Two new glycosides from Sanicula lamelligera

Article abstract of DOI:10.1515/znb-2006-0517

Two new glycosides, 21-O-β-D-glucopyranosyl-olean-12-ene-3β, 16α, 21β, 22α, 28-pentol-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-arabinopyranosyl (1 → 3)]-β-D-glucopyranoside-(1) and 4-O-β-D-glucopyranosyl rosmarinic acid (3), along with the known compound rosmarinic acid (2) were isolated from the whole plants of Sanicula lamelligera Hance. Their structures were elucidated by spectroscopic analysis and chemical means.

Full text of DOI:10.1515/znb-2006-0517

Two New Glycosides from Sanicula lamelligera
Ling-Yun Zhoua, Hai-Yang Liua, Bai-Bo Xiea, Zhi-Hai Liub, and Chang-Xiang Chena
a State Key Laboratory of Phytochemistry and Plant Resources in West China, Kunming Institute of
Botany, The Chinese Academy of Sciences, Kunming, Yunnan 650204, P. R. China
b Guiyang College of Traditional Chinese Medicine, Guiyang, Guizhou 550001, P. R. China
Reprint requests to Prof. C.-X. Chen. Fax: +86-871-5219934. E-mail: cxchen@mail.kib.ac.cn
Z. Naturforsch. 61b, 607 – 610 (2006); received July 14, 2005
Two new glycosides, 21-O-β-D-glucopyranosyl-olean-12-ene-3β, 16α, 21β, 22α, 28-pentol-3-O-
β-D-glucopyranosyl (1 → 2)-[β-D-arabinopyranosyl (1 → 3)]-β-D-glucopyranoside (1) and 4-O-β-
D-glucopyranosyl rosmarinic acid (3), along with the known compound rosmarinic acid (2) were
isolated from the whole plants of Sanicula lamelligera Hance. Their structures were elucidated by
spectroscopic analysis and chemical means.
Key words: Sanicula lamelligera, Triterpenoid Saponin, 4-O-β-D-Glucopyranosyl,
Rosmarinic Acid
Introduction
Sanicula lamelligera Hance (Umbelliferae) is a folk
medicine used for treatment of wounds, mild catarrhs
of the respiratory system and chronic chest pains [1].
It was recently reported that saponins from this genus
were responsible for antimicrobial, hemolytic, and an-
tioxidant activities, whereas hydroxycinnamic deriva-
tives, especially rosmarinic acid, were responsible for
antioxidant activity and acted as active constituents
against HIV-1 [2, 3]. Up to now, the chemical con-
stituents of this plant have not been reported. Our in-
vestigation on this plant led to the isolation of two new
glucosides (1 and 3) (see Figs 1 and 2).
Fig. 1. Chemical structure and key correlations of com-
pound 1.
Results and Discussion
Compound 1 was isolated as a white amorphous
powder and analyzed for C53H88O24 by negative-ion
HRESI-MS. The 1H NMR and 13C NMR spectrum of
1 displayed four anomeric proton signals at δ = 5.70
(d, J = 7.8 Hz), 5.24 (d, J = 7.8 Hz), 4.98 (d, J =
7.8 Hz) and 4.86 (d, J = 7.8 Hz) and four anomeric car-
bon signals at δ = 103.8, 105.3, 106.6 and 105.3. Con-
sequently 1 was assumed to possess four sugar units.
On acid hydrolysis, 1 gave an aglycone, identified as
barringtogenol C (1a) by comparison of its spectral
features with those previously reported [4], and sugar
components which were identified as glucoside, arabi-
nose by comparison with authentic samples on TLC.
The sugar linkages were determined by 2D NMR ex-
Fig. 2. Chemical structures of compounds 2 and 3. glc →
β-D-glucopyranosyl.
periments. In the HMBC spectrum, long-range cor-
relations were observed between H-1 (δ = 4.98, d,
J = 7.8 Hz) of G-3 and C-21 (δ = 92.5) of the agly-
c
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L.-Y. Zhou et al. · Two New Glycosides from Sanicula lamelligera
Table 1. 13C NMR and
1H NMR spectral data for 1,
13C NMR for 1a (in pyridine-
d5, 125 MHz).
1
1a
13C
No
1
13C
1H
13C
105.3
79.6
87.2
69.7
77.8
62.5
103.8
76.6
78.0
72.5
78.7
63.3
105.3
72.9
74.9
69.8
68.0
106.6
75.7
78.3
71.5
78.6
62.6
1H
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
39.6 (t)
26.7 (t)
89.3 (d)
38.9 (s)
55.8 (d)
18.5 (t)
33.1 (t)
40.6 (s)
47.0 (d)
37.3 (s)
23.9 (t)
123.1 (d)
144.0 (s)
42.0 (s)
34.4 (t)
68.0 (d)
48.0 (s)
40.1 (d)
48.0 (t)
1.38, 0.85 (m)
2.16 (br d, 10.5 Hz)
3.27 (dd, 11.5, 4.0 Hz)
39.2
28.2
78.1
39.5
55.9
18.8
33.3
40.1
47.2
37.3
23.9
123.1
144.0
42.1
34.4
67.9
47.4
41.3
48.3
G1-1
4.86 (d, 7.8 Hz)
2
3
4
5
0.71 (d, 11.5 Hz)
1.25, 1.47 (m)
1.22, 1.58 (m)
6
G2-1
5.70 (d, 7.8 Hz)
2
3
4
5
1.75 (m)
1.87 (m)
5.39 (s)
6
ara-1
2
3
4
5.24 (d, 7.8 Hz)
4.98 (d, 7.8 Hz)
1.61, 2.00 (m)
4.0 (t, 9.4 Hz)
5
2.85 (dd, 13.0 Hz, 3.8 Hz)
1.38 (m)
3.14 (t, 13.0)
G3-1
2
3
4
5
6
20
21
22
23
24
25
26
27
28
36.8 (s)
92.5 (d)
74.1 (d)
28.0 (q)
16.8 (q)
15.8 (q)
16.9 (q)
27.6 (q)
67.3 (t)
36.5
78.7
77.3
28.8
16.6
15.8
17.0
27.5
68.4
4.85 (d, 9.2 Hz)
4.61 (d, 9.2 Hz)
1.21 (s)
1.06 (s)
0.80 (s)
0.86 (s)
1.84 (s)
3.65 (d, 10.0 Hz)
3.95 (d, 10.0 Hz)
1.53 (s)
29
30
30.2 (q)
20.6 (q)
30.6
19.5
1.36 (s)
cone, H-1 (δ = 4.86, d, J = 7.8 Hz) of G-1 and C-3 with the data reported [5], including absolute configu-
(δ = 89.3) of the aglycone, H-1 (δ = 5.70, d, J = ration (R) at C-8.
8.0 Hz) of G-2 and C-2 (δ = 79.6) of G-1, H-1 (δ =
5.24, d, J = 7.8 Hz) of terminal arabinosyl and C-3
(δ = 87.2) of G-1. From the NMR data each sugar is
a pyranosyl with β configuration for glucosyl and ara-
binosyl. Therefore, the structure of 1 is assigned as 21-
O-β-D-glucopyranosyl-olean-12-ene-3β, 16α, 21β,
22α, 28-pentol-3-O-β-D-glucopyranosyl(1 → 2)-[β-
D-arabinopyranosyl (1 → 3)]-β-D-glucopyranoside.
Compound 3 was obtained as yellow gum.
Negative-ion HRESI-MS gave an [M-1]− peak at
m/z 521.1287, corresponding to a molecular formular
of C24H26O13. The 1H NMR and 13C NMR spectra of
3 were similar to those of 2, except that signals cor-
responding to the sugar moiety included an anomeric
proton signal [δ = 4.67 (1H, d, J = 7.0 Hz)] and
anomeric carbon signal (δ = 102.5). The absolute con-
Compound 2 was obtained as gray amorphous pow- figuration of 3 deduced as follows. Enzymic hydroly-
der. {[α]2D5 + 91.9 (c 0.42, MeOH).} Negative-ion sis of 3 yielded rosmarinic acid {[α]2D5 + 118.0 (c 0.5,
FAB-MS giving an [M-1]− peak at m/z 359 indicated MeOH)} and D-glucose. The glucosyl unit attached
the molecular formula C18H16O8. The 1H NMR spec- to C-4 was deduced from the heteronuclear multiple-
trum of 2 indicated the presence of two 1, 3, 4-trisubst- bond correlation (HMBC) interactions between the
ituted phenyl proups [δ = 6.98 (1H, d, J = 8.0 Hz), anomeric proton signal (δ = 4.67) and the carbon sig-
6.78 (1H, br d, J = 8.0 Hz), 6.85 (1H, br s), 6.68 nal of C-4 (δ = 144.4). Furthermore, the J value (> 7)
(1H, br s), 6.77 (1H, d, J = 8.0 Hz), 6.65 (1H, br d, of the anomeric proton of the sugar moieties indicated
J = 8.0 Hz)] and one trans double bond [δ = 6.04, the β-orientation at the anomeric center of D-glucose.
7.24 (each 1H, d, J = 16.0 Hz)]. Finally, the structure Thus, 3 was determined to be 4-O-β-D-glucopyranosyl
of 2 was assigned as rosmarinic acid on comparison rosmarinic acid.
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L.-Y. Zhou et al. · Two New Glycosides from Sanicula lamelligera
609
Table 2. 13C NMR and 1H NMR spectral data for 2, 3 (in
D2O, 125 MHz).
specimen is deposited in the Herbarium of Kunming Institute
of Botany, Chinese Academy of Sciences.
No
2
3
13C
1H
13C
1H
Extraction and isolation
1
2
3
4
5
6
7
131.3 (s)
118.1 (d) 6.85 (br s)
144.7 (s)
143.5 (s)
117.1 (d) 6.98 (d, 8.0 Hz)
122.6 (d) 6.78 (br d, 8.0 Hz) 122.6 (d) 6.68 (br d, 8.0 Hz)
37.8 (d) 2.97 37.8 (d) 2.97
(dd, 14.0, 4.3 Hz) (dd, 14.0, 4.3 Hz)
134.5 (s)
118.0 (d) 6.78 (br s)
146.4 (s)
144.4 (s)
117.8 (d) 6.90 (d, 8.0 Hz)
The air-dried plants (5.0 kg) were exhaustively extracted
with 75% MeOH (4 l × 3) at r. t., and the combined ex-
tracts were concertrated under reduced pressure. The crude
extraction (200 g) was subjected to DM 101 resin column
chromatography, eluting with water and 95% EtOH, succes-
sively. Then, the 95% EtOH faction was evaporated (Under
red. press), giving a residue (140 g) that was purified on col-
umn chromatography (silica gel H-60, 15 × 110 cm) with a
CHCl3-MeOH gradient elution. 1 (70 mg) was obtained from
fraction 4 (CHCl3: MeOH: H2O, 7: 3: 0.5 v/v) by repeated
Rp-18 column chromatography. 2 (200 mg) was obtained
from fraction 3 (CHCl3: MeOH, 3: 1 v/v) by repeated col-
umn chromatography 3 (30 mg) was obtained from fraction
5 (CHCl3: MeOH: H2O, 7: 4: 0.5 v/v) by repeated Rp-18
column chromatography.
21-O-β-D-glucopyranosyl-olean-12-ene-3β, 16α, 21β,
22α, 28-pentol-3-O-β-D-glucopyranosyl (1 → 2)-[β-D-
arabinopyranosyl (1 → 3)]-β-D-glucopyranoside (1), white
powder. – M. p. 292 – 294 ◦C. – [α]D25 − 8.3 (c 0.60,
H2O). – UV/vismax (pyridine): λmax(lgε) = 204 nm (0.57). –
IR (KBr) = 3424 (br, OH), 2884, 1632, 1078, 1041 cm−1. –
1H NMR and 13C NMR see Table 1. – MS (FAB−) m/z
(%) = 1107 [M−H]− (100), 946 [M−glucose]− (9). –
MS (HRESI): m/z = 1107.5597 (C53H88O24Na. calcd.
1107.5587).
3.08
3.12
(dd, 14.0, 8.6 Hz)
(dd, 14.0, 8.6 Hz)
8
77.3 (d) 5.18
(dd, 4.3, 8.6 Hz)
177.5 (s)
127.9 (s)
117.0 (d) 6.68 (br s)
145.2 (s)
148.0 (s)
77.1 (d) 5.18
(dd, 4.3, 8.6 Hz)
177.5 (s)
127.9 (s)
117.0 (d) 6.64 (br s)
145.1 (s)
148.0 (s)
9
1’
2’
3’
4’
5’
6’
7’
8’
9’
glc-1
2
116.0 (d) 6.77 (d, 8.0 Hz)
116.0 (d) 6.80 (d, 8.0 Hz)
123.4 (d) 6.65 (br d, 8.0 Hz) 123.4 (d) 6.63 (br d, 8.0 Hz)
146.8 (d) 7.24 (d, 16.0 Hz) 146.9 (d) 7.21 (d, 16.0 Hz)
115.4 (d) 6.04 (d, 16.0 Hz) 115.2 (d) 6.02 (d, 16.0 Hz)
169.7 (s)
169.7 (s)
102.5 (d) 4.67 (d, 7.8 Hz)
73.8 (d)
76.3 (d)
70.1 (d)
3
4
5
6
76.9 (d)
61.3 (d)
The above three compounds showed no anti-tumor
activity against K562, K549, while compound 2 (ros-
marinic acid) showed a weak anti-epiphyte activity.
4-O-β-D-glucopyranosyl rosmarinic acid (3), yellow
gum. – [α]2D5 − 16.7 (c 0.60, H2O). – UV/vismax (H2O):
λ
max(lgε) = 283 nm (3.17). – IR (KBr) ν = 3421 (br,
OH), 1692, 1598, 1510, 1406, 1279 cm−1. – 1H NMR
and 13C NMR see Table 2. – MS (FAB−) m/z (%) =
Experimental Section
521 [M−H]− (100), 359 [M−glucose−H]− (8).
–
General
MS (HRESI): m/z = 521.1287 (C24H26O13Na. calcd.
521.1295).
Melting points were measured on a Kofler melting point
apparatus produced by Sichuan University (China) and are
uncorrected. UV spectra were obtained on a UV-210A spec-
trometer Company apparatus. IR spectra were performed on
a Bio-Rad FTS-135 spectrometer. Optional rotations were
measured on a Japanese Fasco DIP-370 digital polarimeter.
The negative ion FABMS and HR-ESIMS were recorded on
a VG Auto Spec-3000 spectrometer. All NMR experiments
were recorded on a Bruker DRX-500 spectrometer with TMS
as internal standard. The silica gel for TLC and column chro-
matography were obtained from Qingdao Marine Chemical
Inc., China.
Acid hydrolysis of compound 1: A solution of 1 (50 mg)
in 5% H2SO4 was heated at 100 C for 2 h. The reaction
mixture was extracted with EtOAc and purified by MPLC
(CHCl3: MeOH = 30: 1, v/v) to provide compound 1a
(15 mg). 13C NMR see Table 1.
Enzymic hydrolysis of Compound 3: A mixture of com-
pound 3 (20 mg) and complex cellulosic enzyme (1 g) in
AcOH-AcONa buffe◦r (pH 5.5, 10 ml) was incubated with
gentle stirring at 28 C for 72 h. The reaction mixture was
extracted with EtOAc (30 ml×3). The extraction was chro-
matographed on silica gel (CHCl3:MeOH, 7:3 v/v) to give
rosmarinic acid {4 mg, [α]2D5 +118.0 (c 0.5, MeOH)}.
◦
Plant material
Acknowledgements
The whole plants of Sanicula lamelligera Hance were col-
lected from Linglang County of Yunnan Province, P.R. China
The authors are grateful to Mr. Shao-Lian Ma for pro-
in April 2003 and identified by Prof. Y. M. Shui. A voucher viding the sample and the Analytical Group of the Labora-
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610
L.-Y. Zhou et al. · Two New Glycosides from Sanicula lamelligera
tory of Phytochemistry, Kunming Institute of Botany, Chi-
nese Academy of Sciences, for the spectral measurements.
[3] T. H. Schopke, M. Janka, M. Nimtz, V. Wray, K. Hiller,
Planta Med. 64, 83 (1998).
[4] K. Takao, K. H. Lee, J. Nat. Prod. 49, 650 (1986).
[5] J. K. Charles, C. H. Richard, C. Marvin, J. Org. Chem.
41, 449 (1976).
[1] Institutum Botanicum Kunmingense Academiae Sini-
cae Edita, Flora Yunnanica (Science Press, Beijing). To-
mus 7, 376 (1997).
[2] A. Nazli, G. Nezhun, K. Avni, P. Thitima, M. P. John,
S. X. Qiu, A. C. Geoffrey, J. Nat. Prod. 60, 1170 (1997).
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