One new and seven known triterpene glycosides from the aerial parts of Cimicifuga dahurica

Article abstract of DOI:10.1080/10286020.2019.1638370

One new triterpene glycoside, asiaticoside I (1), along with seven known ones (2–8), were isolated from the aerial parts of Cimicifuga dahurica (Turcz.) Maxim. The structure of 1 was elucidated on the basis of extensive spectroscopic methods including 1D-NMR, 2D-NMR and MS data. The structures of known compounds were determined by comparison with the literature data. Compound 1 exhibited moderate cell growth inhibitory activities in vitro against HELF, non-small cell lung cancer A549, and pancreatic cancer PANC-1 cell lines, with IC₅₀ values of 62.97, 43.19, and 60.40 μM, respectively.

Full text of DOI:10.1080/10286020.2019.1638370

Journal of Asian Natural Products Research
ISSN: 1028-6020 (Print) 1477-2213 (Online) Journal homepage: https://www.tandfonline.com/loi/ganp20
One new and seven known triterpene glycosides
from the aerial parts of Cimicifuga dahurica
Yi-Meng Hao, Wen Luo, Guan-Ze Jiang, Chong-Ning Lv & Jin-Cai Lu
To cite this article: Yi-Meng Hao, Wen Luo, Guan-Ze Jiang, Chong-Ning Lv & Jin-Cai Lu (2019):
One new and seven known triterpene glycosides from the aerial parts of Cimicifugaꢀdahurica,
Journal of Asian Natural Products Research, DOI: 10.1080/10286020.2019.1638370
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JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
https://doi.org/10.1080/10286020.2019.1638370
One new and seven known triterpene glycosides from
the aerial parts of Cimicifuga dahurica
Yi-Meng Hao, Wen Luo, Guan-Ze Jiang, Chong-Ning Lv and Jin-Cai Lu
School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang
110016, China
ABSTRACT
ARTICLE HISTORY
Received 19 December 2018
Accepted 27 June 2019
One new triterpene glycoside, asiaticoside I (1), along with seven
known ones (2–8), were isolated from the aerial parts of
Cimicifuga dahurica (Turcz.) Maxim. The structure of 1 was eluci-
dated on the basis of extensive spectroscopic methods including
1D-NMR, 2D-NMR and MS data. The structures of known com-
pounds were determined by comparison with the literature data.
Compound 1 exhibited moderate cell growth inhibitory activities
in vitro against HELF, non-small cell lung cancer A549, and pan-
creatic cancer PANC-1 cell lines, with IC50 values of 62.97, 43.19,
and 60.40 lM, respectively.
KEYWORDS
Ranunculaceae; Cimicifuga
dahurica; the aerial part;
triterpene glycoside;
asiaticoside
1. Introduction
The genus Cimicifuga belongs to Ranunculaceae family, widely distributed in North
American, Europe and Asia [1]. Cimicifuga dahurica is an Asian species of
Cimicifugae, whose dried rhizome is defined as “Shengma” in China Pharmacopoeia
(2015 ed), along with those of Cimicifuga foetida and Cimicifuga heracleifolia [2].
They have been used as cooling, detoxification, antipyretic and analgesic agents for a
long time. In China, the aerial parts of C. dahurica is a kind of edible wild herb and
CONTACT Jin-Cai Lu
jincailu@126.com
Supplemental data for this article is available online at https://doi.org/10.1080/10286020.2019.1638370.
ß 2019 Informa UK Limited, trading as Taylor & Francis Group

2
Y.-M. HAO ET AL.
Figure 1. Chemical structure of compound 1.
is known by the common names “kulongya” or “kuliya”. Phytochemical study docu-
mented that the major chemical constituents of the aerial parts are 9,19-cycloartane
triterpene glycosides, almost in accordance with the roots of the C. dahurica [3]. As
part of an ongoing research to discover structurally unique compounds, the aerial
parts of C. dahurica were targeted and afforded one new triterpene glycoside (1),
together with seven known triterpene glycosides 23-epi-26-deoxycimicifugoside (2)
[4], 23-epi-26-deoxyactein (3) [5], cimiracemoside H (4) [6], cimiracemoside F (5)
[6], 15,16-seco-cimiterpene B (6) [7], 15,16-seco-cycloartane glycoside (7) [8],
(16S,20S,24R)-12b-acetoxy-16,23-epoxy-24,25-dihydroxy-3b-(b-D-xylopyranosyloxy)-
9,19-cyclolanost-22(23)-ene(8) [9] (Figure 1). Herein, the isolation and structural elu-
cidation of these compounds are described, as well as cytotoxicity of compound 1
against three human cell lines.
2. Results and discussion
Compound 1 was obtained as a white amorphous powder. The HR-ESI-MS revealed
a protonated molecular ion [M þ H]þ at m/z 661.3943. Its molecular formula
C37H56O10 was deduced from 13C NMR and MS data.
1
The H NMR spectrum showed a pair of cyclopropane methylene protons at d
0.49 and 1.08 (each 1H, d, J ¼ 3.6 Hz); six tert-methyl groups at d 1.03, 1.06, 1.33,
1.35, 1.59 and 1.64; a secondary methyl group at d 1.14 (3H, d, J ¼ 7.2 Hz) and an
acetyl methyl group at d 2.15. Additionally, an anomeric proton at d 4.84 (1H, d,
J ¼ 7.2 Hz) and two olefinic protons at d 5.22 (1H, d, J ¼ 3.0 Hz) and d 5.12 (1H, dd,
J ¼ 7.2, 2.0 Hz) were also observed. The 13C NMR spectrum showed oxygenated car-
bon signals at d 88.2, 79.8, 77.3, 75.3 and 73.3 and four unsaturated carbon signals at
d 154.3, 148.5, 114.4 and 106.6. An acetyl group at d 171.1 and 21.6 was also showed
in the 13C NMR spectrum. Moreover, compound 1 displayed a very similar 13C NMR
spectrum to those of asiaticoside B except for the signals attributed to the oxygenated
carbon at C-24, C-25 of 1 instead of the keto carbon at d 200.8 (C-24) in asiaticoside
B [10].
In the HMBC spectrum (Figure 2), significant correlations were observed between
d 4.32 (H-24) and d 106.6 (C-22), 154.3 (C-23); d 4.28-4.31 (H-16) and d 106.6 (C-
22), 154.3 (C-23), which suggested that one olefinic bond was present at C-22 and C-

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
3
Figure 2. Key HMBC correlations of compound 1.
23. The other double bond was assigned to C-7 and C-8, which is supported by
HMBC correlations from d 5.12 (H-7) to d 50.8 (C-14), 42.8 (C-5), 22.0 (C-9).
Additionally, HMBC correlations from d 1.59 (H-26) and 1.64 (H-27) to d 79.8 (C-
24) and 73.3 (C-25), together with the data above, revealed the presence of a 16,23-
epoxy-24,25-dihydroxy-22-ene moiety in the side chain. And the correlation between
d 5.41 (H-12) and d 171.1 was also observed in the HMBC spectrum, which indicated
that the acetyl group was located at C-12 position.
Acid hydrolysis of compound 1 afforded one D-xylose moiety, which was detected
from the aqueous fraction by TLC analysis (n-BuOH:AcOH:H2O, 4:1:1) and by com-
paring with authentic sample. Meanwhile, the structure of the sugar was also con-
firmed by comparison of the 13C NMR data with those in the literature. According to
the coupling constants of H-10 (J ¼ 7.2 Hz), the D-xylose moiety should be b-anomer
[11]. Moreover, significant correlations were observed between d 4.85 (1H, d,
J ¼ 7.2Hz, H-10) and d 88.2 (C-3), suggesting that the sugar moiety was located at the
C-3 position.
By comparing 13C NMR data with that reported in the literature [9], the R config-
uration at C-24 on the hydrogen-bonded side chain was then determined. Thus com-
pound 1 was elucidated as (16S,20S,24R)-12b-acetoxy-16,23-epoxy-24,25-dihydroxy-
9,19-cyclolanostane-7(8),22(23)-ene-3-O-b-D-xylopyranoside.
The cytotoxicity of compound 1 in HELF, non-small cell lung cancer A549 and
pancreatic cancer PANC-1 cell lines were tested. The results showed that compound
1 had moderate activities in inhibiting HELF, A549 and PANC-1 cells growth, with
IC50 values of 62.97, 43.19, and 60.40 lM, respectively.
3. Experimental
3.1. General experimental procedures
Optical rotations were determined on an Anton Paar MCP-200 polarimeter (Anton
Paar, Graz, Austria) in MeOH at 20 ꢀC. UV spectra were obtained on a UV-1700 vis-
ꢀ
ible apectrophotometer (Shimadzu Corporation, Kyoto, Japan). The FT-IR spectra
were recorded using a Bruker IFS-55 IR spectrometer (Bruker, Karlsruhe, Germany).
NMR experiments were performed on a Bruker 600 MHz AV ˜ HD spectrometers

4
Y.-M. HAO ET AL.
(Bruker Biospin, Rheinstetten, Germany). HR-ESI-MS (TOF) was carried out on an
Xevo G2-XS Q-TOF apparatus (Waters, Milford, America). Preparative HPLC was
performed on a Shimadzu 20A system with a YMC-pack (ODS-A, 20 ꢁ 250 mm2,
ꢀ
5 lm) (YMC, Kyoto, Japan). Silica gel (100-200 mesh, 200-300 mesh, Qingdao
Marine Chemical, Inc., Qingdao, China.) was used for column chromatography. Silica
GF254 (10-40 lm; Qingdao Marine Chemical, Inc., Qingdao, China) and Silica G (10-
40 lm; Qingdao Marine Chemical, Inc., Qingdao, China) were used for TLC. Spots
were observed by UV light and spraying with 10% H2SO4-EtOH followed by heating.
3.2. Plant material
The aerial parts of Cimicifuga dahurica were collected in Fengcheng County,
Liaoning Province, China, in September 2013 and authenticated by Prof. Jincai Lu,
Department of Pharmacognosy, Shenyang Pharmaceutical University. The voucher
specimen (no. 20130936001) was kept in the Pharmacognosy Laboratory of Shenyang
Pharmaceutical University, Shenyang, China.
3.3. Extraction and isolation
Dried-powdered aerial parts of C. dahurica (3.320 kg) was refluxed with 70% ethanol
(1:10, g/ml) for 3 times. The ethanol extracts were concentrated under reduced pres-
sure to afford a residue (293 g), which was suspended in H2O and partitioned succes-
sively with petroleum ether, dichloromethane, ethyl acetate and n-BuOH. The n-
BuOH extract (45 g) was subjected to silica gel CC (700 g, 8 ꢁ 100 cm) and eluted
with a stepwise gradient of CHCl2/MeOH (1:0 to 0:1, v/v) to afford 9 fractions F1
(0.1 g), F2 (0.5 g), F3 (4.8 g), F4 (11.8 g), F5 (5.3 g), F6 (6.3 g), F7 (4.9 g), F8 (2.1 g), F9
(3.3 g). With the guidance of TLC (developed with the lower layer of CHCl2:CH3OH:
H2O ¼ 9:1:0.125, sprayed with 10% sulfuric acid in ethanol, and heated at 105 ꢀC
until the spots are clear), fraction 3 was divided into two subfractions (F3.1–F3.2).
F3.2 (3.6 g) was recrystallized from MeOH and then purified by preparative HPLC
(eluting with MeOH-H2O, 75:25, v/v, 2.5 ml/min, 210 nm) to yield compounds 2
(5 mg) and 3 (10 mg). Fraction 4 was divided into three subfractions (F4.1–F4.3)
guided by TLC. F4.1 (5.2 g) was separated on a silica gel column, eluted with a
CHCl2-MeOH (gradient from 20:1 to 10:1, v/v) to yield fractions F4.1.1–F4.1.3. F4.1.2
was purified by preparative HPLC (eluting with MeOH-H2O, 70:30, v/v, 3 ml/min,
210 nm) to give compounds 1 (4.5 mg), 4 (150 mg) and 5 (5 mg). Fraction F4.1.3 was
purified by preparative HPLC (eluting with MeOH-H2O, 65:35, v/v, 3 ml/min,
210 nm) to give compounds 6 (2 mg), 7 (3 mg) and 8 (5 mg).
3.3.1. Asiaticoside I (1)
White amorphous powder; [a]20 ꢂ61.0 (c 0.13, MeOH); UV (MeOH) kmax (log e):
D
229 nm; IR (ꢀmax): 3427, 2964, 2930, 2373, 1733, 1628, 1460, 1383, 1243, 1162, 1044,
1
987, 895, 807, 626 cmꢂ1; H NMR and 13C NMR spectral data are shown in Table 1.
HR-ESI-MS: m/z 661.3943 ½M þ Hꢃþ (calcd for C37H57O10, 661.3946).

JOURNAL OF ASIAN NATURAL PRODUCTS RESEARCH
5
Table 1. 1H NMR and 13C NMR spectral data of 1 in pyridine-d5 (600 MHz, d in ppm, J in Hz).
Position
dC
dH
1
2
3
4
5
6
7
8
30.8
29.9
88.2
40.8
42.8
22.2
114.4
148.5
22.0
29.1
36.7
77.3
48.4
50.8
44.8
75.3
53.7
14.9
29.9
24.9
26.1
106.6
154.3
79.8
73.3
27.8
26.3
28.3
26.0
14.6
107.9
76.0
79.0
71.6
67.5
171.1
1.15–1.17 m, 1.55–1.57 m
1.86–1.88 m, 2.26–2.28 m
3.45 dd (11.8, 4.4)
1.19–1.21 m
1.55–1.57 m, 1.82–1.84 m
5.12 dd (7.2, 2.0)
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
10
1.21–1.22 m, 2.84–2.90 m
5.41 dd (9.5, 1.5)
2.19–2.21 m
4.28–4.31 m
2.04–2.07 m
1.33 s
0.49 d (3.6), 1.08 d (3.6)
2.43–2.50 m
1.14 d (7.2)
5.22 d (3.0)
4.32 s
1.59 s
1.64 s
1.06 s
1.35 s
1.03 s
4.85 d (7.2)
4.01–4.06 m
4.14–4.17 m
4.20–4.25 m
3.71–3.75 m, 4.34–4.37 m
20
30
40
50
CH3CO
CH3CO
21.6
2.15 s
3.4 Cytotoxicity assay
The cytotoxicity of compound 1 was tested by MTT method using HELF, non-small
cell lung cancer A549 and pancreatic cancer PANC-1 cell lines. The detailed method-
ology for cytotoxicity has been described in a previous paper [12].
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
The work was financially supported by Doctoral Fund of Ministry of Education of China (no.
2018M633721), Natural Science Foundation of Liaoning Province of China (no. 2018055455)
and Scientific Research Fund of Liaoning Provincial Education Department of China
(no. 2017LQN09).

6
Y.-M. HAO ET AL.
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