Three new triterpenoids from Panax ginseng exhibit cytotoxicity against human A549 and Hep-3B cell lines

Article abstract of DOI:10.1007/s11418-012-0662-y

Three new triterpenoid derivatives, 3-O-β-Dglucopyranosyl- 20(S)-protopanaxtriol (1), 3-formyloxy-20-O-b-D-glucopyranosyl-20(S)- protopanaxtriol (2) and 26-hydroxyl-24(E)-20(S)-protopanaxtriol (3), along with six known ginsenosides, were isolated from lea

Full text of DOI:10.1007/s11418-012-0662-y

J Nat Med (2012) 66:576–582
DOI 10.1007/s11418-012-0662-y
NOTE
Three new triterpenoids from Panax ginseng exhibit cytotoxicity
against human A549 and Hep-3B cell lines
•
Hai-Ying Ma Hui-Yuan Gao Jian Huang
•
•
•
Bo-hang Sun Bo Yang
Received: 13 September 2011 / Accepted: 11 December 2011 / Published online: 12 April 2012
Ó The Japanese Society of Pharmacognosy and Springer 2012
Abstract Three new triterpenoid derivatives, 3-O-b-D-
glucopyranosyl-20(S)-protopanaxtriol (1), 3-formyloxy-
20-O-b-D-glucopyranosyl-20(S)-protopanaxtriol (2) and
26-hydroxyl-24(E)-20(S)-protopanaxtriol (3), along with
six known ginsenosides, were isolated from leaves of
Panax ginseng. Their structures were established on the
basis of spectral analysis (IR, 1D and 2D NMR, HRESI-
MS). Compounds 1–3 exhibited various degree of cyto-
toxicity towards human A549 pulmonary carcinoma cells
and Hep3B hepatoma cells.
Ginsenosides have been shown to interact with numerous
membrane proteins such as ion channels, transporters and
receptors, resulting in a broad range of physiological
activities [2, 3]. In recent years, there is growing evidence
in the literature that ginsenosides possess anti-tumor
activity not only in studies in vitro but also in experiments
in vivo [4–10]. Among these components, 20(R,S)-ginse-
noside-Rg3, as one of the most effective cytostasis ginse-
nosides, can inhibit proliferation and induce cell apoptosis
in mice or human tumor cell lines, and its pharmaceutical
preparation ‘‘Shen Yi Capsule’’ in China [11, 12] is used
for the treatment of lung, liver or other primary cancers in
clinical treatments.
Keywords Panax ginseng ꢀ Dammarane ꢀ Cytotoxicity ꢀ
Human A549 ꢀ Hep3B cells
In order to search for promising anti-tumor agents from
ginseng, in the studies, we aim to find some new structures
from the cheap leaves of Panax ginseng. In this paper,
three new protopanaxtriol (PPT) dammarane derivatives
(1–3) were isolated and their cytotoxicity in vitro against
human A549 and Hep3B cells was evaluated, and their
much stronger inhibitory activity than that of 20(R)-gin-
senoside-Rg3 was determined (Fig. 1).
Introduction
Panax ginseng C.A.Mey. (Ginseng), an ancient and famous
herbal drug in traditional Chinese medicines, has been
widely used in China for more than 4000 years [1]. It is
mainly distributed in the Changbai Mountain regions in
northeast China. Ginseng is characterized by the presence
of ginsenosides, which are triterpene saponins, considered
to be the main bioactive principles of ginseng.
Results and discussion
Structural determination of compounds
H.-Y. Ma (&)
Pharmacy Department, The Fourth Affiliated Hospital of China
Medical University, No. 4, Chongshan Eastern Road,
Yuhong District, Shenyang 110032, China
e-mail: cmu4h-mhy@126.com
Nine compounds (1–9) were obtained from the 95 % EtOH
(v/v) extract of leaves of Panax ginseng. During the process
of chemical study of this plant, three new compounds and six
known ones were isolated and identified as 3-O-b-D-gluco-
pyranosyl-20(S)-protopanaxtriol (1), 3-formyloxy-20-O-b-
D-glucopyranosyl-20(S)-protopanaxtriol (2), 26-hydroxyl-24
(E)-20(S)-protopanaxtriol (3), 20(S)-ginsenoside Rh1 (4)
[13], 20(R)-ginsenoside Rh1 (5) [13], ginsenoside F1 (6) [14],
H.-Y. Gao ꢀ J. Huang ꢀ B. Sun ꢀ B. Yang
Key Laboratory of Structure-Based Drug Design and Discovery
of Ministry of Education, Shenyang Pharmaceutical University,
No. 103, Wenhua Road, Shenhe District, Shenyang 110016,
China
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J Nat Med (2012) 66:576–582
577
Table 1 1H-NMR and 13C-NMR spectral data for 1 and 2 (d: ppm,
J in Hz, C5D5N)
No.
1
2
dH
dC
dH
dC
38.5
1
2
1.56 (m), 0.90 (m)
38.8 1.53 (m), 0.83 (td,
9.8, 3.4)
1.60 (m), 1.43
(overlap.)
27.0 1.58 (m), 1.43
(overlap.)
30.9
3
4
5
6
7
3.50 (dd, 12.0, 4.6)
89.4 4.76 (dd, 11.4, 4.8)
81.4
38.8
61.3
67.4
47.4
–
39.0
–
1.19 (d, 10.2)
4.37 (td, 10.2, 3.0)
2.04 (overlap.)
1.98 (t, 10.8)
–
61.7 1.14 (d, 10.8)
67.5 4.30 (overlap.)
47.4 1.86 (dd, 10.8, 2.4)
1.81 (overlap.)
8
40.5
50.0 1.47 (overlap.)
41.0
–
41.2
49.7
39.1
30.8
9
1.55 (overlap.)
–
10
11
–
2.07 (overlap.), 1.56
(overlap.)
31.3 1.96 (overlap.),
1.45 (overlap.)
12
13
14
15
3.96 (m)
70.9 4.13 (m)
70.2
49.2
51.7
31.0
2.04 (overlap.)
–
48.2 1.92 (overlap.)
51.6
–
1.64 (overlap.), 1.09
(overlap.)
31.4 1.58 (overlap.),
0.97 (m)
16
1.77 (m), 1.35 (m)
26.8 1.78 (overlap.),
1.29 (m)
26.7
17
18
19
20
21
22
2.36 (overlap.)
1.08 (3H, s)
0.95 (3H, s)
–
54.7 2.50 (dd, 10.8, 8.0)
17.4 1.60 (3H, s)
51.4
17.9
17.4
83.4
22.4
36.2
Fig. 1 Structures of compounds 1–3
ginsenoside Rk3 (7) [15], ginsenoside Rh4 (8) [16] and 20(R)-
ginsenoside Rg3 (9) [17], respectively.
17.3 0.93 (3H, s)
72.9
–
Compound 1 was isolated as a white powder (MeOH).
Its molecular formula was determined as C36H62O9 by
(HR) ESI-MS at m/z 661.4284 [M?Na]? (calcd. for
C36H62O9Na, 661.4286). The absorption bands at 3406,
1650 cm-1 in the IR spectrum indicated the presence of
hydroxyl and double-band functions in the structure. The
1H NMR (600 MHz, C5D5N) spectrum (Table 1) exhibited
signals due to eight methyls [dH 2.10, 1.67, 1.64, 1.08,
1.00, 0.95 (each 3H), 1.43 (3H 9 2)], one olefinic proton
[dH 5.34 (1H, t, J = 6.0 Hz)], an anomeric proton [dH 5.03
(1H, d, J = 7.6 Hz)] of a sugar unit and other alkyl groups.
The 13C NMR (150 MHz, C5D5N) spectrum exhibited 36
carbon signals including one glucose unit (dC 107.2, 78.7,
78.3, 75.9, 71.8, 63.0), two olefinic carbons (dC 130.7,
126.2), four O-bearing sp3 carbons (dC 67.5, 70.9, 72.9,
89.4), eight methyls (dC 32.0, 26.5, 25.8, 17.6, 17.4, 17.3,
17.0, 16.9), and other alkyl groups consisting of eight
methylene, four methine and four quaternary carbons. 30
carbon signals of aglycone, especially the signal at dC 61.8
(C-5) as a characteristic of a protopanaxtriol-type aglycone,
suggested that compound 1 was a derivative of proto-
panaxtriol (PPT) [3]; moreover, signals at dC 67.5, 70.9 and
1.43 (3H, s)
26.5 1.60 (3H, s)
2.05 (overlap.), 1.71
(m)
35.8 2.35 (td, 13.8, 3.8),
1.76 (overlap.)
23
2.63 (m), 2.30
(overlap.)
23.0 2.48 (m), 2.22 (m)
23.3
24
5.34 (t, 6.0)
–
126.2 5.21 (t, 6.0)
126.1
131.1
25.9
17.4
31.3
17.1
17.4
162.1
98.4
25
130.7
–
26
1.67 (3H, s)
1.64 (3H, s)
2.10 (3H, s)
1.43 (3H, s)
1.00 (3H, s)
–
25.8 1.59 (3H, s)
17.6 1.11 (3H, s)
32.0 1.60 (3H, s)
16.9 1.29 (3H, s)
17.0 0.95 (3H, s)
27
28
29
30
HCO
–
8.42 (1H, s)
Glc- 5.03 (d, 7.8)
10
107.2 5.16 (d, 7.8)
20
30
40
50
4.11 (m)
75.9 3.97 (t, 8.3)
78.7 4.21 (t, 8.3)
71.8 4.15 (t, 8.3)
75.3
79.5
71.7
78.5
4.28 (overlap.)
4.25 (overlap.)
4.03 (m)
78.3 3.90 (ddd, 8.3 5.3,
2.5)
60
4.62 (dd, 11.5, 2.5),
4.44 (dd, 11.5, 5.4)
63.0 4.30 (overlap.), 4.46
(dd, 5.7, 11.2)
63.0
123

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J Nat Med (2012) 66:576–582
OH
21
6'
4'
21
24
26
HO
O
2'
5'
22
O
HO
HO
OH
25
1'
26
20
23
OH
22
24
3'
25
27
23
20
OH
17
12
11
27
19
13
12
16
15
17
15
18
8
11
13
14
19
18
16
1
14
10
1
9
O
3
10
8
7
30
2
HMBC
5
30
6'
O
7
HO
6
3
5
4
4
5'
6
O
O
H
1'
28
29
HMBC
OH
OH 3'
OH
28
29
OH
OH
H
Fig. 2 Key HMBC correlations of compound 1
C
CH3
CH3
OH
CH3
H
72.9 indicated that the hydroxyl groups were presented at
C-6, C-12 and C-20 respectively. The D-glucose unit was
identified by comparison with an authentic sample of
D-glucose (No. 110833-200302) on TLC after enzymatic
hydrolysis of 1 using b-D-glucosidase which could furnish
the original unit [18]. According to the coupling constant
of the anomeric proton [dH 5.03 (1H, d, J = 7.8 Hz)], the
anomeric configuration was identified as the b form. In
the HMBC spectrum (Fig. 2), a crossing peak between the
anomeric proton and carbon C-3 (dC 89.4) meant that
glucose was linked with the C-3 position. On the basis of
the coincidence of chemical shifts in C-17 (dC 54.7), C-21
(dC 26.5) and C-22 (dC 35.8) with those for 20(S)-ginse-
noside Rh1 (4), the absolute configuration of the C-20
was determined to be S orientation [13]. Finally, compound
1 was elucidated as 3-O-b-D-glucopyranosyl-20(S)-proto-
panaxtriol.
12
10
1
H
O
6
3
H
H3C
H
CH3
HO
H
H
NOESY
Fig. 3 Key HMBC and NOE correlations of compound 2
C-20 was determined to be S orientation. Moreover, the
mutual correlations of CH3-29/CH3-19 and H-6 in the
NOESY spectrum (Fig. 3) agreed with all b-orientations of
CH3-29, CH3-19 and H-6 in the structure, and the corre-
lation between CH3-28 and H-3 confirmed that this for-
myloxy group should be placed with an b-orientation.
Thus, compound 2 was identified as 3-formyloxy-20-O-b-
D-glucopyranosyl-20(S)-protopanaxtriol.
Compound 2 was obtained as a white powder (MeOH).
Its (HR)ESI-MS at m/z 689.4238 [M?Na]? revealed a
molecular formula of C37H62O10 (calcd. for C37H62O10 Na,
689.4235). Its IR spectrum displayed absorption bands
Compound 3 gave the molecular formula C30H52O5 by
its pseudo molecular ion [M?H]? at m/z 493.3888 (calcd.
for C30H53O5, 493.3893) in the (HR)ESI-MS spectrum.
The IR (KBr) spectrum showed hydroxyl absorption at
3406 cm-1 and double bond at 1667 cm-1. In its 1H NMR
(600 MHz, C5D5N) (Table 2) spectra, seven methyls were
observed at dH 0.97, 1.00, 1.10, 1.44, 1.47, 1.83, 2.00 (3H
each, s), respectively. In addition, one olefinic proton sig-
nal at dH 5.86 (1H, t, J = 6.0 Hz) and two protons at dH
4.31 (2H, br s) for one oxomethylene were found. The 13C
NMR (150 MHz, C5D5N) spectrum exhibited 30 carbon
signals. Two signals at dC 125.7 (C-24), 136.2 (C-25) and
five signals at dC 61.8 (C-5), 78.5 (C-3), 67.7 (C-6), 71.1
(C-12), 73.0 (C-20) indicated that compound 3 was also a
derivative of protopanaxtriol (PPT) [19], which was com-
pletely confirmed by its HSQC and HMBC spectral anal-
ysis (Table 2). Moreover, the long-range correlations
between Ha,b-26 (dH 4.31) with C-27 (dC 14.0), and H-24
(dH 5.86) with C-26 (dC 68.2) suggested the presence of a
hydroxyl group at C-26. In addition, the NOE correlation
between Ha,b-26 and H-24 in the ROESY spectrum (Fig. 4)
indicated that the double bond at C-24 was a ‘‘trans-form’’.
arising from hydroxyl (3406 cm-1), carbonyl (1719 cm-1
)
and double bond (1650 cm-1). As well as these data from
IR and HR-MS experiments, compound 2 could be
deduced as a formyl ester derivative of ginsenoside F1(6)
not only because of the similar signals among their spectra
but also because of the presence of a proton signal at dH
8.42 (1H, s) and a carbonyl group at dC 162.1 (600 MHz
1
for H NMR, 150 MHz for 13C NMR, C5D5N) (Table 1).
The HMBC spectrum showed a long-range correlation
between dH 8.42 (1H, s) and a carbon at dC 81.4 (C-3),
which indicted the formyloxy group linked with C-3.
Compound 2 was therefore determined to be 3-formyloxy-
ginsenoside F1. The glucose unit was attached to C-20
(dC 83.4) on the basis of long-range correlation between
C-20 and the anomeric proton [dH 5.16 (1H, d,
J = 7.8 Hz)]. By means of the coincidence of chemical
shifts in C-17 (dC 51.4), C-21 (dC 22.4) and C-22 (dC 36.2)
with those for ginsenoside F1, the absolute configuration of
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J Nat Med (2012) 66:576–582
579
Table 2 1H-NMR and 13C-NMR spectral data for compound 3 (d:
ppm, J in Hz, C5D5N) and carbon spectral data for 27-hydroxyl-
20(S)-protopanaxatriol as reference
21
26
HO
22
OH
24
25
OH
23
20
17
No. Compound 3
27-Hydroxyl-20(S)-
protopanaxatriol
[20]a
12
27
11
19
13
14
16
18
8
15
dH
dC
HMBC
(H ? C)
dC
2
1
4
9
10
30
3
7
1
2
1.68 (overlap.),
1.02 (overlap.)
39.3 C-10, 19
39.3
28.1
5
HMBC
HO
6
28
1.92 (overlap.),
1.88 (overlap.)
28.1 C-1, 3,
4, 28
29
OH
3
4
5
3.52 (dd, 12.0, 6.0)
78.5 C-29
40.3
78.4
40.3
61.8
H
–
26
CH2
24
1.24 (d, 10.5)
61.8 C-6, 7,
19, 29
25
OH
23
6
7
4.42 (td, 15.0, 6.0)
67.7 C-4, 5, 7
47.4 C-8, 14
67.7
47.5
CH3
27
1.99 (dd, 17.5, 4.0),
1.92 (dd, 17.5, 5.0)
8
–
41.1
41.1
50.1
39.3
32.0
NOE effect
9
1.60 (overlap.)
–
50.1 C-1, 10
39.3
Fig. 4 Key HMBC correlations of compound 3, NOE correlation
between Ha,b-26 and H-24 in the ROESY spectrum
10
11 2.17 (dd, 6.6, 3.0),
1.59 (overlap.)
32.0 C-9, 13
Here, an interesting association between 3 and a known
compound 27-hydroxyl-20(S)-protopanaxatriol [20] was
found, and their almost identical signal patterns except for
three carbons at C-24, C-26 and C-27 [dC 125.7, 68.2, 14.0
in compound 3 versus dC 127.9, 21.9, 60.8, respectively, in
27-hydroxyl-20(S)-protopanaxatriol (Table 2)] indicated
that compound 3 was a isoformer of 27-hydroxyl-20(S)-
protopanaxatriol of C-24 double bond. However, according
to detailed information from refs. [21, 22], it was appro-
priate for the known structure to be changed to a cis-form
at C-24 and named as 27-hydroxyl-24(Z)-20(S)-proto-
panaxtriol. This conclusion could be drawn by the coinci-
dent chemical shifts for C-26, 27, which were recorded as
dC 21.8, 60.9 in the similar derivatives with a cis-form (Z-
isomer) at the C-24 position, whereas in the trans-form (E-
isomer) derivatives they were recorded as dC 68.2, 14.0,
respectively. In addition, NOE correlations in the ROESY
spectrum (Fig. 4) showed that compound 3 was a trans-
isomer of 27-hydroxyl-24(Z)-20(S)-protopanaxtriol, and its
structure was finally determined to be 26-hydroxyl-24(E)-
20(S)-protopanaxtriol.
12 3.94 (m)
71.1 C-13
48.2 C-17
51.6
71.0
48.2
51.6
31.3
13 2.04 (overlap.)
14
–
15 1.58 (overlap.),
1.05 (overlap.)
31.3 C-17, 30
16 1.87 (overlap.),
1.38 (overlap.)
26.8 C-15
26.8
17 2.35 (dd, 9.0, 3.0)
18 1.10 (3H, s)
54.7 C-20, 21
54.7
17.6
17.4 C-7, 8,
14
19 1.00 (3H, s)
17.5 C-1, 5,
9, 10
17.5
20
–
73.0
72.9
27.0
21 1.44 (3H, s)
27.1 C-17,
20, 22
22 2.10 (overlap.),
1.75 (overlap.)
35.7 C-21, 24
36.2
23 2.70 (m), 2.38 (m)
24 5.86 (t, 6.0)
22.6 C-20
22.5
125.7 C-21,
23, 26,
27
127.9
25
–
136.2 C-26, 27 136.2
26 1.83 (3H, s)
27 4.31 (2H, br s)
28 2.00 (3H, s)
29 1.47 (3H, s)
30 0.97 (3H, s)
68.2 C-27,
14.0 C-26
32.0 C-2
21.9
60.8
32.0
16.5
17.0
Cell growth inhibition by compounds
Human A549 and Hep3B cells were cultured with
0.1–100 lM of compounds 1–3 and compound 9 [20(R)-
ginsenoside Rg3] for 72 h, and the effects on growth
inhibition were measured. Compound 3 exhibited the most
potent cytotoxicity towards the two cell lines, with IC50
values of 27.9 lM and 31.4 lM, respectively. Compounds
16.4 C-3, 5
17.0 C-8, 14,
15
Bold values indicate an attention on the difference between com-
pound 3 and the reference
a
125 MHz, in pyridine-d5
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J Nat Med (2012) 66:576–582
1 and 2 showed moderate inhibition or weak effect on cell
growth at the same concentrations. It is worth mentioning that
compounds 1–3 showed more effective inhibition than 20(R)-
ginsenoside Rg3. The concentrations of ginsenosides applied
in some in-vitro experiments are as high as 100 lM level in
order to produce any biological effects; moreover, a place for
compound 9 in clinical treatments as an anti-tumor agent
implies potential for compounds 1–3 with much greater
inhibitory activity, but this conclusion must be proven by
further experiments in vivo in the continuing work.
time), and the solution was concentrated by vacuum to
obtain an extract (752 g), which was subjected to a macro-
reticulated absorption resin in AB-8 column and thor-
oughly washed with water, then eluted with 30, 70 and
90 % EtOH (v/v) successively. The 70 % EtOH eluate
(135 g) was chromatographed on a silica gel column with a
gradient CHCl3–MeOH solvent system (100:2, 100:5,
100:10, 100:20, 100:30, 100:50, 100:100, 0:100, each 3 L)
to give seven fractions (Fr. 1–7). Fr. 3 (CHCl3–MeOH
100:10, 13.7 g) was further subjected to reverse-phase
ODS silica gel column chromatography using MeOH–H2O
(45:55–70:30, v/v) to furnish three sub-fractions [SubFr.
3.1 (8.0 g), 3.2 (0.6 g), 3.3 (1.1 g)]. SubFr. 3.2 (0.6 g) was
also applied to HPLC using an ODS column and 73 %
MeOH to give compounds 1 (14 mg), 2 (30 mg), 7 (10 mg)
and 8 (37 mg); 3 g of SubFr. 3.1 was repeatedly purified on
HPLC (MeOH–H2O, UV210 detection) using a ODS col-
umn and 78 % MeOH solution to furnish compound 3
(4 mg), and using 67 % MeOH solution to furnish com-
pounds 4 (15 mg), 5 (15 mg) and 6 (110 mg); Fr. 4
(CHCl3–MeOH 100:20, 27.6 g) was subjected to reverse-
phase silica gel column chromatography using MeOH–
H2O (40:60–70:30, v/v) to give four sub-fractions [SubFr.
4.1 (13.2 g), 4.2 (1.9 g), 4.3 (5.1 g), 4.4 (0.4 g)]. Com-
pound 9 (17 mg) was obtained from SubFr. 4.3 by Si gel
column chromatography with CHCl3–MeOH (8:1) as an
eluate and then recrystallized with mixed solvent (CHCl3–
MeOH 6:1).
Experimental section
General experimental procedures
The infrared ray (IR) spectra were recorded on a Bruker IFS-
55 Fourier transform infrared spectrometer. Optical rotations
were recorded on a PerkinElmer 341 polarimeter at room
temperature. NMR spectra (1H, 13C, and 2D NMR) were
obtained on Bruker Avance 300 and AV-600 instruments
(Bruker, Rheinstetten, Germany) with TMS as internal stan-
dards. The high resolution electrospray ionization mass
spectra (HRESI-MS) were recorded on a Bruker micro-TOF-
Q mass spectrometer. HPLC were carried out on a Yonglin
SP930D liquid chromatograph equipped with a UV730D UV/
VIS detector using
a
Migtysil ODS-RP column
[250 9 20 mm (5 lm); Kanto Chemical Inc.] and monitored
at 210 nm. Column chromatography was performed on
macroreticulated absorption resin of AB-8, Si gel G
(200–300 mesh, Qingdao Ocean Chemical Inc., Qingdao,
China), using Sephadex LH-20 (Pharmacia, Kalamazoo, MI,
USA), and reversed-phase Si gel (Chromatorex C18, Fuji
Silysia Chemical, Kasugai, Japan). A Microplate Reader
(TECAN, Austria) was used for MTT assay. D-Glucose was
purchased from the Natural Institute for the Control of Phar-
maceutical and Biological Products (No. 110833-200302,
China), and b-D-glucosidase was obtained from Sigma
Company (Shanghai, China).
3-O-b-D-glucopyranosyl-20(S)-protopanaxtriol (1)
White powder (MeOH), m.p. 220 °C, a2D1 ?35.0 (c 0.1,
MeOH), IR(KBr) vmax 3406(OH), 2949, 1650 (C=C), 1453,
1385, 1030, 1019 cm-1, HRESI-MS: m/z 661.4284 [M?Na]?
(calcd. for C36H63O9Na, 661.4286); 1H NMR (600 Mz,
C5D5N) and 13C NMR (150 Mz, C5D5N) data see Table 1.
Enzymatic hydrolysis of 1
A solution of compound 1 (5.0 mg) in 0.2 M acetate buffer
(pH 4.5, 3.0 mL) was treated with b-D-glucosidase (15 mg),
and then stirred at 37 °C for 72 h. After treatment of the
reaction mixture with EtOH, the mixture was evaporated to
dryness under reduced pressure and the residue was dissolved
in MeOH and analyzed by TLC with developing agent
CHCl3:MeOH:H2O (30:10:1); spots were visualized by
spraying the plates with 10 % H2SO4 solution.
Plant material
Leaves of Panax ginseng were collected in Jilin province in
August 2008, and were provided by Fusong County Natural
Biotechnology Co., Ltd. The specimen was identified by
Professor Qishi Sun (Shenyang Pharmaceutical University,
Shenyang, China) and a voucher specimen (No. 2008-PG-
09) has been deposited in the same department.
3-Formyloxy-20-O-b-D-glucopyranosyl-20(S)-
protopanaxtriol (2)
Extraction and isolation
White powder (MeOH), m.p. 202–204 °C, a2D1 ?45.5 (c
0.1, MeOH). IR (KBr) vmax 3406 (OH), 2956, 1720 (C=O),
Air-dried leaves of P. ginseng (2.5 kg) were extracted with
95 % ethanol three times at room temperature (15 L each
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J Nat Med (2012) 66:576–582
581
Acknowledgments We thank Mrs. Wen Li, Mr. Yi Sha and Mrs.
Ying Peng of the Analytical Centers of the Shenyang Pharmaceutical
University for recording NMR and HRESI-MS spectra.
Table 3 Growth inhibition of compounds 1–3 and 9 against human
A549 and Hep3B cells
Cells
ICa50 (lM)
1
2
3
9
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Hep3B
a
63.9
48.7
66.5
27.9
31.4
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[100
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