Three new bioactive phenolic glycosides from Liparis odorata

Article abstract of DOI:10.1080/14786419.2014.880916

Chemical investigation of the ethanol extract from the whole plant Liparis odorata (Willd.) Lindl (orchid family) has led to the isolation of three new phenolic glycosides named liparisglycoside A-C (1-3) along with six known compounds (4-9). The structures of compounds were elucidated by means of extensive spectroscopic data analysis and comparison with the literature data. In addition, compounds 3 and 9 revealed inhibitory effects on lipopolysaccharide-stimulated nitric oxide production, and compounds 1, 3 and 9 displayed hypolipidaemic effects in in vitro bioassays.

Full text of DOI:10.1080/14786419.2014.880916

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Three new bioactive phenolic
glycosides from Liparis odorata
a
a
b
a
a
Bin Li , Hongdong Liu , Dongming Zhang , Xuewen Lai , Bo Liu ,
a
a
Xianghong Xu & Peng Xu
a
Pharmacy College, Jiangxi University of Traditional Chinese
Medicine, 818 Xinwan Road, Nanchang 330004, Jiangxi, P.R. China
b
Institute of Materia Medica, Chinese Academy of Medical
Sciences and Peking Union Medical College, Beijing 100050, P.R.
China
Published online: 17 Mar 2014.
To cite this article: Bin Li, Hongdong Liu, Dongming Zhang, Xuewen Lai, Bo Liu, Xianghong Xu &
Peng Xu (2014) Three new bioactive phenolic glycosides from Liparis odorata, Natural Product
Research: Formerly Natural Product Letters, 28:8, 522-529, DOI: 10.1080/14786419.2014.880916
To link to this article: http://dx.doi.org/10.1080/14786419.2014.880916
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Natural Product Research, 2014
Vol. 28, No. 8, 522–529, http://dx.doi.org/10.1080/14786419.2014.880916
Three new bioactive phenolic glycosides from Liparis odorata
a
Bin Li *, Hongdong Liu , Dongming Zhang , Xuewen Lai , Bo Liu , Xianghong Xu
a
b
a
a
a
a
and Peng Xu
a
Pharmacy College, Jiangxi University of Traditional Chinese Medicine, 818 Xinwan Road, Nanchang
3
30004, Jiangxi, P.R. China
Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College,
b
Beijing 100050, P.R. China
(
Received 11 October 2013; final version received 26 December 2013)
Chemical investigation of the ethanol extract from the whole plant Liparis odorata
(
Willd.) Lindl (orchid family) has led to the isolation of three new phenolic glycosides
named liparisglycoside A–C (1–3) along with six known compounds (4–9). The
structures of compounds were elucidated by means of extensive spectroscopic data
analysis and comparison with the literature data. In addition, compounds 3 and 9
revealed inhibitory effects on lipopolysaccharide-stimulated nitric oxide production,
and compounds 1, 3 and 9 displayed hypolipidaemic effects in in vitro bioassays.
Keywords: Liparis odorata (Willd.) Lindl; orchid family; phenolic glycosides; anti-
inflammatory activity; hypolipidaemic effect
1
. Introduction
Liparis odorata (Willd.) Lindl is a plant from the orchid family that has been reported to possess
bioactivities, such as inhibiting inflammation, improving blood flow and dispelling dampness
(
Chinese Materia Medica Editorial Board of State Administration of Traditional Chinese
Medicine 1999), and has been used as an anti-hyperlipidaemic treatment in Jiangxi folk
medicine of China. However, there are currently few chemical or biological reports on this plant.
Thus, to determine its bioactive constituents, our laboratory performed a thorough investigation
of the ethanol extract for the first time. So far, three new phenolic glycosides named
liparisglycoside A–C (1–3) (Figure 1), together with six known compounds, compound 4
(
Huang et al. 2013), succinic acid (5) (Zhu et al. 2011), phloic acid (6) (Wen & Zhou 1998),
batatasin III (7) (Chen et al. 2008) and luteolin (8) (Chai et al. 2004), derivative from enzymatic
hydrolysis of the known compound anodendrosin A (9) (Abe & Yamauchi 1985) have been
isolated. Preliminary evaluation of the effects of the compounds on lipopolysaccharide (LPS)-
stimulated nitric oxide (NO) production and hypolipidaemic activities are also reported.
2
. Results and discussion
The ethanol extract of L. odorata was fractionated by column chromatography and the fractions
were purified by repeated column chromatography to yield compounds 1–9.
Compound 1 was isolated as white needles. Its molecular formula was determined to be
1
13
C H O based on H and C NMR data. The pseudomolecular ion peak in the HRESI- MS at
3
0 44 13
þ
13
m/z 635.2785 [M þ Na] (calcd 635.2674) indicated nine degrees of unsaturation. The
C
*
Corresponding author. Email: libin7077@hotmail.com
q 2014 Taylor & Francis

Natural Product Research
523
11
9
10
O
8
7
O
18
3
O
O
1
7
OH
1
OH
OH
OH
O
O
1
6
O
O
5
1
OH
1
′
OH
2
OH
OH
15
14
O
O
O
O
1′′
OH
1
3
OH
OH
HO
HO
4
1
O
O
O
OH
O
OH
OH
O
O
OH
OH
OH
OH
O
HO
O
HO
O
O
HO
HO
2
Figure 1. Structures of compound 1–4.
3
NMR spectrum revealed 22 carbons, which was 8 carbons less than expected based on the
molecular formula, revealing that compound 1 possessed a symmetrical structure. Based on the
1
literature (Yu & Yang 1999), the H NMR spectrum of 1 displayed two overlapping aromatic
protons at d 7.60 (s, H-2, 6), indicating the existence of a tetrasubstituted benzene ring, two
overlapping olefinic protons at d 5.23 (t, J ¼ 7.2 Hz, H-8, 13), two methylene protons at d 3.51
(
declaring there were two isopentenyl groups, as well as protons at d 3.80 (s, H-18) showing one
d, J ¼ 7.2 Hz, H-7, 12), two sets of methyl protons at d 1.71 (s, H-10, 15) and 1.70 (s, H-11, 16),
1
methoxy. The C NMR, DEPT and HSQC spectra indicated two overlapping aromatic methyne
3
carbons at d 130.3 (C-2, 6), four quaternary aromatic carbons at d 127.8 (C-1), 137.6 (C-3, 5) and
1
58.1 (C-4), two overlapping olefinic methyne carbons at d 124.4 (C-8, 13), two quaternary
olefinic carbons at d 134.3 (C-9, 14), two overlapping methylene carbons at d 29.9 (C-7, 12), two
sets of overlapping methyl carbons at d 26.3 (C-10, 15) and 18.5 (C-11, 16), one carbonyl
1
1
resonance at d 168.9 (C-17) and one methoxy carbon at d 52.8 (C-18). In the H– H COSY
spectrum, the proton at d 3.51 (H-7, 12) was coupled to the proton at d 5.23 (H-8, 13) indicating
that the two methylene groups were connected to two olefinic methynes. In HMBC, correlations
were observed from H-7 to C-2, C-4, C-8, C-9 and from H-12 to C-4, C-6, C-13, C-14 indicating
that there were two isopentenyl groups attached to a tetrasubstituted benzene ring at C-3 and C-
5
, respectively; correlations were observed from H-2, H-6 to C-17 and from H-18 to C-17
indicating that a methyl formate group was attached to C-1 of the benzene ring (Figure 2).
1
13
The H and C NMR spectra of 1 revealed two anomeric protons at d 4.78 (d, J ¼ 7.5 Hz, H-
0
00
0
00
1
) and 4.76 (d, J ¼ 7.5 Hz, H-1 ), two anomeric carbons at d 104.5 (C-1 ) and 105.6 (C-1 ). This
indicated the presence of two sugar moieties, and D-glucose was identified by using GC–MS to
analyse the trimethylsilyl ether derivatives of the acidic hydrolysis products of 1. The HMBC
correlation between the anomeric proton of one D-glucose and C-4 supported the prediction that

5
24
B. Li et al.
11
10
8
18
O
H C
3
O
7
17
1
3
1
6
OH
OH
OH
5
O
O
12
15
14
1′
OH
OH
13
O
O
OH
1′′
HO
Figure 2. Key HMBC ( ! ) and COSY (
) correlations observed of compound 1.
0
0
D-glucose was attached to 4-hydroxyl group with a b configuration. The correlation from H-1 to
0
00
0
C-2 indicated that the two D-glucoses were connected through a 1 to 2 linkage with a b
configuration.
Furthermore, the NMR spectra of compound 1 and known compound 4 (Hung et al. 2013)
were similar, with the exception that 1 had a molecular weight of 30 more than 4. This is caused
by the presence of an additional CH O (d at 3.56, 3.73 and d at 62.9) on a sugar moiety. Only
2
H
C
D-glucose was identified through GC–MS analysis in 1; thus, compound 1 was determined to be
0
2 -O-b-D-glucopyranosyl-4-O-b-D-glucopyranosyl-3,5-bis(3-methyl-2-butenyl)-methyl-17-
benzoate, known as liparisglycoside A.
Compound 2 was isolated as an amorphous powder. Its molecular formula was determined
1
to be C H O based on the H and C NMR data. The pseudomolecular ion peak in the
13
2
3 32 8
þ
HR-ESI-MS at m/z 459.2008 [M þ Na] (calcd 459.1989) indicated eight degrees of
unsaturation. Comparison of the spectral data of 2 with that of 4 indicated that they were similar
1
3
with three differences: (1) there were only arabinose signals in 2, (2) the C NMR spectral data
indicated the presences of an additional oxygenated tertiary carbon at d 71.8 (C-14) and the lack
of an olefinic quaternary carbon in 2, which indicated that there was an additional OH connected
to C-14, which down-shifted carbon 14 of compound 2, (3) there were two trans olefinic protons
at d 7.10 (1H, d, J ¼ 16.0 Hz, H-12) and 6.32 (1H, d, J ¼ 16.0 Hz, H-13) in 2, and in HMBC,
these two protons had correlations with C-14, C-4, C-5, C-6, indicating that these two trans
olefinic carbons were connected to both an aromatic ring and C-14. In addition, in HMBC, the
methyl protons at d 1.36 (3H, s, H-15) and 1.35 (3H, s, H-16) had correlations with C-14 (d
7
1.8), indicating that the two methyl groups were connected to C-14. An anomeric proton of
0
L-arabinose d 4.57 (d, J ¼ 7.5 Hz, H-1 ) had correlations with C-4 (d 157.4), which indicated that
the L-arabinose moiety was attached to the 4-hydroxyl group with an a-configuration. Therefore,
compound 2 was identified as 3-(3-hydroxy-3-methyl-1-butenyl)-5-(3-methyl-2-butenyl)-4-O-
a-L-arabinose-methyl-17-benzoate, also known as liparisglycoside B.
Compound 3 was isolated as white needles. Its molecular formula was determined to
be C H O based on the NMR data. The pseudomolecular ion peak in the HR-ESI-MS
2
9 44 13
þ
at m/z 623.2656 [M þ Na] (calcd 623.2674) indicated eight degrees of unsaturation.
Comparison of the spectral data of 3 with 4 indicated that they contained same two sugar groups,
there were only four differences in aglycone moiety between them: (1) 3 had a molecular weight
of 18 more than 4, (2) the C NMR spectrum indicated that 3 contained an additional
oxygenated tertiary carbon at d 72.1 (C-14) and lacked an olefinic quaternary carbon resonance,
1
3

Natural Product Research
525
1
indicating that there was an OH connected to C-14, (3) the H NMR spectrum indicated there
were six methylene signals at d 3.50 (2H, d, J ¼ 7.0 Hz, H-7), 2.89, 2.81 (2H, dd, J ¼ 12.0,
1
3
5
.0 Hz, H-12), 1.74, 1.69 (2H, dd, J ¼ 12.0, 5.0 Hz, H-13), and these signals appeared in the
C
NMR spectrum at d 29.9 (C-7), 26.6 (C-12) and 45.8 (C-13), (4) one olefinic proton was absent
in 3. In HMBC, the methyl protons at d 1.23 (3H, d, J ¼ 2.0 Hz, H-15) and 1.22 (3H, d,
J ¼ 2.0 Hz, H-16) had correlations with C-14 (d 72.1) and C-13 (d 45.8). In addition, the protons
at d 1.74, 1.69 (H-13) had correlations with the carbons at d 72.1 (C-14) and 26.6 (C-12), the
proton at d 2.80 (2H, dd, J ¼ 12.0, 5.0 Hz, H-12) had correlations with the carbons at d 158.7
(
C-4), 138.8 (C-5), and 130.4 (C-2), all of which indicated that the two methyl fragments were
connected to the oxygenated tertiary carbon C-14 and that the olefinic double bond between
carbons 13 and 14 in compound 4 was hydrogenated in compound 3. Therefore, compound 3 was
0
identified as 2 -O-a-L-arabinose-4-O-b-D-glucopyranosyl-3-(3-hydroxy-3,3-dimethylbutyl)-5-
(
3-methyl-2-butenyl)-methyl-17-benzoate, named liparisglycoside C.
In addition, the effects of the compounds on LPS-stimulated NO production in peritoneal
macrophage cells were determined (Table 1). The results indicated that compounds 3 and 9
inhibited NO production.
A preliminary screening of the hypolipidaemic activities of the compounds was conducted in
an in vitro experiment (Table 2). Compared with the control group, compounds 1, 3 and 9
Table 1. Inhibition of compounds on LPS-stimulated NO production in peritoneal macrophages cells
(
n ¼ 3).
Group
Concentration
–
NO production
Inhibition rate
LPS
Dex
1
2
3
4
5
6
7
38.47
3.48
–
2
2
2
2
2
2
2
2
2
2
5
5
5
5
5
5
5
5
5
5
10
10
10
10
10
10
10
10
10
10
90.95
10.63
9.21
30.73
20.85
10.63
22.29
13.52
7.38
34.38
34.93
26.65
38.80
34.38
39.35
33.27
35.63
20.82
8
9
45.89
Note: Data were expressed as the mean ^ SE.
Table 2. Hypolipaemic effects of compounds on triglyceride content in HepG2.
Group
Results
Model
Simvastatin
0.3410 ^ 0.0113
0.1799 ^ 0.0109**
0.2399 ^ 0.0057*
0.2598 ^ 0.0077
0.2147 ^ 0.0075**
0.2450 ^ 0.0083
0.2488 ^ 0.0265
0.2474 ^ 0.0051
0.2402 ^ 0.0018
0.2414 ^ 0.0123
0.2362 ^ 0.0055*
1
2
3
4
5
6
7
8
9
Note: Data were expressed as mean ^ SE.
*
p , 0.05, **p , 0.01 versus control group.

5
26
B. Li et al.
expressed potential hypolipidaemic effects ( p , 0.05), with compound 3 ( p , 0.01) being
significantly active.
The amount of phenolic glycosides we isolated is sufficient to allow for further
investigations on biological activities, which are warranted based on our preliminary results.
3
3
. Experimental
.1. General procedures
UV spectra were recorded on a JASCO V650 spectrophotometer (Jasco International Co., Ltd.,
Tokyo, Japan). IR spectra were recorded on a Nicolet 5700 FT-IR spectrometer (Thermo
Electron, Waltham, MA, USA) using the microscope transmission method. NMR spectra were
obtained on an INOVA-500 MHz NMR spectrometer (Varian Medical Systems, Palo Alto, CA,
1
13
USA) operating at 500 MHz for H NMR and at 125 MHz for C NMR. Chemical shifts are
given in d (ppm) with solvent (DMSO-d , CDCl or CD OD) peaks used as references. GC–MS
6
3
3
spectra were obtained using an Agilent Technologies 7890A instrument (Agilent, Palo Alto, CA,
USA). ESI-MS spectra were measured on an Agilent 1100 (Agilent Technologies, Palo Alto,
CA, USA) Series LC/MSD ion-trap mass spectrometer. HR-ESI-MS data were recorded on an
Autospec Ultima-TOF mass spectrometer (Micromass Communication, Inc., Manchester,
England). Analytical HPLC was run on an Agilent 1100 series instrument with a UV/diode array
detector using an YMC (YMC Co., Ltd., Kyoto, Japan) column (RP-C18, 4.6 £ 250 mm, 5 mm).
Preparative HPLC was performed on a Shimadzu LC-6AD instrument (Shimadzu Corporation,
Tokyo, Japan) with an SPD-10A detector, using an YMC-Pack ODS-A column (20 £ 250 mm,
5
mm). Octodecylcyclohexane silane (ODS) (45–70 mm, Merck KGaA, Darmstadt, Germany)
and silica gel (200–30 mesh, Qingdao Marine Chemical, Inc., Qingdao P.R. China) were used
for column chromatography, glass pre-coated with silica gel GF254 (200–300 mesh, Qingdao
Marine Chemical, Inc.) was used for TLC.
3
.2. Plant material
L. odorata (Willd.) Lindl was collected in the Jiangxi province of China in 2010, identified by
Professor Xuewen Lai in Jiangxi University of Traditional Chinese Medicine in China, and a
voucher specimen (No. 002017) has been deposited in the Herbarium of Jiangxi University of
Traditional Chinese Medicine.
3
.3. Extraction and isolation
The air-dried and powdered plant of L. odorata (50.0 kg) was extracted with 95% EtOH under
reflux three times (each time for 1.0 h). Then, the EtOH extract was concentrated under reduced
pressure to give a brown solid material (980 g), which was dissolved in 6000 mL of 0.2 M HCl.
The HCl-soluble fraction was basified with NH zH O to a pH of 10, and extracted three times
3
2
with ethyl acetate. The ethyl acetate-soluble fraction was concentrated under reduced pressure to
give a brown solid material (95 g). Next, the total ethyl acetate fraction (95 g) was subjected to
neutral silica gel column chromatography, eluted with a CHCl /MeOH (10:1–1:1) to give eight
3
fractions (A1–A8). Fraction A2 (20 g) was subjected to a neutral silica gel column eluted with
CH Cl /MeOH (40:1–1:1) to give 49 subfractions. Subfractions A2-5–A2-10 (3 g) were
2
2
subjected to neutral silica gel column chromatography eluted with petroleum ether/acetone
10:1–1:1). Fraction 7 contained crystals that were recrystallised in hexane to obtain compound
(50 mg). Fraction 9 contained crystals that were recrystallised in petroleum ether to obtain
(
7
compound 5 (100 mg). Fraction 15 contained crystals that were recrystallised in petroleum ether
to obtain compound 6 (50 mg). Subfractions A2-16–A2-21 (1.3 g) were subjected to neutral

Natural Product Research
527
silica gel column chromatography eluted with CHCl /CH COCH (1:1), continually purified by
3
3
3
preparative HPLC using 60% MeOH to give, 1 (100 mg) and 4 (90 mg). Subfractions A2-26–
A2-29 (0.6 g) were subjected to ODS column chromatography eluted with 60% MeOH,
compounds 2 (30 mg) and 8 (60 mg) were obtained by continue preparative HPLC eluted with
25% MeOH. Fraction A3 (17.7 g) was subjected to neutral silica gel column eluted with CH Cl /
2 2
MeOH (40:1–0:1) to give 39 subfractions. Subfractions A3-32–A3-36 (4.1 g) were subjected to
neutral silica gel column chromatography eluted with CHCl /MeOH (20:1–0:1), fractions
3
2
4–29 were continually purified with another neutral silica gel column to give 9 (30 mg).
3
.4. Liparisglycoside A (1)
2
D
1
0
White needles, m.p. 185.3–186.5, ½aꢀ 2 7:23 (c ¼ 0.1, MeOH); UV (MeOH) l
(log) 207
3.53), 240 (3.02); IR(KBr) nmax (cm ) 3322, 1723, 1437, 1320, 1286, 1078, 1024; HR-ESI-MS
max
2
(
m/z 635.2785 (calcd 635.2674 for C H O Na). H NMR(500 MHz, CD OD): d 1.70 (s, 6H,
1
3
0
44 13
3
H
Me-11, 16), 1.71 (s, 6H, Me-10, 15), 3.51 (d, J ¼ 7.2 Hz, 4H, H-7, 12), 3.80 (s, 3H, OMe-18),
0
00
4
1
3
1
6
.78 (d, J ¼ 7.5 Hz, 1H, H-1 ), 4.76 (d, J ¼ 7.5 Hz, 1H, H-1 ), 5.23 (t, J ¼ 7.2 Hz, 2H, H-8,
3 C
1
3),7.60 (s, 2H, H-2, 6). C NMR (125 MHz, CD OD): d 127.8 (C-1), 130.3 (C-2, 6), 137.6 (C-
3
, 5), 158.1 (C-4), 29.9 (C-7, 12), 124.4 (C-8, 13), 134.3 (C-9, 14), 26.3 (C-10, 15), 18.5 (C-11,
0
0
0
0
0
6), 168.9 (C-17), 52.8 (C-18), 104.5 (C-1 ), 83.1 (C-2 ), 78.4 (C-3 ), 71.6 (C-4 ), 78.7 (C-5 ),
00
0
00
00
00
00
00
2.9 (C-6 ), 105.6 (C-1 ), 76.3 (C-2 ), 78.3 (C-3 ), 72.0 (C-4 ), 78.4 (C-5 ), 63.2 (C-6 ).
3
.5. Liparisglycoside B (2)
2
0
White amorphous powder, ½aꢀ –11.57 (c ¼ 0.1, MeOH); UV (MeOH) l
(log) 209 (3.55),
D
max
-
1
2
m/z 459.2008 (calcd 459.1989 for C H O Na). H NMR (500 MHz, CD OD): d 1.35 (s, 3H,
45 (3.03); IR (KBr) n
(cm ) 3393, 2972, 1719, 1599, 1435, 1081, 1015, 909; HR-ESI-MS
max
1
2
3
32
8
3
H
Me-16), 1.36 (s, 3H, Me-15), 1.67 (s, 3H, Me-11), 1.69 (s, 3H, Me-10), 3.44 (dd,
J ¼ 7.0,11.0 Hz, 1H, H-7a), 3.57 (t, J ¼ 7.0 Hz, 1H, H-7b), 3.83 (s, 3H, OMe-18), 4.57 (d,
0
J ¼ 7.5 Hz, 1H, H-1 ), 5.24 (t, J ¼ 7.0 Hz, 1H, H-8), 6.32 (d, J ¼ 16.0 Hz, 1H, H-13), 7.10 (d,
1
3
J ¼ 16.0 Hz, 1H, H-12), 7.63 (s, 1H, H-6), 7.94 (s, 1H, H-2). C NMR (125 MHz, CD OD): d
3
C
1
26.8 (C-1), 127.4 (C-2), 132.5 (C-3), 157.4 (C-4), 138.2 (C-5), 130.6 (C-6), 29.8 (C-7), 123.6
C-8), 134.0 (C-9), 25.9 (C-10), 18.0 (C-11), 123.0 (C-12), 140.7 (C-13), 71.8 (C-14), 29.7 (C-
(
0
0
0
0
0
1
5,16), 168.4 (C-17), 52.6 (C-18), 106.7 (C-1 ), 73.1 (C-2 ), 74.4 (C-3 ), 71.8 (C-4 ), 69.8 (C-5 ).
3
.6. Liparisglycoside C (3)
2
0
Colourless needles, m.p. 123.3–135.58C, ½aꢀ –13.64 (c ¼ 0.1, MeOH); UV (MeOH) l
D
max
2
1
(
log) 209 (3.50), 245(3.01); IR(KBr) n
(cm ) 3379, 2966, 1713, 1086, 776; HR-ESI-MS m/z
max
1
6
23.2656 (calcd 623.2674 for C H O Na). H NMR (500 MHz, CD OD): d 1.22 (d,
29 44 13 3 H
J ¼ 2.0 Hz, 3H, Me-16), 1.23 (d, J ¼ 2.0 Hz, 3H, Me-15), 1.68 (s, 3H, Me-11), 1.69 (dd, J ¼ 5.0,
2.0 Hz, 1H, H-13a), 1.74 (dd, J ¼ 5.0, 12.0 Hz, 1H, H-13b), 1.71 (s, 3H, Me-10), 2.81 (dd,
J ¼ 5.0, 12.0 Hz, 1H, H-12a), 2.89 (dd, J ¼ 5.0, 12.0 Hz, 1H, H-12b), 3.50 (d, J ¼ 7.0 Hz, 2H,
1
0
0
0
H-7), 3.81 (s, 3H, OMe-18), 4.66 (d, J ¼ 8.0 Hz, 1H, H-1 ), 4.75 (d, J ¼ 7.0 Hz, 1H, H-1 ), 5.24
1
3
(
t, J ¼ 6.5 Hz, 1H, H-8), 7.59 (s, 1H, H-2), 7.67 (s, 1H, H-6). C NMR (125 MHz, CD OD): d
3
C
1
27.9 (C-1), 130.4 (C-2), 137.8 (C-3), 158.7 (C-4), 138.8 (C-5), 130.6 (C-6), 29.9 (C-7), 124.3
C-8), 134.6 (C-9), 26.4 (C-10), 18.7 (C-11), 26.6 (C-12), 45.8 (C-13), 72.1 (C-14), 30.1 (C-15),
(
0 0 0 0
2
9.7 (C-16), 169.0 (C-17), 53.0 (C-18), 105.1 (C-1 ), 81.6 (C-2 ), 74.3 (C-3 ), 69.5 (C-4 ), 67.2
0 00 00 00 00 00 00
(
C-5 ), 106.1 (C-1 ), 76.2 (C-2 ), 78.5 (C-3 ), 71.9 (C-4 ), 78.6 (C-5 ), 63.1 (C-6 ).

5
28
B. Li et al.
3
.7. Acidic hydrolysis and sugar analysis of compound 1
Compound 1 (5 mg) was refluxed in 2 M HCl (3 mL) at 608C for 3 h and concentrated to give a
brown material, which was dissolved in water (3 mL) and partitioned with EtOAc (3 mL £ 3).
The aqueous layer was evaporated to dryness under reduced pressure to give one
monosaccharide residue and the sugars were identified as D-glucose by comparison with
retention times of authentic sample of D-glucose (19.84 min) (Kinijo et al. 1992; Wang et al.
2
013).
3
.8. Effect of compounds on LPS-stimulated NO production in peritoneal macrophage cells
The LPS-stimulated NO production effects of the compounds were valued by measuring the
peritoneal macrophage cells in vitro (Ding et al. 1988; Chen et al. 2006).
3
.9. Hypolipidaemic activity assays
The hypolipidaemic activities of compounds 1–9 were assayed by measuring the triglyceride
content in HepG2 cells in vitro (G o´ mez-Lech o´ n et al. 2007; Wang et al. 2013).
4
. Conclusions
Three new phenolic glycosides named liparisglycoside A–C, together with six known
compounds, were isolated from L. odorata for the first time. In addition, new compound
liparisglycoside C and known compound 9 revealed potential anti-inflammatory activities, and
new compounds liparisglycoside A, liparisglycoside C, known compound 9 expressed potential
hypolipidaemic effects. Further investigations are ongoing in our laboratory.
Supplementary material
Supplementary material relating to this article is available online, alongside Figures S1–S12.
Acknowledgements
The project was supported by the Chinese National Natural Scientific fund (No. 81260629), Jiangxi
Province Jinggang Star fund (No. 2008-222), Jiangxi Province Medical Ministry fund (No. 2009A056),
Jiangxi Province Major Educational Ministry fund (No. GJJ12515).
References
Abe F, Yamauchi T. 1985. Glycosyl nervogenic acid esters of carbohydrates from Anodendron affine (Anodendron. VI).
Chem Pharm Bull. 33:2712–2720.
Chai XY, Wang L, Song Y, Chen J, Li P. 2004. Study on the flavonoids from Lonicera confuse DC. J Chin Pharm Univ.
35:299–302.
Chen W, Lin XH, Huang LY, Luo HB, Chen J. 2006. Effects of polysaccharide of Aloe barbadensis on nitric oxide
production by mouse peritoneal macrophages. Chin J Mod Appl Pharm. 23:17–19.
Chen YG, Li JT, Wang LQ, Liu Y. 2008. Aromatic compounds from Den drobiumaphyllum. Biochem Syst Ecol.
36:458–461.
Chinese Materia Medica Editorial Board of State Administration of Traditional Chinese Medicine. 1999. Chinese materia
medica (II). Shanghai: Shanghai Science Press; p. 737.
Ding AH, Nathan CF, Stuehr DJ. 1988. Release of reactive nitrogen intermediates and reactive oxygen intermediates
from mouse peritoneal macrophages. J Immunol. 141:2407–2412.
G o´ mez-Lech o´ n MJ, Donato MT, Mart ´ı nez-Romero A, Jim e´ neza N, Castella JV, O’Connorb J-E. 2007. A human
hepatocellular in vitro model to investigate steatosis. Chem Biol Interact. 165:106–116.
Huang S, Zhou XL, Wang CJ, Wang HY, Wang YS, Shan LH, Weng J. 2013. New nervogenic acid derivatives from
Liparis nevosa. Planta Med. 79:281–287.

Natural Product Research
529
Kinijo J, Araki K, Fukui K, Higuchi H, Ikeda T, Nohara T, Ida Y, Takemoto N, Miyakoshi M, Shoji J. 1992. Six new
triterpenoidal glycosides including two new sapogenols from Albizziae cortex V. Chem Pharm Bull.
40:3269–3273.
Wang ZW, Li B, Yu SS, Lai XW, Liu B, Chen J, Hayashi T. 2013. Eight new monoterpene acylglucosides from Sibiraea
angustata. Phytochem Lett. 6:461–466.
Wen J, Zhou FX. 1998. Studies on the chemical constituents from the leaves of Malus dourneri (Bois) Chev. J Chin Med
Mater. 21:572.
Yu DQ, Yang JS. 1999. Handbook of analytical chemistry. Vol. 7. 2nd ed. Beijing: Chemical Industry Press; p. 839, 820,
9
09.
Zhu HL, Wei WX, Zhou M, Yang D, Fan XW. 2011. Chemical constituents of Phyllanthus niruri L. Nat Prod Res Dev.
3:401–403.
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