Arylnaphthalene lignan glycosides and other constituents from Phyllanthus reticulatus

Article abstract of DOI:10.1080/10286020.2012.712040

Two new arylnaphthalene lignan glycosides, named reticulatusides A (1) and B (2), together with eight known compounds were isolated from the 95% EtOH extract of the whole plant of Phyllanthus reticulatus. The structures of the new compounds were elucidated by spectroscopic methods.

Full text of DOI:10.1080/10286020.2012.712040

This article was downloaded by: [University of Tennessee, Knoxville]
On: 24 August 2013, At: 23:19
Publisher: Taylor & Francis
Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered
office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Asian Natural Products
Research
Publication details, including instructions for authors and
subscription information:
http://www.tandfonline.com/loi/ganp20
Arylnaphthalene lignan glycosides and
other constituents from Phyllanthus
reticulatus
a b c
c
a
a
Jian-Xiong Ma
, Ming-Sheng Lan , Shi-Jin Qu , Jun-Jie Tan
a
a
a
,
a
Hong-Feng Luo , Chang-Heng Tan & Da-Yuan Zhu
Department of Natural Medicinal Chemistry, Shanghai Institute of
Materia Medica, Chinese Academy of Sciences, Shanghai, 201203,
China
b
Guangxi Medical University, Nanning, 530021, China
c
Guangxi Institute of Medicinal Plant, Nanning, 530023, China
Published online: 28 Aug 2012.
To cite this article: Jian-Xiong Ma , Ming-Sheng Lan , Shi-Jin Qu , Jun-Jie Tan , Hong-Feng Luo ,
Chang-Heng Tan & Da-Yuan Zhu (2012) Arylnaphthalene lignan glycosides and other constituents
from Phyllanthus reticulatus , Journal of Asian Natural Products Research, 14:11, 1073-1077, DOI:
10.1080/10286020.2012.712040
To link to this article: http://dx.doi.org/10.1080/10286020.2012.712040
PLEASE SCROLL DOWN FOR ARTICLE
Taylor & Francis makes every effort to ensure the accuracy of all the information (the
“
Content”) contained in the publications on our platform. However, Taylor & Francis,
our agents, and our licensors make no representations or warranties whatsoever as to
the accuracy, completeness, or suitability for any purpose of the Content. Any opinions
and views expressed in this publication are the opinions and views of the authors,
and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content
should not be relied upon and should be independently verified with primary sources
of information. Taylor and Francis shall not be liable for any losses, actions, claims,
proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or
howsoever caused arising directly or indirectly in connection with, in relation to or arising
out of the use of the Content.
This article may be used for research, teaching, and private study purposes. Any
substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing,
systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &

Conditions of access and use can be found at http://www.tandfonline.com/page/terms-
and-conditions

Journal of Asian Natural Products Research
Vol. 14, No. 11, November 2012, 1073–1077
Arylnaphthalene lignan glycosides and other constituents from
Phyllanthus reticulatus
abc1
Jian-Xiong Ma , Ming-Sheng Lan , Shi-Jin Qu , Jun-Jie Tan , Hong-Feng Luo ,
c1
a
a
a
a
a
Chang-Heng Tan * and Da-Yuan Zhu
Department of Natural Medicinal Chemistry, Shanghai Institute of Materia Medica, Chinese
a
b
Academy of Sciences, Shanghai 201203, China; Guangxi Medical University, Nanning 530021,
c
China; Guangxi Institute of Medicinal Plant, Nanning 530023, China
(Received 8 May 2012; final version received 11 July 2012)
Two new arylnaphthalene lignan glycosides, named reticulatusides A (1) and B (2),
together with eight known compounds were isolated from the 95% EtOH extract of the
whole plant of Phyllanthus reticulatus. The structures of the new compounds were
elucidated by spectroscopic methods.
Keywords: reticulatuside A; reticulatuside B; arylnaphthalene lignan glycoside;
Phyllanthus reticulatus
1
.
Introduction
and C [10], (þ)-lyoniresinol [11],
3S,5R,6S,9R)-megastigman-3,9-diol 3-O-
(
Phyllanthus reticulatus Poir. (Euphorbia-
ceae) is a climbing shrub distributed in
tropic area. In the Guangxi Zhuang national
area of China, it is used as a folk medicine
for the treatment of inflammation and
rheumatism [1]. Pharmacological studies
on the extracts of this plant showed
antiplasmodial [2], antidiabetic [3], anti-
microbial [4], cytotoxic [4], and hepato-
protective [5] bioactivities. Several
phytochemical investigations on this plant
revealed various constituents such as
triterpenoids, phytosterols, coumarin [4],
flavonoids, and other phenols [6]. As a
continuous interest in Zhuang national
medicines [7], we investigated the 95%
EtOH extract of the title plant, which led to
the isolation of two arylnaphthalene lignan
glycosides, reticulatusides A (1) and B (2)
a-L-arabinofuranosyl-(1 ! 6)-b-D-gluco-
pyranoside, and 7-megastigmen-3-ol-9-
one 3-O-a-L-arabinofuranosyl-(1 ! 6)-b-
D-glucopyranoside [12]. The above-men-
tioned compounds were reported from this
plant for the first time. In this paper, we
reported the isolation and structure eluci-
dation of 1 and 2.
2. Results and discussion
Compound 1 showed the quasi-molecular
þ
ion peaks at m/z 697 [M þ Na] , 1371
þ
2
[
2M þ Na] , and 719 [M þ HCOO] in
the positive and negative ESI-MS, respect-
ively, suggesting the molecular formula
C H O , which was further confirmed
3
2 34 16
by the HR-ESI-MS. Two ESI-MS fragment
þ
peaks at m/z 543 [M þ H 2 132] and 381
þ
(
Figure 1), together with eight known
compounds, spruceanol [8], cleistanthol
8], syringaresinol [9], phyllanthusmins B
[M þ H 2 132 2 162] indicated the
existence of one pentose and one hexose
in compound 1. The sugar moiety of 1 was
[
*Corresponding author. Email: chtan@mail.shcnc.ac.cn
ISSN 1028-6020 print/ISSN 1477-2213 online
q 2012 Taylor & Francis
http://dx.doi.org/10.1080/10286020.2012.712040
http://www.tandfonline.com

1
074
J.-X. Ma et al.
established to be a-L-arabinofuranosyl-
OR2
7
8
9
(1 ! 6)-b-D-glucopyranosyl based on the
1
ing NMR data: (i) H signals at d 4.75 (1H,
R O
5
4
1
1
acid hydrolysis experiment and the follow-
O
9'
8'
'
MeO
7
1
'
O
d, J ¼ 7.7 Hz) and 4.86 (1H, overlapped by
3
'
H O peak, HSQC cross-peak indicated as a
2
1
3
4
'
O
br s); (ii) C signals at d 107.1 (d), 78.4 (d),
77.2 (d), 75.7 (d), 72.3 (d), and 69.1 (t), as
well as 110.3 (d), 85.9 (d), 83.7 (d), 78.9
(d), and 63.0 (t); (iii) HMBC correlations
O
1
0'
1
2
R = Me, R = Glu (6 1) Ara(f)
1 2
R = Ara (p), R = Glu
1
2
0
00
00
between H-1 (d 4.86) and C-6 (d 69.1).
C
H
Figure 1. Structures of compounds 1 and 2.
1
The H NMR spectrum (Table 1) of the
1 13
Table 1. H and C NMR spectral data of 1 and 2 (400 and 100 MHz, in CD
3
OD).
1
2
1
3
1
13
1
H
Site
C
H
C
1
2
3
4
5
6
7
8
9
132.2
129.1
107.35/107.32
152.0
153.7
103.03/103.02
146.6
132.80/132.71
69.6
128.3
130.2
108.0
151.9
151.5
106.8
146.2
132.0
69.6
a
a
7.05/7.04 s
a
7.13/7.12 s
a
a
8.14/8.13 s
8.25 s
a
a
5.81/5.80 d (15.0)
a
5.62/5.61 d (15.0)
a
.41/5.40 d (15.0)
a
5.464/5.459 d (15.0)
5
0
a
1
130.39/130.35
112.2/112.0
149.3
149.3
109.31/109.24
130.2
112.0
149.3
149.3
109.3
125.0
137.8
120.9
172.1
102.6
0
a
a
6.83/6.77 br s
a
6.81/6.80 d (1.5)
2
0
3
0
4
0
a
5
0
6.92 d 7.7
6.79/6.76 br d (7.7)
6.97 d (7.9)
6.77 dd (7.9, 1.5)
a
a
6
0
125.1/124.9
7
0
137.9
120.5
172.6
102.6
57.0
8
0
9
0
a
6.03/6.02 s
1
0
6.04/6.06 s
MeO-5
MeO-4
Glu-1
4.00 s
3.70 s
4.75 d (7.7)
3.75 dd (9.1, 7.7)
3.48 t (9.1)
3.36 t-like (9.4)
3.49 m
3.61 dd (10.6, 2.9)
.03 dd (10.6, 1.6)
4.86 br s (overlapped)
3.94 br s
3.80 dd (6.3, 2.9)
3.78 m
3.57 dd (11.9, 4.2)
56.3
56.5
107.3
75.1
78.3
71.7
78.2
62.9
3.75 s
4.77 d (7.6)
0
0
107.1
75.7
78.4
0
0
0
0
0
0
0
0
0
0
2
3
4
5
6
3.59 t-like (7.6)
3.69 dd (9.0, 7.6)
3.40 t-like (9.0)
3.72 ddd (9.0, 3.4, 2.8)
3.70 dd (12.0, 3.4)
3.97 dd (12.0, 2.8)
5.29 d (7.2)
3.98 dd (9.2, 7.2)
3.64 dd (9.2, 3.5)
3.86 br s
3.92 dd (12.5, 2.2)
3.51 br d (12.5)
a
72.34/72.38
77.24/77.21
69.13/69.09
a
a
4
0
00
a
Ara-1
110.29/110.26
83.78/83.74
101.5
73.0
74.5
70.0
67.9
0
0
0
0
00
00
00
00
a
2
3
4
5
78.9
85.9
63.0
3
.55 dd (11.9, 2.8)
a
Splitted signals caused by atropisomerism.

Journal of Asian Natural Products Research
1075
HOH C
2
OH
O
O
HO
HO
HO
OH
OH
O
HO
OH
O
O
O
HO
HO
O
OH
O
MeO
OH
O
O
MeO
MeO
O
O
O
O
O
O
1
2
Figure 2. Selected HMBC correlations (H to C) of 1 with 2.
aglycone of 1 showed signals for one 1,3,4-
trisubstituted aromatic ring at d 6.92 (1H, d,
J ¼ 7.7 Hz), 6.83 (1H, br s), and 6.79 (1H,
br d, J ¼ 7.7 Hz); two isolated olefinic
protons at d 8.14 and 7.05 (each 1H, s); one
dioxymethylene at d 6.03 (2H, s); two
methoxyls at d 4.00 and 3.70 (each 3H, s);
and an isolated oxygenated methylene at dH
the slow rotation of the sugar unit at
room temperature (rt) around the glucosi-
dic linkage with the naphthalide ring
system.
Reticulatuside B (2) was obtained as
yellowish amorphous powder. Its molecu-
lar formula was determined to be
C H O by the HR-ESI-MS. After acid
31
32 16
5
.81 and 5.41 (each d, J ¼ 15.0 Hz). The
hydrolysis, 2 gave D-glucose and L-
1
3
1
arabinose as sugar moiety. The H and
13
C NMR spectra (Table 1) of 2 displayed
C NMR spectrum (Table 1) of 1 displayed
2
1 carbons (12 sp C, 5 sp CH, 2 OMe, 1
2
2
OCH O, and 1 CH ) for the aglycone
signals for an arylnaphthalene skeleton
which was nearly superposed with that of 1.
Two sugar units were identified to be b-
glucopyranosyl and a-arabinopyranosyl on
2
2
moiety of 1. The above evidences indicated
1
to be an arylnaphthalene glycoside [13].
In the HMBC spectrum of 1 (Figure 2),
obvious cross-peaks of MeO (d 3.70)/C-4
H
the basis of NMR data at d 4.77 (1H, d,
H
(
d 152.0), MeO (d 4.00)/C-5 (d 153.7),
C
J ¼ 7.6 Hz) and d 107.3 (d), 78.3 (d), 78.2
H
C
C
0
0
and H-1 (dH 4.75)/C-7 (dC 146.6)
were observed, demonstrating that the
two methoxyls and the sugar linkage
were on C-4, C-5, and C-7 of the
arylnathalene skeleton, respectively. Fur-
thermore, HMBC correlations between
(d), 75.1(d), 71.7 (d), and 62.9 (t), as well as
at d 5.29 (1H, d, J ¼ 7.2 Hz) and d 101.5
H
C
(d), 74.5 (d), 73.0 (d), 70.0 (d), and 67.9 (t)
1
13
[14,15]. Comparing the H and C NMR
spectral data of 2 with those of 1, the most
important differences were that 2 lacked
signals for the second methoxyl, and 2 had a
terminal glucosyl (dC-600: 62.9 of 2 vs. 69.1
0
0
0
H -10 /C-3 and C-4 , and H -9/C-7, C-8,
2
2
0
and C-9 , substantiated further the struc-
0
0
ture 3 ,4 -methylenedioxy-4,5-dimethoxy-
0
of 1) and a pyran-configuration arabinosyl.
The structure of reticulatuside B was
0
0
7
-hydroxy-2,7 -cycloligna-7,7 -dien-9 ,9-
0
0
olide 7-O-a-L-arabinofuranosyl(1 ! 6)-
finally elucidated to be 3 ,4 -methylene-
dioxy-4-methoxy-5-a-L-arabinofuranosy-
b-D-glucopyranoside for reticulatuside A
(
0
1). As indicated by Al-Abed et al. [13],
loxy-7-b-D-glucopyranosyloxy-2,7 -cyclo-
0 0
1
13
some H and C signals of 1 exist in pairs
due to the equilibrium between two
conformational isomers resulting from
ligna-7,7 -dien-9 ,9-olide (2) by the HMBC
0
0
correlations (Figure 2) between H-1 /C-7,
0
00
H-1 /C-5, and MeO/C-4.

1
076
J.-X. Ma et al.
The EtOAc fraction (15 g) was subjected
3
.
Experimental
to silica gel CC with gradient CHCl3–
3
.1 General experimental procedures
MeOH (100:1 ! 5:1, v/v) as eluant to
Optical rotations were determined on a
PerkinElmer 341 polarimeter (Perkin
Elmer, Inc., Wellesley, MA, USA). UV
spectra were obtained on a Shimadzu UV-
gain subfractions E1–E6. Fr. E2 (1.0 g)
was purified by silica gel CC (CHCl3–
MeOH, 30:1), following Sephadex LH-
20 CC (CHCl –CH OH, 1:1) to achieve
3 3
2
550 spectrophotometer (Shimadzu Corp.,
spruceanol (20 mg) and syringaresinol
9 mg). By the similar procedure, cleis-
tanthol (18 mg) and phyllanthusmin B
15 mg) were obtained from Fr. E4. Fr.
Kyoto, Japan). IR spectra were recorded on
a Thermo Nicolet Magna-IR 750 FTIR
spectrometer (Nicolet Instrument, Inc.,
Madison, WI, USA) with KBr tablet.
(
(
E5 was separated into Frs E5.1–5.4 by
silica gel CC (CHCl –MeOH, 10:1).
1
13
H (400 MHz) and C NMR (100 MHz)
3
spectra were acquired on a Bruker AM-400
spectrometer with tetramethylsilane (TMS)
as an internal standard. ESI-MS and HR-
ESI-MS data were measured on a Bruker
Esquire 3000plus and a Finnigan LCQ
DECA mass spectrometers, respectively.
Silica gel (200–300 meshes, Qingdao
Haiyang Chemical Group Co., Qingdao,
China), Sephadex LH-20 (Pharmacia Bio-
tech AB, Uppsala, Sweden), and ODS-A
gel (Mitsubishi Chemical Industries Co.,
Ltd., Tokyo, Japan) were used for column
chromatography (CC), and precoated
plates of silica gel (HSGF254; Qingdao
Haiyang Chemical Group Co., Qingdao,
China) were used for TLC.
Phyllanthusmin C (8 mg) and (þ)-
lyoniresinol (17 mg) were isolated from
Frs E5.2 and E5.3, respectively, after the
purification of Sephadex LH-20 CC
(MeOH). The n-BuOH fraction (100 g)
was subjected to silica gel CC (CHCl3–
MeOH, 20:1, 10:1, 6:1, 3:1, finally
MeOH) to afford Frs B1 2 B5. Fr. B3
(
CC (CHCl –MeOH–H O, 5:1:0.1) to
10.0 g) was further isolated by silica gel
3
2
get Frs B3.1 2 3.5. Fr. B3.3 furnished 1
9 mg) and 7-megastigmen-3-ol-9-one
(
3-O-a-L-arabinofuranosyl-(1 ! 6)-b-D-
glucopyranoside (9 mg) after purification
by ODS-A gel CC (MeOH–H O, 30–
2
70%) and Sephadex LH-20 CC (MeOH).
By the same procedure, Fr. B3.4
afforded 2 (12 mg) and (3S,5R,6S,9R)-
megastigman-3,9-diol 3-O-a-L-arabino-
furanosyl-(1 ! 6)-b-D-glucopyranoside
3
.2 Plant material
The fresh whole plants of P. reticulatus
were collected in Yongning County,
Guangxi Province, China. The plant was
identified by Prof. D.-Y. Zhu of Shanghai
Institute of Materia Medica. A voucher
specimen (No. 09-108) is deposited in
the Herbarium of Shanghai Institute of
Materia Medica.
(6 mg).
3.3.1 Reticulatuside A (1)
2
5
Yellowish amorphous powder; ½aꢀ 2 82
D
(
c ¼ 0.065, MeOH). UV l
(MeOH) nm
log1): 263 (4.69). IR (KBr) nmax: 3423,
max
(
2
1
923, 1734, 1623, 1508, 1434, 1265, 1230,
3
.3 Extraction and isolation
21
1
170, 1074, 1039, and771 cm . For H and
1
3
The air-dried and powdered whole plants
of P. reticulates (5 kg) were extracted
three times with 20 liters of 95% EtOH
at rt. The concentrated extract was
CNMRspectraldata, seeTable1.ESI-MS
þ
m/z: (positive ion model) 697 [M þ Na] ,
þ
þ
1371 [2M þ Na] , 543 [M þ H 2 132] ,
2
þ
and381[M þ H 2 132 2 162] ;(negative
2
[2M 2 H] . HR-ESI-MS m/z: 697.1748
partitioned between H O and petroleum
2
ion model) 719 [M þ HCOO] and 1347
ether, EtOAc and n-BuOH, respectively.

Journal of Asian Natural Products Research
1077
þ
[
M þ Na] (calcd for C H O Na,
Project ‘Key New Drug Creation and Manu-
facturing Program’ (No. 2011ZX09307-002-
03) and the National Natural Science Foun-
dation of China (No. 81072545).
3
2
34 16
6
97.1745).
3
.3.2 Reticulatuside B (2)
2
D
5
Yellowish amorphous powder, ½aꢀ 2 22
Note
(
(
c ¼ 0.045, MeOH). UV l
(MeOH) nm
log 1): 259 (4.69). IR (KBr) nmax: 3415,
1. These authors contributed equally to this
work.
max
2
1
921, 1749, 1618, 1508, 1384, 1257, 1074,
2
1
1
13
037, and 775 cm . For H and C NMR
References
spectral data, see Table 1. ESI-MS m/z:
þ
[
1] The Health Bureau of Guangxi Province
(Ed.), Compilation of Medicinal Herbs in
Guangxi (II) (Guangxi People Press,
Nanning, 1974), p. 1588.
(
positive ion model) 661 [M þ H] , 683
þ
þ
[
M þ Na] , 1343 [2M þ Na] ; (negative
2
ion model) 705 [M þ HCOO] , 1319
2
2M 2 H] . HR-ESI-MS m/z: 683.1586
[2] E. Omulokoli, B. Khan, and C. Chhabra,
J. Ethnopharmacol. 56, 133 (1997).
[3] S. Kumar, D. Kumar, R.R. Deshmukh,
P.D. Lokhande, S.N. More, and
V.D. Rangari, Fitoterapia 79, 21 (2008).
[
þ
[
M þ Na] (calcd for C H O Na,
3
1
32 16
6
83.1588).
[
4] T. Begum, M.S. Rahman, and M.A.
Rashid, Dhaka Univ. J. Pharm. Sci. 5,
3
.4 Analysis of the sugar moiety
2
1 (2006).
Compounds 1 and 2 (1 mg each) were
refluxed in 2 N HCl–dioxane (1:1 v/v,
[
5] B.K. Das, S. Bepary, B.K. Datta, A.A.
Chowdhury, M.S. Ali, and S. Rouf, Pak.
J. Pharm. Sci. 21, 333 (2008).
[6] S.H. Lam, C.Y. Wang, C.K. Chen, and
S.S. Lee, Phytochem. Anal. 18, 251
2
neutralized with NaHCO . After extrac-
ml) for 2 h. On cooling, the mixture was
3
tion with AcOEt, the aqueous layer was
concentrated by blowing with N . The
(
2007).
2
[
7] M.S. Lan, J.X. Ma, C.H. Tan, S. Wei, and
D.Y. Zhu, Helv. Chim. Acta 93, 2276
(2010).
[8] H.C. Krebs, H. Duddeck, S. Malik,
W. Beil, P. Rasoanaivo, and M. Andria-
narijaona, Z. Naturforsch. B 59, 58
residue was purified by CC of Sephadex
LH-20 (MeOH–H O ¼ 1:1, v/v) to give
2
the sugar moiety. The purified sugar and
standard D-glucose and L-arabinose
(
Sigma, St. Louis, MO, USA) were treated
with L-cysteine methyl ester hydrochloride
2 mg) in pyridine (1 ml) at 608C for 1 h.
(
2004).
[
9] Y. Nakasone, K. Takara, K. Wada,
J. Tanaka, S. Yogi, and N. Nakatani,
Biosci. Biotech. Biochem. 60, 1714
(
Then, the solution was treated with N,O-
bis(trimethylsilyl)trifluoroacetamide
(
1996).
10] S.J. Wu and T.S. Wu, Chem. Pharm. Bull.
4, 1223 (2006).
[
[
(0.02 ml) at 608C for 1 h. The supernatant
was applied to gas chromatography (GC)
5
11] H. Shibuya, Y. Takeda, R.S. Zhang,
A. Tanitame, Y.L. Tsai, and I. Kitagawa,
Chem. Pharm. Bull. 40, 2639 (1992).
12] G.P. Li, J.F. Zhao, Y.Q. Tu, X.D. Yang,
H.B. Zhang, and L. Li, J. Integr. Plant
Biol. 47, 362 (2005).
13] Y. Al-Abed, S. Sabri, M. Abu Zarga,
Z. Shah, and Atta-Ur-Rahman, J. Nat.
Prod. 53, 1152 (1990).
analysis (Supelco; 2308C, N ). D-Glucose
2
and L-arabinose were detected from 1 and
2
[
[
by comparing its retention time with
those of the authentic samples (t ¼ 16.8
R
and 10.2 min for D-glucose and L-arabi-
nose, respectively).
[
[
14] J.M. Tian, X.J. Hao, and H.P. He, Helv.
Chim. Acta 89, 291 (2006).
15] J.M. Tian, H.P. He, Y.T. Di, X.W. Yang,
Z.L. Gao, and X.J. Hao, J. Asian Nat.
Prod. Res. 10, 239 (2008).
Acknowledgments
This study was financially supported by grants
from the National Science & Technology Major