignans stereoisomers from Uncaria sinensis (Oliv. )
Havil.
Guangli SUN1.2, Xiaopo ZHANG1, Xudong XU1*, Junshan YANG1, Lixun LV2 andMingliang
ZHONG1,2
1
Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education,
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College,
100193, Beijing, China.
2
Hebei United University, 063000, Hebei Tangshan, China.
Accepted 3 February, 2012

The study of the chemical constituents from Uncaria sinensis (oliv.) Havil. has resulted in the isolation
and identification of four stereosturcture of lignans, which were identified as: (2R,3R,4S)-lyoniresinol-
3-O--D-glucopyranoside (1); (2S,3S,4R)-lyoniresinol-3-O--D-glucopyranoside (2); (2S,3R,4S)-
lyoniresinol-3-O--D-glucopyranoside (3); (2R,3S,4R)-lyoniresinol-3-O--D-glucopyranoside (4). All
these four compounds were obtained from the Uncaria genus for the first time. The compounds were
individually identified by one-dimensional (1D), two-dimensional (2D) nuclear magnetic resonance
(NMR) and mass spectrometry (MS) spectral analyses and comparisons with reported data, and their
absolute stereosturcture were identified by the helicity rule of exciton coupled circular dichroism for
the first time. The study also constitutes a method of the identification of absolute stereosturcture by
the helicity rule of exciton coupled circular dichroism for the first time.

Key words: Uncaria sinensis (oliv.) Havil., Rubiaceae, natural products, lignans, the helicity rule.

INTRODUCTION

The Uncaria genus in Rubiaceae comprises more than absolute configuration with the method of the helicity rule
30 species widely distributed in tropical regions, including of exciton coupled circular dichroism. In this paper, four
Asia, Africa, South America (Risdale, 1978). 12 species stereoisomers of lignans (1-4) were isolated and
of them are distributed in China, which have been found identified.
widespread use in traditional medicine. Uncaria sinensis
(Oliv.) Havil. has been used for the treatment of
hypertension, headache and fever in Chinese EXPERIMENTAL
Pharmacopoeia, and found growing in Sichuan, Guangxi,
Collection and preparation of plant material
Yunnan, Hubei, Guizhou, Hunan, Shanxi, Gansu
Province, etc., in China (Luo, 1999; State Pharmacopeia The medicinal material was collected at Province Guangxi in 2007,
Committee of China, 2010). The isolation of alkaloids was and identified by Dr. Jing Quan Yuan at the Institute of Medicinal
brought into focus in previous study of the Uncaria genus; Plant Development, Chinese Academy of Medical Sciences and
the previous study also led to isolation of some flavonoids Peking Union Medical College. A voucher specimen has been
and triterpenies (Heitzman et al., 2005; Laus, 2004). deposited there (Voucher specimen #20070026).
However, few information is available on lignans of U.
sinensis, and no information about the identification of
Extraction and isolation

The dried and powdered bard and branch with curve


*Corresponding author. E-mail: xdxu@implad.ac.cn. hooks of the plant (8.5 kg) of U. sinensis were
Sun et al. 2201

Figure 1. Compounds 1-4 isolated from U. sinensis and significant NOESY correlations ().

exhaustively extracted with 70% ethanol. The extract was dimensional (2D) nuclear magnetic resonance (NMR)
filtered and concentrated on reduced pressure until only and mass spectrometry (MS) spectral data compared
H2O remained. The remaining solution was sequentially with the data reported (Lee, 2005). And the absolute
partitioned with petroleum ether, CHCl3, EtOAc and n- stereosturcture were identified by nuclear overhauser
BuOH. The n-BuOH partion was repeatedly subjected to effect spectroscopy (NOESY) and the helicity rule of
column chromatography on silica gel, MCI gel, and exciton coupled circular dichroism (Nakanishi et al.,
reversed phase C18 gel, to yield Compounds 1 to 4 1997).
(Figure 1). Compoud 1: A white amorphous solid, ESI-MS (m/z):
+ 1
583[M+H] . Formula, C28H38O13. HNMR (600 MHz,
MeOH) : 1.71 (1H, m, H-2), 2.09 (1H, m, H-3), 2.62 (1H,
RESULTS AND DISCUSSION m, Ha-1), 2.72 (1H, dd, J=4.2, 15.0 Hz, Hb-1), 3.35 (3H, s,
5-OCH3), 3.54 (1H, m, Ha-6''), 3.65 (1H, m, Hb-6''), 3.75
Planar structure elucidation (6H, s, 3'-OCH3, 5'-OCH3), 3.86 (3H, s, 7-OCH3), 4.28
(1H, d, J=7.8Hz, H-1''), 4.42 (1H, d, J=6.0Hz, H-4), 6.43
13
These four compounds (1 to 4) were stereoisomers of (2H, s, H-2',6'), 6.58 (1H, s, H-8). CNMR (150 MHz,
lignans. Their sturcture were identified as (2R, 3R, 4S)- MeOH) : 34.0 (C-1), 41.4 (C-2), 46.8 (C-3), 43.4 (C-4),
lyoniresinol-3-O--D-glucopyranoside (1), (2S, 3S, 4R)- 147.7 (C-5), 139.1 (C-6), 148.9 (C-7), 108.0 (C-8), 126.4
lyoniresinol-3-O--D-glucopyranoside (2), (2S, 3R, 4S)- (C-9), 130.4 (C-10), 66.4 (C-2), 72.2 (C-3), 134.8 (C-
lyoniresinol-3-O--D-glucopyranoside (3), (2R, 3S, 4R)- 1'), 107.3 (C-2'), 149.2 (C-3'), 139.6 (C-4'), 149.2 (C-5'),
lyoniresinol-3-O--D-glucopyranoside (4). Their planar 107.3 (C-6'), 104.1 (C-1''), 75.3 (C-2''), 78.2 (C-3''), 71.7
structure were identified by one-dimensional (1D), two- (C-4''), 78.4 (C-5''), 62.9 (C-6''), 60.3 (5-OCH3), 56.8 (7-
2202 J. Med. Plants Res.

Compoud 4: A white amorphous solid, ESI-MS (m/z):


+ 1
583[M+H] . Formula, C28H38O13. HNMR (600 MHz,
MeOH) : 1.68 (1H, m, H-2), 2.13 (1H, m, H-3), 2.67 (1H,
m, Ha-1), 2.68 (1H, m, Hb-1), 3.35 (3H, s, 5-OCH3), 3.54
(1H, m, Ha-6''), 3.65 (1H, m, Hb-6''), 3.75 (6H, s, 3'-OCH3,
5'-OCH3), 3.85 (3H, s, 7-OCH3), 4.13 (1H, d, J=7.8 Hz, H-
1''), 4.22 (1H, d, J=6.0Hz, H-4), 6.41 (2H, s, H-2',6'), 6.58
13
(1H, s, H-8). CNMR (150 MHz, MeOH) : 34.0 (C-1),
41.4 (C-2), 46.8 (C-3), 43.4 (C-4), 147.7 (C-5), 139.1 (C-
6), 148.9 (C-7), 108.0 (C-8), 126.4 (C-9), 130.4 (C-10),
66.4 (C-2), 72.2 (C-3), 134.8 (C-1'), 107.3 (C-2'), 149.2
(C-3'), 139.6 (C-4'), 149.2 (C-5'), 107.3 (C-6'), 104.4 (C-
1''), 75.3 (C-2''), 78.2 (C-3''), 71.7 (C-4''), 78.4 (C-5''), 62.9
(C-6''), 60.3 (5-OCH3), 56.8 (7-OCH3), 57.1 (3-OCH3),
57.1 (5-OCH3). The aforementioned data are consistent
Figure 2. Planar structure of compounds 1-4 with the reported data (Lee, 2005) of lyoniresinol-3-O--
D- glucopyranoside (Figure 2).

OCH3), 57.1 (3- OCH3), 57.1 (5- OCH3). The


aforementioned data are in agreement with the reported Stereostructure elucidation
data (Lee, 2005) of lyoniresinol-3-O--D-
glucopyranoside (Figure 2). Compound 2: A white In the NOESY experiments, the correlations between H-2
+
amorphous solid, ESI-MS (m/z): 583[M+H] . Formula, and H-4, between H-2, 6 and H-3 (Figure 1) on
1
C28H38O13. HNMR (600 MHz, MeOH) : 1.71 (1H, m, H- Compouds 1 and 2, between H-2 and H-3, between H-2,
2), 2.09 (1H, m, H-3), 2.62 (1H, m, Ha-1), 2.72 (1H, dd, 6 and H-3 (Figure 1) on compouds 3 and 4, indicated
J=4.2, 15.0 Hz, Hb-1), 3.35 (3H, s, 5-OCH3), 3.54 (1H, m, that the relative configuration of the aglycone of
Ha-6''), 3.65 (1H, m, Hb-6''), 3.75 (6H, s, 3'-OCH3, 5'- Compound 1 was the same as the aglycone of
OCH3), 3.86 (3H, s, 7-OCH3), 4.28 (1H, d, J=7.8 Hz, H- Compound 2, the relative configuration of the aglycone of
1''), 4.42 (1H, d, J=6.0 Hz, H-4), 6.43 (2H, s, H-2',6'), 6.58 Compound 3 was the same as the aglycone of
13
(1H, s, H-8). CNMR (150 MHz, MeOH) : 34.0 (C-1), Compound 4.
41.4 (C-2), 46.8 (C-3), 43.4 (C-4), 147.7 (C-5), 139.1 (C- The absolute configurations were determined by
6), 148.9 (C-7), 108.0 (C-8), 126.4 (C-9), 130.4 (C-10), examinations of the circular cichroism (CD) spectra of
66.4 (C-2), 72.2 (C-3), 134.8 (C-1'), 107.3 (C-2'), 149.2 Compounds 1 to 4. The CD curves of the four
(C-3'), 139.6 (C-4'), 149.2 (C-5'), 107.3 (C-6'), 104.4 (C- compounds should be spiltted into two parts, for the two
1''), 75.3 (C-2''), 78.2 (C-3''), 71.7 (C-4''), 78.4 (C-5''), 62.9 chromophores (two benzene rings) of -* transition of
(C-6''), 60.3 (5-OCH3), 56.8 (7- OCH3), 57.1 (3-OCH3), each of the four compounds. The CD curve at 244 nm
57.1 (5-OCH3). The aforementioned data are in (+11.5473) and 224 nm (-2.45488) of Compound 1
agreement with the reported data (Lee, 2005) of indicated the CD spectrum was a positive chiral spectrum
lyoniresinol-3-O--D- glucopyranoside (Figure 2). (Figure 3) (Nakanishi et al., 1997). According to the
Compoud 3: A white amorphous solid, ESI-MS (m/z): helicity rule of exciton coupled circular dichroism, the
+ 1
583[M+H] . Formula, C28H38O13. HNMR (600 MHz, exciton coupling of the two identical chromophores with
MeOH) : 1.71 (1H, m, H-2), 2.08 (1H, m, H-3), 2.62 (1H, positive chirality when Compound 1 was accepted as the
m, Ha-1), 2.70 (1H, dd, J=4.2, 15.0 Hz, Hb-1), 3.35 (3H, s, stereochemistry of C4-S (Nakanishi et al., 1997).
5-OCH3), 3.54 (1H, m, Ha-6''), 3.65 (1H, m, Hb-6''), 3.74 Consequently, Compound 1 was determined to be (2R,
(6H, s, 3'-OCH3, 5'-OCH3), 3.85 (3H, s, 7-OCH3), 4.28 3R, 4S)-lyoniresinol-3-O--D-glucopyranoside. By the
(1H, d, J=7.8 Hz, H-1''), 4.42 (1H, d, J=6.0Hz, H-4), 6.43 same rule, Compound 2 was determined to be (2S, 3S,
13
(2H, s, H-2',6'), 6.58 (1H, s, H-8). CNMR (150 MHz, 4R)-lyoniresinol-3-O--D-glucopyranoside, for the CD
MeOH) : 33.0 (C-1), 40.8 (C-2), 46.9 (C-3), 43.0 (C-4), curve of Compound 2 was a negative chiral spectrum at
147.8 (C-5), 139.1 (C-6), 148.8 (C-7), 108.0 (C-8), 126.6 224 nm (-5.52452) and 217 nm (+7.61588) (Figure 4).
(C-9), 130.4 (C-10), 66.4 (C-2OCH3), 71.7 (C-3OCH3), Compound 3 was determined to be (2S, 3S, 4R)-
134.7 (C-1'), 107.1 (C-2'), 149.2 (C-3'), 139.5 (C-4'), lyoniresinol-3-O--D-glucopyranoside, for the CD curve
149.2 (C-5'), 107.1 (C-6'), 105.0 (C-1''), 75.4 (C-2''), 78.1 of Compound 3 was a positive chiral spectrum at 244 nm
(C-3''), 71.8 (C-4''), 78.4 (C-5''), 63.0 (C-6''), 60.4 (5- (+0.689788) and 220 nm (-0.895488) (Figure 5).
OCH3), 56.8 (7-OCH3), 57.1 (3-OCH3), 57.1(5-OCH3). Compound 4 was determined to be (2S, 3S, 4R)-
The aforementioned data are in agreement with the lyoniresinol-3-O--D-glucopyranoside, for the CD curve
reported data (Lee, 2005) of lyoniresinol-3-O--D- of Compound 4 was a negative chiral spectrum at 243.5
glucopyranoside (Figure 2). nm (-1.11085) and 222 nm (+0.437642) (Figure 6).
Sun et al. 2203

Figure 3. CD spectrum of Compound 1.

Figure 4. CD spectrum of Compound 2.


2204 J. Med. Plants Res.

Figure 5. CD spectrum of Compound 3.

Figure 6. CD spectrum of Compound 4.


Sun et al. 2205

DISCUSSION REFERENCES

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convenience. The four lignans were isolated from both
Lyoniresinol-3-O--D-glucopyranoside isolated from the root bark of
Uncaria genus and U. sinensis for the first time. And the Lycium Chinense miller against human pathogenic microorganisms.
lignans also have the widely activities, such as anti- Arch. Pharm. Res., 28(9): 1031-1036.
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ACKNOWLEDGEMENTS

Financial support from Chinese Traditoinal Medicine


Researches of Special Projects (Grant No. 200707007);
the technological large platform for comprehensive
research and development of newdrugs in the Twelfth
Five-Year Significant New Drugs Created Science and
Technology Major Projects (No. 2012ZX09301-002-001-
026)the chemical composition of the digital library of
traditional Chinese medicine for drug discovery in the
Twelfth Five-Year Significant New Drugs Created (No.
2011ZX09307-002 -01) are gratefully acknowledged. We
are also grateful to Dr. Jing Quan Yuan (the Institute of
Medicinal Plant Development, Chinese Academy of
Medical Sciences and Peking Union Medical College) for
kindly providing help.