Phenolic constituents from the stems of Acanthopanax senticosus

Article abstract of DOI:10.1007/s10600-011-9773-z

A new phenolic glycoside was isolated from the stems of Acanthopanax senticosus together with sixteen known compounds. The structure of the new compound was determined to be 2,6-dimethoxy-4-[(1E)-3,3-dimethoxy-1-propenyl] phenyl β-D-glucopyranoside (1) by means of physical, chemical, and spectroscopic methods. Of the known compounds, salvadoraside (7), (7R,8S)-dihydrodehydrodiconiferyl alcohol 4,9′-di-O-β-D- glucopyranoside (8), 3-(4-O-β-D-glucopyranosylferuloyl)quinic acid (15), rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl) -3-methyl-2Z,4E-pentadienoic acid (16), and lycoperodine-l (17) were first reported from the title plant. The inhibitory activities of the isolated compounds against α-glucosidase from rat intestine were also reported.

Full text of DOI:10.1007/s10600-011-9773-z

Chemistry of Natural Compounds, Vol. 46, No. 6, 2011
PHENOLIC CONSTITUENTS FROM THE STEMS
OF Acanthopanax senticosus
1,2
1*
1
1,3
Guan-E Yang, Wei Li, Chao Huang, Lin Lin,
UDC 547.566.1
3
1
Qingbo Zhang, and Kazuo Koike
A new phenolic glycoside was isolated from the stems of Acanthopanax senticosus together with sixteen
known compounds. The structure of the new compound was determined to be 2,6-dimethoxy-4-[(1E)-3,3-
dimethoxy-1-propenyl]phenyl ꢀ-D-glucopyranoside (1) by means of physical, chemical, and spectroscopic
methods. Of the known compounds, salvadoraside (7), (7R,8S)-dihydrodehydrodiconiferyl alcohol 4,9ꢁ-di-
O-ꢀ-D-glucopyranoside (8), 3-(4-O-ꢀ-D-glucopyranosylferuloyl)quinic acid (15), rel-5-(1R,5S-dimethyl-
3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid (16), and
lycoperodine-l (17) were first reported from the title plant. The inhibitory activities of the isolated compounds
against ꢂ-glucosidase from rat intestine were also reported.
Keywords: Acanthopanax senticosus, Araliaceae, stems, phenolic compound.
Acanthopanax senticosus (Rupr. et Maxim.) Harms (Araliaceae) is a shrub that is commonly distributed in the northeast
of Asia. Various parts of the plant, including roots, rhizomes, stems, and leaves, have been widely used as Chinese traditional
medicines for the treatment of a variety of human diseases, such as ischemic heart diseases, neurasthenia, hypertension,
arthritis, and tumors [1]. As for the chemical constituents of these parts, although triterpenoid saponins were reported as the
main constituents in the leaves, fruits, seeds, roots, and rhizome [2–6], it was found that the saponins did not occur in the
stems. In the stems, lignoids and phenylpropanoid glycosides were reported as the main constituents [7, 8]. The stems were
reported to exert several pharmacological effects, such as antioxidant, anti-inflammatory, antiaging, antifatigue,
immunoregulatory, anti-stress, etc. [8, 9]. As a part of our continuing studies on A. senticosus, we have investigated the
chemical constituents of the leaves and fruits of A. senticosus [5, 6]. Herein we reported the isolation and structural determination
of a new phenolic glycoside (1) together with sixteen known compounds from the stems. Of the known compounds, salvadoraside
(7), (7R,8S)-dihydrodehydrodiconiferyl alcohol 4,9ꢁ-di-O-ꢀ-D-glucopyranoside (8), 3-(4-O-ꢀ-D-glucopyranosylferuloyl)-quinic
acid (15), rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-yl)-3-methyl-2Z,4E-pentadienoic acid
(16), and lycoperodine-l (17) were first reported from the title plant. The inhibitory activities of the isolated compounds
against ꢂ-glucosidase from rat intestine were also reported.
OCH
3
H CO
3
4
7
9
OCH
3
HO
HO
HO
O
1
O
OCH
3
OH 1'
1
1) Faculty of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba 274-8510, Japan, e-mail:
liwei@phar.toho-u.ac.jp; 2) School of Pharmaceutical Sciences, Shanxi Medical University, Taiyuan 030001, P. R. China;
3) Department of Chinese Traditional Medicine, Heilongjiang Provincial Institute for Drug Control, Harbin 150001, P. R. China.
Published in Khimiya Prirodnykh Soedinenii, No. 6, pp. 744–746, November–December, 2010. Original article submitted
September 10, 2009.
876
0009-3130/11/4606-0876 02011 Springer Science+Business Media, Inc.

Compound 1 was isolated as a colorless amorphous solid. The molecular formula of 1 was determined as C H O
19 28 10
1
13
by HR-ESI-MS. In the H and C NMR spectra, it showed the presence of a ꢀ-glucopyranosyl moiety with the anomeric
signals at ꢃ 4.89 (1H, d, J = 7.5 Hz) and ꢃ 105.3. Theꢄꢀ-glucopyranosyl moiety was confirmed to be of the D-form by acid
H
C
1
13
hydrolysis of 1. In the H NMR and C NMR spectra, it also showed the presence of a tetrasubstituted aromatic ring with
symmetric structure by the characteristic proton signal at ꢃ 6.78 (2H, s) and carbon signals at ꢃ 134.3, 105.9 ꢅ 2, 154.5 ꢅ 2, and
1
136.5. The H NMR spectrum further showed two trans-relational olefinic protons at ꢃ 6.67 (1H, br.d, J = 16.0 Hz) and 6.12
(1H, dd, J = 16.0, 5.3 Hz), a methine proton at 4.92 (1H, dd, J = 5.3, 1.2 Hz), and the protons due to four methoxyl moieties at
3.85 (6H, s) and 3.35 (6H, s). The correlation between the olefinic proton at ꢃ 6.12 and the methine proton at 4.92 in the
1
1
H– H COSY spectrum suggested the presence of a propenyl moiety. The positions of the ꢀ-D-glucopyranosyl moiety at C-1,
the propenyl moiety at C-4, and two methoxyl moieties at C-2 and C-6 were suggested by the HMBC correlations of ꢃ 4.89
H
(H-1ꢁ)/ꢃ 136.5 (C-1), ꢃ 6.67 (H-7)/ꢃ 105.9 (C-3,5), and ꢃ 3.85 (2,6-OCH )/ꢃ 154.5 (C-2,6). The two methoxyl moieties
C
H
C
H
3
C
at C-9 were suggested by the HMBC correlation between ꢃ 3.35 (9-OCH )/ꢃ 104.7 (C-9). Thus, the structure of 1 was
H
3
C
determined to be 2,6-dimethoxy-4-[(1E)-3,3-dimethoxy-1-propenyl]phenyl ꢀ-D-glucopyranoside.
The known compounds were identified as (+)-syringaresinol (2) [10], (+)-syringaresinol 4ꢁ-O-ꢀ-D-glucoside (3)
[11], (+)-syringaresinol di-O-ꢀ-D-glucoside (4) [12], (+)-medioresinol di-O-ꢀ-D-glucopyranoside (5) [13], (+)-pinoresinol
di-O-ꢀ-D-glucopyranoside (6) [13], salvadoraside (7) [14], (7R,8S)-dihydrodehydrodiconiferyl alcohol 4,9ꢁ-di-O-ꢀ-D-
glucopyranoside (8) [15], isofraxidin (9) [16], isofraxidin-7-O-ꢀ-D-glucoside (10) [17], syringin (11) [17], sinapaldehyde
4-O-ꢀ-D-glucopyranoside (12) [18], glucoferulic acid (13) [7], 3-feruloylquinic acid (14) [19], 3-(4-O-ꢀ-D-
glucopyranosylferuloyl) quinic acid (15) [20], rel-5-(1R,5S-dimethyl-3R,4R,8S-trihydroxy-7-oxa-6-oxobicyclo[3,2,1]oct-8-
yl)-3-methyl-2Z,4E-pentadienoic acid (16) [21], and lycoperodine-l (17) [22] by interpretation of their physical and spectroscopic
data and comparison with values reported in the literature. The known compounds 7, 8, 15, 16, and 17 were first reported from
the title plant.
The isolated compounds 1–17 were subjected to anꢄꢂ-glucosidase inhibition assay. Compounds 12 and 17 exhibited
IC values to rat intestinal sucrase at 2.0 and 1.9 mg/mL, respectively. Compounds 12 and 16 exhibited IC values to rat
50
50
intestinal maltase at 2.0 and 1.6 mg/mL, respectively.
It was reported that A. senticosus could be used to treat diabetic peripheral neuropathy [23, 24] and saponin isolated
from the leaves of A. senticosus could decrease various cases of experimental hyperglycemias induced by injection of adrenaline,
glucose, and alloxan [25]. But this is the first report on ꢂ-glucosidase inhibition assay of compounds from the stems of
A. senticosus. Further research is needed in order to understand the materials foundation and action mechanism of A. senticosus
in diabetes treatment.
EXPERIMENTAL
Generals. Optical rotations were measured with a JASCO DIP-370 digital polarimeter in a 0.5 dm cell. UV spectra
were recorded on a Shimadzu UV-260 spectrometer. IR spectra were measured with a Perkin–Elmer 683 infrared spectrometer
1
13
(by the KBr disk method). The H and C NMR measurements were recorded using a JEOL ECP-500 NMR spectrometer
with TMS as the internal reference, and chemical shifts are expressed in ꢃ (ppm). ESI-MS and HR-ESI-MS were conducted
using a JEOL JMS-T100LP AccuTOF LC-plus mass spectrometer. Diaion HP-20 resin (Mitsubishi Chemical Corporation,
Tokyo, Japan) and ODS (Chromatorex, 100–200 mesh, Fuji Sylisia Chemical, Ltd., Aichi, Japan) were used for column
chromatography. For HPLC, a JASCO PU-1580 HPLC system, equipped with a Shodex RI-71 differential refractometer
detector, was used. TLC was conducted in Kieselgel 60 F plates (Merck).
254
Plant Material. The stems of A. senticosus were collected from Hulin city, Heilongjiang Province, P. R. China in
September, 2007, and identified by one of the authors (W. L.). A specimen of the plant is kept in the Herbarium of the Faculty
of Pharmaceutical Sciences, Toho University.
Extraction and Isolation. The air-dried stems (1 kg) were extracted twice with MeOH at room temperature. Evaporation
of the solvent under reduced pressure gave the MeOH extract (36 g). The MeOH extract was applied to a Diaion HP-20
column and eluted with H O, 30%, 70%, 100% MeOH and acetone to give five fractions (Fr.1–Fr.5). Fraction 2 (1.75 g) was
2
separated by ODS column chromatography using 50% and 100% MeOH to give two fractions (Fr.21–Fr.22). Fraction 21 was
purified by HPLC (15% MeCN containing 0.06% TFA) to give compounds 13 (31 mg), 14 (12 mg), and 15 (3 mg).
Fraction 3 (5.25 g) was separated by ODS column chromatography using 30%, 50%, and 100% MeOH to give seven fractions
877

(Fr. 31–Fr. 37). Fraction 32 was purified by HPLC (15% MeCN containing 0.06% TFA) to give compounds 10 (83 mg), 11
(67 mg), 12 (4 mg), and 16 (3 mg). Fraction 33 was purified by HPLC (30% MeCN containing 0.06% TFA) to give compounds
1 (6 mg), 4 (57 mg), 5 (4 mg), 6 (10 mg), 7 (2.5 mg), 8 (6 mg), and 17 (1 mg). Fraction 34 was purified by HPLC (40% MeCN
containing 0.06% TFA) to give compounds 2 (2 mg), 3 (6 mg), and 9 (16 mg).
2,6-Dimethoxy-4-[(1E)-3,3-dimethoxy-1-propenyl]phenyl ꢀ-D-glucopyranoside (1). Colorless amorphous solid;
[ꢂ]1D9
–1
–1.65ꢆ (c 0.31, MeOH). UV/Vis (MeOH, ꢇ , nm) (log ꢈ): 230 (1.47), 314 (1.40). IR (KBr, ꢉ, cm ): 3386, 1671, 1634,
max
1
1590, 1506, 1455, 1383, 1312, 1273, 1161, 1074, 1026. H NMR (500 MHz, CD OD, ꢃ, ppm, J/Hz): 6.78 (2H, s, H-3,5), 6.67
3
(1H, br.d, J = 16.0, H-7), 6.12 (1H, dd, J = 16.0, 5.3, H-8), 4.92 (1H, dd, J = 5.3, 1.2, H-9), 4.89 (1H, d, J = 7.5, H-1ꢁ), 3.48–3.40
(3H, m, H-2ꢁ, H-3ꢁ and H-4ꢁ), 3.21 (1H, m, H-5ꢁ), 3.66 (1H, dd, J = 12.0, 5.3, H-6ꢁ), 3.78 (1H, dd, J = 12.0, 2.3, H-6ꢁ), 3.85 (6H,
13
s, 2 and 6-OCH ), 3.35 (6H, s, 9-OCH ). C NMR (125 MHz, CD OD, ꢃ): 154.5 (C-2 and 6), 136.5 (C-1), 134.6 (C-7), 134.3
3
3
3
(C-4), 126.8 (C-8), 105.9 (C-3 and 5), 105.3 (C-1ꢁ), 78.4 (C-3ꢁ), 77.9 (C-5ꢁ), 104.7 (C-9), 75.8 (C-2ꢁ), 71.5 (C-4ꢁ), 62.7 (C-6ꢁ),
+
+
57.1 (2 and 6-OCH ), 49.5 (2 ꢅ 9-OCH ). ESI-MS: m/z 439 [M + Na] . HR-ESI-MS: m/z 439.1571 [M + Na] , calcd for
3
3
C H NaO , 439.1580.
19 28
10
Acid Hydrolysis of 1. A solution of 1 (0.5 mg) in 1 M HCl (dioxane–H O, 1:1, 0.2 mL) was heated at 100ꢆC for 1 h
2
under an Ar atmosphere. After the dioxane was removed, the solution was partitioned between EtOAc and H O. The aqueous
2
layer was concentrated under reduced pressure to dryness. The residue was dissolved in pyridine (0.1 mL), to which 0.08 M
L-cysteine methyl ester hydrochloride in pyridine (1.5 mL) was added. The mixture was kept at 60ꢆC for 1.5 h. After the
mixture was dried in vacuo, the residue was trimethylsilylated with 1-trimethylsilylimidazole (0.1 mL) for 2 h. The mixture
was partitioned between n-hexane and H O (0.3 mL each) and the n-hexane extract was analyzed by GC-MS under the
2
following conditions: capillary column, Equity TM-1 (30 m ꢅ 0.25 mm ꢅ 0.25 ꢊm, Supelco), column temperature, 230ꢆC;
injection temperature, 250ꢆC; carrier N gas. Based on the acid hydrolysate of 1, D-glucose was confirmed by comparison of
2
the retention times of their derivatives with those of the D-glucose and L-glucose derivatives prepared in a similar way.
ꢂ-Glucosidase Inhibition Assay. ꢂ-Glucosidase was prepared from rat intestine acetone powder (Sigma-Aldrich
Japan Co., Tokyo, Japan). Rat intestine acetone powder (100 mg) was dissolved in 56 ꢊM maleate buffer (pH 6.0, 0.9 mL),
sonicated at 4ꢆC for 20 min, and then centrifuged at 15,000 g at 4ꢆC for 60 min to obtain the supernatant. The supernatant,
diluted by adding a twofold volume of 56 ꢊM maleate buffer, was used as sucrase solution, and by adding a fortyfold volume
of 56 ꢊM maleate buffer was used as maltase solution. Sucrose or maltose in 56 ꢊM maleate buffer (20 mg/mL) was used as
substrate solution. The above enzyme solution (0.1 mL) was added to a mixture of substrate solution (0.1 mL) and various
concentrations of samples in MeOH (0.01 mL) and 56 ꢊM maleate buffer (0.04 mL). The reaction mixture was incubated for
60 min at 37ꢆC in a final volume of 0.25 mL, then heated at 102ꢆC for 10 min to stop the reaction. The glucose released in the
solution was determined in a 96-well plate using a glucose assay kit (Glucose CII-test Wako, Wako Pure Chem. Co., Osaka,
Japan) based on the glucose oxidase/peroxidase method. The negative control was prepared by adding 56 ꢊM maleate buffer
instead of the sample in the same way as the test. Acarbose was used as the positive control. The blank was prepared by adding
56 ꢊM maleate buffer instead of the enzyme solution using the same method.
The inhibition rates (%) = [(OD
– OD
) – (OD – OD
)]/(OD
– OD
) ꢅ 100%.
negative control
blank
test
test blank
negative blank
blank
IC values of the sample were calculated using IC calculation software.
50
50
ACKNOWLEDGMENT
This research was partially supported by the Ministry of Education, Science, Sports, and Culture of Japan and the
Grant-in-Aid for Young Scientists (B), 18790102, 2008. One of the author, Guan-E Yang, is grateful to The Talented Man
Culturing Project of Japan loan in the inland region of China, 2007–2008.
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