Five new triterpene bisglycosides with acyclic side chains from the rhizomes of Cimicifuga foetida L.

Article abstract of DOI:10.1248/cpb.58.729

Five new triterpene bisglycosides, foetidinosides A (1), B (2), C (3), D (4) and E (5), including three of the cycloartane type, one of its derivative, and one of the lanostane type were isolated from the rhizomes of Cimicifuga foetida. Their structures were elucidated on the basis of spectroscopic data and chemical evidence. They were the first bisglycosides with acyclic side chains which were different from the typical triterpenes with side chains epoxidized with ring D in Cimicifuga species.

Full text of DOI:10.1248/cpb.58.729

May 2010
Note
Chem. Pharm. Bull. 58(5) 729—733 (2010)
729
Five New Triterpene Bisglycosides with Acyclic Side Chains from the
Rhizomes of Cimicifuga foetida L.
,a
Lu LU,a,b Jian-Chao CHEN,a He-Jiao SONG,c Yan LI,a Yin NIAN,a,b and Ming-Hua QIU
*
a State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of Botany, Chinese
Academy of Sciences; Kunming 650204, P. R. China: b Graduate School of Chinese Academy of Sciences; Beijing 100039,
P. R. China: and c College of Life Science and Technology, Kunming University of Science and Technology; Kunming
650224, P. R. China. Received November 17, 2009; accepted January 21, 2010; published online February 25, 2010
Five new triterpene bisglycosides, foetidinosides A (1), B (2), C (3), D (4) and E (5), including three of the cy-
cloartane type, one of its derivative, and one of the lanostane type were isolated from the rhizomes of Cimicifuga
foetida. Their structures were elucidated on the basis of spectroscopic data and chemical evidence. They were the
first bisglycosides with acyclic side chains which were different from the typical triterpenes with side chains
epoxidized with ring D in Cimicifuga species.
Key words Cimicifuga foetida; bisglycoside; lanostane; cycloartane; Ranunculaceae
Our group focuses on the chemical diversity and bioactivi- troscopy (TOCSY) was applied to determine the ambiguous
ties of the genus Cimicifuga (Ranunculaceae) growing in the assignments of the sugar moieties. The HMBC correlations
Yunnan-Guizhou Plateau.1—4) In our previous study, Cimi- of H-16 [dH 4.74 (dd, Jꢁ12.7, 7.4 Hz)] with C-13 (dC 44.8,
cifuga foetida L. distributed at altitudes higher than 2000 m C) and C-14 (dC 45.0, C); and H-15 (dH 1.78, 2.09) and H-17
were found containing a series of cycloartane bisglycosides (dH 1.80) with C-16 (dC 71.3, CH) positioned one hydroxyl
with disaccharide chains which were rarely reported be- group at C-16. The HMBC associations of H3-26 (dH 1.33)
fore.2,3) Curiosity concerning the potential existence of cy- and H3-27 (dH 1.42) with C-25 (dC 72.1, C) indicated that the
cloartane glycosides with more saccharides led to the unex- other hydroxyl group was at C-25. The olefinic bond was lo-
pected isolation of one lanostane bisglycoside, three cycloar- cated at C-9 and C-11 due to the HMBC cross-peaks of H-11
tane bisglycosides, and one cycloartane derivative (1—5). [dH 5.28 (d, Jꢁ5.3 Hz)] with C-10 (dC 39.5, C) and C-13 (dC
These new compounds 1—5 obtained in low yields pos- 44.8, C); H-12 (dH 1.99, 2.10) with C-9 (dC 149.0, C) and C-
sessed highly oxidized acyclic side chains other than the typ- 11 (dC 115.2, CH) while H3-19 (dH 1.07) also showed a
ical triterpenes in Cimicifuga species with side chains epoxi- strong correlation with C-9. Hydrolysis of 1 afforded D-xy-
dized with ring D.4) This paper deals with the isolation and lose and D-glucose, which were confirmed by GC analysis.
structrual elucidation of these compounds.
The linkages of glycosides were determined by the clear
HMBC associations of H-1ꢂ [dH 4.79 (d, Jꢁ7.6 Hz)] with C-
3 and H-1ꢃ [dH 5.07 (d, Jꢁ7.8 Hz)] with C-24. The anomeric
Results and Discussion
The roots of C. foetida collected in Lijiang county, Yunnan centre of both sugars are b oriented due to the J value of the
province, were extracted with 80% MeOH/H2O, and the anomeric protons. In rotating frame Overhauser enhance-
extract was partitioned between the CHCl3-soluble and n- ment spectroscopy (ROESY), the crosspeak between H-16
BuOH-soluble portions. The remaining water-soluble portion and H3-28 indicated that the hydroxyl group at C-16 has a b
was separated by silica gel and octadecyl silica gel column configuration. The 13C-NMR data of C-24 were comparable
chromatographies, and finally afforded five new triterpene to analogue compounds having a 24R configuration.5—7) Con-
bisglycosides (1—5).
sequently, the structure of 1 was elucidated as lanosta-9(11)-
The molecular formula of compound 1 was inferred as en-3b,16b,24R,25-tetraol 3-O-b-D-xylopranosyl-24-O-b-D-
C41H70O13 by negative high-resolution (HR)-electrospray ion- glucopyranoside.
ization (ESI)-MS showing a [C41H69O13]ꢀ ion peak at m/z
769.4742. In the H-NMR spectrum (Table 1), there were C41H62O14 due to the molecular ion peak at m/z 777.4040 in
seven tertiary methyl groups at dH 0.75, 1.04, 1.07, 1.12, the negative HR-ESI-MS. In the H-NMR spectrum (Table
The molecular formula of compound 2 was established as
1
1
1.32, 1.33 and 1.42, and a secondary methyl group at dH 1.06 1), six quaternary methyls at dH 0.68, 0.91, 1.02,1.43, 1.59
(partially overlaped). Additionally, two anomeric protons at and 1.72, and a secondary methyl at dH 1.04 (d, Jꢁ6.8 Hz)
dH 4.79 (d, Jꢁ7.6 Hz) and 5.07 (d, Jꢁ7.8 Hz) displayed were observed. However, the conventional cyclopropane
downfield as well as an olefinic proton signal at dH 5.28 (d, methylene upfield was not found. There were also two
Jꢁ5.3 Hz). The 13C-NMR spectrum showed 30 resonances anomeric protons downfield at dH 4.79 (d, Jꢁ7.5 Hz) and
attributed to the aglycon part along with signals of a xylose 5.20 (d, Jꢁ7.8 Hz) which suggested the b-oriented anomeric
and a glucose (Table 2). Then the planar structure of 1 was centres of both sugars. Hydrolysis of 2 also afforded D-xy-
established as lanosta-9(11)-en-3,16,24,25-tetraol 3-O-xylo- lose and D-glucose, which were confirmed by GC analysis.
syl-24-O-glucoside (Fig. 1) by analysis of distortionless en- Besides, three olefinic protons displayed at dH 5.37 (d,
hancement by poarization transfer (DEPT), heteronuclear Jꢁ5.9 Hz), 5.40 (d, Jꢁ6.3 Hz) and 5.50 (s). The 41 signals in
single quantum coherence (HSQC), heteronuclear multiple the 13C-NMR spectrum were assigned to a xylose, a glucose
bond correlation (HMBC) and 1H–1H correlation spec- while the rest 30 ones to the aglycone (Table 2). By analysis
troscopy (COSY) experiments. HSQC-total correlation spec- of DEPT, HSQC, HMBC and 1H–1H COSY, the planar struc-
© 2010 Pharmaceutical Society of Japan
∗ To whom correspondence should be addressed. e-mail: mhchiu@mail.kib.ac.cn

730
Vol. 58, No. 5
Table 1. 1H-NMR Spectral Data of Compounds 1—5 (Pyridine d5)
1b)
2a)
3a)
4b)
5a)
1
2
1.48, 1.81
1.98,
5.50 s
2.33,
1.23, 1.54
1.94,
1.22, 1.54
1.93,
1.19, 1.50
1.92,
2.34
2.78 d (16.7)
3.68 dd (8.2, 5.5)
2.35 d (10.1)
3.50 dd (11.1, 3.7)
2.34 dd (12.4, 3.0)
3.49 dd (11.6, 4.2)
2.32 dd (12.5, 3.1)
3.48 dd (11.6, 4.3)
3
4
5
6
3.37 dd (12.0, 4.0)
0.96 d (10.8)
1.38,
1.63
1.29,
1.62
2.40
2.32,
2.59 d (15.3)
5.40 d (6.3)
1.29
0.50 q (11.9),
1.40
0.93 dd (23.6, 12.4),
1.14
1.34
1.29
0.61 q (12.8),
1.44
0.92,
1.08
1.29
0.70 dd (24.6, 12.3),
1.50
1.05,
1.29
1.68
7
8
9
2.32
1.43
10
11
12
5.28 d (5.3)
1.99,
2.10
5.37 d (5.9)
2.14 dd (17.4, 5.8),
2.27
1.04, 1.92
1.60,
1.65
1.09, 1.94
1.69 t (7.25),
2H
1.05, 1.92
1.68,
1.73
13
14
15
1.78,
2.09
4.74 dd (12.7, 7.4)
1.80
1.12 s
1.07 s
2.07 d (17.8),
2.33
2.02 d (16.0),
2.19 dd (13.8, 7.7)
4.32
1.89
1.23 s
0.22 s,
0.36 s
2.73
1.14 d (6.1)
1.34,
1.91,
2.17 dd (13.9, 8.0)
4.43
2.70 dd (10.6, 8.2)
1.29 s
0.25 d (3.6),
0.44 d (3.1)
2.46 dd (11.8, 4.8)
1.22 d (6.8)
4.72 d (10.4)
1.82 dd (13.1, 4.2),
2.21 dd (13.1, 7.9)
5.07
3.00 dd (10.7, 7.3)
1.44 s
0.24 d (3.9),
0.50 d (3.4)
2.92 dd (10.6, 6.7)
1.47 d (6.6)
4.91 s
16
17
18
19
2.36
0.68 s
3.11 d (13.5),
3.16 d (14.2)
2.67 d (5.6)
1.04 d (6.8)
3.11 dd (19.6, 8.9),
4.02 dd (17.4, 9.2)
20
21
22
2.32
1.06 d
1.67,
2.44
1.25,
1.83
2.65 t (12.7)
4.66 d (9.4)
23
1.86,
2.51 d (12.3)
4.37
4.33
4.33
24
25
26
27
28
29
30
Xyl-c)
1ꢂ
3.92
4.65 s
3.56 s
1.33 s
1.42 s
0.75 s
1.32 s
1.04 s
1.72 s
1.59 s
1.02 s
1.43 s
0.91 s
1.62 s
1.59 s
0.87 s
1.34 s
1.06 s
1.46 s
1.50 s
0.87
1.33 s
1.06 s
1.70 s
1.68 s
1.11 s
1.33 s
1.05 s
4.79 d (7.6)
4.03 t (8.2)
4.17 t (8.8)
4.23
4.79 d (7.5)
4.01
4.16 t (8.7)
4.22
4.88 d (7.5)
4.04
4.17 t (8.7)
4.20
4.86 d (7.5)
4.02 t (8.2)
4.17 t (8.7)
4.22
4.87 d (7.6)
4.03
4.15
2ꢂ
3ꢂ
4ꢂ
4.22
5ꢂ
3.79 t (10.7),
4.41 dd (11.2, 5.2)
3.73 t (10.6),
4.34 dd (11.2, 5.6)
3.79 t (10.7),
4.41 dd (11.2, 5.2)
3.73 t (10.7),
4.35
3.75 t (10.6),
4.35
Glc-c)
1ꢃ
2ꢃ
5.07 d (7.8)
4.07 t (8.3)
4.21
5.20 d (7.8)
4.07
4.26
4.76 d (7.5)
4.04
4.20
4.80 d (7.9)
3.99 t (7.7)
4.21
5.03 d (7.9)
4.05
4.13
3ꢃ
4ꢃ
4.20
4.23
4.20
4.22
3.95
5ꢃ
3.93
3.96
3.90
3.75
3.93
6ꢃ
4.30 dd (11.6, 5.6),
4.52 dd (11.6, 2.1)
4.36 dd (11.4, 5.5),
4.51 d (10.1)
4.34,
4.52 d (11.2)
4.34,
4.43
4.12,
4.59 d (10.5)
a) Recorded at 400 Hz. b) Recorded at 500 Hz. c) Assigned by HSQC-TOCSY.
ture was determined as 9,10-seco-cycloarta-1(10),7,9-trien- with C-8, C-12 (dC 36.7, CH2) and C-13 (dC 43.4, C) and H-
3,24,25-triol-16,23-dione 3-O-xylosyl-25-O-glucoside (Fig. 12 with C-9, C-11 (dC 121.3, CH), C-13, C-14 and C-18 (dC
2). HSQC-TOCSY was used to clarify the ambiguous part of 16.7, CH3) located the three olefinic bonds at C-1–C-10,
the assignments in the sugar moieties. The HMBC correla- C-7–C-8 and C-9–C-11 respectively. The distribution of
tions of H-1 [dH 5.50 (s)] with C-3 (dC 83.9, CH) and C-19 olefinic bonds were confirmed by UV. The experiment value
(dC 43.6, CH2); H-2 [dH 2.78 (d Jꢁ16.7 Hz)] with C-1 (dC of 248 (3.8) nm of the UV (MeOH) lmax (log e) matched the
120.6, CH), C-3, C-4 (dC 39.0, C) and C-10 (dC 138.7, C); calculated value of lmax of 244 nm derived from the Wood-
H-7 [dH 5.40 (d, Jꢁ6.3 Hz)] with C-5 (dC 51.3, CH), C-6 (dC ward–Fieser rules.8) Two carbonyl groups were positioned at
25.1, CH2), C-9 (dC 142.6, C) and C-14 (dC 45.9, C); H-6 C-16 and C-23 due to the HMBC associations of H-15 [dH
[dH 2.32, 2.59 (d, Jꢁ6.3 Hz)] with C-5, C-7 (dC 125.3, CH), 2.07 (d, Jꢁ17.8 Hz), 2.33] and H-17 (dH 2.36) with C-16 (dC
C-8 (dC 137.9, C) and C-10; H-11 [dH 5.37 (d, Jꢁ5.9 Hz)] 218.5, C) and H-22 [dH 3.11 (dd, Jꢁ19.6, 8.9 Hz), 4.02 (dd,

May 2010
731
Table 2. 13C-NMR Spectral Data with DEPT of Compounds 1—5 (Pyri-
dine d5)
1b)
2a)
3a)
4b)
5a)
1
2
3
4
5
6
7
8
9
36.6 t
27.3 t
88.8 d
39.9 s
53.2 d
21.5 t
28.5 t
42.1 d
149.0 s
39.5 s
115.2 d
37.6 t
44.8 s
45.0 s
47.6 t
71.3 d
56.2 d
15.8 q
22.6 q
30.1 d
17.9 q
32.5 t
29.4 t
90.6 d
72.1 s
27.2 q
25.0 q
19.4 q
28.4 q
17.1 q
120.6 d
32.4 t
83.9 d
39.0 s
51.3 d
25.1 t
125.3 d
137.9 s
142.6 s
138.7 s
121.3 d
36.7 t
43.4 s
45.9 s
47.9 t
218.5 s
59.9 d
16.7 q
43.6 t
27.0 d
20.4 q
47.2 t
212.4 s
81.4 d
79.6 s
23.2 q
23.7 q
25.1 q
24.9 q
15.4 q
32.2 t
30.0 t
88.5 d
41.3 s
47.5 d
21.0 t
26.1 t
47.8 d
19.9 s
26.3 s
26.4 t
32.9 t
45.7 s
47.0 s
48.1 t
83.2 d
57.5 d
19.5 q
30.1 t
26.5 d
17.6 q
42.0 t
68.5 d
80.0 d
73.7 s
26.8 q
28.3 q
20.3 q
25.8 q
15.5 q
32.2 t
30.1 t
88.5 d
41.3 s
47.6 d
21.1 t
26.2 t
48.1 d
20.0 s
26.3 s
26.5 t
32.9 t
45.4 s
47.2 s
48.4 t
83.1 d
53.1 d
19.6 q
30.1 t
37.1 d
11.7 q
68.7 d
38.6 t
76.5 d
72.7 s
25.7 q
26.4 q
20.4 q
25.7 q
15.5 q
32.2 t
30.1 t
88.6 d
41.4 s
47.6 d
21.1 t
26.6 t
48.2 d
20.1 s
26.4 s
26.4 t
33.2 t
45.9 s
47.3 s
49.1 t
72.3 d
52.4 d
19.4 q
30.0 t
36.4 d
14.6 q
85.8 d
75.6 d
76.3 d
74.4 s
24.7 q
28.7 q
20.6 q
25.8 q
15.5 q
Fig. 1. HMBC and 1H–1H COSY Correlations of 1
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
Xyl-c)
1ꢂ
Fig. 2. HMBC and 1H–1H COSY Correlations of 2
Fig. 3. HMBC and 1H–1H COSY Correlations of 3
107.8 d
75.6 d
78.7 d
71.1 d
67.2 t
107.3 d
75.5 d
78.6 d
71.2 d
67.2 t
107.6 d
75.6 d
78.7 d
71.3 d
67.2 t
107.5 d
75.6 d
78.6 d
71.2 d
67.1 t
107.6 d
75.1 d
78.7 d
71.3 d
67.2 t
2ꢂ
3ꢂ
4ꢂ
5ꢂ
at dH 0.87, 1.06, 1.23, 1.34, 1.59 and 1.62 and a secondary
1
methyl at dH 1.14 (d, Jꢁ6.1 Hz) were observed in the H-
Glc-c)
1ꢃ
NMR spectrum (Table 1). Two anomeric protons downfield
at dH 4.88 (d, Jꢁ7.5 Hz) and 4.76 (d, Jꢁ7.5 Hz) revealed the
existence of two sugars with the b configuration. Hydrolysis
experiment with GC detection also proved that 3 possessed
the same type of sugar moieties (D-xylose and D-glucose) as
the compounds above. However, in the 13C-NMR spectrum,
there were no characteristc signals of hemiketal or acetal
carbons.9—12) The planar structure was elucidated to be
cycloarta-3,16,23,24,25-pentaol 3-O-xylosyl-24-O-glucoside
105.7 d
75.6 d
78.5 d
71.7 d
78.5 d
62.6 t
97.8 d
75.4 d
78.8 d
71.5 d
78.6 d
62.6 t
107.0 d
76.0 d
78.4 d
71.8 d
78.4 d
62.6 t
106.5 d
75.8 d
78.8 d
71.8 d
78.2 d
62.8 t
106.5 d
75.1 d
78.7 d
72.0 d
78.8 d
63.2 t
2ꢃ
3ꢃ
4ꢃ
5ꢃ
6ꢃ
a) Recorded at 100 Hz. b) Recorded at 125 Hz. c) Assigned by HSQC-TOCSY.
Jꢁ17.4, 9.2 Hz)] and H-24 [dH 4.65, (s)] with C-23 (dC by comprehending the information of DEPT, HSQC, HMBC
1
212.4, C). The HMBC cross-peaks of H-24 with C-23, C-25 and H–1H COSY experiments (Fig. 3). The HMBC correla-
(dC 79.6, C), C-26 (dC 23.2, CH3) and C-27 (dC 23.7, CH3) tions of H-22 [dH 1.34, 2.65 (d, Jꢁ12.7 Hz)] with C-23 (dC
resulted in the hydroxyl group at C-24. The linkages of 68.5, CH); H-24 [dH 3.56 (s)] with C-22 (dC 42.0, CH2), C-
glycosides were deduced by the HMBC associations of H-1ꢂ 25 (dC 73.7, C), C-26 (dC 26.8, CH3) and C-27 (dC 28.3,
[dH 4.79 (d, Jꢁ7.5 Hz)] with C-3 and H-1ꢃ [dH 5.20 (d, CH3) and H3-26 [dH 1.62 (s)] and H3-27 [dH 1.59 (s)] with C-
Jꢁ7.8 Hz)] with C-25. The ROESY experiment displayed 24 and C-25 indicated the location of the three hydroxyl
normal cross-peaks of H-3 with H-5, and H3-28 with H-17 groups at C-23, C-24 and C-25. H-20 (dH 2.73), H-21 [dH
indicating that there was no configuration change in the cy- 1.14 (d, Jꢁ6.1 Hz)], H-22, H-23 [dH 4.66 (d, Jꢁ9.4 Hz)] and
1
cloartane skeleton except for the cleavage of the C-9–C-10 H-24 also correlated with neighbouring protons in H–1H
bond. The configuration of C-24 was not determined. There- COSY. The glycosidation positions were proved at C-3 and
fore, compound 2 was characterized as 9,10-seco-cycloarta- C-16 considering the HMBC cross-peaks btween H-1ꢂ [dH
1(10),7,9-trien-3b,24z,25-triol-16,23-dione 3-O-b-D-xylo- 4.88 (d, Jꢁ7.5 Hz)] and C-3 (dC 88.5, CH) as well as be-
pranosyl-25-O-b-D-glucopyranoside.
tween H-1ꢃ [dH 4.76 (d, Jꢁ7.5 Hz)] and C-16 (dC 83.2, C).
The molecular formula of compound 3 was C41H70O14 de- The ROESY associations of H-15a, which concluded by the
duced by the molecular ion peak appearing at m/z 785.4665 cross-peaks of H-15a with H3-28, and H-17 with H-16 sug-
in the negative HR-ESI-MS. The typical cycloproprane gested that the glycosidated hydroxyl group at C-16 has the b
methylene at dH 0.22 (s) and 0.36 (s); six quaternary methyls form. Comparing the NMR data of C-22—C-25 with refer-

732
Vol. 58, No. 5
Table 3. Comparison of NMR Data of Total Four Configurations of C-23 and C-24
3
Cumingianoside
Q18)
23,24,25-Trihydroxy
cycloartan-3-one19)
Alisol A13)
40.0
Alisol P14)
39.9
Alisol E15)
Orbicoside16,17)
C-22
C-23
H-23
C-24
H-24
C-25
42.0
68.5
4.66 d (br 9.4)
80.0
3.56 s (br)
73.7
38.5
43.0
42.0
40.0
69.5
3.77 dd (9.6, 3.6)
77.6
3.00 s (br)
74.1
69.6
3.77 dd (9.3, 3.5)
77.4
3.00 s (br)
74.4
73.2
3.47 (t-like)
79.1
3.27 d (6)
71.6
73.2
4.32 td (8.4, 3)
79.1
3.78 d (8.4)
74.3
69.4
4.52 br t (7)
77.0
3.57 br s
73.8
69.7
4.13 dd (9.4, 4.8)
75.0
3.18 s (br)
74.3
ence data13—19) (Table 3), the configurations of C-23 and C-
24 in 3 were 23R and 24S, respectively. In sum, 3 was estab-
lished as cycloarta-3b,16b,23R,24S,25-pentaol 3-O-b-D-xy-
lopyranosyl-16-O-b-D-glucopyranoside.
Compound 4 is an isomer of 3 (molecular ion peak at m/z
785.4696 in the negative HR-FAB-MS). The major changes
in their NMR data were the hydroxylation of C-22 and C-24
in 4 instead of C-23 and C-24 in 3 (Tables 1, 2). The HMBC
experiment showed correlations of H-23 [dH 1.86, 2.51 (d,
Jꢁ12.3 Hz)] with C-22 (dC 68.7, CH); H3-26 [dH 1.46 (s)]
and H3-27 [dH 1.50 (s)] with C-24 (dC 76.5, CH) and C-
25 (dC 72.7, C). Furthermore, their cross-peaks of H-22
[dH 4.72 (d, Jꢁ10.4 Hz)], H-23 and H-24 (dH 4.37) also
Fig. 4. HMBC and 1H–1H COSY Correlations of 4
1
can be observed in H–1H COSY spectrum (Fig. 4). Conse-
quently, the structure of 4 was concluded as cycloarta-
3b,16b,22z,24z,25-pentaol 3-O-b-D-xylopyranosyl-16-O-b-
D-glucopyranoside.
The NMR data showed that 5 was also a cycloartane bis-
glycoside (Tables 1, 2). Hydrolysis reaction with GC detec-
tion confirmed that the sugars were D-xylose and D-glucose
as in the other compounds. The negative HR-ESI-MS in-
ferred a molecular formula of C41H70O15 (molecular ion peak
Fig. 5. HMBC and 1H–1H COSY Correlations of 5
m/z 801.4625) which suggested that 5 was an analogue of 4 diamminedichloroplatinum (DDP) as a positive control.
with an extra hydroxyl group. The HMBC correlations of H- Compound 1 exerted moderate inhibition against HL-60 and
15 [dH 1.82 (dd, Jꢁ13.1, 7.9 Hz), 2.51 (dd, Jꢁ13.1, 4.2 Hz)] SMMC-7721 cell growth with IC50 values of 12.64 and
and H-17 [dH 3.00 (dd, Jꢁ10.7, 7.3 Hz)] with C-16 (dC 72.3, 30.59 mM respectively. The rest compounds were inactive
CH) and H3-26 [dH 1.70 (s)] and H3-27 [dH 1.68 (s)] with C- with IC50 values ꢄ40 mM.
25 (dC 74.4, C) and C-24 (dC 76.3, CH) implied hydroxy-
Compounds 1—5 widened our outlook of triterpenoids in
lated positions at C-16, C-24 and C-25. The H–1H COSY Cimcifuga. The large quantity of cycloartane type triter-
associations of H-20 [dH 2.92 (dd, Jꢁ10.6, 6.7 Hz)], H-22 penoids have their side chains epoxidized with ring D in
[dH 4.91 (s)] and H-23 (dH 4.33) exhibited an other hydroxyl Cimicifuga. However, these five trace triterpenoid bisglyco-
group at C-23. The glycosidated hydroxyls were located sides seem to have C-16 or side chain glycosylated so as to
at C-3 and C-22 due to the HMBC correlations of H-1ꢂ keep from epoxidation. Their roles in the biosynthesis
[dH 4.87 (d, Jꢁ7.6 Hz)] with C-3 (dC 88.6, CH) and H-1ꢃ process are interesting to seek after as well as their possible
[dH 5.03 (d, Jꢁ7.9 Hz)] with C-22 (dC 85.8, CH). The ano- bioactivities.
1
meric centres of both sugars were b oriented due to the J
value of the anomeric protons mentioned above. The struc-
Experimental
General Procedure Optical rotations were recorded on a Horiba SEPA-
300 polarimeter; UV spectra on a Shimadzu UV-2401A spectrophotometer;
ture of
5 was determined accordingly as cycloarta-
3b,16b,22z,23z,24z,25-hexaol 3-O-b-D-xylopyranosyl-22-
1
and IR spectra on a Bio-Rad FTS-135 infrared spectrophotometer. H-, 13C-
O-b-D-glucopyranoside.
NMR and 2D-NMR spectra were recorded on a Bruker AV-400 MHz or
a DRX-500 spectrometer with TMS as an internal standard. MS were
Compounds 1—3 and 5 were tested for cytotoxicity
against human promyelocytic leukemia (HL-60) cells, human recorded using VG Autospec-300, Finnigan MAT-90 and API Qstar-Plusar-1
spectrometers. Column chromatography was carried out on silica gel (200—
300 mesh, Qingdao Haiyang Chemical Ltd.) and Lichroprep RP-18 (40—
63 mm, Merck, 4.5 cmꢅ20 cm). TLC was performed on GF254 pre-coated
plates (0.20—0.25 mm, Qingdao Haiyang Chemical Co., Ltd.) and detected
by dipping with 10% H2SO4 followed by heating. GC experiments were
achieved by using a GC-17A with an H2 flame ionization detector (FID)
hepatocellular carcinoma (SMMC-7721) cells, carcinomic
human alveolar basal epithelial (A-549) cells, human breast
cancer (SK-BR-3) cells and human pancreatic adenocarci-
noma (PANC-1) cells by the 3-(4,5)-dimethylthiahiazo(-z-
y1)-3,5-di-phenytetrazoliumromide (MTT) method with cis-

May 2010
733
D-glucose while compared with the authentic D- and L-xylose (tR 7.414 and
7.759 min respectively) and glucose (tR 9.821 and 10.023 min respectively).
(Shimadzu). The GC column was a DB-1 capillary column (15 mꢅ
0.25 mm). Sample D-glucose and D-xylose were purchased from Sinopharm
Chemical Reagent Co., Ltd. as well as hexamethyldisilazane (HMDS) and
trimethylchlorosilane (TMCS). Sample L-xylose and L-glucose were com-
mercially obtained from Acros Organics along with L-cysteine methyl easter
hydrochloride.
Plant Material The roots of C. foetida were collected in Lijiang county,
Yunnan province in July 2004 and identified by Prof. Pei Shengji (Kunming
Institute of Botany, Chinese Academy of Sciences). A voucher specimen
(KIB 04072601) has been deposited at the State Key Laboratory of Phyto-
chemistry and Plant Resources in West China, Kunming Institute of Botany,
Chinese Academy of Sciences.
Extraction and Isolation The air-dried and powdered roots of C.
foetida (10 kg) were extracted three times with MeOH under reflux. After re-
moval of the solvent by evaporation, the residue (950 g) was suspended in
H2O and partitioned sequentially with CHCl3 and n-BuOH. The remaining
water-soluble portion (105 g) was subjected to silica gel chromatography and
eluted with CHCl3–MeOH (10 : 1, 8 : 1, 5 : 1, 1 : 1) to give four fractions (fr.
1—4). Fr. 2. (0.5 g) was chromatographed repeatedly over RP-18 (60—70%
MeOH–H2O) to yield 2 (13 mg), 1 (25 mg) and 3 (14 mg) successively, while
fr. 3. (0.3 g) afforded 5 (13 mg) and 4 (17 mg) in the same polarity of elution
as above.
Acknowledgments This work was supported by the Natural Science
Foundation of Yunnan (No. 2005C0010Z) and Natural Science Foundation
of China (No. 30772636), as well as Foundation of Key State Lab. of Phyto-
chemistry and Plant Resources in West China (P2008ZZ05), Innovative Key
Projects of Chinese Academy of Sciences (CAS) (KSCX2-YW-R-194) and
CAS action-plan for West Development (29KZCX2-XB2-15-03).
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Compound 1: A white powder, [a]D19 ꢆ16.3° (cꢁ0.12, C5H5N); HR-ESI-
MS (769.4742; Calcd for C41H69O13, 769.4738). IR (KBr) cmꢀ1: 3425, 2942,
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Compound 3: A white powder; [a]D19 ꢀ4.8° (cꢁ0.10, C5H5N); HR-ESI-
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Compound 4: A white powder; [a]D19 ꢆ6.8° (cꢁ0.07, MeOH); HR-FAB-
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Sugar Analysis Compounds 1—5 (each 3 mg) were separately refluxed
with 2 N HCl/1,4-dioxane 1 : 1 (2 ml) at 100°C for 2 h. After neutralizing
with Ag2CO3 (300 mg), CHCl3 (2 mlꢅ3) was used for extraction. The filtrate
of the aqueous layer was concentrated to dryness under reduced pressure and
then dissolved in pyridine (100 ml). After that, L-cysteine methyl ester hy-
drochloride (1.1 mg) was added, and the mixture was kept at 60 °C for 1 h.
Then the trimethylsilylation reagents HMDS (100 ml) and TMCS (50 ml)
were added successively, and the mixture was kept at 4—8 °C for 8 h. The
filtrate was subjected to GC analysis under the following conditions: column
temperature, 180 °C→250 °C, 8 °C/min; column pressure, 80 kPa; injector
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