Two new glycosides from Viburnum plicatum Thunb. ex Murray var. plicatum f. plicatum

Article abstract of DOI:10.1007/s11418-010-0455-0

Two new glycosides, named plicatumoside A (1) and (+)-neomedioresinol 4,4′-di-O-β-d-glucopyranoside (2), together with 13 known compounds, were isolated from the leaves of Viburnum plicatum Thunb. ex Murray var. plicatum f. plicatum. Their structures were established on the basis of NMR, MS, and chemical data.

Full text of DOI:10.1007/s11418-010-0455-0

J Nat Med (2011) 65:202–205
DOI 10.1007/s11418-010-0455-0
NOTE
Two new glycosides from Viburnum plicatum Thunb.
ex Murray var. plicatum f. plicatum
•
•
•
Masao Kikuchi Rie Onoguchi Yasunori Yaoita
Koichi Machida
Received: 9 June 2010 / Accepted: 22 July 2010 / Published online: 22 September 2010
Ó The Japanese Society of Pharmacognosy and Springer 2010
Abstract Two new glycosides, named plicatumoside A
(1) and (?)-neomedioresinol 4,40-di-O-b-D-glucopyrano-
side (2), together with 13 known compounds, were isolated
from the leaves of Viburnum plicatum Thunb. ex Murray
var. plicatum f. plicatum. Their structures were established
on the basis of NMR, MS, and chemical data.
Results and discussion
The leaves of V. plicatum Thunb. ex Murray var. plicatum
f. plicatum were extracted with MeOH. The MeOH extract
was fractionated among CHCl3, AcOEt, n-BuOH, and
H2O. The n-BuOH- and H2O-soluble fractions were each
subjected to separation by a combination of chromato-
graphic procedures. From the n-BuOH-soluble fraction,
compounds 1 and 3–6 were isolated, while compounds 2
and 7–15 were isolated from the H2O-soluble fraction. The
known compounds 3–15 were identified as henryoside (3)
[2], 5-O-caffeoyl shikimic acid (4) [3], 1-O-caffeoyl
b-D-glucopyranoside (5) [4], chlorogenic acid (6) [5],
(?)-medioresinol 4,40-O-di-b-D-glucopyranoside (7) [6, 7],
(?)-pinoresinol 4,40-O-di-b-D-glucopyranoside (8) [7],
(?)-syringaresinol 4,40-O-di-b-D-glucopyranoside (9) [7],
kaempferol 3-O-rutinoside (10) [8], kaempferol 3-O-ro-
biobioside (11) [8], quercetin 3-O-rutinoside (12) [8],
quercetin 3-O-robiobioside (13) [8], secoxyloganin (14)
[9], and loganic acid (15) [10] by comparison of their
spectroscopic data with those previously described in the
literature (Fig. 1).
Keywords Viburnum plicatum ꢀ Caplifoliaceae ꢀ
Glycoside
Introduction
We have reported the isolation of three new glycosides and
seven known ones from the leaves of Viburnum plicatum
Thunb. var. tomentosum Miq. (Capriforiaceae), known as
´
´
`
the Hu die shu in Chinese herbal medicine [1]. It has been
used for lymphadenitis, ringworm, and infantile tantrum. In
a continuation of our investigation of the chemical con-
stituents from plants of the genus Viburnum species, we
have now examined the chemical constituents of the leaves
of V. plicatum Thunb. ex Murray var. plicatum f. plicatum.
As far as we know, there are no reports regarding the
chemical constituents of this plant. This paper describes the
structural elucidation and identification of 2 new glyco-
sides, named plicatumoside A (1) and (?)-neomedioresinol
4,40-O-di-b-D-glucopyranoside (2), isolated along with 13
known compounds (3–15) from this plant.
Compound 1 was obtained as a hygroscopic amorphous
powder, [a]D -40.3° (MeOH). The molecular formula of 1
was determined as C26H32O15 on the basis of a pseudo
molecular ion peak at m/z 607.1636 [M ? Na]? in the
1
high-resolution (HR)-FAB-MS. The H-NMR spectrum of
1 showed the signals due to one 1,2,6-trisubstituted aro-
matic ring at dH 6.56 (1H, br d, J = 8.4 Hz, H-5), 6.68
(1H, br d, J = 8.1 Hz, H-3), and 7.25 (1H, dd, J = 8.4,
8.1 Hz, H-4), one ortho-disubstituted aromatic ring at dH
7.40 (2H, br t, J = 7.7 Hz, H-40, H-50) and 7.51 (2H, dd,
J = 7.7, 1.5 Hz, H-30, H-60), one CH2 as an AB system at
dH 5.37 (1H, d, J = 13.2 Hz, Ha-7) and 5.39 (1H, d,
J = 13.2 Hz, Hb-7), and two anomeric protons at dH 4.44
M. Kikuchi (&) ꢀ R. Onoguchi ꢀ Y. Yaoita ꢀ K. Machida
Tohoku Pharmaceutical University,
4-4-1 Komatsushima, Aoba-ku, Sendai,
Miyagi 981-8558, Japan
e-mail: mkikuchi@tohoku-pharm.ac.jp
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J Nat Med (2011) 65:202–205
203
means of chiral detection in HPLC analysis. The coupling
constants of the two anomeric protons indicated that the
glycosyl linkages are of b-configuration. These spectral
features, including the molecular formula, were very sim-
ilar to those of henryoside (3), except for the 13C-NMR
chemical shifts due to the ortho-disubstituted aromatic ring
and the CH2 as an AB system (Table 1). The chemical shift
variations noted in both 1 and 3 could be explained by the
absence of a b-D-glucopyranosyl unit at C-20 in 1. This
b-D-glucopyranosyl unit was determined to be involved in
a glycosyl linkage at C-200 of the other b-D-glucopyranosyl
group bonded to C-2, because the signal due to C-200 was
markedly downfield shifted at dC 83.4 (?8.6 ppm), when
comparing the 13C-NMR spectrum of 1 with that of 3. This
finding was supported by the HMBC experiment between
H-1000 and C-200 (Fig. 2). From the above data, the structure
of 1 was established as 2-hydroxybenzyl 2-(2-O-b-D-glu-
copyranosyl-b-D-glucopyranosyloxy)-6-hydroxybenzoate,
and named plicatumoside A.
Fig. 1 Structures of 1–15
Table 1 13C-NMR data for compounds 1 and 3 (150 MHz, CD3OD)
Position
1
3
Position
1
3
Compound 2 was obtained as colorless needles, [a]D
?82.0° (MeOH). The molecular formula of 2 was deter-
mined as C32H42O17 on the basis of a pseudo molecular ion
peak at m/z 699.2481 [M ? H]? in the HR-FAB-MS. In
the 1H-NMR spectrum of 2, two methoxyl groups [dH 3.84
(3H, s, OMe-5), 3.87 (3H, s, OMe-30)], two anomeric
proton signals [dH 4.65 (1H, d, J = 7.6 Hz, H-100), 4.87
(1H, d, J = 7.6 Hz, H-1000)], a 1,3,4-trisubstituted benzene
ring [dH 6.91 (1H, dd, J = 8.2, 2.0 Hz, H-60), 7.02 (1H, d,
J = 2.0 Hz, H-20), 7.14 (1H, d, J = 8.2 Hz, H-50)], and a
set of meta-coupled doublets of an asymmetrically 1,3,4,
5-tetrasubstituted benzene ring [dH 6.54 (1H, d,
J = 2.0 Hz, H-2), 6.57 (1H, d, J = 2.0 Hz, H-6)] were
observed. Acid hydrolysis of 2 in the above described
1
110.2
157.2
106.3
133.9
111.1
160.0
169.8
99.7
110.5
157.2
106.7
134.2
111.7
160.1
170.1
102.9
74.8
10
20
30
40
50
60
70
1000
2000
3000
4000
5000
6000
129.5
137.3
129.7
129.6
129.6
129.7
68.2
126.8
157.0
117.1
130.0
123.8
130.8
63.7
2
3
4
5
6
7
100
200
300
400
500
600
104.5
76.0
102.6
74.9
83.4
77.4
78.3
77.8
78.3
70.7
71.2
70.8
71.3
77.6
78.2
77.5
78.1
61.4
62.6
62.4
62.6
1
manner gave only D-glucose. The H-NMR data was clo-
sely related to those of (?)-medioresinol 4,40-O-di-b-D-
glucopyranoside (7), except for the disappearance of an
OMe group. Furthermore, compound 2 possesses a set of
meta-coupled doublets (H-2 and H-6) instead of a chemi-
cally equivalent singlet of 1,3,4,5-tetrasubstituted benzene
ring in 7. The molecular formula of 2 represented a loss of
a CH2 unit compared to that of 7. These findings indicated
that an OMe group of 1,3,4,5-tetrasubstituted benzene ring
in 7 was replaced by an OH group in 2. This deduction was
supported by the HMBC and the NOE difference (NOED)
experiments as follows (Fig. 3): the anomeric proton at
H-100 [dH 4.65 (1H, d, J = 7.6 Hz)] showed HMBC
cross-peak with dC 134.5, which is also correlated to the
meta-coupled doublet proton signal [dH 6.57 (1H, d,
J = 2.0 Hz)]. In the NOED experiment, irradiation of the
OMe proton [dH 3.84 (3H, s, 5-OMe)] showed an NOE on
the signal at dH 6.57. Thus, the signals at dC 134.5 and dH
6.57 were assigned to C-4 and H-6, respectively. Further-
more, 2 was treated with (trimethylsilyl) diazomethane to
Fig. 2 The main HMBCs of 1. Heavy lines indicate partial structures
1
inferred from H-1H COSY
(1H, d, J = 7.7 Hz, H-1000) and 5.28 (1H, d, J = 7.3 Hz,
H-100). Acid hydrolysis of 1 yielded only D-glucose, which
was identified by its retention time and optical rotation by
123

204
J Nat Med (2011) 65:202–205
temperature for 3 weeks. The MeOH extract was concen-
trated under reduced pressure, and the residue (74.5 g)
was suspended in water. This suspension was succes-
sively extracted with CHCl3, AcOEt, n-BuOH, and H2O.
The n-BuOH-soluble fraction was concentrated under
reduced pressure to produce a residue (14.4 g). The extract
was chromatographed on a silica gel column using
CHCl3-MeOH-H2O (40:10:1), and the eluate was separated
into 20 fractions (frs. 1–20). Fraction 7 was chromato-
graphed on a Sephadex LH-20 column using 50% MeOH,
and the eluate was separated into ten fractions (frs. 7-1 to
7-10). Fraction 7-3 was subjected to preparative HPLC
[column, TSK gel ODS 120T; mobile phase, MeOH-H2O
(1:2); UV detector, 205 nm; flow rate, 1.0 mL/min; column
temperature, 40°C] to give 1 (20.5 mg) and 3 (12.5 mg).
Fraction 13 was chromatographed on a Sephadex LH-20
column using 50% MeOH, and the eluate was separated into
six fractions (frs. 13-1 to 13-6). Fraction 13-2 was subjected
to preparative HPLC [column, TSK gel ODS 120T; mobile
phase, MeOH-H2O (1:4); UV detector, 205 nm; flow rate,
1.5 mL/min; column temperature, 40°C] to give 4 (15.8 mg)
and 5 (22.5 mg). Fraction 17 was subjected to preparative
HPLC [column, TSK gel ODS 120T; mobile phase,
MeOH-H2O (1:5); UV detector, 205 nm; flow rate, 1.5 mL/
min; column temperature, 40°C] to give 6 (8.5 mg). The H2O
layer was passed through a Mitsubishi Diaion HP-20 col-
umn, and the adsorbed material was eluted with H2O and
MeOH. The MeOH eluate-fraction from the Diaion HP-20
column was concentrated. The residue (2.4 g) was chro-
matographed on a silica gel column using CHCl3-MeOH-
H2O (30:10:1), and the eluate was separated into 10 fractions
(frs. 1–10). Fraction 5 was subjected to preparative HPLC
[column, TSK gel ODS 120T; mobile phase, MeOH-H2O
(1:2); UV detector, 205 nm; flow rate, 1.5 mL/min; column
temperature, 40°C] to give 2 (18.0 mg), 7 (15.0 mg), 8
(16.5 mg), 9 (25.5 mg), 10 (23.0 mg), 11 (16.0 mg), 12
(20.0 mg), and 13 (12.0 mg). Fraction 7 was subjected to
preparative HPLC [column, TSK gel ODS 120T; mobile
phase, MeOH-H2O (1:2); UV detector, 205 nm; flow rate,
1.5 mL/min; column temperature, 40°C] to give 14
(15.0 mg) and 15 (11.5 mg).
Fig. 3 The main HMBCs and NOE correlation of 2. Heavy lines
1
indicate partial structures inferred from H-1H COSY
give 7, which was identified by comparison of the spectral
data including the optical rotation, suggesting that the
absolute configuration of 2 was determined to be the same
as 7 [6, 7]. From these data, the structure of 2 was
elucidated as shown and named (?)-neomedioresinol
4,40-O-di-b-D-glucopyranoside.
Experimental
General
Optical rotations were measured with a JASCO DIP-360
digital polarimeter. UV spectra were recorded with a
1
Beckman DU-64 spectrometer. H- and 13C-NMR spectra
were recorded on JEOL JNM-LA 600 (600, 150 MHz,
respectively) spectrometers. Chemical shifts are given on a
d (ppm) scale, with tetramethylsilane (TMS) as internal
standard. FAB-MS were recorded on a JEOL JMS-DX 303
mass spectrometer. Column chromatography was carried
out on Kieselgel 60 (230–400 mesh, Merck, Darmstadt,
Germany), Sephadex LH-20 (Pharmacia Fine Chemicals,
Sweden), and Diaion HP-20 (Mitsubishi Chemical). HPLC
was carried out on a Tosoh HPLC system [pump, CCPS;
detector, UV-8020; column, TSK gel ODS 120T (7.8 mm
i.d. 9 30 cm, Tosoh, Tokyo, Japan)], TSK gel Amide-80
(7.8 mm i.d. 9 30 cm, Tosoh), and Cosmosil 5SL (10 mm
i.d. 9 25 cm, Nacalai, Tokyo, Japan)].
Plant material
Plicatumoside A (1)
Leaves of V. plicatum Thunb. ex Murray var. plicatum f.
plicatum were collected in May 2007 in Sendai, Miyagi
prefecture, Japan, and identified by one of the authors
(M.K.). A voucher specimen (2007-5-KM3) is held in the
laboratory of M. Kikuchi.
An amorphous powder; [a]2D5 -40.3° (c = 0.31, MeOH);
UV kmax (MeOH) nm (log e): 205 (4.6), 247 (3.9); FAB-MS
m/z: 607 [M ? Na]?; HR-FAB-MS m/z: 607.1636
[M ? Na]? (calc’d for C26H32O15Na: 607.1639); 1H-NMR
(600 MHz, CD3OD) d: 3.08 (1H, ddd, J = 9.8, 5.8, 2.6 Hz,
H-5000), 3.15 (1H, dd, J = 9.2, 7.7 Hz, H-2000), 3.17 (1H, dd,
J = 12.0, 5.8 Hz, Ha-6000), 3.28–3.41 (4H, m, H-400, H-500,
H-3000, H-4000), 3.47 (1H, dd, J = 12.0, 2.6 Hz, Hb-6000), 3.50
(1H, dd, J = 8.5, 7.3 Hz, H-200), 3.63 (1H, t, J = 9.2 Hz,
Extraction and isolation
Fresh leaves of V. plicatum Thunb. ex Murray var. plicatum
f. plicatum (860 g) were extracted with MeOH at room
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J Nat Med (2011) 65:202–205
205
Table 2 13C-NMR data for compounds 2 and 7 (150 MHz, CD3OD)
0.5 mL). The reaction mixture was stirred at room tem-
perature for 2 h. The mixture evaporated to give a residue
that purified by prep. HPLC [column, Cosmosil 5SL;
CH2Cl2-MeOH-H2O (60:10:1)] to afford 7 (1.6 mg), which
was identified by comparison of the spectral data including
the optical rotation [6, 7].
Position
2
7
Position
2
7
1
140.2
108.0
152.0
134.5
154.4
102.7
87.1
135.7
105.4
154.5
139.6
154.5
105.4
87.2
10
20
30
40
50
60
70
137.5
111.7
151.0
147.6
118.1
119.8
87.1
137.5
111.7
151.1
147.6
118.1
119.8
87.1
2
3
4
5
Acid hydrolysis of 1 and 2
6
7
Each of the compounds (ca. 1.0 mg) was refluxed with 1 M
HCl (1 mL) for 5 h. The reaction mixture was neutralized
with Ag2CO3 and filtered. The solution was concentrated in
vacuo and dried to give a sugar fraction. The sugar fraction
was analyzed by HPLC under the following conditions: col-
umn, TSK gel Amide-80; column temperature, 45°C; mobile
phase, CH3CN-H2O (4:1); flow rate, 1.0 mL/min; chiral
detection (JASCO OR-2090). Identification of D-glucose
present in the sugar fractionwas carriedout bythe comparison
of the retention time and optical rotation with that of authentic
sample; tR (min) 39.0 (D-glucose, positive optical rotation).
8
55.7
55.7
80
90
55.5
55.5
9
70.9
71.4
72.9
72.9
OCH3
100
200
300
400
500
600
56.79
106.9
75.4
57.6, 57.6
104.9
75.8
OCH3
1000
2000
3000
4000
5000
6000
56.77
102.9
75.0
56.8
102.9
75.0
77.7
77.9
78.3
78.3
73.0
73.0
71.4
71.4
78.4
78.4
77.9
77.9
62.1
62.6
62.5
62.6
H-300), 3.66 (1H, dd, J = 12.1, 5.1 Hz, Ha-600), 3.83 (1H, dd,
J = 12.1, 1.8 Hz, Hb-600), 4.44 (1H, d, J = 7.7 Hz, H-1000),
5.28 (1H, d, J = 7.3 Hz, H-100), 5.37 (1H, d, J = 13.2 Hz,
Ha-70), 5.39 (1H, d, J = 13.2 Hz, Hb-70), 6.56 (1H, br d,
J = 8.4 Hz, H-5), 6.68 (1H, br d, J = 8.1 Hz, H-3), 7.25
(1H, dd, J = 8.4, 8.1 Hz, H-4), 7.40 (2H, br t, J = 7.7 Hz,
H-40, H-50), 7.51 (2H, dd, J = 7.7, 1.5 Hz, H-30, H-60);
13C-NMR (CD3OD): Table 1.
Acknowledgments The authors are grateful to Mrs. S. Sato and
T. Matsuki of Tohoku Pharmaceutical University for conducting the
NMR and MS measurements.
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To a solution of 2 (2.0 mg) in MeOH (2 mL) was added
(trimethylsilyl) diazomethane (2.0 M solution in n-hexane,
123