Two new triterpenoid saponins from Caragana microphylla seeds

Article abstract of DOI:10.1007/s11418-011-0624-9

Two new triterpenoid saponins, caraganins A and B (1 and 2), structurally characterized by a 22-oxo group, were isolated from the seeds of Caragana microphylla Lam., together with their n-butyl esters as artifacts (1a and 2a). Their structures and configurations were elucidated by detailed spectroscopic analyses on the basis of NMR, IR, and MS data. Compounds 1a and 2a exhibited potent antibacterial activity against Staphylococcus aureus and Bacillus subtilis with minimum inhibitory concentration values ranging from 3.125 to 6.25 μg/ml.

Full text of DOI:10.1007/s11418-011-0624-9

J Nat Med (2013) 67:190–195
DOI 10.1007/s11418-011-0624-9
NOTE
Two new triterpenoid saponins from Caragana microphylla seeds
•
Cheng-Jian Zheng Gui-Lin Jin Jing-Ping Zou
•
•
•
Yi-Ping Jiang Pei-Xin Sun Lu-Ping Qin
•
Received: 14 November 2011 / Accepted: 28 December 2011 / Published online: 14 January 2012
Ó The Japanese Society of Pharmacognosy and Springer 2012
Abstract Two new triterpenoid saponins, caraganins
A and B (1 and 2), structurally characterized by a 22-oxo
group, were isolated from the seeds of Caragana micro-
phylla Lam., together with their n-butyl esters as artifacts
(1a and 2a). Their structures and configurations were elu-
cidated by detailed spectroscopic analyses on the basis of
NMR, IR, and MS data. Compounds 1a and 2a exhibited
potent antibacterial activity against Staphylococcus aureus
and Bacillus subtilis with minimum inhibitory concentra-
tion values ranging from 3.125 to 6.25 lg/ml.
rheumatoid arthritis effects [7]. The seeds of the plant have
been widely used in folk medicine for insecticidal and
antipruritic purposes [8]. Our previous phytochemical
studies on the seeds revealed the presence of flavonoids,
triterpenoids, and phytosterols [6]. In continuation of our
investigations on this medicinal seed, we isolated two new
triterpenoid saponins, caraganins A and B (1 and 2),
together with their n-butyl ester derivatives (1a and 2a),
whose structures (Fig. 1) were unequivocally determined
by extensive 1D and 2D NMR spectroscopy as well as by
mass spectrometry and by comparison with literature data.
All the isolated compounds were evaluated for their anti-
bacterial activity against Escherichia coli, Staphylococcus
aureus, Pseudomonas aeruginosa, Bacillus subtilis, and
Klebsislla pneumoniae.
Keywords Caragana microphylla ꢀ Triterpenoids ꢀ
Caraganin
Introduction
Caragana microphylla Lam. (Leguminosae) is a perennial
shrub widely distributed in northern and western China,
and a popular plant for restoring vegetation on desertified
sandy land [1]. It has also exhibited a wide variety of
biological activities including antioxidant [2], anti-inflam-
matory [3, 4], antitumor [5], analgesic [6], and anti-
Results and discussion
Repeated column chromatography of the ethanol extract of
C. microphylla seeds yielded two new triterpenoid sapo-
nins, caraganins A and B (1 and 2), together with their
n-butyl esters (1a and 2a), structurally illustrated by 1H-, 13C-,
and 2D-NMR data.
Compound 1 was obtained as a amorphous powder and
analyzed for the molecular formula C36H54O10 by negative
HRESIMS [M–H]- at m/z 645.3644 (calc. 645.3639),
which was supported by its NMR data. Absorptions for
Cheng-Jian Zheng and Gui-Lin Jin contributed equally to this work.
C.-J. Zheng ꢀ G.-L. Jin ꢀ Y.-P. Jiang ꢀ P.-X. Sun ꢀ
L.-P. Qin (&)
Department of Pharmacognosy, School of Pharmacy,
Second Military Medical University, 325 Guohe Road,
Shanghai 200433, People’s Republic of China
e-mail: qinsmmu@126.com
hydroxyl (3440 cm-1) and carboxyl (1734 and 1703 cm-1
)
1
groups were observed in its IR spectrum. The H-NMR
spectrum of 1 (Table 1) was characterized by the presence
of a trisubstituted olefinic proton signal at dH 5.40 (1H, br s),
seven methyl singlets assignable to the aglycone, one dd
ascribable to H-18 at dH 2.63 (1H, dd, J = 13.7, 3.2 Hz),
and one dd due to H-3 at dH 3.45 (1H, dd, J = 11.5,
J.-P. Zou
No. 6 Sanatorium of Retired Cadres of Beijing Military Region,
23 Maliandao Road, Xicheng District, Beijing 100055,
People’s Republic of China
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J Nat Med (2013) 67:190–195
191
COOH
O
Fig. 1 Structure of compounds
1, 2 and their n-butyl esters
(1a and 2a)
Compound 2 was obtained as a colorless amorphous
powder. The negative HRESIMS of 2 exhibited its [M-H]-
peak at m/z 661.3612 (calc. 661.3588), consistent with the
molecular formula C36H54O11. Its 1H and 13C NMR spectra
were closely analogous to those of 1, except for the
appearance of a pair of doublets due to a hydroxymethyl
group (dH 3.76, d, J = 11.3 Hz; dH 4.45, d, J = 11.3 Hz).
The molecular weight of 2 is only one more oxygen atom
than that of 1. Therefore, compound 2 was considered to be
a 24-hydroxy derivative of 1, deduced from the crosspeaks
of H-24 (dH 3.76, 4.45) coupled with the carbon signal C-3
(dC 89.2), C-4 (dC 44.4), and C-23 (dC 23.5), observed in
the HMBC experiment (Fig. 2). The aglycone moiety of 2
was therefore identified as melilotigenin [9, 10] and com-
pound 2 was finally established as 3b,24-dihydroxy-22-
oxo-12-oleanen-29-oic acid 3-O-b-D-glucuropyranoside
and was named caraganin B.
H
R1OOC
HO
HO
H
O
O
H
OH
R2
1. R1 = H
R2 = CH3
R2 = CH3
1a. R1 = n-butyl
2. R1 = H
R2 = CH2OH
R2 = CH2OH
2a. R1 = n-butyl
3.7 Hz) suggested the presence of a b-oxygenated substi-
tution at C-3. In addition, a group of sugar-proton signals
occurred in the narrow spectral region of dH 5.1 to 4.0,
of which one anomeric proton signal at dH 5.10
(d, J = 7.5 Hz) indicated b-D-linkage of the sugar moiety
in 1. The 13C-NMR spectrum of 1 revealed 6 carbons for
the glycone portion and 30 carbon signals for the aglycone
portion. Combined with the analysis of the DEPT and
HMQC spectra of 1, the aglycone portion showed the
presence of a ketone carbon at dC 215.0, two carboxyl
carbon at dC 179.2 and 173.6, a tri-substituted olefinic
group (dC 141.6 and 124.9), an oxymethine at dC 89.1,
seven methyl carbons at dC 28.3, 25.7, 21.8, 21.1, 17.2,
17.1, and 15.8, nine methylene carbons at dC 46.7, 41.8,
38.9, 33.0, 27.5, 26.8, 25.6, 24.0, and 18.6, three methine
carbons at dC 55.8, 47.9, and 47.2, and six quaternary
carbons at dC 48.4, 44.8, 42.2, 40.1, 39.7, and 37.0. The
above-mentioned evidence indicated an oleanane triterpe-
noid aglycon for 1. Further analysis of the 13C-NMR data
of glycone [dC 173.6 (s), 107.4 (d), 78.3 (d), 78.0 (d), 75.7
(d), and 73.6 (d)] indicated glucuronic acid as the sugar
moiety of 1. The D-configuration for glucuronic acid was
determined by the measurement of optical rotation after
separation from the crude sugar residue. The point of
attachment of the sugar chain was deduced by the long-
range 1H–13C correlation observed from the anomeric
proton (dH 5.10) to C-3 (dC 89.1) of the aglycone (Fig. 2).
Moreover, the typical signal of ketone carbon at dC 215.0
was located at the C-22 position by the correlations with dH
1.30 (3H, s, H-28), dH 3.52 (d, J = 14.5 Hz, H-21a), and
2.75 (d, J = 14.5 Hz, H-21b) in the HMBC spectrum. The
NOESY correlations (Fig. 3) were detected between H-3/
H-5, H-3/H-23, H-5/H-9, H-24/H-25, H-25/H-26, H-9/H-
27, H-18/H-30, and H-18/H-28, indicating a b-orientation
for H-18, H-24, H-25, H-26, H-28, and H-30, and an
a-orientation for H-3, H-5, H-9, H-23, and H-27. The 1H–1H
COSY spectrum revealed the existence of fragment –CH2-
CH2-CH-, from C-1 to C-3, fragment –CH-CH2-CH2-,
from C-5 to C-7, and fragment –CH-CH2-CH-, from C-9 to
C-12 (Fig. 2). The aglycone moiety of 1 was therefore
identified as subprogenin D [9] and compound 1 was finally
established as 3b-hydroxy-22-oxo-12-oleanen-29-oic acid
3-O-b-D-glucuropyranoside and was named caraganin A.
Compound 1a was obtained as a colorless amorphous
powder. The positive HRESIMS of 3 exhibited its
[M?Na]? peak at m/z 725.4263 (calc. 725.4235) consistent
with the molecular formula C40H62O10. The IR spectrum of
3 also indicated the existence of hydroxyl (3445 cm-1) and
carbonyl groups (1732, 1704 cm-1). The 1H- and 13C-
NMR data of 3 are summarized in Tables 1 and 2, sug-
gesting the presence of most structural features in common
with compound 1 except for the presence of an oxygenated
n-butyl group (one triplet methyl and three methylene
groups), which led to the assumption that 3 is an n-butyl
ester derivative of 1. In the HMBC spectrum, the oxy-
genated methylene protons at dH 4.38 correlated with the
carboxyl carbon of the glucuronic acid at dC 170.5. This
means glucuronic acid has been n-butyl esterified. Fur-
thermore, all of the correlations supporting the structure of
1 were observed in the 2D NMR spectroscopic data of 1a,
suggesting that the configuration of 1a was the same as 1.
Thus, 1a was established as 3b-hydroxy-22-oxo-12-olea-
nen-29-oic acid 3-O-b-D-glucuronopyranosyl-60-O-n-butyl
ester, which might be an artifact produced from 1 during
the extraction procedure with n-BuOH.
Compound 2a was obtained as a colorless amorphous
powder. The positive HRESIMS of 2a exhibited its
[M?Na]? peak at m/z 741.4163 (calc. 741.4184) and gave
the molecular formula C40H62O11. Further comparison of the
NMR data of 2a with those of 2 revealed that 2a is an n-butyl
ester derivative of 2, shown by the presence of an oxygenated
n-butyl group (one triplet methyl and three methylene
groups). HMBC correlation observed from the oxygenated
methylene protons at dH 4.36 to the carboxyl carbon of the
glucuronic acid at dC 170.4 indicated that the glucuronic acid
has been n-butyl esterified. Compound 2a was therefore
established as 3b,24-dihydroxy-22-oxo-12-oleanen-29-oic
acid 3-O-b-D-glucuronopyranosyl-60-O-n-butyl ester, which
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Table 1 1H-NMR (400 MHz) data for compounds 1, 2 and their n-butyl esters (1a and 2a) in C5D5N
No.
dH (d, ppm; J, Hz)
1
2
1a
2a
1
1.51 m, 0.99 m
1.31 m, 1.36 m
3.45 dd (11.5, 3.7)
0.89 m
1.55 m, 0.94 m
1.28 m, 1.35 m
3.64 m
1.61 m, 1.02 m
1.31 m, 1.36 m
3.45 dd (11.5, 3.7)
0.88 m
1.57 m, 0.99 m
1.30 m, 1.36 m
3.66 m
2
3
5
0.97 m
0.99 m
6
1.41 m, 1.60 m
1.51 m, 1.35 m
1.67 m
1.49 m, 1.74 m
1.51 m, 1.37 m
1.67 m
1.41 m, 1.63 m
1.52 m, 1.38 m
1.68 m
1.51 m, 1.77 m
1.51 m, 1.35 m
1.67 m
7
9
11
12
15
16
18
19
21
1.90 m
1.91 m
1.94 m
1.93 m
5.40 s
5.40 s
5.44 s
5.40 s
1.77 m, 1.10 m
2.31 m, 1.92 m
2.63 dd (13.7, 3.2)
2.96 t (13.5), 2.05 m
3.52 d (14.5)
2.75 d (14.5)
1.40 s
1.77 m, 1.10 m
2.17 m, 2.01 m
2.64 dd (13.7, 3.2)
2.96 t (13.5), 2.06 m
3.52 d (14.5)
2.80 d (14.5)
1.58 s
1.32 m, 1.08 m
2.24 m, 1.95 m
2.64 dd (13.7, 3.2)
2.96 t (13.8), 2.04 m
3.55 d (14.5)
2.75 d (14.5)
1.38 s
1.77 m, 1.10 m
2.14 m, 2.09 m
2.64 dd (13.7, 3.2)
2.96 t (13.5), 2.09 m
3.54 d (14.5)
2.80 d (14.5)
1.62 s
23
24
25
26
27
28
30
10
20
30
40
50
1.10 s
4.45 d (11.3), 3.76 d (11.3)
0.93 s
1.09 s
4.45 d (11.3), 3.76 d (11.3)
0.93 s
0.97 s
0.96 s
1.02 s
0.99 s
1.02 s
0.99 s
1.35 s
1.34 s
1.32 s
1.36 s
1.30 s
1.27 s
1.30 s
1.22 s
1.48 s
1.50 s
1.50 s
1.50 s
5.10 d (7.5)
4.21 t (8.4)
4.40 t (8.9)
4.67 t (9.2)
4.70 d (9.7)
5.19 d (7.8)
4.17 t (8.5)
4.36 m
5.08 d (7.4)
4.18 t (8.4)
4.38 t (8.9)
4.60 t (9.4)
4.68 t (9.7)
4.38 m
5.19 d (7.8)
4.17 t (8.7)
4.36 m
4.60 t (9.2)
4.72 d (9.7)
4.60 t (9.2)
4.72 d (9.7)
4.36 m
100
200
300
400
1.66 m
1.67 m
1.48 m
1.48 m
0.86 t (7.4)
0.86 t (7.5)
COOH
O
COOH
O
Me
H
O
COOH
HOOC
HO
HO
HOOC
HO
HO
O
O
Me
H
O
HOOC
O
OH
CH2OH
OH
HO
HO
Me
O
Me
2
1
Me
H
O
H
1
OH
Fig. 2 Key HMBC (rightwards arrow) and H–1H COSY (thin line)
correlations of caraganins A (1) and B (2)
Me
Me
might also be an artifact produced from 2 during the
extraction procedure with n-BuOH.
Fig. 3 Key NOESY (double-ended arrow) correlations of caraganin
A (1)
All the isolated compounds were assessed in vitro for
their antibacterial activities against Escherichia coli,
Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus
subtilis, and Klebsislla pneumoniae. Compound 1a, the
most active compound, exhibited a minimum inhibitory
concentration (MIC) value of 3.125 lg/ml against Staphy-
lococcus aureus and Bacillus subtilis (Table 3). Compound
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193
Table 2 13C-NMR (100 MHz) data for compounds 1, 2 and their n-butyl esters (1a and 2a) in C5D5N
No.
daC (d, ppm)
No.
daC (d, ppm)
1
2
1a
38.9 t
2a
38.7 t
1
2
1a
46.8 t
2a
1
38.9 t
27.5 t
89.1 d
39.7 s
55.8 d
18.6 t
33.0 t
40.1 s
47.9 d
37.0 s
24.0 t
124.9 d
141.6 s
42.2 s
25.6 t
26.8 t
48.4 s
47.2 d
41.8 t
44.8 s
38.7 t
27.5 t
89.2 d
44.4 s
56.2 d
18.8 t
33.2 t
40.1 s
47.9 d
36.9 s
24.1 t
21
22
23
24
25
26
27
28
29
30
10
46.7 t
215.0 s
28.3 q
17.2 q
15.8 q
17.1 q
25.7 q
21.1 q
179.2 s
21.8 q
107.4 d
75.7 d
78.3 d
73.6 d
78.0 d
173.6 s
46.8 t
46.6 t
215.0 s
23.4 q
63.4 t
2
28.3 t
89.2 d
39.7 s
55.8 d
18.6 t
33.0 t
40.6 s
47.9 d
37.0 s
24.0 t
124.8 d
141.7 s
42.1 s
25.5 t
26.8 t
48.4 s
47.2 d
41.8 t
44.6 s
27.5 t
89.2 d
44.5 s
56.2 d
18.9 t
33.3 t
40.0 s
47.7 d
36.7 s
24.2 t
124.8 d
141.6 s
42.1 s
25.5 t
27.0 t
48.4 s
47.2 d
41.8 t
44.7 s
215.0 s
23.5 q
63.4 t
215.0 s
28.2 q
17.1 q
15.8 q
17.0 q
25.6 q
21.1 q
179.1 s
21.9 q
107.5 d
75.6 d
78.2 d
73.2 d
77.5 d
170.5 s
65.1 t
3
4
5
15.6 q
17.0 q
25.5 q
21.1 q
179.1 s
21.8 q
106.8 d
75.5 d
78.2 d
73.4 d
77.8 d
174.4 s
15.6 q
16.9 q
25.5 q
21.1 q
179.1 s
21.8 q
106.7 d
75.4 d
78.1 d
73.3 d
77.6 d
170.4 s
65.2 t
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
a
124.9 d
141.6 s
42.1 s
25.5 t
26.8 t
48.4 s
47.2 d
41.8 t
44.7 s
20
30
40
50
60
100
200
300
400
31.0 t
31.0 t
19.4 t
19.4 t
13.9 q
13.9 q
Multiplicity was determined by DEPT experiments (s = quaternary, d = methine, t = methylene, q = methyl)
2a also displayed selective activity against Staphylococcus
aureus and Bacillus subtilis, with MIC values of 3.125 and
6.25 lg/ml, respectively. However, both caraganins A (1)
and B (2) showed no activity (MIC values[100 lg/ml). A
comparison of the chemical structures showed that intro-
duction of an n-butyl ester group plays a vital role in the
potency of these compounds and formation of an ester group
from a carboxyl group may therefore produce beneficial
changes in medicinal parameters, such as solubility, polar
surface area, molecular dipole, and intestinal absorption
[11].
chromatography were silica gel (100–200 mesh; Huiyou
Silical Gel Development Co. Ltd. Yantai, China), silica gel
H (10–40 lm; Yantai), Sephadex LH-20 (40–70 lm;
Amersham Pharmacia Biotech AB, Uppsala, Sweden), and
YMC-GEL ODS-A (50 lm; YMC, Milford, MA).
HSGF254 silica gel TLC plates (Yantai) were used for
analytical TLC.
Plant material
The seeds of C. microphylla, which were identified by Prof.
Han-Chen Zheng, School of Pharmacy, Second Military
Medical University, were obtained from the middle of
Inner Mongolia Autonomous Region, People’s Republic of
China, in July, 2008. A voucher specimen (#2008-116) has
been deposited in the herbarium of the Department of
Pharmacognosy, School of Pharmacy, Second Military
Medical University, Shanghai, China.
Experimental
General
Optical rotations were acquired with a Perkin-Elmer 341
polarimeter. IR spectra were recorded on a Bruker Vector
22 spectrometer with KBr pellets. NMR spectra were
recorded on a Bruker Avance 400 NMR spectrometer with
TMS as an internal standard. ESIMS were measured on
an Agilent LC/MSD Trap XCT mass spectrometer,
whereas HRESIMS were measured using a Q-TOF micro
mass spectrometer (Waters, USA). Materials for column
Extraction and isolation
The dried seeds (10 kg) were powdered and extracted with
hot 80% EtOH (50 L) three times for 2 h each time. After
removal of solvent under reduced pressure, the concen-
trated EtOH extract (1.2 kg) was suspended into water
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J Nat Med (2013) 67:190–195
Table 3 Antimicrobial activities of compounds 1, 2 and their n-butyl esters (1a and 2a) as MIC values (lg/ml)
Compounds
Bacteria
Escherichia
coli
Staphylococcus
aureus
Pseudomonas
aeruginosa
Bacillus
subtilis
Klebsiella
pneumoniae
1
[100
[100
[100
[100
3.125
[100
[100
3.125
3.125
3.125
[100
[100
[100
[100
50
[100
[100
3.125
6.25
[100
[100
[100
[100
6.25
2
1a
2a
Chloramphenicol
3.125
Compound 2a
(2 L) and extracted successively with petroleum ether
(4 9 2 L), CH2Cl2 (4 9 2 L), EtOAc (4 9 2 L), and
n-BuOH (4 9 2 L). The n-BuOH fraction (156.3 g) was
chromatographed on a silica gel (200–300 mesh) column
eluting with a step gradient of CH2Cl2–MeOH (30:1, 15:1,
10:1, 5:1, 2:1, 1:1) to give six fractions (Fr1–Fr6). Fr3
(11.6 g) was subjected to a RP-18 silica gel column using
H2O–MeOH (60/40; flow rate: 3 mL/min) as the eluent to
yield three subfractions designated Fr3-1, Fr3-2, and Fr3-3.
Fr3-1 (1.1 g) was further purified by Sephadex LH-20
[CHCl3–MeOH (1:1)] to afford 1 (86 mg). Fr3-2 was re-
chromatographed on a silica gel column with CH2Cl2–
MeOH (15:1) to give 1a (118 mg) as a colorless powder.
Fr4 (18.8 g) was chromatographed on a silica gel column
eluting with CH2Cl2–MeOH–H2O (9:1:0.1) to yield 2
(20 mg) and 2a (58 mg).
20
Colorless powder; ½aꢁD ?20.8° (c 0.16, MeOH); positive
HRESIMS [M?Na]? peak at m/z 741.4163 (calcd for
C40H62O11Na, 741.4184); IR (KBr) vmax 3451, 2952, 1733,
1704, 1384; 1H-NMR (C5D5N, 400 MHz, Table 1) and
13C-NMR (C5D5N, 100 MHz, Table 2) spectra.
Acid hydrolysis
Compounds 1 (12 mg) and 2 (10.0 mg) were hydrolyzed in
5.0 mL of 5% H2SO4 under reflux for 1 h. The reaction
solution was neutralized by BaCO3 then filtered to remove
the precipitate. The hydrolysate was extracted with ether.
The ether extract was evaporated and chromatographed on
silica gel with CHCl3–MeOH (30:1) to yield the agly-
20
cone subprogenin D [8.4 mg; mp 222–224°C; ½aꢁD ?6.8°
Compound 1
(c 0.2, CH3OH)] from 1 and melilotigenin [6.5 mg; mp
20
20
319–320°C; ½aꢁD ?47.4° (c 0.2, CH3OH)] from 2, which
Colorless powder; ½aꢁD ?8° (c 0.16, MeOH); negative
HRESIMS [M-H]- at m/z 645.3644 (calcd for C36H53O10,
645.3639); IR (KBr) vmax 3440, 2948, 1734, 1703, 1384;
1H-NMR (C5D5N, 400 MHz, Table 1) and 13C-NMR
(C5D5N, 100 MHz, Table 2) spectra.
was identified by direct comparison with reported data
[9, 10]. The sugars from the water layer were identified as
glucuronic acid by TLC comparison with an authentic
sample developed with CHCl3–MeOH–H2O (1: 1: 0.2) at
the same Rf (0.4) value. The absolute configuration of
glucuronic acid was further determined to be D by the
measurement of optical rotation after separation from the
Compound 2
20
aqueous phase of the hydrolysate mixture: ½aꢁD ?11.9°
20
Colorless powder; ½aꢁD ?16.7° (c 0.16, MeOH); negative
HRESIMS [M-H]- at m/z 661.3612 (calcd for C36H53O11,
661.3588); IR (KBr) vmax 3450, 2950, 1735, 1705, 1384;
1H-NMR (C5D5N, 400 MHz, Table 1) and 13C-NMR
(C5D5N, 100 MHz, Table 2) spectra.
(c 0.1, H2O) [12, 13].
Antibacterial assay
Antibacterial assays of compounds 1-4 were individually
tested against pathogenic bacteria including Escherichia
coli, Staphylococcus aureus, Pseudomonas aeruginosa,
Bacillus subtilis, and Klebsislla pneumoniae, using the
microtitre plate broth dilution method [14]. Chloram-
phenicol was used as a positive control and MIC values of
the tested compounds were determined from three indi-
vidual experiments.
Compound 1a
20
Colorless powder; ½aꢁD ?10° (c 0.16, MeOH); positive
HRESIMS [M?Na]? peak at m/z 725.4263 (calcd for
C40H62O10Na, 725.4235); IR (KBr) vmax 3445, 2953, 1732,
1704, 1384; 1H-NMR (C5D5N, 400 MHz, Table 1) and
13C-NMR (C5D5N, 100 MHz, Table 2) spectra.
123

J Nat Med (2013) 67:190–195
195
Acknowledgments The authors are grateful for help from Shanghai
Institute of Organic Chemistry Chinese Academy of Sciences for the
NMR measurements.
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