A new phenolic glycoside and cytotoxic constituents from Celosia argentea

Article abstract of DOI:10.1080/10286020.2010.503652

A new phenolic glycoside, 4-O β- D-apifuranosyl-(12) β- D-glucopyranosyl-2-hydroxy-6-methoxyacetophenone (2) and 11 known compounds were isolated from the MeOH extract of the plant Celosia argentea. The structures of the compounds were elucidated on the basis of spectroscopic analysis and chemical methods. Among the isolated compounds, stigmasterol (10) showed moderate inhibitory activities against SGC-7901 and BEL-7404 cells.

Full text of DOI:10.1080/10286020.2010.503652

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A new phenolic glycoside and cytotoxic
constituents from Celosia argentea
Shuo Shen a b , Xiao Ding c , Ming-An Ouyang c , Zu-Jian Wu a &
Lian-Hui Xie a
a Institute of Plant Virology, Fujian Agriculture and Forestry
University, Fuzhou, 350002, China
b Institute of Biotechnology of Qinghai Academy of Agriculture and
Forestry, Xining, 810016, China
c Key Laboratory of Bio-Pesticide and Chemistry-Biology, Ministry
of Education, Fujian Agriculture and Forestry University, Fuzhou,
350002, China
Published online: 13 Sep 2010.
To cite this article: Shuo Shen , Xiao Ding , Ming-An Ouyang , Zu-Jian Wu & Lian-Hui Xie (2010): A
new phenolic glycoside and cytotoxic constituents from Celosia argentea , Journal of Asian Natural
Products Research, 12:9, 821-827
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Journal of Asian Natural Products Research
Vol. 12, No. 9, September 2010, 821–827
NOTE
A new phenolic glycoside and cytotoxic constituents from
Celosia argentea
Shuo Shenab, Xiao Dingc, Ming-An Ouyangc*, Zu-Jian Wua and Lian-Hui Xiea
aInstitute of Plant Virology, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
bInstitute of Biotechnology of Qinghai Academy of Agriculture and Forestry, Xining 810016, China;
cKey Laboratory of Bio-Pesticide and Chemistry-Biology, Ministry of Education, Fujian Agriculture
and Forestry University, Fuzhou 350002, China
(Received 26 March 2010; final version received 21 June 2010)
A new phenolic glycoside, 4-O-b-D-apifuranosyl-(1 ! 2)-b-D-glucopyranosyl-2-
hydroxy-6-methoxyacetophenone (2) and 11 known compounds were isolated from
the MeOH extract of the plant Celosia argentea. The structures of the compounds were
elucidated on the basis of spectroscopic analysis and chemical methods. Among the
isolated compounds, stigmasterol (10) showed moderate inhibitory activities against
SGC-7901 and BEL-7404 cells.
Keywords: Amaranthaceae; Celosia argentea; phenolic glycoside; cytotoxicity; SGC-
7901 cells; BEL-7404 cells
1. Introduction
important roles in mitosis, cell signaling,
and motility in eucaryotes [8].
Celosia argentea (Amaranthaceae) is a
kind of annual herbaceous plant. Its seeds
are used as a Chinese herbal medicine [1].
Early researches on C. argentea indicated
that its seeds were pharmaceutically active
on eyes and displayed hepatoprotective [2],
protecting oxidative injury of lens [3], anti-
hyperglycemic [4], and anti-metastatic
effects [5]. The water and alcoholic
extracts of the leaves of the title plant
also exhibited antimicrobial activities [6].
Betalains [7] and peptides [8] are found
from the seeds of C. argentea. Betalains
[9], indole derivatives, were suggested to
have antioxidant activity in vivo and
radioprotective activity in vivo and
in vitro. Cyclic peptides were reported to
display antimitotic activity, which played
In this paper, we describe the isolation
and structural elucidation of a new
phenolic glycoside, 4-O-b-D-apifurano-
syl-(1 ! 2)-b-D-glucopyranosyl-2-hydro-
xy-6-methoxyacetophenone (2), along
with 11 known compounds: 4-O-b-D-
glucopyranosyl-2-hydroxy-6-methoxyace-
tophenone (1) [10], eugenyl-O-b-D-gluco-
pyranoside (3) [11], sucrose (4) [12],
quercetin-3-O-b-D-glucopyranoside (5)
[13], isorhamnetin-3-O-b-D-glucopyrano-
side (6) [14], rhamnazin-3-O-b-D-gluco-
pyranoside (7) [15], isorhamnetin-3-O-a-
L-rhamnopyranosyl-(1 ! 2)-b-D-gluco-
pyranoside (8) [16], b-sitosterol (9) [17],
stigmasterol (10) [18], stigmasterol-3-O-
b-D-glucopyranoside (11) [19], and
*Corresponding author. Email: maouyang@hqu.edu.cn
ISSN 1028-6020 print/ISSN 1477-2213 online
q 2010 Taylor & Francis
DOI: 10.1080/10286020.2010.503652
http://www.informaworld.com

822
S. Shen et al.
OH
6
8
9
2
6
HO
O
4
1
HO
O
6'
1
4
7
O
O
CH3
8
C
7
OH
OH
O
R
3
OH
3
4'
1'
H3CO
OH
2'
OCH3
OH
1 R = OH
2 R = D-apiose
Figure 1. Structures of compounds 1–3.
oleanolic acid (12) [20] (Figure 1). Then,
we test their cytotoxicities against SGC-
7901 and BEL-7404 cells. It was found
that compound 10 had moderate cytotox-
icity against both cells at the measured
concentration.
163.6, 107.3, 97.5, and 92.4 (Table 1).
Because of the intramolecular hydrogen
bond, the ketone position should be
presumed to be located ortho to the
hydroxyl group of the aromatic ring.
1
Comparison of H and 13C NMR spectral
data of 2 with those reported for compound
1 indicated that a D-apiose moiety was
located at C-20 in 2 instead of the hydroxy
group in 1 (Figure 1). In the HMBC
experiment, the hydrogen-bonded pheno-
lic hydroxyl proton afforded all of the
possible two- and three-bond correlations,
leading to the assignment of the carbon
signals for C-1, C-2, and C-3. Further-
more, the aryl proton H-3 correlated with
C-1, C-2, C-4, and C-5; the aryl proton H-5
with C-1, C-3, C-4, and C-6; the methyl
proton H-8 with C-1 and C-7; the methoxy
protons with C-6; the anomeric proton of
Glc with C-4; and the anomeric proton of
Api with C-2 of Glc (Figure 2). The above
HMBC correlations revealed the structure
of 2 as 4-O-b-D-apifuranosyl-(1 ! 2)-b-
D-glucopyranosyl-2-hydroxy-6-methoxy-
acetophenone, named as argentoside A. In
the NOESY experiment (Figure 2), the key
correlations between H-5 and OCH3 at
C-6, between H-1 of Glc and H-3 and H-5
were observed. Thus, the structure of 2
was determined unambiguously as shown
in Figure 1.
2. Results and discussion
Compound 2 was obtained as a colorless
amorphous powder, whose molecular
formula was determined to be C20H28O13
(seven unsaturation degrees) by obser-
vation of a quasi-molecular ion peak at m/z
475.1450 [M 2 H]2 in the positive ion
HR-FAB mass spectrum. The IR spectrum
indicated the presence of a hydroxyl
group (3390 cm21), a conjugated ketone
(1693 cm21), and
a
benzene ring
(1601 cm21). Acid hydrolysis afforded a
mixture of sugars, which were identified as
D-glucose and D-apiose detected by
HPTLC. The 1H NMR spectrum of 2
showed aromatic protons at dH 6.75 (1H, d,
J ¼ 1.8 Hz) and 6.51 (1H, d, J ¼ 1.8 Hz),
a methyl signal at dH 2.50 (3H, s), a
methoxyl group at dH 3.76 (3H, s), and two
anomeric protons at dH 6.57 (1H, s) and
1
5.59 (1H, d, J ¼ 7.8 Hz). The H NMR
spectrum of 2 also revealed the presence of
an intramolecularly hydrogen-bonded,
phenolic hydroxyl group (dH 14.29) and
permitted the assignment of a tetra-
substituted benzene ring. The 13C NMR
spectrum of 2 gave a carbonyl resonance at
dC 203.7, a methyl signal at dC 33.1, two
anomeric carbons at dC 110.5, 99.9, and an
aromatic ring signal at dC 167.4, 164.9,
Table 2 showed the cytotoxic activities
of all compounds. Among these, compound
10 displayed significant inhibition, 86.96%
against SGC-7901 cells and 82.74%
against BEL-7404 cells, respectively.

Journal of Asian Natural Products Research
823

824
S. Shen et al.
OH
OH
O
O
C
HO
HO
O
C
CH3
O
CH3
O
O
O
O
OH
O
OH
O
OCH3
OCH3
OH
OH
HO
HO
NOESY
HMBC
OH
OH
OH
OH
Figure 2. Key HMBC and NOESY correlations of compound 2.
Table 2. Cytotoxic activities of compounds 1–12.
Compound no.
Inhibition ratea (%)
Inhibition rateb (%)
c
1
2
3
4
5
6
7
8
9
10
11
12
–
–
14.73 ^ 0.05
14.82 ^ 0.25
–
21.65 ^ 0.04
16.26 ^ 0.03
18.51 ^ 0.07
29.20 ^ 0.06
28.48 ^ 0.01
82.74 ^ 0.11
–
–
–
10.80 ^ 0.19
19.36 ^ 0.17
27.00 ^ 0.14
–
14.44 ^ 0.13
–
86.96 ^ 0.02
–
49.05 ^ 0.07
85.59 ^ 0.41
84.77 ^ 0.98
31.09 ^ 0.06
83.45 ^ 3.28
89.79 ^ 1.78
5-FUd
CDDPd
Notes: Compounds’ concentration is 200 mg/ml.
a Inhibition rate of compounds against SGC-7901 cells.
b Inhibition rate of compounds against BEL-7404 cells.
c No inhibitory activity.
d 5-FU diluted in 50 mg/ml and CDDP diluted in 20 mg/ml, served as positive controls.
Table 3. The cytotoxic activity of compound 10 (IC50).
a
IC50 (mg/ml ^ SD)
b
IC50 (mg/ml ^ SD)
Compound no.
10
5-FUc
CDDPc
44.00 ^ 0.16
1.01 ^ 0.04
1.37 ^ 0.26
52.06 ^ 3.28
1.45 ^ 0.21
0.72 ^ 0.11
Notes: aCytotoxicity against SGC-7901 cells.
b Cytotoxicity against BEL-7404 cells.
c Positive controls.

Journal of Asian Natural Products Research
825
But compounds 9 and 11, possessing
similar structures of their aglycones,
were hardly active against both cells. The
reason should be due to a glucose linking
at position C-3 in compound 11, and a
CvC double bond at the position C-20 in
compound 9. Compounds 5–8, flavonoid
glycosides, with the same skeleton but
differing mainly in their substituted
groups, showed inhibitory activities from
14.44 to 29.20%, except for compound 7,
which showed no inhibitory activity
against SGC-7901 cells. The IC50 values
of compound 10 were 44.00 mg/ml against
SGC-7901 cells and 52.06 mg/ml against
BEL-7404 cells (see Table 3).
layer 9 þ 1 ml HOAc for sugars. Com-
pounds on TLC were detected by spraying
with 5% H2SO4, followed by heating.
Sugars were detected by spraying with
aniline–phthalate reagent.
50-Fluorouracial
(Sigma-Aldrich, Inc., St Louis, MO,
USA), cis-Platin (CDDP, $ 99%)
(5-FU, $ 99%)
(Sigma-Aldrich, Inc.), RPMI-1640 med-
ium (Hyclone Biochemical Products Co.
Ltd, Logan, UT, USA), penicillin–strep-
tomycin solution (Hyclone Biochemical
Products Co. Ltd), Hepes (Sigma-Aldrich,
Inc.), methylthiazolyldiphenyl-tetrazolium
bromide (MTT) (Sigma-Aldrich, Inc.), and
fetal bovine serum (Hyclone, Thermo
Scientific, Tauranga, New Zealand) were
all purchased from Xiamen Lulong Bio-
tech Co. Ltd, Xiamen, China.
3. Experimental
3.1 General experimental procedures
The 1H, 13C, DEPT, 1H–1H COSY,
NOESY, HMQC, and HMBC NMR spectra
were performed using a Bruker AM-400
and DRX-500 spectrometer. FAB mass
spectra were recorded on a Jeol JMS-HX
110 instrument. IR spectra were recorded
with a Perkin-Elmer 1750 FT-IR spec-
trometer and the films of all the samples
were measured on KBr disks. Optical
rotations were measured with a Jasco DIP-
180 digital polarimeter spectrophotometer.
Chromatographic stationary phase used
RP-18 (40–60mm; Merck & Co. Inc.,
Darmstadt, Germany), silica gel (160–
200 mesh; Qingdao Oceanic Chemical
Co., Qingdao, China), Sephadex LH-20
(25–100 mm; Pharmacia Fine Chemical
Co. Ltd, Tokyo, Japan), and MCI-gel
CHP20P (75–150 mm; Mitsubishi Chemi-
cal Industries, Ltd, Tokyo, Japan). HPLC
was performed on a P-230-UV-230 (Dalian
Elite Analytical Instruments Co., Ltd,
Dalian, China) and HPLC column (YMC-
Pack ODS-A, 250–10 mm; Kyoto, Japan).
The following solvent systems were used:
(a) CHCl3 –MeOH–H2O (80:20:3),
CHCl3 –MeOH–H2O (70:30:5), and
MeOH–H2O (0–100%) for the glycosides
and (b) CHCl3–MeOH–H2O (7:3:1) lower-
3.2 Plant material
C. argentea was collected from the suburb
of Quanzhou, Fujian in 2003. The speci-
men was identified by Prof. Ke-Cuo He,
College of Plant Protection, Fujian Agri-
culture and Forestry University. A voucher
specimen (No. 4316) has been deposited in
the College of Plant Protection, Fujian
Agriculture and Forestry University.
3.3 Extraction and isolation
The fresh C. argentea (10 kg) was
extracted with MeOH (2 £ 15 liters) at
room temperature. The extract was evap-
orated in vacuo to yield a residue, which
was dissolved in water and filtered. The
water soluble fraction was passed through
a Diaion column and eluted with water
(4 £ 3 liters) and methanol (4 £ 1 liters).
Evaporation of the methanol eluate
yielded 386 g of a brown fraction (A).
Fraction A was subjected to dry column
chromatography (DCC) on silica gel
(3.0 kg), eluted with CHCl3 –MeOH–
H2O (10:2:0.2) to afford 13 fractions.
Each fraction was purified by Sephadex
LH-20 (MeOH–H2O, 5–50%), RP-18 gel
column chromatography (MeOH–H2O,

826
S. Shen et al.
10–70%), then purified by a silica gel
column with CHCl3 –MeOH–H2O
(100:10:1–70:30:5), and finally repeatedly
purified by RP-HPLC with MeOH–H2O
(60–80%) as the solvent to yield 1
(300 mg), 2 (50 mg), 3 (120 mg), 4
(31 mg), 5 (38 mg), 6 (24 mg), 7 (30 mg),
and 8 (47 mg). Water insoluble fraction
was treated by DCC on silica gel (1.0 kg),
eluted with CHCl3–MeOH (30:1–10:2) to
afford 9 (23 mg), 10 (41 mg), 11 (23 mg),
and 12 (19 mg).
grown in media containing equivalent
amounts of DMSO without compound
treatment served as a negative control.
3.6 MTT assay
Cancer cells were plated onto 96-well
culture dishes at a density of 1 £ 103
cells/well in a 180 ml medium. After
plating for 24 h, cells were treated with
compounds ranging from 12.5 to
200 mg/ml (using DMSO as the vehicle at
a maximum concentration of 0.1%). Cells
were incubated with various concen-
trations of the agents for 72 h, at which
time 20 ml of 2 mg/ml MTT was added,
and the absorbance at 570 nm was
determined by a microtiter plate reader
[21]. 5-FU at a concentration of 50 mg/ml
and CDDP at a concentration of 20 mg/ml
were used as the positive controls.
Experiments were conducted in triplicate.
Cytotoxicity was expressed as IC50 (con-
centration of 50% cytotoxicity, which was
extrapolated from linear regression anal-
ysis of the experimental data).
3.3.1 Compound (2)
A colorless amorphous powder, [a ]2D1 217
(c ¼ 0.22, MeOH); IR nmax (liquid)/cm21
:
3390, 1693, 1601, 1450, 1012, 903; 1H and
13C NMR spectral data (see Table 1); FAB-
MS: m/z 475 [M 2 H]2; HR-FAB-MS:
m/z 475.1450 [M 2 H]2 (calcd for
C20H27O13, 475.1452).
3.4 Acid hydrolysis
A solution of compound 2 (50 mg) was
heated under reflux at 1008C in 2 M
aqueous CF3COOH (5 ml) on a water
bath for 3 h. The reaction mixture was then
diluted with H2O (15 ml) and extracted
with CH2Cl2 (3 £ 5 ml). The aqueous layer
was neutralized with Amberlite IR-45 and
concentrated in vacuo to dryness. The
sugars were analyzed by comparison with
authentic sample (solvent system b,
CHCl3–MeOH–H2O (7:3:1) lower-layer
9 þ 1 ml HOAc) on silica gel HPTLC.
Acknowledgements
This research was supported by the Fujian
Provincial Administration of Education (Grant
Nos. 20093515120005 and JA09100) and
Fujian Provincial Scientific and Technological
Program (Grant Nos. 2006NZ0002C4,
2008J0073, and 2009N0016).
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