Three new phenolic glucosides from the roots of Rheum palmatum

Article abstract of DOI:10.1248/cpb.60.241

A novel naphthalene glucoside, rheumone A (1), with an unprecedented skeleton containing a seven-membered lactone, and two new compounds, 1-O-phloroglucinyl-2-O-galloyl-6-O-cinnamoyl-β-d-glucoside (2) and chrysophanol 1-O-β-d-(6′-O-malonyl)glucoside (3), together with three known compounds (4-6) were isolated from the roots of Rheum palmatum. Their structures were elucidated mainly by spectroscopic analysis. These compounds were evaluated in vitro for their cytotoxicities towards human hepatocellular cancer cell lines Bel-7402 and Bel-7402/5Fu, and human gastric carcinoma cell line BGC-823. None of them showed cytotoxicity with IC₅₀ far beyond 50 μm.

Full text of DOI:10.1248/cpb.60.241

February 2012
Note
Chem. Pharm. Bull. 60(2) 241—245 (2012)
241
Three New Phenolic Glucosides from the Roots of Rheum palmatum
Zhi-Wei Wang, Jun-Song Wang, Jun Luo, Dan-Dan Wei, and Ling-Yi Kong*
Department of Natural Medicinal Chemistry, China Pharmaceutical University; 24 Tong Jia Xiang, Nanjing 210009,
People’s Republic of China.
Received June 18, 2011; accepted October 25, 2011; published online November 24, 2011
A novel naphthalene glucoside, rheumone A (1), with an unprecedented skeleton containing a seven-
membered lactone, and two new compounds, 1-O-phloroglucinyl-2-O-galloyl-6-O-cinnamoyl-β-d-glucoside
(
2) and chrysophanol 1-O-β-d-(6ꢀ-O-malonyl)glucoside (3), together with three known compounds (4—6) were
isolated from the roots of Rheum palmatum. Their structures were elucidated mainly by spectroscopic analy-
sis. These compounds were evaluated in vitro for their cytotoxicities towards human hepatocellular cancer
cell lines Bel-7402 and Bel-7402/5Fu, and human gastric carcinoma cell line BGC-823. None of them showed
cytotoxicity with IC50 far beyond 50μm.
Key words Rheum palmatum; phenolic glucoside; cytotoxicity
Rheum palmatum is one of the three genuine species of rhu- Results and Discussion
1)
barb listed in Chinese Pharmacopoeia. Its roots and rhizomes
Compound 1, obtained as a yellow powder, had a mo-
were generally used in traditional Chinese medicine (TCM) lecular formula of C H O as established by the high
2
1
22
9
2
)
to treat renal disorders and promote blood circulation. The resolution-electrospray ionization-mass spectra (HR-ESI-MS)
major constituents of rhubarb are a variety of phenolic com- pseudo-molecular ion at m/z 441.1161 [M+Na]+ (Calcd for
3,4)
5)
−1
pounds such as anthraquinones,
naphthalene glucosides
C H O Na: 441.1156). The absorption band at 1649cm in
and tannins. Strong purgative, antibacterial and antitumor the IR spectrum suggested the presence of a lactone group.
activities have been reported for these principles. In con- The H-NMR spectrum of 1 displayed five aromatic proton
tinuation of our searching bioactive natural products from signals at δH 7.75 (1H, d, J=7.5Hz), 7.43 (1H, d, J=8.5Hz),
TCMs, we investigated the constituents of Rheum palmatum. 7.36 (1H, t, J=7.5Hz), 7.01 (1H, s), 6.24 (1H, d, J=1.5Hz),
As a result, three new compounds, rheumone A (1), 1-O- a methylene resonances at δH 3.29 and 3.21 (each 1H, d,
21 22 9
6
)
7,8)
1
phloroglucinyl-2-O-galloyl-6-O-cinnamoyl-β-D-glucoside (2), J=13.5Hz), a methyl signal at δ 1.61 (3H, d, J=1.5Hz).
H
13
and chrysophanol 1-O-β-D-(6′-O-malonyl)glucoside (3), along Analysis of its C-NMR and heteronuclear single-quantum
with three known ones (4—6)9
—11)
were isolated. In this pa- coherence (HSQC) spectral data revealed that 1 contained 21
per, details for the isolation and structural elucidation of these carbon resonances, including six typical glycosyl moiety sig-
isolates, and the evaluation on their cytotoxicities against nals at δC 105.2, 80.1, 78.9, 75.2, 71.7, 62.9, an ester carbonyl
2
Bel-7402, Bel-7402/5Fu and BGC-823 cancer cell lines were carbon at 165.4, twelve sp carbons, one methylene and one
reported.
methyl group. The above evidence suggested 1 to be a naph-
thalene glycoside containing a lactone in its structure.
Fig. 1. Chemical Structures of Compounds 1—6
*
To whom correspondence should be addressed. e-mail: cpu_lykong@126.com
© 2012 The Pharmaceutical Society of Japan

2
42
Vol. 60, No. 2
Fig. 2. Key HMBC (→) and ROESY (↔) Correlations of 1
1
13
Long-range
HMBC) correlations (Fig. 2) from H-5 to C-5a, C-11a, C-4,
C-3, and CH -12; from CH -12 to C-4; and from H-3 to CH -
heteronuclear
multiple-bond
correlation Table 1. H- (500MHz) and C-NMR (125MHz) Data of 1 in C D N
5 5
(
No.
δH (J in Hz)
δC
3
3
3
1
2, and the ester carbon C-1 further confirmed the existence
1
3
4
5
165.4 (s)
of a seven-membered lactone. Meanwhile, the HMBC cor-
relations from H-6 to C-5, C-6a, C-11 (a weak J correlation),
C-10a, C-11a, C-1 (a weak J correlation); from H-7 to C-6,
C-6a, C-9; from H-8 to C-6a, C-9, C-10; and from H-9 to
C-10, C-10a established the naphthalene moiety. In the rotat-
ing-frame Overhauser effect spectroscopy (ROESY) spectrum
of 1, the correlations (Fig. 2) of CH -12, H-5, and H-7 with
H-6, and of H-3 and H-5 with CH -12 further confirmed the
6.24 (d, 1.5)
135.5 (d)
128.4 (s)
36.5 (t)
4
4
3.29 (d, 13.5)
3.21 (d, 13.5)
7.01 (s)
6
7
8
9
115.7 (d)
123.2 (d)
130.0 (d)
112.3 (d)
157.1 (s)
158.8 (s)
141.7 (s)
138.2 (s)
115.8 (s)
112.8 (s)
17.8 (q)
105.2 (d)
75.2 (d)
80.1 (d)
71.7 (d)
78.9 (d)
62.9 (t)
7.43 (d, 8.5)
7.36 (t, 7.5)
7.75 (d, 7.5)
3
3
10
structure of aglycone.
11
The sugar moiety was proved to be glucose by acid hy-
drolysis of 1 followed by co-TLC with authentic sugars. The
anomeric signal at δH 5.69 (1H, d, J=7.5Hz) indicated a
β-configuration for the glucose. The absolute configuration of
the sugar was determined to be D-glucose based on GC-MS
analysis of its chiral derivative. The HMBC correlation arising
from this anomeric proton signal to C-10 located the glucosyl
moiety at C-10, which was also confirmed by the ROESY cor-
relation between H-1′ and H-9. Consequently, the structure
of 1 was unambiguously established and named rheumone A,
the first naphthalene glucoside fused with a seven-membered
lactone ring.
5
a
a
6
1
0a
1a
-CH3
1
4
1.61 (d, 1.5)
5.69 (d, 7.5)
4.33 (*)
1′
2′
3′
4′
5′
6′
4.20 (m)
4.30 (*)
4.33 (*)
4.60 (dd, 2.0, 12.0)
4.41 (dd, 5.5, 12.0)
Compound 2 was obtained as a off-white amorphous
powder with a molecular formula of C H O as deduced
*
Refers to signal pattern unclear due to overlapping.
2
8
26 13
from the HR-ESI-MS pseudo-molecular ion at m/z 593.1258
+
[
M+Na] (Calcd for C H O Na: 593.1266).
connectivities of them. The HMBC correlations (Fig. 3) of
The presence of a galloyl group in 2 was evidenced by H-6, H-2′, and H-3′ with C-1′, of anomeric proton H-1 with
NMR signals at δH 7.15 (2H, s) and at δC 166.1, 146.0 (2C), C-1″, and of H-2 with C-1‴ and anomeric carbon C-1 posi-
38.9, 121.8, 110.1 (2C); the existence of a phloroglucinyl tioned the cinnamoyl, phloroglucinyl, and galloyl groups at
group in 2 was supported by the observed signals at δH 6.07 C-6, C-1, and C-2 position, respectively. Thus, the structure
2H, d, J=2.0Hz) and 6.02 (1H, t, J=2.5Hz), and at δ 160.4, of compound 2 was determined to be 1-O-phloroglucinyl-2-O-
2
8
26 13
1
(
1
C
59.9 (2C), 98.1, 96.7 (2C); a trans-cinnamoyl moiety was galloyl-6-O-cinnamoyl-β-D-glucoside.
indicated by resonances at δH 7.72 (2H, overlapped),7.45 (3H,
Compound 3 was isolated as a yellow powder, and its mo-
overlapped), trans-olefinic proton signals at δH 6.67 and 7.73 lecular formula of C H O was established by the observed
24
22 12
+
(
1
each 1H, d, J=16.0Hz), and by carbon resonances at δ 167.2, pseudo-molecular ion at m/z 525.1003 [M+Na] (Calcd for
45.7, 135.5, 131.1, 129.8 (2C), 129.1 (2C), 118.9. Carbon reso- C H O Na: 525.1003) in the HR-ESI-MS. It showed a posi-
nances at δ 100.2, 75.8, 75.1, 74.6, 71.5, 64.4 in the C-NMR tive Borntrager reaction, indicative of a hydroxyl anthraqui-
spectrum indicated the presence of a sugar moiety. The sugar none nature. Analysis of its C-NMR and HSQC spectral data
C
24 22 12
13
C
13
moiety was also identified as D-glucose in a same way to that revealed that 3 contained two typical carbonyl signals of an-
of 1. The relatively large coupling constant (J=8.0Hz) for the thraquinone at δ 189.2 and 183.1, and two other carbonyl sig-
C
anomeric proton suggested a β-configuration for the glucose.
nals at δ 170.0 and 168.5, twelve aromatic carbon resonances,
C
The presence of these units were further confirmed by one methyl group at δ 22.6, one methylene signal at δ 43.2,
C
C
HSQC and HMBC spectra, which also helped to establish the and a sugar moiety at δ 103.1, 78.7, 76.3, 75.3, 71.7, and 66.1.
C

February 2012
243
Fig. 3. Key HMBC Correlations of 2
Fig. 4. Key HMBC Correlations of 3
1
13
1
13
Table 2. H- (500MHz) and C-NMR (125MHz) Spectral Data of 2 in Table 3. H- (500MHz) and C-NMR (125MHz) Spectral Data of 3 in
Acetone-d6
C D N
5
5
No.
δH (J in Hz)
5.22 (d, 8.0)
δC
No.
δH (J in Hz)
δC
1
2
3
4
5
6
100.2 (d)
74.6 (d)
75.8 (d)
71.5 (d)
75.1 (d)
64.4 (t)
1
2
3
4
5
6
7
8
9
159.9 (s)
124.0 (d)
148.3 (s)
122.9 (d)
119.2 (d)
136.6 (d)
125.1 (d)
163.1 (s)
189.2 (s)
183.1 (s)
136.4 (s)
133.8 (s)
118.1 (s)
120.0 (s)
22.6 (q)
103.1 (d)
75.3 (d)
78.7 (d)
71.7 (d)
76.3 (d)
66.1 (t)
5.18 (m)
7.86 (s)
3.91 (m)
3.65 (dd, 9.0, 9.5)
3.91 (m)
7.91 (s)
7.86 (d, 7.5)
7.56 (dd, 7.5, 8.0)
7.34 (d, 8.0)
4.65 (dd, 2.0, 12.0)
4.37 (dd, 6.5, 12.0)
1′
167.2 (s)
118.9 (d)
145.7 (d)
135.5 (s)
129.1 (d)
129.8 (d)
131.1 (d)
129.8 (d)
129.1 (d)
160.4 (s)
96.7 (d)
159.9 (s)
98.1 (d)
159.9 (s)
96.7 (d)
166.1 (s)
121.8 (s)
110.1 (d)
146.0 (s)
138.9 (s)
146.0 (s)
110.1 (d)
2′
6.67 (d, 16.0)
7.73 (d, 16.0)
3′
4′
5′
6′
10
4a
10a
8a
7.72 (*)
7.45 (*)
7.45 (*)
7.45 (*)
7.72 (*)
7′
8′
9′
9a
3-CH3
2.44 (s)
1′
2′
3′
4′
5′
6′
5.78 (d, 8.0)
1″
4.55 (dd, 8.5, 8.5)
4.39 (*)
2″
3″
4″
5″
6″
6.07 (d, 2.0)
6.02 (t, 2.5)
6.07 (d, 2.0)
4.19 (dd, 9.5, 9.5)
4.39 (*)
5.24 (dd, 1.5, 13.0)
4.83 (dd, 7.5, 11.8)
1‴
1″
2″
168.5 (s)
43.2 (t)
2‴
3‴
4‴
5‴
6‴
7‴
3.83 (d, 3.0)
7.15 (s)
3″-COOH
170.0 (s)
*
Refers to signal pattern unclear due to overlapping.
7.15 (s)
same protocol for 1. Moreover, the observed anomeric
proton resonance at δH 5.78 (1H, d, J=8.0Hz) suggested a
β-configuration for the glucose. The down-field shifted methy-
*
Signal pattern unclear due to overlapping.
1
The H-NMR spectrum of 3 showed the presence of one meth- lene carbon signal of the glucose indicated that the glucose
yl group at δH 2.44 (3H, s), one methylene signal at δH 3.83 was substituted at C-6′. The substituent was determined to be
2H, d, J=3.0Hz), and five aromatic protons: two singlet at δH a malonyl group by the HMBC correlations between H-6′ and
.91 (1H, s) and 7.86 (1H, s), and three mutually ortho-coupled one of the carbonyl carbon at δC 168.5 (C-1″), which and the
at δH 7.86 (1H, d, J=7.5Hz), 7.56 (1H, dd, J=7.5, 8.0Hz) and second such type of carbon at δC 170.0 both correlated with
the methylene protons at δH 3.83 (2H, d, J=3.0Hz). Finally,
In the HMBC spectrum (Fig. 4), the correlations of H-2 the HMBC correlation between the anomeric proton and C-1
with C-1, C-3, 3-CH , C-4, and C-4a (a weak J correlation), located the glucosyl moiety at C-1. Thus, the structure of 3
(
7
7
.34 (1H, d, J=8.0Hz).
4
3
of H-4 with 3-CH , C-9a, and C-10, of H-5 with C-7, C-10, was established as chrysophanol 1-O-β-D-(6′-O-malonyl)glu-
3
and C-8a, and of H-6 with C-8 and C-10a established the agly- coside.
cone as chrysophanol.
All of the isolated compounds were evaluated in vitro for
The sugar unit was identified as D-glucose followed the their cytotoxicity against Bel-7402, Bel-7402/5Fu and BGC-

2
44
Vol. 60, No. 2
8
23 cancer cell lines. None of them showed cytotoxicity with
6-Hydroxymusizin Glucoside (4): Pale yellow needles
1
IC50 far beyond 50μM.
(MeOH); H-NMR (500MHz, acetone-d ) δ : 7.09 (1H, d,
6
H
J=2.0Hz, H-5), 6.94 (1H, s, H-4), 6.77 (1H, d, J=2.5Hz, H-7),
Experimental
2.53 (3H, s, 2-COCH ), 2.26 (3H, s, 3-CH ); Glc: 5.16 (1H,
3
3
General Procedures NMR spectra were recorded on d, J=7.5Hz, H-1′), 3.48—3.66 (4H, m, H-2′, 3′, 4′, 5′), 3.77
1
13
Bruker ACF-500 NMR instruments ( H: 500MHz, C: (1H, dd, J=6.0, 12.0Hz, H-6′a), 4.00 (1H, dd, J=2.0, 12.0Hz,
13
1
25MHz), with tetramethylsilane (TMS) as internal standard. H-6′b); C-NMR (125MHz, acetone-d ) δ : 204.7 (2-COCH ),
6 C 3
Mass spectra were obtained on a MS Agilent 1100 series LC/ 157.8 (C-1), 157.0 (C-6), 153.3 (C-8), 138.8 (C-4a), 135.0 (C-3),
MSD ion-trap mass spectrometer (ESI-MS) and a Mariner 123.3 (C-2), 119.2 (C-4), 109.3 (C-8a), 105.0 (C-5), 104.4 (C-7),
ESI-time-of-flight (TOF) spectrometer (HR-ESI-MS), respec- 32.5 (2-COCH ), 20.2 (3-CH ); Glc: 104.0 (C-1′), 78.5 (C-3′),
3
3
tively. IR (KBr disks) spectra were recorded by Bruker Tensor 77.8 (C-5′), 74.6 (C-2′), 71.3 (C-4′), 62.6 (C-6′).
2
7 spectrometer. UV spectra were measured on a Shimadzu
Torachrysone Glucoside (5): Pale yellow needles (MeOH);
H-NMR (500MHz, acetone-d ) δ : 7.07 (1H, s, H-4), 7.04
1
UV-2501 PC spectrophotometer. Optical rotations were con-
6
H
ducted on a JASCO P-1020 polarimeter. RP-C18 (40—63μm, (1H, d, J=2.0Hz, H-5), 6.89 (1H, d, J=2.5Hz, H-7), 3.89 (3H,
Fuji) were used for column chromatography. Preparative s, 6-CH ), 2.53 (3H, s, 2-COCH ), 2.28 (3H, s, 3-CH ); Glc:
3
3
3
HPLC was carried out using an Agilent 1100 Series instru- 5.21 (1H, d, J=7.5Hz, H-1′), 3.51—3.66 (4H, m, H-2′, 3′, 4′,
ment with a shim-park RP-C18 column (20×200mm) and a 5′), 3.76 (1H, dd, J=6.0, 12.0Hz, H-6′a), 3.97 (1H, dd, J=2.5,
1
3
1
100 Series multiple wavelength detector. The GC-MS system 12.0Hz, H-6′b); C-NMR (125MHz, acetone-d ) δ : 204.7
6 C
consisted of a Varian CP-3800 gas chromatograph interfaced (2-COCH ), 159.8 (C-1), 156.8 (C-6), 152.9 (C-8), 138.5 (C-4a),
3
with a Varian Saturn 2200 mass spectrometer.
135.2 (C-3), 124.2 (C-2), 119.8 (C-4), 110.0 (C-8a), 104.0 (C-5),
Plant Material The roots of Rheum palmatum were col- 103.9 (C-7), 55.8 (6-OCH ), 32.4 (2-COCH ), 20.1 (3-CH );
3
3
3
lected from Li county, Gansu province, China, in August 2010, Glc: 102.2 (C-1′), 78.6 (C-3′), 77.8 (C-5′), 74.6 (C-2′), 71.2 (C-
and were authenticated by Professor Min-Jian Qin of Research 4′), 62.4 (C-6′).
Department of Pharmacognosy, China Pharmaceutical
University. The voucher specimen (No. RP-201008) was de- powder; H-NMR (500MHz, DMSO-d ) δ : 12.84 (1H,
Chrysophanol 8-O-β-D-(6′-O-Malonyl)glucoside (6): Yellow
1
6
H
posited at the Department of Natural Medicinal Chemistry, s, 1-OH), 12.78 (1H, brs, 3″-COOH), 7.88 (1H, dd, J=7.5,
China Pharmaceutical University.
7.5Hz, H-6), 7.88 (1H, d, J=7.5Hz, H-5), 7.65 (1H, dd, J=2.0,
Extraction and Isolation The air-dried and powdered 8.0Hz, H-7), 7.51 (1H, brs, H-4), 7.2 (1H, brs, H-2), 3.39
roots of R. palmatum (500g) were extracted with EtOH under (2H, d, J=9.0Hz, H-2″), 2.43 (3H, s, 3-CH ); Glc: 5.19 (1H,
3
reflux and then filtered by filter paper. The EtOH extract was d, J=8.0Hz, H-1′), 3.25—3.73 (4H, m, H-2′, 3′, 4′, 5′), 4.12
concentrated by evaporation; the residue (52g) was suspended (1H, dd, J=7.0, 12.0Hz, H-6′a), 4.43 (1H, dd, J=1.5, 12.0Hz,
1
3
in H O and partitioned with EtOAc. The EtOAC fraction H-6′b); C-NMR (125MHz, DMSO-d ) δ : 187.5 (C-9), 182.1
2
6
C
(
13g) was subjected to MCI gel and eluted with MeOH/H O (C-10), 167.9 (C-3″), 166.8 (C-1″), 161.6 (C-1), 157.9 (C-8),
2
(
0:10, 2:8, 5:5, 10:0) to give four further fractions, E1—E4. 147.6 (C-3), 136.0 (C-6), 134.8 (C-10a), 132.1 (C-4a), 124.1
Fraction E3 was subsequently subjected to ODS open column (C-4), 122.4 (C-5), 120.7 (C-7), 120.6 (C-2), 119.4 (C-8a), 114.8
chromatography (MeOH/H O, 30:70 to 50:50) to afford (C-9a), 41.5 (C-2″), 21.5 (3-CH ); Glc: 100.2 (C-1′), 76.3 (C-3′),
2
3
fractions EO1 to EO3. Fraction EO1 was further purified by 73.9 (C-5′), 73.2 (C-2′), 69.6 (C-4′), 64.1 (C-6′).
preparative HPLC (MeOH/H O, 35:65) to yield 4 (18mg).
Sugar Analysis Compounds 1—3 (4mg each) dissolved in
Fraction EO2 was purified by preparative HPLC (MeOH/H O, acetonitrile (3mL) and 2M HCl (3mL) were refluxed at 90°C
2
2
4
0:60) to give 1 (14mg), 2 (23mg) and 5 (45mg). Fraction for 3h. The reactant solution was concentrated and the residue
EO3 was purified by preparative HPLC(acetonitrile/H O, was suspended with H O and extracted with EtOAc (3mL×3).
2
2
2
8:72) to afford 3 (25mg) and 6 (35mg).
Rheumone A (1): Yellow powder, [α]D −38.4 (c=0.03, with authentic sugars developed with CH Cl –MeOH–H O
The aqueous layer was concentrated and examined by co-TLC
2
5
2
2
2
pyridine); UV λmax (acetonitrile) nm (logε): 350 (3.03), 234 (6:4:1). A portion of the aqueous layer was evaporated to
−1
(
3.77); IR (KBr) cm : 3496, 2961, 2918, 1649, 1629, 1571; dryness and then dissolved in anhydrous pyridine (0.3mL), to
1
13
H- and C-NMR spectral data: see Table 1; ESI-MS m/z: which L-cysteine methyl ester hydrochloride (5mg) was added.
−
+
4
17.0 [M−H] ; HR-ESI-MS m/z: 441.1161 [M+Na] (Calcd for The mixture stayed warm at 60°C for 2h, to which hexameth-
C H O Na: 441.1156).
yldisilazan (0.3mL) and trimethylsilyl chloride (0.3mL) were
21
22
9
1
-O-Phloroglucinyl-2-O-galloyl-6-O-cinnamoyl-β-D- added, kept warm at 60°C for another 1h. The final reactant
2
5
glucoside (2): Off-white amorphous powder, [α] : −27.2 was partitioned between H O and n-hexane (each 0.3mL),
D
2
(
c=0.07, acetone); UV λ
(MeOH) nm (logε): 276 (4.56), and the n-hexane extract was subjected to GC-MS (70eV)
05 (4.90); IR (KBr) cm : 3422, 2963, 1703, 1608; H- and analysis under the following conditions: column, CP-sil 5 CB
max
−
1
1
2
13
C-NMR spectral data: see Table 2; ESI-MS m/z: 569.1 (15m×0.25mm×0.25µm); column temperature, 150—260°C
−
+
[
M−H] ; HR-ESI-MS m/z: 593.1258 [M+Na] (Calcd for with an increase rate of 8°C/min; carrier gas, He (1mL/min).
In the acid hydrolysates of 1—3, only D-glucose was detected
Chrysophanol 1-O-β-D-(6′-O-Malonyl)glucoside (3): Yellow by comparision of the retention time (tR 15.24min) of their
C H O Na: 593.1266).
2
8
26 13
2
5
powder, [α] : −47.9 (c=0.07, acetone); UV λ
(MeOH) nm derivatives with that of the authentic D-glucose derivative pre-
D
max
−1
(
2
logε): 409 (3.66), 258 (4.16), 222 (4.30); IR (KBr) cm : 3421, pared in the same way.
921, 1733, 1672, 1632, 1599; H- and C-NMR spectral data:
see Table 3; ESI-MS m/z: 500.5 [M−H] ; HR-ESI-MS m/z: their in vitro cytotoxicities using human hepatocellular can-
25.1003 [M+Na] (Calcd for C H O Na: 525.1003).
1
13
Cytotoxic Activity All these isolates were evaluated for
−
+
5
cer cell lines Bel-7402 and Bel-7402/5Fu, and human gastric
2
4
22 12

February 2012
245
Phytochemistry, 31, 1063—1065 (1992).
Okabe H., Matsuo K., Nishioka I., Chem. Pharm. Bull., 21, 1254—
260 (1973).
Tsuboi M., Minami M., Nonaka G., Nishioka I., Chem. Pharm.
Bull., 25, 2708—2712 (1977).
carcinoma cell line BGC-823 by standard 3-(4,5-dimethylthi-
azol-2-yl)2,5-diphenyltetrazolium bromide (MTT) colorimetric
method with 5-fluorouracil as a positive control.
4
)
1
12)
5
6
)
)
Acknowledgements This research work was financially
supported by the Key Project of National Natural Science
Foundation of China (30830116), the National Key Scientific
and Technological Special Projects (2009ZX09502-011;
Kashiwada Y., Nonaka G. I., Nishioka I., Yamagishi T.,
Phytochemistry, 27, 1473—1477 (1988).
7) Oshio H., Naruse Y., Tsukui M., Chem. Pharm. Bull., 26, 2458—
2464 (1978).
2
009ZX09308-005), and the Project Funded by the Priority
8) Komatsu K., Nagayama Y., Tanaka K., Ling Y., Basnet P., Meselhy
M. R., Chem. Pharm. Bull., 54, 941—947 (2006).
Academic Program Development of Jiangsu Higher Education
Institutions.
9
)
Lemli J., Toppet S., Cuveele J., Janssen G., Planta Med., 43, 11—17
1981).
0) Yang X. W., Zhao J., Zhang Y., Li J. X., Ma C. M., Chin. Tradit.
Herb. Drugs, 29, 5—11 (1998).
(
1
References
1
)
Chinese Pharmacopoeia Commission, “Chinese Pharmacopoeia I,”
Chemical Industry Press, Beijing, 2005, p. 17.
1
1) Yang X. W., Ma C. M., Masao H., Tsuneo N., Chin. Tradit. Herb.
Drugs, 29, 289—293 (1998).
2) Zhang Y., Wang J. S., Wei D. D., Wang X. B., Luo J., Luo J. G.,
2
)
Shibutani S., Nagasawa T., Oura H., Nonaka G., Nishioka I., Chem.
Pharm. Bull., 31, 2378—2385 (1983).
1
Kong L. Y., Phytochemistry, 71, 2199—2204 (2010).
3
)
Kubo I., Murai Y., Soediro I., Soetarno S., Sastrodihardjo S.,