A New Dammarane-Type Saponin from the Rhizomes of Panax japonicus

Article abstract of DOI:10.1007/s10600-018-2452-6

A new dammarane-type saponin, named yesanchinoside R₃ (1), along with three known compounds, has been isolated from the rhizomes of Panax japonicus. Their structures were elucidated on the basis of spectroscopic evidence.

Full text of DOI:10.1007/s10600-018-2452-6

DOI 10.1007/s10600-018-2452-6
Chemistry of Natural Compounds, Vol. 54, No. 4, July, 2018
A NEW DAMMARANE-TYPE SAPONIN FROM THE RHIZOMES
OF Panax japonicus
Tong-Fei Zhu, Qing-Hua Deng,1,2 Ping Li, Lin-Feng Hu,1,2
1
,2
1,2
1
,2*
1,2*
Zhi-Hui Yan,
and Yuan-Juan Yang
A new dammarane-type saponin, named yesanchinoside R (1), along with three known compounds, has
3
been isolated from the rhizomes of Panax japonicus. Their structures were elucidated on the basis
of spectroscopic evidence.
Keywords: Panax japonicus, dammarane-type saponin, yesanchinoside R3.
Panax japonicus (T. Nees) C. A. Mey. grows wild throughout Japan and the region from Central to Southwest China.
Its rhizome is used for enhancing immunity, diminishing inflammation, and transforming phlegm [1]. This paper describes the
structural elucidation and identification of a new dammarane-type saponin, named yesanchinoside R (1), and the known
3
compounds 2–4, which were identified as three dammarane-type triterpenoid saponins, i.e., yesanchinoside R (2) [2],
1
notoginsenoside R (3) [3], and ginsenoside Rd (4) [4].
2
Compound 1 was obtained as a white amorphous powder. Its molecular formula was assigned as C H O
3
6 62 10
+
from the HR-ESI-MS peak at m/z 677.4239 ([M + Na] ; calcd 677.4240). The IR spectrum of 1 showed absorption bands
–
1
1
at ꢀmax 3420, 1063, and 1652 cm due to hydroxy groups, a glycosidic linkage, and a double bond. The H NMR
spectrum of 1 (Table 1) exhibited seven Me singlets at ꢁ 0.82 (H-30), 1.03 (H-19), 1.20 (H-18), 1.34 (H-21), 1.48 (H-29),
1
ꢁ
3
.83 (H-27), and 2.11 (H-28), and the signals of one CH OH group at ꢁ 4.25 (2H, m, H-26), of three CH–O groups at
3.55 (1H, dd, J = 12.0, 4.8 Hz, H-3), 3.95 (1H, ddd, J = 12.0, 10.0, 4.2 Hz, H-12), and 4.46 (1H, ddd, J = 12.0, 10.0,
2
.2 Hz, H-6), of an olefinic H-atom at ꢁ 5.77 (1H, br.t, J = 6.4 Hz, H-24), as well as of an anomeric H-atom at ꢁ 5.04
1
3
(
1H, d, J = 7.8, H-1ꢂ). The C NMR spectrum of 1 showed 36 carbon signals, including a set of six signals (ꢁ 106.0,
5.5, 79.7, 72.0, 78.4, 63.1) contributing to a ꢃ-D-glucopyranosyl unit and 30 remaining ones of a sapogenol moiety.
7
The signal of C-5 at ꢁ 61.4 is characteristic of a protopanaxatriol-type aglycone common among dammarane-type
saponins in Panax ginseng with variations in its side chain. The key HMBC (Fig. 1) correlation between H-1ꢂ and C-6
1
13
demonstrated that the glucoside was connected to the C-6. Furthermore, the H and C NMR data of 1 were similar to
those of yesanchinoside R (2) [2] except for the signals belonging to the side-chain part (C-22–C-27) of the aglycone.
1
HO 21
24
26
HO
23
OH
H 1 20
7
25
H
1
1
18
27
19
13
1
9
10
5
15
H
3
30
HO
6
H
2
9
28
O
OH
6'
5'
O
HO
HO
1'
3
'
OH
1
1
) Chongqing Medical and Pharmaceutical College, 401331, Chongqing, P. R. China, fax: +86 23 61969190,
e-mail: chuanqichuanqi@163.com; 2) Chongqing Engineering Research Center of Pharmaceutical Sciences, 401331, Chongqing,
P. R. China, e-mail: yang_1889@sina.com. Published in Khimiya Prirodnykh Soedinenii, No. 4, July–August, 2018,
pp. 605–607. Original article submitted January 27, 2017.
0
714
0009-3130/18/5404-0714 2018 Springer Science+Business Media, LLC

1
13
TABLE 1. H (400 MHz) and C NMR (100 MHz) Data of 1 and 2 (CD OD, ꢁ, ppm, J/Hz)
3
1
2
C atom
ꢁH
ꢁC
ꢁH
ꢁC
1
2
3
4
5
6
7
1.75 (m)*; 1.03 (m)
1.97 (m); 1.88 (m)
3.55 (dd, J = 12.0, 4.8)
–
39.7
27.9
78.8
40.3
61.4
80.0
45.4
1.74 (m)*; 1.03 (m)
1.98 (m); 1.85 (m)
3.52 (dd, J = 11.8, 5.0)
–
39.7
27.9
78.7
40.4
61.5
80.1
45.3
1.44 (d, J = 10.0)
1.45 (d, J = 10.0)
4.45 (ddd, J = 12.3, 10.0, 3.8)
2.67 (dd, J = 12.3, 3.2);
1.97 (t, J = 10.0)
–
4.46 (ddd, J = 12.0, 10.0, 3.2)
2.68 (dd, J = 12.0, 3.2);
1
.98 (t, J = 10.0)
8
9
–
41.3
50.3
39.7
32.0
71.2
48.3
51.7
31.2
26.8
54.8
17.4
17.7
73.1
27.1
36.3
22.6
125.6
136.5
68.3
14.3
31.8
16.8
16.9
106.0
75.5
79.7
72.0
78.4
63.1
41.2
50.3
39.6
32.0
71.1
48.3
51.7
31.1
26.8
54.7
17.4
17.7
73.1
27.1
36.2
22.6
128.0
136.2
60.9
21.9
31.8
16.4
16.9
106.1
75.5
79.6
72.0
78.3
63.2
1.58 (dd, J = 12.0, 2.8)
1.58 (dd, J = 12.5, 3.0)
–
1
0
1
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
7
8
9
0
–
2.13 (m)*; 1.45 (m)
3.95 (ddd, J = 12.0, 10.0, 4.2)
–
1
1
1
1
1
1
1
1
1
2
2
2
2
2
2
2
2
2
2
3
2.14 (m)*; 1.46 (m)
3.96 (ddd, J = 12.4, 10.0, 4.5)
–
–
–
1.64 (m)*; 1.08 (m)
1.81 (m); 1.38 (m)
2.33 (dd, J = 10.4, 7.2)
1.20 (s)
1.61 (m)*; 1.10 (m)
1.82 (m); 1.39 (m)
2.31 (dd, J = 10.0, 7.2)
1.19 (s)
1.03 (s)
1.03 (s)
–
–
1.34 (s)
1.35 (s)
2.10 (m)*; 1.75 (m)
2.72 (m)*
5.77 (br.t, J = 6.4)
–
2.06 (m)*; 1.74 (m)
2.73 (m)*
5.47 (br.t, J = 6.4)
–
4.25 (m)*
1.83 (s)
2.11 (s)
1.48 (s)
4.56 (m)*
2.03 (s)
2.12 (s)
1.44 (s)
0.82 (s)
0.80 (s)
1
2
3
4
5
6
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
ꢂ
5.04 (d, J = 7.8)
4.09 (t, J = 7.8)
4.30 (m)*
5.03 (d, J = 8.0)
4.10 (t, J = 8.0)
4.32 (m)*
4.26 (m)*
3.98 (m)*
4.37 (dd, J = 12.0, 6.0);
4.27 (m)*
3.99 (m)
4.40 (dd, J = 12.0, 6.0);
4
.56 (br.d, J = 12.0)
4.57 (br.d, J = 12.0)
_
*
_____
Signal pattern unclear due to overlapping.
H
HO
OH
OH
H
H
H
HO
O
OH
O
HO
HO
OH
1
HMBC
ROESY
Fig. 1. Key HMBC and ROESY correlations of 1.
715

The configuration of the double bond at C-24 of 1 was inferred to be (E) from the differences in the 13C NMR data between 1 and
[2: ꢁ 128.0 (C-24), 60.9 (C-26), 21.9 (C-27); 1: ꢁ 125.6 (C-24), 68.3 (C-26), 14.3 (C-27)] and the NOE correlation between H-26
at ꢁ 4.25 (2H, m) and H-24 at ꢁ 5.77 (1H, br.t, J = 6.4 Hz) in the ROESY spectrum of 1 (Fig. 1). Consequently, the structure of
2
yesanchinoside R (1) was identified as (20S,24E)-3ꢃ,6ꢄ,12ꢃ,20ꢃ,26-pentahydroxydammar-24-ene 6-O-ꢃ-D-glucopyranoside (1).
3
EXPERIMENTAL
General Experimetal Procedures. FTIR spectra were measured on a PerkinElmer 157G infrared spectrophotometer.
1
13
Optical rotations were obtained using a PerkinElmer 241-MC polarimeter. H (400 MHz) and C NMR (100 MHz) spectra
were obtained on a Bruker AV 400 spectrometer with CD OD as the solvent and TMS as an internal standard. HR-ESI-MS
3
data were measured with a Bruker AOEXIII 7.0 TESLA FTMS. HPLC was conducted using reverse-phase columns
(
Mightysil RP-18 and 8, Kantho Chemical Co. Ltd) with the MeOH–H O solvent system. Colum chromatography was carried
2
out on silica gel (Qingdao Marine Chemical Company, China; 200–300 mesh) and Sephadex LH-20 (Amersham Pharmacia
Biotech AB). Silica GF254 for TLC was produced by Qingdao Marine Chemical Company, China and Merck.
Plant Material. The rhizomes of Panax japonicus C. A. Meyer were collected in July 2014 in Hubei Province, China
and identified by Prof. Qing-wen Sun (Guiyang College of Traditional Chinese Medicine). A voucher specimen has been
deposited in the Chongqing Medical and Pharmaceutical College, China.
Extraction and Isolation. The rhizomes of Panax japonicus (T. Nees) C.A. Mey. (1.0 kg) were successively extracted
three times with 90% EtOH under reflux. After removal of the solvent in vacuo, the residue (138 g) was suspended in H O and
2
then extracted successively with EtOAc and n-BuOH. The n-BuOH layer was concentrated in vacuo to give a viscous residue
(
108 g), which was then dissolved in water (500 mL) and subjected to macroporous D-101 resin column chromatography and
eluted successively with water and ethanol (water, 30%, 50%, and 100% ethanol). The 30% EtOH eluted fraction was evaporated
in vacuo to yield a residue (33 g) that was subjected to silica gel column chromatography eluting with mixtures of
CHCl –MeOH–H O(10:3:0.4) of increasing polarity to give 10 fractions (Frs.1–10).
3
2
Fraction 5 (1.3 g) was purified by Sephadex LH-20 chromatography and further separated by reversed-phase HPLC
using MeOH–H O (55:45–52:48) as the mobile phase to yield compounds 1 (13.2 mg) and 2 (18.3 mg). Fraction 7 (2.2 g) was
2
purified by Sephadex LH-20 chromatography and further separated by reversed-phase HPLC using MeOH–H O (53:47–48:52)
2
as the mobile phase to yield compounds 3 (18.7 mg) and 4 (41.6 mg).
2
5
–1
Compound 1. White amorphous powder. [ꢄ] +18.3ꢅ (c 0.10, MeOH). IR (KBr, ꢀ , cm ): 3420, 1652, 1250,
D
max
1
13
+
1
063. For H and C NMR spectra, see Table 1. HR-ESI-MS m/z 677.4239 [M + Na] (calcd for C H O Na, 677.4240).
36 62 10
Acid Hydrolysis of 1. A solution of compound (1) (6 mg) in 2 N TFA (3 mL) was refluxed at 100ꢅC for 3 h. Then,
the reaction mixture was concentrated in vacuo to dryness. The residue was extracted with EtOAc and H O (5 mL each,
2
3
times). D-glucose was found to be the only sugar present in the water part following the procedure of Oshima, Yamauchi
and Kumanotani [5].
ACKNOWLEDGMENT
This study was supported by the Chongqing City Health and Family Planning Committee of Traditional Chinese
Medicine Science and Technology Project (No. zy201402159).
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1
.
Institute of Materia Medica, Chinese Academy of Medical Sciences, Zhong Yao Zhi (The Traditional Chinese
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M. Zhou, M. Xu, D. Wang, H. T. Zhu, C. R. Yang, and Y. J. Zhang, Helv. Chim. Acta, 94, 2010 (2011).
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2
844 (1981).
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.
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S. Sanada, N. Kondo, J. Shoji, O. Tanaka, and S. Shibata, Chem. Pharm. Bull., 22, 421 (1974).
R. Oshima, Y. Yamauchi, and J. Kumanotani, Carbohydr. Res., 107, 169 (1982).
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16
7