A new triterpenoid saponin from Clinopodium chinense (Benth.) O. Kuntze

Article abstract of DOI:10.1080/14786419.2015.1095745

A new triterpene saponin, 3β,16β,23,28β,30β-pentahydroxyl-olean-11,13(18)-dien-3β-yl-[β-d-glucopyranosyl-(1→2)]-[β-d-glucopyranosyl-(1→3)]-β-d-fucopyranoside, was named Clinoposaponin D (1), together with six known triterpene saponins, buddlejasaponin IVb (2), buddlejasaponin IVa (3), buddlejasaponin IV (4), clinopodisides D (5), 11,16β,23,28-Tetrahydroxyolean-12-en-3β-yl-[β-d-glucopyranosyl-(1→2)]-[β-d-glucopyranosyl-(1→3)]-β-d-fucopyranoside (6) and prosaikogenin A (7), and two known triterpenes, saikogenin A (8) and saikogenin F (9) were isolated from Clinopodium chinense (Benth.) O. Kuntze. Their structures were elucidated on the basis of 1D, 2D NMR and MS analysis. Meanwhile, the effects of all compounds on rabbit platelet aggregation and thrombin time (TT) were investigated in vitro. Compounds 4 and 7 had significant promoting effects on platelet aggregation with EC50 value at 53.4 and 12.2 M, respectively. In addition, the highest concentration (200 M) of compounds 2 and 9 shortened TT by 20.6 and 25.1%, respectively.

Full text of DOI:10.1080/14786419.2015.1095745

Natural Product Research
Formerly Natural Product Letters
ISSN: 1478-6419 (Print) 1478-6427 (Online) Journal homepage: http://www.tandfonline.com/loi/gnpl20
A new triterpenoid saponin from Clinopodium
chinense (Benth.) O. Kuntze
Biao Zeng, Guang-Da Liu, Bao-Bao Zhang, Shan-shan Wang, Rui Ma, Bei-Shan
Zhong, Bai-qiu He, Yan Liang & Fei-Hua Wu
To cite this article: Biao Zeng, Guang-Da Liu, Bao-Bao Zhang, Shan-shan Wang, Rui
Ma, Bei-Shan Zhong, Bai-qiu He, Yan Liang & Fei-Hua Wu (2015): A new triterpenoid
saponin from Clinopodium chinense (Benth.) O. Kuntze, Natural Product Research, DOI:
10.1080/14786419.2015.1095745
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Date: 15 November 2015, At: 23:47

Natural Product research, 2015
http://dx.doi.org/10.1080/14786419.2015.1095745
A new triterpenoid saponin from Clinopodium chinense
(Benth.) O. Kuntze
Biao Zenga,b,‡, Guang-Da Liua,‡, Bao-Bao Zhanga, Shan-shan Wanga, Rui Maa,
Bei-Shan Zhonga, Bai-qiu Hea, Yan Liangc and Fei-Hua Wua
aschool of traditional chinese Pharmacy, china Pharmaceutical university, Nanjing, china; bGuangzhou
Baiyun shan Ming Xing Pharmaceutical co. ltd., Guangzhou, china; cNanjing sanhome Pharmaceutical
co. ltd., Nanjing, china
ARTICLE HISTORY
ABSTRACT
received 2 February 2015
accepted 7 august 2015
A new triterpene saponin, 3β,16β,23α,28β,30β-pentahydroxyl-olean-
11, 13(18)-dien-3β-yl-[β-
d
-glucopyranosyl-(1→2)]-[β-d-glucopyranosyl-
(1→3)]-β- -fucopyranoside, wasnamedClinoposaponinD(1), together with
d
KEYWORDS
six known triterpene saponins, buddlejasaponin IVb (2), buddlejasaponin
IVa (3), buddlejasaponin IV (4), clinopodisides D (5), 11α,16β,23,28-
Tetrahydroxyolean-12-en-3β-yl-[β-d-glucopyranosyl-(1→2)]-[β-d-
glucopyranosyl-(1→3)]-β-d-fucopyranoside (6) and prosaikogenin A (7),
and two known triterpenes, saikogenin A (8) and saikogenin F (9) were
isolated from Clinopodium chinense (Benth.) O. Kuntze. Their structures
were elucidated on the basis of 1D, 2D NMR and MS analysis. Meanwhile,
the effects of all compounds on rabbit platelet aggregation and thrombin
time (TT) were investigated in vitro. Compounds 4 and 7 had significant
promoting effects on platelet aggregation with EC50 value at 53.4 and
12.2 μM, respectively. In addition, the highest concentration (200 μM)
of compounds 2 and 9 shortened TT by 20.6 and 25.1%, respectively.
Clinopodium chinense
(Benth.) o. Kuntze; triterpene
saponin; platelet aggregation;
thrombin time
CONTACT Yan liang
yanl66@sina.cn; Fei-hua Wu
fhwu2000@sina.com
‡Both authors have contributed equally to the paper.
supplemental data for this article can be accessed at http://dx.doi.org/10.1080/14786419.2015.1095745.
© 2015 taylor & Francis

2
B. ZENG ET AL.
1. Introduction
The genus Clinopodium comprises nearly 20 species and is mainly distributed in Asia and
Europe. Clinopodium chinense (Benth.) O. Kuntze (Lamiaceae), a traditional Chinese Medicine
(TCM) is popularly used for the treatment of haematuria, allergic dermatitis, dysentery,
influenza, heliosis and trauma, etc. (Wang et al. 2013; Zhong et al. 2014). Furthermore, C.
chinense is one of two medicinal herbs in‘Duan Xueliu’, which has long been used as a TCM
with an astrictive remedy property, and is officially listed in the Chinese Pharmacopoeia
(Pharmacopeia Committee of P. R. China 2010). In recent years, its aqueous or ethanol extract
has been found to exhibit haemostatic activities (Zhong et al. 2012). In the course of our
research for bioactive constituents from C. chinense, one new triterpene saponin named
Clinoposaponin D (1), together with six known triterpene saponins (2–7) and two triter-
penes (8–9) were obtained. In the present study, we reported the isolation and structural
identification of compounds 1–9 and their effects on platelet aggregation and thrombin
time (TT) in vitro.
2. Results and discussion
Compound 1 was isolated as a white amorphous powder. Its molecular formula was estab-
lished as C48H78O19 on the basis of HR-ESI-MS (m/z 981.5050 [M + Na]+, Calcd. for 981.5035; m/z
957.5072 [M − H]−, Calcd. for 957.5059). A positive reaction with the Liebermann–Burchard
and Molisch reagents indicated that it might be a saponin (Tsuyoshi et al. 2004; Liu et al.
2013).
1H-NMR (500 MHz, pyridine-d5) spectrum showed the presence of five methyl signals [δH
0.87 (3H, s), 0.96 (3H, s), 1.08 (3H, s), 1.09 (3H, s), 1.12 (3H, s)], three anomeric proton signals
[δH 4.95 (1H, d, J = 7.8 Hz), 5.34 (1H, d, J = 7.8 Hz) and 5.62 (1H, d, J = 7.8 Hz)] and two olefinic
protons mutually coupled [δH 6.60 (1H, dd, J = 11.0, 2.9 Hz) and 5.68 (1H, d, J = 11.1 Hz)]. 13C-
NMR (125 MHz, pyridine-d5) spectrum showed 48 carbons including five methyl carbons [δC
13.2, 19.1, 17.4, 22.4 and 21.2], three anomeric carbons signals [δC 104.4, 104.5 and 105.5],
four olefinic carbons [δC 127.4, 126.2, 137.1 and 133.6], which were characteristic olefinic
carbons of Δ11,13(18)-Olean diene. The data above suggested that 1 was a highly oxygenated
oleanolic triterpenoid saponin derivative with three sugar moieties (Yamamoto, Miyase,
et al. 1993; Liu et al. 1995; Bi et al. 2012; Zhang et al. 2012). The assignment of 1H-NMR and
1
13C-NMR spectroscopic data of 1 (Table S1) was based on H–1H COSY (Figure S7), HSQC
1
(Figure S8) and HMBC spectrum (Figure S9). The H-NMR and 13C-NMR features of 1 were
similar to those of the known compound buddlejasaponin IVb (2) (Table S1), except for the
proton signal (H-30) of methylene group that connected to hydroxy group in compound 1.
The correlation between δH 6.13 (1H, brs, 30-OH) and δH 3.67 (2H, s, H-30) in the 1H–1H COSY
spectrum confirmed their connection. In the HMBC spectrum, the correlations between
30-OH and C-30, H-29 and C-20 proved the linkage of C-30/C-20/C-29. Specific HMBC cor-
relations were given in Figure S10.
Further correlations between 30-OH/H-28 and H-29/H-22α in the ROESY spectrum (Figure
S11) proved that the C-30 hydroxy group was β-oriented and C-29 methyl group was α-ori-
ented. Specific ROESY correlations are given in Figure S12.
The sugar moiety was located at C-3 on the basis of the correlation between the ano-
meric proton signal at δH 4.95 (d, J = 7.8 Hz) and carbon signal at δC 83.0 (C-3) in the HMBC

NATuRAL PRODuCT RESEARCH
3
spectrum. Furthermore, the correlations between anomeric proton signals at δH 5.62 (d,
J = 7.8 Hz, Glu-H-1″) and carbon signal at δC 77.6 (Fucose-C-2′), and correlation from δH 5.34
(d, J = 7.8 Hz, Glu-H-1ꢀꢀꢀ) to δC 85.2 (Fucose-C-3′) proved that sugar–sugar linkages were
the same as buddlejasaponin IVb. The type and absolute configuration of the sugars were
identified as β-d-fucose and β-d-glucose on the basis of the TLC method comparison with
authentic monosaccharides (CH2Cl2:MeOH:H2O = 3:2:0.2, visualisation with ethanol–5%
H2SO4 spraying) and both coupling constants of anomeric protons were 7.8 Hz. Therefore,
the structure of 1 was elucidated as 3β, 16β, 23α, 28β, 30β-pentahydroxyl-olean-11,13(18)-
dien-3β-yl-[β-
named Clinoposaponin D.
By comparing physical and spectroscopic data (MS, H NMR, 13C NMR) with published
literatures, the eight known compounds (2–9) were identified as buddlejasaponin IVb (2)
(Yamamoto, Miyase, et al. 1993), buddlejasaponin IVa (3) (Yamamoto, Suzuki, et al. 1993), bud-
dlejasaponin IV (4) (Yamamoto, Suzuki, et al. 1993), clinopodisides D (5) (Liu et al. 1995), 11α,
d
-glucopyranosyl-(1→2)]-[β-
d
-glucopyranosyl-(1→3)]-β-
d
-fucopyranoside and
1
16β, 23, 28-tetrahydroxyolean-12-en-3β-yl-[β-
d
-glucopyranosyl(1→2)]-[β-
d
-glucopyranosyl
(1→3)]-β-d-fucopyranoside (6) (Lee et al. 2012), prosaikogenin A (7) (Shimizu et al. 1985),
saikogenin A (8) (Shimizu et al. 1985) and saikogenin F (9) (Shimizu et al. 1985), respectively
(Figure 1).
Blood platelets play an essential role in haemostasis by involving in the formation of
thrombi to prevent blood loss and maintain vascular integrity (Huang et al. 2014). The pre-
vious study showed that total saponins of C. chinense significantly increased the rat platelet
aggregation (Liu et al. 1985). Therefore, the platelet aggregation effect of isolated com-
pounds was evaluated. The results showed that compounds 1, 2, 4, 6, 7 and 9 exhibited
varying degrees to promote rabbit platelet aggregation at concentrations up to 100 μM.
Additionally, compounds 4 and 7 exhibited potent platelet aggregation activity and their
EC50 value were 53.4 and 12.2 μM, respectively. Interestingly, their effects on promoting plate-
let aggregation activity were stronger than polyphyllins II at the concentration of 100 μM
(Table S2). To determine the effect of the isolated compounds on haemostasis in vitro, TT were
measured. As shown in Table S3, 200 μM of compounds 2 and 9 remarkably shortened TT by
20.6 and 25.1%, respectively. Similarly, thrombin (0.006 u) remarkably shortenedTT by 22.0%.
3. Experimental
3.1. General experimental procedures
NMR spectra were measured on Bruker AV-500 NMR (Bruker Corporation, Switzerland) with
TMS as an internal standard. Mass spectra were obtained on a Mariner API-TOF mass spec-
trometer (Applied Biosystems, uSA).uV spectra (Shelton, Connecticut) were obtained on
a Shimadzu uV-2401 uV/vis spectrometer (Shimadzu, Japan). All solvents were analytical
grade (Tianjin Chemical Plant). Column chromatography was performed on silica gel (100–
200 mesh, 200–300 mesh, Qingdao Marine Chemical Co., Ltd., China), macroporous resin
D101 (Mitsubishi Chemical Holdings Corp. Japan), Sephadex LH-20 (40–75 μm, Pharmacia,
Sweden) and MCI GEL CHP 20P (75–150 μm, Mitsubishi Chemical Co., Japan). Thin-layer
chromatography was performed on silica gel GF254 (Qingdao Marine Chemical Co., Ltd.
China). YMC C-18 reversed-phase silica gel (40–63 μm, Beijing H&E Co., Ltd. China).

4
B. ZENG ET AL.
Figure 1. structures of compounds 1–9.
3.2. Plant material
The herbs of C. chinense (Benth.) O. Kuntze were collected from Putian city, Fujian province,
China, and identified by Prof. Sheban Pu from Department of Pharmacognosy, School of
Traditional Chinese Pharmacy, China Pharmaceutical university, Nanjing, China. The voucher
specimen (CPu-CC20100820) was deposited in the Herbarium of China Pharmaceutical
university.

NATuRAL PRODuCT RESEARCH
5
3.3. Isolation and identification
Dried aerial parts of C. chinense (6 kg) were extracted with 80% EtOH (3 × 60 L) under reflux.
After removal of solvent under vacuum, the extract was first suspended in H2O and then
partitioned with petroleum ether, EtOAc and n-BuOH saturated with water, successively.
The n-BuOH soluble (226 g) was subjected to a column chromatography of macroporous
resin D101 (2000 g, 10 × 100 cm column), washing with H2O, ethanol–H2O mixtures with
increasing amount of ethanol to give four fractions (Fr. I–IV). Fr. III (50 g) was subjected to
YMC RP-18 column chromatography, eluting with CH3OH–H2O (30:70–100:0) to yield four
fractions (Fr. A–D). Fr. B (12 g) was firstly subjected to YMC RP-18 CC using a step gradient of
CH3OH–H2O (50:50–70:30), to obtain four fractions (Fr. B1–Fr. B4). Fr. B2 (1.4 g) was further
purified byYMC RP-18 CC with CH3OH–H2O (50:50), and Sephadex LH-20 (methanol) to yield
compound 1 (64 mg). Fr. C (26 g) was fractionated by silica gel column chromatography with
CH2Cl2–CH3OH (20:1–1:1) to afford 6 fractions (Fr. C1–C6). Fr. C1 (230 mg) was subjected to
silica gel CC using a step gradient of petroleum ether–acetic ether (2:1–1:1) to yield com-
pound 9 (32 mg). Fr. C2 (160 mg) was subjected to silica gel CC using a step gradient of
CH2Cl2–CH3OH (20:1–10:1) and Sephadex LH-20 (methanol) to yield compound 7 (5 mg). Fr.
C3 (15 g) was subjected toYMC RP-18 CC using a step gradient of CH3OH–H2O (40:60–100:0)
to obtain compounds 3 (3200 mg), 4 (3600 mg), 5 (32 mg), 6 (220 mg) and 2 (3500 mg). Fr. D
(7 g) was firstly subjected to silica gel CC using a step gradient of CH2Cl2–CH3OH (20:1–10:1)
and further purified by preparativeTLC with CH2Cl2–EtOAc (1:2) to yield compound 8 (16 mg).
All the pure compounds were structurally elucidated through 1D NMR, 2D NMR, HR-ESI-MS,
ESI-MS, uV spectra and literature data.
Clinoposaponin D (1): White amorphous powder: [ꢀ]2D0 = −22.74(c = 0.1, MeOH), m.p.
268–270 °C; uV (MeOH, λmax, nm): 240, 251, 260 nm; HR-ESI-MS: m/z 981.5050 (Calcd. for
C48H78O19 Na [M + Na]+, 981.5035), 957.5072 (Calcd. for C48H77O19 [M − H]−, 957.5059). 1H-NMR
(500 MHz, pyridine-d5) δ: 1.84 (1H, m, H-1), 1.08 (1H, m, H-1), 2.28 (1H, m, H-2), 2.03 (1H, m,
H-2), 4.19 (1H, dd, J = 12.1, 4.6 Hz, H-3), 1.66 (1H, d, J = 9.8 Hz, H-5), 1.84 (1H, m, H-6), 1.45 (1H,
m, H-6), 1.45 (1H, m, H-7), 1.35 (1H, d, J = 8.7 Hz, H-7), 2.09 (1H, s, H-9), 6.60 (1H, dd, J = 10.7,
2.1 Hz, H-11), 5.68 (1H, d, J = 10.7 Hz, H-12), 1.59 (1H, dd, J = 3.7, 12.7 Hz, H-15), 2.23 (1H, d,
J = 12.7 Hz, H-15), 4.31 (1H, m, H-16), 2.75 (1H, d, J = 14.5 Hz, H-19), 2.27 (1H, d, J = 14.1 Hz,
H-19), 2.16 (1H, td, J = 13.3, 3.8 Hz, H-21), 1.50 (1H, m, H-21), 3.07 (1H, dt, J = 14.7, 3.8 Hz,
H-22), 1.76 (1H, td, J = 13.8, 3.6 Hz, H-22), 3.73 (1H, d, J = 10 Hz, H-23), 4.42 (1H, dd, J = 10.5,
4.75 Hz, H-23), 1.08 (3H, s, H-24), 0.96 (3H, s, H-25), 0.87 (3H, s, H-26), 1.12 (3H, s, H-27), 4.28
(2H, m, H-28), 1.09 (3H, s, H-29), 3.67 (2H, s, H-30), δ4.95 (1H, d, J = 7.8 Hz, H-1′), 4.70 (1H, dd,
J = 9.7, 7.8 Hz, H-2′), 4.12 (1H, dd, J = 9.7, 3.1 Hz, H-3′), 4.22 (1H, m, H-4′), 3.63 (1H, m, H-5′),
1.44 (3H, d, J = 6.3 Hz, H-6′), δ5.62 (1H, d, J = 7.8 Hz, H-1″), 4.12 (1H, m, H-2″), 4.21 (1H, m,
H-3″), 4.21 (1H, m, H-4″), 3.65 (1H, m, H-5″), 4.35 (1H, m, H-6″), 4.31 (1H, m, H-6″), δ5.34 (1H,
d, J = 7.8 Hz, H-1ꢀꢀꢀ), 4.03 (1H, t, J = 8.3 Hz, H-2ꢀꢀꢀ), 4.28 (1H, m, H-3ꢀꢀꢀ), 4.27 (1H, m, H-4ꢀꢀꢀ), 3.98
(1H, m, H-5ꢀꢀꢀ), 4.50 (1H, dd, J = 11.8, 2.5 Hz, H-6ꢀꢀꢀ), 4.35 (1H, m, H-6ꢀꢀꢀ); 13C-NMR (125 MHz,
pyridine-d5), δ: 38.7 (C-1), 26.4 (C-2), 83.0 (C-3), 44.2 (C-4), 48.2 (C-5), 18.7 (C-6), 32.8 (C-7), 40.8
(C-8), 54.9 (C-9), 36.9 (C-10), 127.4 (C-11), 126.2 (C-12), 137.1 (C-13), 44.7 (C-14), 35.4 (C-15),
76.8 (C-16), 45.3 (C-17), 133.6 (C-18), 33.8 (C-19), 38.7 (C-20), 30.3 (C-21), 29.8 (C-22), 65.0
(C-23), 13.2 (C-24), 19.1 (C-25), 17.4 (C-26), 22.4 (C-27), 64.6 (C-28), 21.2 (C-29), 73.5 (C-30),
104.4 (C-1′), 77.6 (C-2′), 85.2 (C-3′), 72.0 (C-4′), 70.9 (C-5′), 17.6 (C-6′), 104.5 (C-1″), 76.6 (C-2″),

6
B. ZENG ET AL.
79.2 (C-3″), 72.6 (C-4″), 77.8 (C-5″), 63.6 (C-6″), 105.5 (C-1ꢀꢀꢀ), 75.8 (C-2ꢀꢀꢀ), 78.8 (C-3ꢀꢀꢀ), 72.4
(C-4ꢀꢀꢀ), 78.9 (C-5ꢀꢀꢀ), 63.0 (C-6ꢀꢀꢀ).
3.4. Acid hydrolysis of compound 1
Compound 1 (1.4 mg) was heated in 3 mol L−1 HCl–CH3OH (5 mL) for 3 h in a sealed tube
at 90 °C. The mixture was then extracted with EtOAc. The aqueous layer was evaporated
to dryness with ethanol in vacuo at 50 °C until neutral. The residues were determined in
comparison with authentic monosaccharides using HPTLC (CHCl3:CH3OH:H2O = 3:2:0.2, vis-
ualisation with ethanol–5% H2SO4 spraying). For the sugars of compound 1, the Rf values of
glucose and fucose by TLC were 0.32 and 0.40, respectively.
3.5. Effect of compounds on platelet aggregation
The blood platelet aggregation test was performed according to the method of Born
et al. (1963). Briefly, whole blood samples were collected from the carotid of urethane-
anesthetised rabbits and mixed with 3.8% sodium citrate (9:1 v/v), followed by centrifuging at
1000 rpm for 10 min. The supernatants were obtained and used as platelet rich plasma (PRP).
While the remaining blood sample was centrifuged at 3000 rpm for10 min and the super-
natants were collected as platelet poor plasma (PPP). The effect of individual compounds
on the platelet aggregation was measured by the Born’s Light Transmission Aggregometry
method using Agg-RAM-Aggregometer (Helena Laboratories, Beaumont, Texas). Briefly, the
amount of platelet in PRP with PPP was adjusted to 3 × 1011 L−1. The light transmission was
set at 0% with PRP and at 100% with PPP. PRP (225 μL) was incubated with 25 μL of vehicle
(final concentration 0.3% DMSO) or different concentrations of individual compounds in
turbidimetric cup at 37 °C for 5 min. The changes in the light transmittance of the reaction
mixture were continuously recorded for 5 min and the maximal aggregation was recorded.
Polyphyllins II and ADP were used as positive control.
3.6. TT assay
PPP was prepared referring to the method in Section 3.5. TT was determined by a coagu-
lation-analysis instrument (LG-PABER-I, Steellex Co., China) (Liu et al. 2010). A reagent kit
according to the manufacturer’s recommendations was used to assay TT. Briefly, 50 μL of
PPP and 10 μL of individual compounds solution (100, 200 μM) was incubated for 3 min at
37 °C, followed by addition of 100 μL of thrombin solution. Shortly after adding thrombin
solution, the coagulation-analysis instrument was started up to record TT.
3.7. Statistical analysis
All data are expressed as mean SD. Statistical differences among groups were analysed by
the one-way analysis of variance (ANOVA) and two-tailed Student’s t-test. A value of p < 0.05
was considered significant.

NATuRAL PRODuCT RESEARCH
7
4. Conclusions
In this study, seven triterpenoid saponins and two triterpenes including a new triterpenoid
saponin were isolated and elucidated from C. chinense (Benth.) O. Kuntze. Among them,
compounds 4 and 7 markedly promoted rabbit platelet aggregation and compounds 2 and
9 significantly shortened plasma TT, which indicated that compounds 4 and 7 could acceler-
ate the thrombosis through promoting platelet aggregation so as to stop the bleeding and
compounds 2 and 9 could affect the function of fibrinogen in plasma. Such results suggest
that compounds 2, 4, 7 and 9 might be the main haemostatic ingredients of C. chinense.
Furthermore, the Δ11,13(18)-Olean diene saponins and 13, 28-epoxy-Olean-11-ene saponins
exhibited better haemostatic activity when compared to the Δ12-Oleanene saponins in C.
chinense (Benth.) O. Kuntze.
Supplementary material
Supplementary material relating to this article is available online, alongside Figures S1–S12, Tables
S1 and S3.
Disclosure statement
No potential conflict of interest was reported by the authors.
Funding
This research work was supported by National Natural Science Foundation of China [grant number
81102862]; National New Drug Innovation Major Project of China [grant number 2011ZX09307-002-02];
Qing Lan Project of Jiangsu Province, the Priority Academic Program Development of Jiangsu Higher
Education Institutions; National College Students Innovation Project for the R&D of Novel Drugs [grant
number J1030830].
References
Bi LL, Tian XR, Dou F, Hong LJ, Tang HF, Wang SW. 2012. New antioxidant and antiglycation active
triterpenoid saponins from the root bark of Aralia taibaiensis. Fitoterapia 83:234–240.
Born G, Cross M. 1963. The aggregation of blood platelets. J Physiol. 168:178–195.
Huang LF, Shi HL, Gao B, Wu H, Yang L, Wu XJ, Wang ZT. 2014. Decichine enhances hemostasis of
activated platelets via AMPA receptors. Thromb Res. 133:848–854.
Lee IK, Choi Su, Lee KR. 2012. Triterpene saponins from Pleurospermum kamtschaticum and their
biological activity. Chem Pharm Bull. 60:1011–1018.
Liu L, Ma HY, Yang NY, TangYP, Guo JM, Tao WW, Duan JA. 2010. A series of natural flavonoids as thrombin
inhibitors: structure–activity relationships. Thromb Res. 126:e365–e378.
Liu QY, Peng DY, Lu M, Dai M. 1985. Experimental study on the effects of TSCP, TSCC on blood platelet
function and their mechanism. Chin Pharmacol Bull. 3:175–178.
Liu YR, Wu ZJ, Li CT, Xi FM, Sun LN, Chen WS. 2013. Heracleifolinosides A–F, new triterpene glycosides
from Cimicifuga heracleifolia, and their inhibitory activities against hypoxia and reoxygenation. Planta
Med. 79:301–307.
Liu Z, Li D, Owen NL, Grant DM, Cates RG, Jia ZJ. 1995. Oleanane triterpene saponins from the Chinese
medicinal herb Clinopodium chinensis. J Nat Prod. 58:1600–1604.
Pharmacopeia Committee of P. R. China. 2010. Pharmacopoeia of People’s Republic of China. Beijing:
China Medical Science and Technology Press; p. 1103–1104.

8
B. ZENG ET AL.
Shimizu K, Amagaya S, Ogihara Y. 1985. New derivatives of saikosaponins. Chem Pharm Bull. 33:3349–
3355.
Tsuyoshi I, Hidetsugu T, Masafumi O, Toshihiro N. 2004. Pregnane- and furostane-type oligoglycosides
from the seeds of Allium tuberosum. Chem Pharm Bull. 52:142–145.
Wang SN, Ma GX, Zhong ML, Yu SC, Xu XD, Hu YX, Zhang YZ, Wei H, Yang JS. 2013. Triterpene saponins
from Tabellae Clinopodii. Fitoterapia 90:14–19.
Yamamoto A, MiyaseT, ueno A, MaedaT. 1993. Clinoposaponins I–V, new oleanane–triterpene saponins
from Clinopodium gracile O. Kuntze. Chem Pharm Bull. 41:1270–1274.
Yamamoto A, Suzuki H, Miyase T, ueno A, Maeda T. 1993. Clinoposaponins VI and VIII, two oleanane–
triterpene saponins from Clinopodium micranthum. Phytochemistry 34:485–488.
Zhang Y, Ma ZQ, Hu C, Wang LH, Li LZ, Song SJ. 2012. Cytotoxic triterpene saponins from the leaves of
Aralia elata. Fitoterapia 83:806–811.
Zhong ML, Sun GB, Zhang XP, Sun GG, Xu XD, Yu SC. 2012. A new prenylated naphthoquinoid from the
aerial parts of Clinopodium chinense (Benth.) O. Kuntze. Molecules 17:13910–13916.
Zhong ML, Wu HF, Zhang XP, Sun GL, Sun GB, Yu SC, Xu XD. 2014. A new diterpene from Clinopodium
chinense. Nat Prod Res. 28:467–472.