Three new cycloartenol triterpenoid saponins from the roots of cimicifuga simplex wormsk

Article abstract of DOI:10.3390/molecules16064348

Three new cycloartenol triterpene saponins, named shengmaxinsides A-C, have been isolated from the ethyl acetate soluble fraction of an ethanol extract of Cimicifuga simplex Wormsk roots. Their structures were established by chemical tests and detailed spectroscopic analysis as 25-O-acetyl-7,8-didehydrocimigenol- 3-O-β-D-galactopyranoside (1), 7,8-didehydrocimigenol-3-O-β-D- galactopyranoside (2) and 7,8-didehydro-24S-Oacetylhydroshengmanol- 3-O-β-D-galactopyranoside (3), respectively.

Full text of DOI:10.3390/molecules16064348

Molecules 2011, 16, 4348-4357; doi:10.3390/molecules16064348
OPEN ACCESS
molecules
ISSN 1420-3049
www.mdpi.com/journal/molecules
Article
Three New Cycloartenol Triterpenoid Saponins from the Roots
of Cimicifuga simplex Wormsk
Haixue Kuang *, Yang Su, Bingyou Yang, Yonggang Xia, Qiuhong Wang, Zhibin Wang and
Zhengfan Yu
Key Laboratory of Chinese Materia Medica, Heilongjiang University of Chinese Medicine, Ministry of
Education, Harbin 150040, China
*
Author to whom correspondence should be addressed; E-Mail: hxkuang@hotmail.com;
Tel.: +86-451-82193001; Fax: +86-451-82110803.
Received: 3 May 2011; in revised form: 23 May 2011 / Accepted: 23 May 2011 /
Published: 25 May 2011
Abstract: Three new cycloartenol triterpene saponins, named shengmaxinsides A-C, have
been isolated from the ethyl acetate soluble fraction of an ethanol extract of Cimicifuga
simplex Wormsk roots. Their structures were established by chemical tests and detailed
spectroscopic analysis as 25-O-acetyl-7,8-didehydrocimigenol-3-O-β-D-galactopyranoside
(
1), 7,8-didehydrocimigenol-3-O-β-D-galactopyranoside (2) and 7,8-didehydro-24S-O-
acetylhydroshengmanol-3-O-β-D-galactopyranoside (3), respectively.
Keywords: Cimicifuga simplex Wormsk; 9,19-cyclanstane triterpenic glycosides;
cycloartenol triterpenoid saponins
1
. Introduction
The Ranunculaceae is a small family with five genera and around 19 species found throughout the
World. Currently, about nine Cimicifuga species grow in China. C. simplex (Shengma in Chinese) is a
deciduous perennial herb widely distributed in China. Traditionally, the root of C. simplex has been
used in oriental countries as an anti-inflammatory and anti-viral agent [1-3] and the beneficial
ingredients responsible for the anti-inflammatory effects are ferulic acid and isoferulic acid [4,5]. This
herb has also been used for the treatment of human immunodeficiency virus (HIV), and its more
general analgesic, antipyretic, antidiabetes, antimalaria and vasoactive properties [3-5]. Its chemical

Molecules 2011, 16
4349
constituents have been extensively investigated and the main constituents are 9,19-cyclolanostane
triterpenoid glycosides, flavonoids, alkaloids, and chromones [6-9]. More than 200 uncommon
cycloartane-type triterpenoid saponins have been isolated from Cimicifuga plants [6]. Genjiro and his
team have isolated more than fifty cycloartane-type triterpenoids from C. simplex grown in Japan
[
10-22]. It was reported that 9,19-cyclolanostane triterpene glycosides exhibited antiosteoporosis,
anti-tumor and anti-complement activities [23-25]. Furthermore, triterpenoids may be useful
candidates for the development of new drugs for cardiovascular disorders due to their antioxidant and
anti-inflammatory activity [3]. In continuation of our search for pharmacological and structurally
interesting substances from Chinese traditional herbal drugs, we investigated the chemical constituents
of C. simplex. Fractionation of the ethyl acetate soluble extract of the roots of C. simplex by column
chromatography afforded three new cycloartane-type triterpenoid saponins (Figure 1). We report here
on the isolation and structural elucidation of these compounds by chemical and spectroscopic analysis.
Figure 1. Structures of 1-3.
2
. Results and Discussion
Compound 1, named shengmaxinside A, was obtained as colorless needles and gave positive results
for the Liebermann-Burchard reaction and Molish reagents, indicating it to be a triterpenoid glycoside.
+
Its molecular formula was established as C38
H
58
O
11 by the positive HRESIMS from the [M-H
2
O+H]
+
and [M+Na] signals at m/z 673.3964 (calc. for C38
H
57
O
10, 673.3952) and 713.3891 (calc. for
C H O Na, 713.3877), respectively, indicating ten degrees of unsaturation.
3
8
58 11
1
The H-NMR spectrum (Table 1) showed the presence of cyclopropane methylene groups at δ
H
0.47
1.04, 1.17, 1.33, 1.47, 1.53 and 1.64,
0.96 (1H, d, J = 5.6 Hz), an acetyl methyl at δ 2.01, one anomeric proton at
4.89 (1H, d, J = 7.6 Hz), four oxygenated methine protons (δ 3.51, 3.77, 4.53, 4.60) and a series of
and 1.06 (each 1H, d, J = 3.6 Hz), six tertiary methyl groups at δ
a secondary methyl at δ
H
H
H
δ
H
H
13
overlapped signals suggesting a cycloartane-type triterpene glycoside. The C-NMR spectrum (Table 1)
displayed a total of thirty eight carbon signals due to the aglycon moiety, along with a sugar unit and
1
3
an acetyl unit. The C-NMR spectrum exhibited anomeric carbons at δ 107.5. All the above evidence
C
suggested that 1 was a highly oxygenated 9,19-cycloartane triterpene glycoside. Moreover, δ
suggests 1 to be a cimigenol type saponin [26].
C
112.8
After acid hydrolysis and derivatization as alditol acetates, the gas chromatography (GC) analysis
revealed the presence of D-galacose. The presence of a galacose was further confirmed by its NMR

Molecules 2011, 16
4350
data [16], and the galactose linkage was assigned as β from observation of the anomeric proton
coupling constant at δ 4.89 (1H, d, J = 7.6 Hz). The residual three further signals at δ 4.60 (1H, ddd,
H
H
J = 2.0, 4.3, 9.2 Hz), 4.53 (1H, d, J = 7.6 Hz) and 3.77 (1H, d, J = 4.4 Hz) in the region of aglycon
moiety suggest three additional oxygen-bearing carbons on the aglycone. This hypothesis was
confirmed by the HMBC spectrum, which showed cross-peaks between proton signal at δ 4.53 (1H,
H
d, J = 7.6 Hz) with C-14 and C-16, C-13 and C-17, between proton signal at δ
H
4.60 (1H, ddd, J = 2.0,
4
.3, 9.2 Hz) with C-24 and C-22, and between proton signal at δ 3.77 (1H, d, J = 4.4 Hz) with C-23
H
and C-25. This unambiguously iindicated that the oxygen-bearing carbons are C-15, C-23 and C-24. In
the HMBC spectrum, significant correlations between δ 4.89(H-1') and 88.4(C-3) suggested that the
H
galactopyranosyl was located at the C-3 position. Furthermore, the long-range correlations between an
acetyl proton (δ 2.01) with C-25 (δ 79.8) indicated that the acetyl unit locating at C-25. Other key
H
C
long-range correlations were observed for H-19/C-19, H-1'/C-3, H-24/C-25, H-23/C-22 and C-24, and
an acetyl methyl proton and an acetyl carbon and C-25.
1
3
Comparison of the C-NMR spectral data of 1 with those of the known compound 25-O-acetyl-7,8-
didehydrocimigenol-3-O-β-D-xyloside [19] showed that the aglycone of 1 was very similar to that of
the known compound, except for the signals of the sugar moieties. This suggested that 1 had the same
1
1
aglycone as 25-O-acetyl-7,8-didehydrocimigenol-3-O-β-D-xyloside. Thus, from the above the H - H
COSY, HSQC, DEPT and HMBC we concluded that the planar structure of 1 corresponded to
2
5-O-acetylcycloartane-7-en-3-O-β-D-galactopyranoside.
Compound 2, named shengmaxinside B, was obtained as colorless needles and gave positive results
for the Liebermann-Burchard reaction and Molish reagent, which was considered evidence of a
triterpenoid glycoside. Its molecular formula was established as C36 10 by the positive HRESIMS
O+H] and [M+Na] signals at m/z 631.3884 (calc. for C36 631.3846) and
71.3802 (calc. for C36 10Na 671.3771), respectively, indicating nine degrees of unsaturation.
In the C-NMR spectrum (Table 1) a total of thirty six carbon signals due to the aglycon moiety
were observed, along with a sugar unit. Compared to 1, there is no acetyl unit signal. In the meantime,
only the chemical shifts of C-25, C-26 and C-27, located at δ 68.6, 30.7, 25.9, respectively, were
56
H O
+
+
from the [M-H
2
55 9
H O
6
56
H O
13
C
1
3
changed compared to 1. The comparison of the C-NMR data of 2 to those of the moieties of the
ether-linkage and ester-linkage sugar chains of 1 suggested that 2 possessed the same sugar chains as
1
. This deduction was confirmed by the HMBC experiment. On the basis of these data, 2 was
elucidated as 7,8-didehydrocimigenol-3-O-β-D-galactopyranoside.
Compound 3, named shengmaxinside C, was obtained as a white amorphous powder, which was
considered to be a triterpenoid glycoside due to the positive results with the Liebermann-Burchard
reaction and Molish reagents. Its molecular formula was determined as C38
H
60
O
12 according to the
positive HRESIMS from the [M-2H O+Na] signals at m/z 673.3975
59
calc. for C38 10, 673.3952), 691.4102 (calc. for C38H O11, 691.4057) and 713.3837 (calc. for
+
+
+
2 2 2
O+H] , [M-H O+H] and [M-H
(
H
57
O
C H O Na, 713.3877), respectively, indicating nine degrees of unsauration.
3
8
58 11
1
The H-NMR spectrum (Table 1) showed the presence of cyclopropane methylene groups at δ
H
0.44
and 1.03 (each 1H, d, J = 4.0 Hz), six tertiary methyl groups at δ
H
1.02, 1.18, 1.29, 1.37, 1.49, and
1
.74, a secondary methyl at δ 0.95 (1H, d, J = 6.4 Hz), an acetyl methyl at δ 2.01, one anomeric
H
H
proton at δ
H
4.84 (1H, d, J = 10.4 Hz), four oxygenated methine protons (δ
H
3.50, 4.21, 4.32, 4.85) and
1
3
a series of overlapped signals suggesting a cycloartane-type triterpene glycoside. The C-NMR

Molecules 2011, 16
4351
spectrum (Table 1) showed a total of thirty eight carbon signals due to the aglycon moiety, along with
1
3
C
a sugar unit and an acetyl unit. The C-NMR spectrum exhibited anomeric carbons at δ 107.5. All the
above evidence suggested that 3 was a highly oxygenated 9,19-cycloartane triterpene glycoside.
Moreover, δ 106.7 indicated 3 to be a hydroshengmanol type saponin [26].
C
After acid hydrolysis and derivatization as alditol acetates, the gas chromatography (GC) analysis
revealed the presence of D-galacose. This was further confirmed by its NMR data [16], and the
galactose linkage was assigned as β form on the basis of the anomeric proton coupling constant at
δ 4.84 (1H, d, J = 10.4 Hz). Three further signals at δ 4.32 (1H, d, J = 10.4 Hz), 4.85 (1H, d,
H
H
J = 2.0 Hz) and 4.21 (t, J = 8.8 Hz) in the region of the aglycon moiety suggested the presence of three
additional oxygen-bearing carbons. This hypothesis was confirmed by the HMBC spectrum, which
showed cross-peaks between the proton signal at δ 4.32 (1H, d, J = 10.4 Hz) with C-14, C-16, C-13
H
H
and C-17, between the proton signal at δ 4.85 (1H, d, J = 2.0 Hz) with C-23 and C-25, and between
the proton signal at δ 4.21 (1H, t, J = 8.8 Hz) with C-22 and C-24, thus unambiguously identifying
H
the oxygen-bearing carbons as C-15, C-23, and C-24. In the HMBC spectrum, significant correlations
between δ 4.84(H-1') and 88.4(C-3) suggested that the galactopyranosyl was located at the C-3
H
position. Moreover, the long-range correlations between H-24 (δ
170.7) indicated that the acetyl unit locating at C-24. Other key long-range correlations were found
for H-1/C-3, H-19/C-19, 23-H/C-22 and C-24, H-24/C-25 (Figure 2).
H
4.85) with an acetyl carbon
(δ
C
1
1
Figure 2. Key HMBC and H- H COSY correlations of 3.
O
O
H
O
OH
OH
OH
OH
OH
O
O
HO
OH
1
1
H- H COSY
HMBC
According to the literature, the configuration of C-24 is R when C-16 chemical shift in the
C-NMR spectrum should be 102.9~103.7, while for S it appears to be 106.1~106.8 [19] In the case of
1
3
1
13
3
, the C-16 chemical shift is 106.7. The H- and C-NMR spectrum of 3 were similar to those of 7,8-
didehydro-24S-O-acetylhydroshengmanol-3-O-xyloside [27], respectively, except for the sugar moiety
Table 1). Thus, compound 3 was elucidated as 7,8-didehydro-24S-O-acetylhydroshengmanol-3-O-β-
D-galactopyranoside.
(

Molecules 2011, 16
4352
5
Table 1. NMR data for 1-3 in pyridine-d (J in Hz).
H/C
1
2
3
1
2
1.25 (m),1.80 (m)
2.43 (m),1.94 (m)
30.3
29.5
1.24 (m), 1.77 (m)
.43 (dd,13.2, 4.0),
.96 (m)
30.4
29.5
1.25 (m), 1.60 (m)
30.3
2
2.42 (m), 1.93 (m)
29.5
1
3
4
5
6
7
8
9
3.51 (dd, 11.6, 4.0)
88.4
40.4
42.7
21.2
114.4
148.0
21.8
28.2
3.52 (dd, 11.6, 4.0)
88.5 3.50 (dd, 11.6, 4.0)
88.4
40.4
42.7
21.8
114.0
148.4
21.2
28.2
---
---
40.4
42.8
---
1.25 (m)
1.33 (m)
1.21 (m)
1.65 (m),1.96 (m)
1.62 (m),1.98 (m)
21.8
1.60 (m), 1.88 (m)
6.11 (d, 6.4)
6.07 (d, 6.4)
114.3
148.1
21.2
6.11 (d, 6.0)
---
---
---
---
---
---
---
---
---
1
0
1
28.2
1
.13 (m), 2.18
1.11 (m), 2.08
(overlapping)
1.12 (m), 2.12
(overlapping)
1
25.6
25.5
25.6
(
overlapping)
1
1
1
1
1
1
1
2
3
4
5
6
7
8
1.66 (m),1.83 (m)
34.2
41.1
50.8
78.4
112.8
60.5
21.6
1.66 (m),1.71 (m)
34.0
41.1
50.8
78.6
112.5
60.7
21.6
1.63 (m),1.75 (m)
34.2
40.2
50.0
80.8
106.7
61.2
22.1
---
---
---
---
---
---
4.53 (d, 7.6)
---
4.51 (d, 7.6)
---
4.32 (d, 10.4)
---
1.71 (m)
1.17 (s)
1.72 (m)
1.17 (s)
1.49 (m)
1.18 (s)
0
1
.47 (d, 3.6),
.06 (d, 3.6)
0.44 (d, 3.6),
1.06 (d, 3.6)
0.44 (d, 4.0),
1.03 (d,4.0)
1
9
28.2
28.4
28.3
2
2
0
1
1.70 (m)
23.0
19.6
1.68 (m)
23.4
19.7
1.74 (m)
25.8
20.6
0.96 (d,5.6)
0.97 (d,5.6)
0.95 (d,6.4)
1
.97(m), 2.66
2
2
2
3
1.60 (m), 2.0 (m)
30.5
73.3
29.6
73.9
1.57 (m), 1.90 (m)
4.21 (dd, 2.0, 8.8)
33.9
72.8
(
t, 22.0,12.0)
4
.6 (ddd, 2.0, 4.4,
.2)
4.62 (ddd, 2.0, 4.4,
9
9.2)
2
2
2
2
2
2
3
4
5
6
7
8
9
0
3.77 (d, 4.4)
---
84.1
79.8
24.6
23.2
18.5
25.9
14.3
3.72 (d, 4.4)
---
84.1
68.6
30.7
25.9
18.5
26.0
14.3
4.85 (d, 2.0)
---
80.3
75.5
32.8
27.2
18.8
25.8
14.3
1.64 (s)
1.53 (s)
1.47 (s)
1.33 (s)
1.04 (s)
1.41 (s)
1.33 (s)
1.27 (s)
1.28 (s)
1.03 (s)
1.49 (s)
1.74 (s)
1.37 (s)
1.29 (s)
1.02 (s)

Molecules 2011, 16
4353
Table 1. Cont.
1
2
3
4
5
'
'
'
'
'
4.89 (d, 7.6)
4.49 (m)
107.5
73.2
75.5
70.3
76.9
4.88 (d,8.0)
4.47 (m)
107.5
73.2
75.5
70.3
76.8
4.84 (d, 10.4)
4.46 (dd, 9.2, 4.0)
4.16 (dd, 9.4, 3.4)
4.59 (d, 3.2)
107.5
73.2
75.2
70.3
76.8
4.17 (dd, 9.6, 3.2)
4.60 (overlapping)
4.09 (t, 12.4,8.4)
4.17 (dd, 9.2,3.2)
4.59 (overlapping)
4.08 (t, 12.4, 6.0)
4.08 (t, 10.0, 6.2)
4
4
.48 (overlapping),
.50 (overlapping)
4.47 (overlapping),
4.48 (overlapping)
4.42 (t, 9.2, 4.0),
4.46(t,9.2 ,4.0)
6'
62.5
62.5
62.5
-
CO
CH
CO
CH3
-
--
169.8
22.6
---
---
---
---
---
170.7
21.0
3
-
2
.01(s)
2.01 (s)
3. Experimental Section
3.1. General
The optical rotations were recorded on a Perkin-Elmer 341 polarimeter. IR spectra were taken on a
Shimadzu FTIR-8400 S. The NMR spectra were recorded on a Bruker DPX 400 instrument (400 MHz
1
13
5
for H-NMR and 100 MHz for C-NMR). Samples were prepared in pyridine-d with TMS as an
internal standard and coupling constants J are given in Hz. The UV spectra were recorded on a
Shimadzu UV-1601 instrument and GC analysis was carried out on an Agilent HP 6890N gas
chromatograph using an HP-5 capillary column. The HRESIMS was determined on an IonSpec Ultima
7.0 T FTICR. Preparative HPLC (Waters, Delta 600-2487) was performed on Hypersil-ODS II
(
(
10 μm, 20×300 mm, Yilite, Da Lian, China). Column chromatography was performed with silica gel
200-300 mesh, Qingdao Haiyang Chemical Group Co. Ltd, Qingdao, P. R. China), ODS-A (120A,
5
0μm, YMC Co.) and Sephadex LH-20 (25-100 μm, Pharmacia). Analytical TLC spots were detected
2 4
on silica gel 60 F254 (Merck, Germany) by spraying with 10% ethanolic H SO reagent followed
by heating.
3
.2. Plant Material
Root of C. simplex was collected in Heilong Jiang province of China, on September 2009, and
identified by Prof. Wang (Heilongjiang University of Chinese Medicine). The voucher specimen
20090065) was deposited at the Herbarium of Heilongjiang University of Chinese Medicine,
Harbin, China.
(
3.3. Extraction and Isolation
The roots of C. simplex (2.6 kg) was extracted under reflux conditions with 75% ethanol (3L×3×2 h
each). The ethanolic solution was concentrated in vacuo to yield a syrup-like extract (225 g), which
was dissolved in H O (1500 mL) and then partitioned with different solvents to give petroleum
2

Molecules 2011, 16
4354
ether–soluble (7.6 g), ethyl acetate-soluble (75 g) and n-butyl alcohol-soluble (19g) portions. The ethyl
acetate-soluble portion was subjected to silica gel column chromatography (CHCl /MeOH, 20:1→1:1)
3
to afford Fractions A-H. Fraction D (6 g) was re-chromatographed on silica gel (200-300 mesh, 150 g),
eluted with CHCl -MeOH (20:1) as solvent, to afford three sub-fractions. Sub-fraction D (3.6 g) was
3
2
further separated by ODS (MeOH/H
2
O, 6:4→9:1) to afford five fractions. Fraction D2-3 was followed
2
by Sephadex LH-20 and purified by preparative HPLC with MeOH/H O 7:3 to afford compound 1 (23
mg). Fraction D2-4 was purified by preparative HPLC with MeOH/H O 6:4 to furnish 2 (28 mg).
2
2
Fraction E (3.3 g) was further chromatographed on OSD (MeOH/H O, 1:1→9:1) to afford three
fractions. Compound 3 (23 mg) was purified from the Fraction E
2
by repeated ODS and
HPLC methods.
2
5
Shengmaxinside A (1). Colorless needles; [α] D: +0.02 (MeOH); IR (KBr): 3431.13, 3423.41,
2
956.67, 2937.38, 2871.81, 1730.03, 1367.44, 1240.14, 1151.42, 1070.42, 1058.85, 1043.42,
−
1
+
+
9
75.91 cm ; HRESIMS from the [M-H O+H] and [M+Na] signals at m/z 673.3964 (calc. for
2
1
13
C
38 57 58
H O10, 673.3952) and 713.3894 (calc. for C38H O11Na, 713.3877); H-NMR and C-NMR data
are shown in Table 1.
2
5
Shengmaxinside B (2). Colorless needles; [α]
: +0.03 (MeOH); IR (KBr): 3431.13, 3421.48,
D
−1
2
960.53, 2931.6, 2871.81, 2358.78, 2341.42, 2331.78, 1155.28, 1056.92, 987.49, 977.84 cm ;
+
+
HRESIMS from the [M-H
2
O+H] and [M+Na] signals at m/z 631.3887 (calc. for C36
H
55
O
9
631.3846)
1
13
and 671.3804 (calc. for C36H O10Na 671.3771); H-NMR and C-NMR data are shown in Table 1.
56
2
5
Shengmaxinside C (3). White amorphous powder; [α]
: +0.02 (MeOH); IR (KBr): 3411.84, 2952.81,
D
-1
2
935.46, 1718.46, 1379.01, 1244.00, 1163.00, 1151.42, 1062.7, 1031.85, 981.7 cm ; HRESIMS from
+
+
+
the [M-2H
2
O+H] , [M-H
2
O+H] and [M-H
2
O+Na] signals at m/z 673.3975 (calc. for C38
H
57
O
10
,
6
73.3952), 691.4102 (calc. for C H O , 691.4057) and 713.3837 (calc. for C H O Na, 713.3877);
3
8
59 11
38 58 11
1
13
H-NMR and C-NMR data are shown in Table 1.
3.4. Acid hydrolysis
Acid hydrolysis was performed by a previously described method [28]. For this purpose, each
compound (10 mg) was heated in an ampule with aqueous 12% HCl (5 mL) at 90 °C for 2h. The
aglycone was extracted with chloroform, and each aqueous residue was adjusted to pH 7.0 with 12%
NaOH and reduced with NaBH (40 mg), followed by acidification with dilute CH COOH, and then
4
3
co-distilled with pure CH
:1 pyridine-Ac O in a boiling water bath for 2 h to give the corresponding alditol acetates, which
were analyzed by GLC on a HP 6890 N gas chromatograph (Agilent) equipped with a flame ionization
detector FID) using N as carrier gas. The instrument was fitted with a HP-5 capillary column
30 m×0.32mm×0.25 μm). The injector temperature was set at 250 °C and the column temperature
3
OH to remove excess boric acid. The reduced sugars were acetylated with
1
2
2
(
program was as follows: the initial temperature of 120 °C was increased by 3°/min to the final
temperature of 210 °C, then was held 4 min. The detector temperature was set at 300 °C. The standard

Molecules 2011, 16
4355
monosaccharides were subjected to the same reaction and GC analysis under the same conditions
D-galacose, t , 30.8 min)
(
R
4
. Conclusions
It has been reported that 9,19-cyclolanostane triterpene glycosides exhibit varied biological
activities, including antiosteoporosis, antitumor, anti-complement, antioxidant and anti-inflammatory
effects [23-25]. As a part of our chemical investigation on C. simplex, three new cycloartenol
triterpene saponins with galactopyranosyl moieties, shengmaxinsides A-C, were isolated. Their
structures were established on the basis of spectroscopic analysis and chemical evidence. Their
biological activities will be further researched in our laboratory.
Acknowledgments
We appreciate the kind help of Weiguo Zhu of Zhengzhou University for measurement of NMR
spectra. We are grateful to Zhenyue Wang in College of Pharmacy, Heilongjiang University of
Chinese Medicine, for the plant identification.
References and Notes
1
.
Schmid, D.; Woehs, F.; Svoboda, M.; Thalhammer, T.; Chiba, P.; Moeslinger, T. Aqueous
extracts of Cimicifuga racemosa and phenolcarboxylic constituents inhibit production of
proinflammatory cytokines in LPS-stimulated human whole blood. Can. J. Physiol. Pharmacol.
2
009, 87, 963-972.
2
.
.
Liu, I.M.; Chi, T.C.; Hsu, F.L.; Chen, C.F.; Cheng, J.T. Isoferulic acid as active principle from the
rhizoma of Cimicifuga dahurica to lower plasma glucose in diabetic rats. Planta Med. 1999, 65,
7
12-714.
3
Kim, H.Y.; Shin, H.S.; Park, H.; Kim, Y.C.; Yun, Y.G.; Park, S.; Shin, H.J.; Kim, K. In vitro
inhibition of coronavirus replications by the traditionally used medicinal herbal extracts,
Cimicifuga rhizoma, Meliae cortex, Coptidis rhizoma, and Phellodendron cortex. J. Clin. Virol.
2
008, 41, 122-128.
4
5
Hirabayashi, T.; Ochiai, H.; Sakai, S.; Nakajima, K.; Terasawa, K. Inhibitory effect of ferulic acid
and isoferulic acid on murine interleukin-8 production in response to influenza virus infections
in vitro and in vivo. Planta Med. 1995, 61, 221-226.
.
Sakai, S.; Kawamata, H.; Kogure, T.; Mantani, N.; Terasawa, K.; Umatake, M.; Ochiai, H.
Inhibitory effect of ferulic acid and isoferulic acid on the production of macrophage inflammatory
protein-2 in response to respiratory syncytial virus infection in RAW264.7 cells. Mediators
Inflamm. 1999, 8, 173-175.
6
.
.
Li, J.X.; Yu, Z.Y. Cimicifugae rhizoma: from origins, bioactive constituents to clinical outcomes.
Curr. Med. Chem. 2006, 13, 2927-2951.
7
Cao, P.; Pu, X.F.; Peng, S.L.; Zhang, X.R.; Ding, L.S. Chemical constituents from Cimicifuga
foetida. J. Asian Nat. Prod. Res. 2005, 7, 145-149.

Molecules 2011, 16
4356
8
9
.
.
Li, C.J.; Li, Y.H.; Xiao, P.G.; Mabry, T.J.; Watson, W.H.; Krawiec, M. A unusual cycloartane
triterpenoid from Cimicifuga foetida. Phytochemistry 1996, 42, 489-494.
Sun, L.R.; Qing, C.; Zhang, Y.L.; Jia, S.Y.; Li, Z.R.; Pei, S.J.; Qiu, M.H.; Gross, M.L.; Qiu, S.X.
Cimicifoetisides A and B, two cytotoxic cycloartane triterpenoid glycosides from the rhizomes of
Cimicifuga foetida, inhibit proliferation of cancer cells. Beilstein J. Org. Chem. 2007, 3, 3-8.
1
1
0. Genjiro, K.; Sachiko, H.; Yoshikazu, K.; Tsunematsu, T. Studies on the constituents of
Cimicifuga spp. XIII. Structure of Cimicifugoside. Chem. Pharm. Bull. 1977, 25, 3182-3189.
1. Genjiro, K.; Mayumi, I.; Yoshie, S.; Makio, S.; Akiko, K.; Takashi, I. Studies on the constituents
of Cimicifuga species. XIV. A new xyloside from the aerial parts of Cimicifuga simplex Wormsk.
Chem. Pharm. Bull. 1994, 42, 1106-1110.
1
1
1
1
1
1
1
1
2
2
2. Akiko, K.; Makio, S.; Satoshi, K.; Genjiro, K.; Shigeo, N.; Shinji, F. Studies on the constituents of
Cimicifuga species. XV. Two new diglycosides from the aerial parts of Cimicifuga simplex
Wormsk. Chem. Pharm. Bull. 1994, 42, 1940-1943.
3. Akiko, K.; Kei, S.; Mayumi, I.; Makio, S.; Katsuhiko, M.; Genjiro, K. Studies on the constituents
of Cimicifuga species. XVI. Three new cycloartane xylosides from the aerial parts of Cimicifuga
simplex Wormskjord. Chem. Pharm. Bull. 1995, 43, 279-283.
4. Akiko, K.; Makio, S.; Genjiro, K. Studies on the constituents of Cimicifuga species. XVII. Four
new glycosides from the aerial parts of Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1994,
4
3, 1167-1170.
5. Akiko, K.; Makio, S.; Genjiro, K. Studies on the constituents of Cimicifuga species. XVIII. Four
new xylosides from the aerial parts of Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1996, 44,
1
67-172.
6. Akiko, K.; Makio, S.; Genjiro, K.; Toshio, M. Studies on the constituents of Cimicifuga species.
XIX. Eight new glycosides from Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1996, 44,
2
078-2085.
7. Akiko, K.; Masayuki, T.; Makio, S.; Yasuko, I.; Toshimasa, I.; Toshio, M.; Genjiro, K. Studies on
the constituents of Cimicifuga species. XX. Absolute stereostructures of cimicifugoside and actein
from Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1998, 46, 467-472.
8. Akiko, K.; Masayuki, T.; Makio, S.; Toshio, M.; Genjiro, K. Studies on the constituents of
Cimicifuga species. XXI. Two new cyclolanostanol xylosides, Bugbanosides A and B from
Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1998, 46, 1001-1007.
9. Akiko, K.; Masayuki, T.; Makio, S.; Toshio, M.; Genjiro, K. Studies on the constituents of
Cimicifuga species. XXVI. Twelve new cyclolanostanol glycosides from the underground parts of
Cimicifuga simplex Wormsk. Chem. Pharm. Bull. 1999, 47, 511-516.
0. Toshihiro, A.; Ryuichi, H.; Kazuo, K.; Yumiko, K.; Tamotsu, N. Cycloarta-16,24-dien-3β-ol:
revised structure of cimicifugenol, a cycloartane triterpenoid. Chem. Pharm. Bull. 1999, 47,
1
157-1160.
1. Akiko, K.; Makio, S.; Genjiro, K. Studies on the constituents of Cimicifuga species. XXVII.
Malonyl cyclolanostanol glycosides from the underground parts of Cimicifuga simplex Wormsk.
Chem. Pharm. Bull. 1999, 47, 1175-1179.

Molecules 2011, 16
4357
2
2. Akiko, K.; Makio, S.; Daisuke, T.; Genjiro, K. Studies on the constituents of Cimicifuga species.
XXVIII. Four new cycloart-7-enol glycosides from the underground parts of Cimicifuga simplex
Wormsk. Chem. Pharm. Bull. 2001, 49, 437-441.
2
2
2
3. Qiu, M.; Kim, J.H.; Lee, H.K.; Min, B.S. Anticomplement activity of cycloartane glycosides from
the rhizome of Cimicifuga foetida. Phytother. Res. 2006, 20, 945-948.
4. Tian, Z.; Pan, R.L.; Si, J.Y.; Xiao, P.G. Cytotoxicity of cycloartane triterpenoids from aerial part
of Cimicifuga foetida. Fitoterapia 2006, 77, 39-42
5. Tian, Z.; Pan, R.L.; Chang, Q.; Xiao, P.G.; Wu, E. Cimicifuga foetida extract inhibits proliferation
of hepatocellular cells via induction of cell cycle arrest and apoptosis. J. Ethnopharmacol. 2007,
1
14, 227-233.
2
2
6. Jianhua, J.; Dong, L.; Junshan, Y. The spectroscopic features of natural 9,19-cyclolanstane
triterpenic glycosides. Chin. J. Magn. Reson. 2001, 18, 79-90.
7. Jian, X.L.; Shigetoshi, K.; Masao, H.; Sadao, Y.; Motoo, S.; Naoko, O.; Hiroshi, M.; Tsuneo, N.
Constituents of Cimicifugae Rhizoma. I. Isolation and Characterization of Ten New Cycloartenol
Triterpenes from Cimicifuga heracleifolia Komarov. Chem. Pharm. Bull. 1993, 41, 832-841.
8. Li, G.Y.; Zhang, Y.H.; Yang, B.Y.; Xia, Y.G.; Zhang, Y.L.; Lv, S.W.; Kuang, H.X.
Leiyemudanosides A-C, three new bidesmosidic triterpenoid saponins from the roots of
Caulophyllum robustum. Fitoterapia 2010, 81, 200-204.
2
Sample Availability: Samples of the compounds 1-3 are available from the authors.
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