Glycosides from the methanol extract of Notopterygium incisum

Article abstract of DOI:10.1055/s-0031-1279995

Five new (15) and twelve known (617) different types of glycosides together with a known sesquiterpene triol (18) were isolated from the methanol extract of the rhizomes of Notopterygium incisum. The new structures were elucidated by means of spectroscopic and chemical methods to be pregn-5-en-3,20(S)-diol-3-O- bis - D-glucopyranosyl-(l2,16) - D-glucopyranoside (1), oleuropeic aldehyde 8-O - D-glucopyranoside (2), 2(R)-(3,4-dimethoxyphenyl)-propane-1,3-diol-1-O - D-glucopyranoside (3), eudesman-3,4,11-triol-11-O - D-glucopyranoside (4), and marmesin-11-O - D-glucopyranosyl-(16) - D-glucopyranoside (5). The absolute configuration of the aglycone in compound 3 was assigned by application of Klyne's rule. Georg Thieme Verlag KG Stuttgart - New York.

Full text of DOI:10.1055/s-0031-1279995

Letters 1939
Key words
Notopterygium incisum · Umbelliferae · structure elucidation ·
Glycosides from the Methanol
glycosides
Extract of Notopterygium incisum
Supporting information available online at
http://www.thieme-connect.de/ejournals/toc/plantamedica
Mei You1, Juan Xiong2, Yun Zhao1, Lei Cao1, Shi-Biao Wu1,
Gang Xia1, Jin-Feng Hu1,2
1
Department of Natural Products for Chemical Genetic Research,
Key Laboratory of Brain Functional Genomics, Ministry of Education
(MOE), East China Normal University, Shanghai, P.R. China
Department of Natural Products Chemistry, School of Pharmacy,
The traditional Chinese medicine “Qianghuo” is composed of the
rhizomes of two species of Notopterygium (Umbelliferae),
N. incisum Ting ex H.T. Chang and N. forbesii De Boiss [1]. Chemi-
cally, only a few polar constituents (coumarin glycosides, amino
acids, chlorogenic acid, ferulic acid, and daucosterol) were previ-
ously characterized from this herb medicine [2–6]. In an earlier
work on the chloroform extract of the rhizomes of N. incisum, a
number of lipophilic constituents (including antiproliferative fu-
rocoumarins) have been identified [7]. After being exhaustively
extracted with chloroform, the rhizomes were further extracted
with methanol. The methanol extract was found to show a pre-
liminary cytotoxic effect against the human MCF-7 breast cancer
cell line. During a reinvestigation of polar constituents from this
plant and in a continuation of our ongoing project towards the
discovery of novel antitumor agents from natural products [8],
seventeen (1–17) different types of glycosides and a polar sesqui-
2
Fudan University, Shanghai, P.R. China
Abstract
!
Five new (1–5) and twelve known (6–17) different types of glyco-
sides together with a known sesquiterpene triol (18) were iso-
lated from the methanol extract of the rhizomes of Notoptery-
gium incisum. The new structures were elucidated by means of
spectroscopic and chemical methods to be pregn-5-en-3β,20(S)-
diol-3-O-bis-β-D-glucopyranosyl-(l → 2,1 → 6)-β-D-glucopyrano-
side (1), oleuropeic aldehyde 8-O-β-D-glucopyranoside (2), 2(R)-
(3,4-dimethoxyphenyl)-propane-1,3-diol-1-O-β-D-glucopyrano-
side (3), eudesman-3α,4α,11-triol-11-O-β-D-glucopyranoside (4),
and marmesin-11-O-β-D-glucopyranosyl-(1 → 6)-β-D-glucopy-
ranoside (5). The absolute configuration of the aglycone in com-
pound 3 was assigned by application of Klyneʼs rule.
"
terpene hedytriol (18) [9] (l Fig. 1) were obtained from the
methanol extract. These include a new pregnane glycoside (1), a
new monoterpenoid glycoside (2), a new phenylpropanoid glyco-
side (3), a new eudesmane glycoside (4), and a new furanocou-
Fig. 1 Chemical structures of compounds 1–18.
You M et al. Glycosides from the… Planta Med 2011; 77: 1939–1943

1940 Letters
Table 1 1H NMR (500 MHz, J in Hz) and 13C NMR (125 MHz) data of compounds 1 and 6.
No.
1a
6a
No.
1a
6a
δH
δC
37.2
δH
δC
37.1
δH
δC
δH
δC
1
2
1.72 br d, 13.1
0.95 m
1.69 br d, 12.8
0.92 m
2.16 br d, 11.0
1.82 m
3.91 m
2.84 dd, 13.4, 1.2
2.69 dd, 13.4, 10.8
–
1′
2′
3′
4′
5′
6′
5.10 d, 7.8
4.17 dd, 9.0, 7.8
4.38 ddb
100.7
83.3
77.4
71.0e
76.9
69.6
5.11 d, 7.6
4.18 dd, 8.7, 8.4
4.39 ddb
100.7
83.3
77.4
71.1e
76.8
69.7
2.16 m
29.9
29.8
1.82 m
4.30 dd, 9.4, 8.9
4.06 m
4.30 dd, 9.3, 9.0
4.07 m
3
4
3.92 m
79.1
39.0
79.0
39.0
2.84 dd, 13.2, 1.2
2.69 dd, 13.2, 12.1
–
4.81 br d, 11.0
4.39 ddb
4.82 br d, 11.0
4.41 dd, 11.0, 4.6
5.31 d, 7.6
4.08 ddc
4.22 ddd
4.23 ddd
5
6
7
140.6
121.7
31.9
140.7
121.4
31.7
1″
2″
3″
4″
5″
6″
5.29 d, 7.7
4.06 ddc
4.22 ddd
4.23 ddd
105.2
75.9
77.9f
70.9e
77.9f
62.2g
105.1
75.8
77.9f
71.0e
77.9f
62.2g
5.46 br s
1.96 m
5.44 br s
1.91 m
1.52 m
1.32 m
0.88 m
–
1.55 m
8
9
1.39 m
31.5
50.0
36.7
20.8
31.6
49.8
36.6
21.0
3.92 m
3.91 m
0.89 m
4.51 br d, 11.0
4.36 ddb
4.52 br d, 11.8
4.38 ddb
10
11
–
1.39 m
1.43 m
1.26 m
1.91 m
1.23 m
–
1‴
2‴
3‴
4‴
5‴
6‴
5.05 d, 7.8
4.06 ddc
4.22 ddd
4.23 ddd
105.2
74.8
78.1f
71.2e
77.7f
62.4g
5.07 d, 7.6
4.08 ddc
4.22 ddd
4.23 ddd
105.1
74.7
78.1f
71.3e
77.6f
62.4g
12
1.88, m
1.06 m
–
38.8
38.5
13
14
15
41.3
56.6
24.2
43.5
56.5
24.3
3.92 m
3.91 m
0.95 m
1.57 m
1.09 m
2.16 m
1.88 m
1.51 m
0.71 s
0.92 m
1.49 m
1.08 m
2.28 m
1.54 m
2.42 t, 8.8
0.57 s
4.51 br d, 11.0
4.36 ddb
4.52 br d, 11.8
4.38 ddb
16
26.5
22.8
17
18
19
20
21
59.0
12.3
19.4
68.9
24.4
63.3
12.9
1.02 s
0.97 s
19.2
3.92 m
1.43 d, 6.0
–
208.0
31.0
2.06 s
a Recorded in C5D5N. b–d Signals were overlapped within the same superscript in the same column. e–g Assignments may be interchangeable within the same superscript in the same
column
marin glycoside (5). In this paper, we report the isolation and
structure elucidation of the new compounds and evaluation of
their cytotoxic effects on a small panel of human cancer cell lines.
Comparing their MS and NMR data and their physical properties
with the literature, or by direct comparison with authentic sam-
ples, the known compounds were identified as pregn-5-en-3β-
ol-20-one-3-O-bis-β-D-glucopyranosyl-(l → 2,1 → 6)-β-D-gluco-
pyranoside (6) [10], celerioside E (7) [11], ananosmoside A (also
named pumilaside A) (8) [12,13], marmesinin (9) [14], 6′-O-
trans-feruloyl-nodakenin (10) [2], 5-methoxy-8-O-β-D-glucosy-
loxypsoralene (11) [15], bergaptol-O-β-D-glucopyranoside (12)
[2], fraxin (13) [16], methyl α-D-fructofuranoside (14) [17], meth-
yl β-D-fructofuranoside (15) [17], 2-{(6-Ο-[β-D-apiofuranosyl]-β-
D-glucopyranosyl)oxy}propane (16) [18], β-daucosterol (17), and
hedytriol (18) [9].
very similar to those of pregn-5-en-3β-ol-20-one-3-O-bis-β-D-
glucopyranosyl-(l → 2,1 → 6)-β-D-glucopyranoside (6), a known
compound previously obtained from an apocynaceous plant Ne-
rium odorum [10]; however, its 1H and 13C NMR data were not
"
completely assigned until this study (l Table 1). The most ob-
vious difference between these two compounds was that a meth-
yl singlet at δ 2.06 in 6 was replaced by one methyl doublet at δ
1.43 (Me-21) in 1, indicating that the ketone carbonyl group at C-
20 of 6 was reduced to a hydroxyl group in 1. This secondary hy-
droxyl group at C-20 was supported by the HMBC correlations be-
tween H-20 (δ 3.92) and C-16 (δ 26.5)/C‑17 (δ 59.0), between Me-
21 (δ 1.43) and C-20 (δ 68.9)/C‑17 (δ 59.0).
The glycosidic linkage position at C-3 was determined by the
HMBC NMR experiment. A clear 3J correlation was found between
the anomeric proton H-1′ resonating at δ 5.10 and C-3 at δ 79.1.
The interglycosidic linkage positions (1 → 2, 1 → 6) were unam-
biguously confirmed by HMBC correlations from H-1″ (δ 5.29) to
C-2′ (δ 83.3) and H-1‴ (δ 5.05) to C-6′ (δ 69.6). The observed cou-
pling constants of the anomeric protons [H-1′ (J = 7.8 Hz), H-1″
(J = 7.8 Hz), H-1‴ (J = 7.7 Hz)] were characteristic for β-glucosidic
linkage in glucopyranosyl units. In addition, acid hydrolysis of 1
with 2 N HCl yielded a monosaccharide and the aglycone (1a)
(Supporting Information). The sugar was ascertained as D-glu-
cose by direct comparison with an authentic sample according
to HPLC analysis and optical rotation detection. The S absolute
configuration at C-20 in 1a was determined by comparing the
proton chemical shift (in CDCl3) of Me-21 (δ 1.22, see Supporting
The molecular weight of compound 1 and its chemical formula
C
39H64O17 were determined from the positive mode HR-ESIMS.
1
"
The H NMR spectrum (l Table 1) of 1 displayed signals assign-
able to two methyl singlets at δ 0.71 (3H, s) and 1.02 (3H, s), one
methyl doublet at 1.43 (3H, d, J = 6.0 Hz), three anomeric protons
at δ 5.05 (1H, d), 5.10 (1H, d), and 5.29 (1H, d), and an olefinic pro-
ton resonating at δ 5.46 (1H, br s). The 13C and DEPT NMR spectra
"
(l Table 1) of 1 revealed twenty-one carbon signals classified as
three sp3 methyls, eight sp3 methylenes, six sp3 (two oxygenated),
and one sp2 methines, two sp3 and one sp2 quaternary carbons in
addition to eighteen carbon signals attributed to three glucopy-
ranosyl units. These NMR data showed that 1 has general features
You M et al. Glycosides from the… Planta Med 2011; 77: 1939–1943

Letters 1941
Fig. 2 Application of Klyneʼs rule to compound 3.
Information) with those of C-20 epimers of pregn-5-en-3β,20-di-
ol [20(R): δ 1.14; but 20(S): δ 1.21] [19]. Consequently, 1 was elu-
cidated to be pregn-5-en-3β,20(S)-diol-3-O-bis-β-D-glucopyran-
osyl-(l → 2,1 → 6)-β-D-glucopyranoside.
gave a pentaacetate derivative [3a: [α]2D0 = − 23.0 (c 0.15, CHCl3)].
However, we failed to obtain the desired aglycone (3b) by acid
hydrolysis in the presence of 2 N HCl. Significantly, enantiomers
of
9-acetoxy-7-(2,5-dimethoxy-4-methylphenyl)propan-8-ol
Compound 2 exhibited an [M + Na]+ ion peak at m/z 353.1572 in
the positive mode HR-ESIMS, indicating its molecular formula to
be C16H26O7. The 13C and DEPT NMR spectra of 2 exhibited ten
carbon signals assignable to a monoterpene moiety in addition
[(7R)-3c: [α]D = + 11.8; (7S)-3 d: [α]D = − 17.3] have been previ-
ously reported in a synthetic study [20]. The absolute configura-
tion at C-7 in 3a could be deduced by the application of Klyneʼs
"
rule [21]. As shown in l Fig. 2, the calculated [M]D (− 68.4) of 3b
1
to six carbon signals attributed to a glucopyranosyl unit. The H
(subtracting the [M]D of methyl tetraacetyl-β-D-glucopyranoside
(− 65.9) [22] from the calculated [M]D − 134.3 of 3a) was in good
correspondence to the 7S configuration. Therefore, compound 3
was assigned as 2(R)-(3,4-dimethoxyphenyl)-propane-1,3-diol-
1-O-β-D-glucopyranoside.
NMR spectrum of 2 revealed the presence of two tertiary methyl
groups [δ 1.23, 1.26 (each 3H, s)], an olefinic proton [δ 6.94 (1H,
brdd, J = 2.7, 2.3 Hz)], and an aldehyde proton [δ 9.38 (1H, s)] in
the aglycone moiety, which was further deduced to be oleuropeic
aldehyde (a menthane-type monoterpene) by a spin system
(-CH2CH2CHCH2CH=) in its COSY NMR spectrum. Thus, com-
pound 2 was determined to be oleuropeic aldehyde 8-O-β-D-glu-
copyranoside. The bulky substituent at C-4 was in an equatorial
orientation due to the large coupling constant (J = 11.3 Hz) be-
tween H-4ax and H-3ax. An attempt to obtain the aglycone of 2
by hydrolysis with 2 N HCl failed probably due to sample decom-
position, and the absolute configuration at C-4 hence remains un-
known.
The positive mode HR-ESIMS of compound 4 showed an [M + Na]
+
ion peak at m/z 441.2468, corresponding to the molecular for-
mula C21H38O8. The assignments of 1H and 13C NMR data of 4
were made by a combination of 1D and 2D NMR techniques
(COSY, HSQC, and HMBC). These NMR data showed general fea-
tures very similar to celerioside E (7), a eudesmane glucoside
previously isolated from the polar extract of fruits of Apium
graveolens L. (Umbelliferae) [11]. The major difference between
these two compounds was that the secondary hydroxyl group at
C-1 in 7 was relocated at C-3 in 4 of ring A, which was confirmed
by HMBC correlations of Me-14 (δ 1.08) with C-3 (δ 75.7), C-4 (δ
74.4), and C-5 (δ 48.1), and correlations of Me-15 (δ 0.91) with C-
The molecular formula C17H26O9 of compound 3 was determined
by HR-ESIMS, which gave an [M + Na]+ ion peak at m/z 397.1452.
1
Detailed analyses of H, 13C NMR data of 3 with the aid of COSY
3
and HSQC NMR experiments established the presence of a ben-
zene ring with an ABX system [δ 6.94 (1H, brs), 6.88 (1H, brd,
J = 8.0 Hz), 6.84 (1H, d, J = 8.0 Hz)], two methoxyl groups [δ 3.83,
3.80 (each 3H, s)], one 1,3-propanediol group, together with a
glucopyranosyl unit. The above data indicated that 3 was a
(1,3,4-trisubstituted phenyl)propanoid glucoside. In the HMBC
NMR spectrum of 3, the anomeric proton H-1′ at δ 4.31 exhibited
a correlation with C-8 (δ 72.2), whereas H-7 at δ 3.03 showed cor-
relations with C-1 (δ 135.1), C-2 (δ 113.6), and C-6 (δ 113.1), re-
spectively. The β-orientation of the glucosidic linkage was also
deduced from the characteristic coupling constant (J = 7.9 Hz) of
the anomeric proton.
1 (δ 34.9), C-5 (δ 48.1), C-9 (δ 45.9), and C-10 (δ 35.2). A clear J
HMBC correlation was also observed between the anomeric pro-
ton H-1′ at δ 4.46 (1H, d, J = 7.7 Hz) and C-11 at δ 81.6, indicating
that C-11 was the glycosidic linkage position. The relative stereo-
chemistry at C-3, C-4, C-5, C-7, and C-10 in 4 was characterized
through analyses of the coupling patterns of the protons bonded
to the cyclohexane ring and the NOE correlations in the NOESY
NMR experiment. The small coupling constant (br s) found for
H-3 resonating at δ 3.51 indicated that it is in an equatorial orien-
tation. Clear NOE correlations were observed between Me-14 (δ
1.08) and Me-15 (δ 0.91), between Me-14 and H-3 (δ 3.51), be-
tween Hax-9 (δ 1.24) and H-5 (δ 1.57), as well as between H-5
and H-7 (δ 1.55). As indirect evidence, the 13C NMR data of C-6
(δ 22.1), C-7 (δ 49.6), C-8 (δ 24.3), C-11 (δ 81.6), C-12 (δ 23.5),
The determination of the absolute configuration at C-7 has been
challenging. Acetylation of 3 with anhydrous Ac2O in pyridine
You M et al. Glycosides from the… Planta Med 2011; 77: 1939–1943

1942 Letters
and C-13 (δ 25.5) of compound 4 resembled those of 7, indicating
H-2′), 2.59 (1H, brd, J = 19.2 Hz, Ha-3), 2.43 (1H, m, Ha-6), 2.23
(1H, brdd, J = 19.1, 11.3 Hz, Hb-3), 2.14 (1H, m, Ha-5), 2.01 (1H,
m, Hb-6), 1.80 (1H, m, H-4), 1.28 (3H, s, Me-9), 1.26 (3H, s, Me-
10), 1.21 (1H, m, Hb-5). 13C NMR (125 MHz, CD3OD): δ 142.7 (C-
1), 153.9 (C-2), 29.4 (C-3), 45.1 (C-4), 23.1 (C-5), 23.7 (C-6), 195.9
(C-7), 80.4 (C-8), 24.7 (C-9), 23.4 (C-10), 98.5 (C-1′), 75.3 (C-2′),
78.3 (C-3′), 71.8 (C-4′), 77.5 (C-5′), 62.9 (C-6′). LR-ESIMS: m/z 353
[M + Na]+, 683 [2M + Na]+; HR-ESIMS: m/z 353.1572 [M + Na]+
(calcd. for C16H26O7Na: 353.1571, Δ = − 0.3 ppm).
2(R)-(3,4-Dimethoxyphenyl)-1,3-propanediol-1-O-β-D-glucopyran-
oside (3): Colorless oil; [α]2D4 − 18.2 (c 0.79, MeOH); 1H NMR
(500 MHz, CD3OD): δ 6.94 (1H, brs, H-2), 6.88 (1H, brd, J = 8.0 Hz,
H-6), 6.84 (1H, d, J = 8.0 Hz, H-5), 4.31 (1H, d, J = 7.9 Hz, H-1′), 4.13
(1H, dd, J = 10.0, 6.0 Hz, Ha-8), 3.89 (1H, dd, J = 11.5, 6.5 Hz, Ha- 9),
3.86 (1H, brd, J = 12.0 Hz, Ha-6′), 3.84 (1H, dd, J = 10.0, 6.5 Hz, Hb-
8), 3.83 (3H, s, OMe), 3.80 (3H, s, OMe), 3.78 (1H, dd, J = 11.5,
6.5 Hz, Hb-9), 3.67 (1H, dd, J = 12.0, 5.4 Hz, Hb-6′), 3.34 (1H, dd,
J = 8.3, 6.5 Hz, H-3′), 3.27 (2H, m, H-4′, H-5′), 3.20 (1H, dd, J = 8.5,
8.0 Hz, H-2′), 3.03 (1H, quint, J = 6.0 Hz, H-7). 13C NMR (125 MHz,
CD3OD): δ 135.1 (C-1), 113.6 (C-2), 149.3 (C-3), 150.3 (C-4), 121.7
(C-5), 113.1 (C-6), 49.0 (C-7), 72.2 (C-8), 64.8 (C-9), 56.5 (OMe×2),
104.7 (C-1′), 75.1 (C-2′), 78.2 (C-3′), 71.7 (C-4′), 78.0 (C-5′), 62.8 (C-
6′). LR-ESIMS: m/z 397 [M + Na]+, 771 [2M + Na]+, 409 [M + Cl]−,
783 [2M + Cl]−; HR-ESIMS: m/z 397.1452 [M + Na]+ (calcd. for
that these two compounds have the same configuration at C-7.
Consequently, compound 4 was determined to be eudesman-
3α,4α,11- triol-11-O-β-D-glucopyranoside.
The 1H and 13C NMR data revealed that 5 is structurally related to
the known coumarin glucosides marmesinin (9) [14] and decuro-
side I (5b) [23]. Similar to 5b, the carbon chemical shift (δ 62.6)
[15] of C-6′ in 9 was shifted downfield to δ 68.3 in 5, indicating
that the terminal glucopyranosyl unit is linked to C-6′. Acid hy-
drolysis of compound 5 gave D-glucose. Meanwhile, the mea-
sured optical rotation ([α]2D4 = + 15.6) of the aglycone (5a) was
consistent with the previous reported data for marmesin (lit.
[24]: [α]D = + 23) rather than nodakenetin (lit. [2]: [α]D = − 26.0).
Interestingly, marmesin was previously isolated as a major com-
ponent from the chloroform extract of this plant [7]. Therefore,
the structure of 5 was established as marmesin-11-O-β-D-gluco-
pyranosyl-(1 → 6)-β-D-glucopyranoside.
The known glucosides 6–8, 13, 16, and the polyhydroxylated ses-
quiterpene 18 were isolated for the first time from the genus No-
topterygium. Until now, phytochemical investigation of the rhi-
zomes of N. incisum has been accomplished in our group. The re-
sults would give a broad spectrum of naturally occurring com-
pounds from N. incisum. In agreement with our previous findings
(especially coumarins and sesquiterpenoids) from the chloro-
form extract [7], six coumarin glucosides (5, 9–13) and three ses-
quiterpenoid glucosides (4, 7, 8) were isolated from the methanol
extract in this study. In the case of compounds 1, 2, 5, 9, and 10,
we extensively proved the sugar moiety to be β-D-glucopyrano-
side (Supporting Information). In accordance to this result and
with the assumption of a common biosynthetic pathway for the
rest new and known glycosides, the sugar unit should also be D-
glucose.
All the isolates (except 2 and 17) were evaluated for their in vitro
cytotoxic effects against a small panel of human cancer cell lines
(SNU739, NUGC-3, MCF-7, SH-SY5Y) using the CellTiter GloTM lu-
minescent cell viability assay. But none of them appear to be ac-
tive (IC50 > 100 µM). It is worthy of note that the inactivity of fur-
anocoumarin glucosides (5, 9–12) against MCF-7 cells might sup-
port our previous hypothesis that a lipophilic side chain bearing a
free hydroxyl is essential for the cytotoxic effect of linear furano-
coumarins [8].
C
17H26O9Na: 397.1469, Δ = + 4.4 ppm).
Eudesman-3α,4α,11-triol-11-O-β-D-glucopyranoside (4): White
amorphous powder; [α]2D4 − 3.3 (c 0.33, MeOH); 1H NMR
(500 MHz, CD3OD): δ 4.46 (1H, d, J = 7.7 Hz, H-1′), 3.85 (1H, brd,
J = 11.9 Hz, Ha-6′), 3.59 (1H, dd, J = 11.9, 5.6 Hz, Hb-6′), 3.51 (1H,
brs, H-3), 3.35 (1H, dd, J = 8.9, 8.0 Hz, H-3′), 3.22 (2H, m, H-4′, H-
5′), 3.13 (1H, dd, J = 8.9, 7.7 Hz, H-2′), 2.14 (1H, brd, J = 12.3 Hz, Ha-
6), 1.84 (1H, brdd, J = 14.5, 13.8 Hz, Ha-2), 1.68 (1H, brd,
J = 14.5 Hz, Hb-2), 1.59 (1H, m, Ha-8), 1.57 (1H, dd, overlapped,
H-5), 1.55 (1H, m, H-7), 1.53 (1H, m, Ha-1), 1.41 (1H, brd,
J = 11.7 Hz, Ha-9), 1.28 (1H, m, Hb-8), 1.21 (1H, m, Hb-9), 1.05
(1H, m, Hb-1), 1.02 (1H, ddd, overlapped, Hb-6), 1.26, 1.21, 1.08,
0.91 (each 3H, s, Me-13, Me-12, Me-14, Me-15, respectively). 13
C
NMR (125 MHz, CD3OD): δ 34.9 (C-1), 26.7 (C-2), 75.7 (C-3), 74.4
(C-4), 48.1 (C-5), 22.1 (C-6), 49.6 (C-7), 24.3 (C-8), 45.9 (C-9), 35.2
(C-10), 81.6 (C-11), 23.5 (C-12), 25.5 (C-13), 21.7 (C-14), 18.9 (C-15),
98.4 (C-1′), 75.3 (C-2′), 78.2 (C-3′), 72.0 (C-4′), 77.7 (C-5′), 63.0 (C-6′).
LR-ESIMS: m/z 441 [M + Na]+, 859 [2M + Na]+, 453 [M + Cl]−; HR-
ESIMS: m/z 441.2468 [M + Na]+ (calcd. for C21H38O8Na: 441.2459,
Δ = − 2.1 ppm).
Materials and Methods
!
For general experimental procedure, collection, and identifica-
tion of the plant material, see a preceding paper [7]. For extrac-
tion and isolation of compounds 1–18 from the methanol extract
of the rhizomes of N. incisum, see Supporting Information.
Pregn-5-en-3β,20(S)-diol-3-O-bis-β-D-glucopyranosyl-(l → 2,1 →
6)-β-D-glucopyranoside (1): White amorphous powder; [α]2D4
− 30.4 (c 0.85, MeOH); IR νmaxKBr · cm−1: 3359 (br), 2929, 2857,
Marmesin-11-O-β-D-glucopyranosyl(1 → 6)-β-D-glucopyranoside
(5): White amorphous powder; [α]2D4 − 33.7 (c 0.43, MeOH); 1H
NMR (500 MHz, DMSO-d6): δ 7.92 (1H, d, J = 9.5 Hz, H-4), 7.49
(1H, s, H-5), 6.82 (1H, s, H-8), 6.21 (1H, d, J = 9.5 Hz, H-3), 4.86
(1H, dd, J = 9.0, 8.5 Hz, H-10), 4.42 (1H, d, J = 8.0 Hz, H-1″), 4.22
(1H, d, J = 7.5 Hz, H-1′), 3.79 (1H, brd, J = 11.0 Hz, Ha-6″), 3.67
(1H, brd, J = 11.5 Hz, Ha-6′), 3.52 (1H, dd, J = 11.0, 6.5 Hz, Hb-6″),
3.43 (1H, dd, J = 11.5, 4.5 Hz, Hb-6′), 3.29 (1H, dd, J = 16.0, 8.0 Hz,
Ha- 9), 3.19 (1H, dd, J = 16.0, 8.5 Hz, Hb- 9), 3.15 (2H, m, H-3′, H-
3″), 3.04 (4H, m, H-4′, H-5′, H-4″, H-5″), 2.93 (1H, dd, J = 8.0,
8.0 Hz, H-2″), 2.87 (1H, dd, J = 8.0, 7.5 Hz, H-2′), 1.25 (3H, s, Me-
12), 1.22 (3H, s, Me-13). 13C NMR (125 MHz, DMSO-d6): δ 160.5
(C-2), 111.3 (C-3), 144.7 (C-4), 112.2 (C-4a), 124.0 (C-5), 125.6 (C-
6), 163.1 (C-7), 96.8 (C-8), 155.0 (C-8a), 28.9 (C-9), 90.0 (C-10),
77.1 (C-11), 22.9 (C-12), 21.8 (C-13), 97.2 (C-1′), 73.5 (C-2′), 76.7
(C-3′), 70.1 (C-4′), 75.6 (C-5′), 68.3 (C-6′), 103.2 (C-1″), 73.4 (C-2″),
76.9 (C-3″), 69.9 (C-4″), 76.6 (C-5″), 61.1 (C-6″). LR-ESIMS: m/z 593
1653, 1074; 1H NMR and 13C NMR data, see l Table 1. LR-ESIMS:
"
m/z 827 [M + Na]+, 1631 [2M + Na]+, 863 [M + CH3COO−]−, 1607
[2M − H]−; HR-ESIMS: m/z 827.4046 [M + Na]+ (calcd. for
C
39H64O17Na: 827.4036, Δ = − 1.2 ppm)
Oleuropeic aldehyde 8-O-β-D-glucopyranoside (2): Colorless oil;
[α]2D4 − 2.0 (c 0.13, MeOH); 1H NMR (500 MHz, CD3OD): δ 9.38
(1H, s, CHO), 6.94 (1H, brdd, J = 2.7, 2.3 Hz, H-2), 4.49 (1H, d, J =
7.8 Hz, H-1′), 3.81 (1H, dd, J = 11.9, 2.2 Hz, Ha-6′), 3.64 (1H, dd, J =
11.9, 5.4 Hz, Hb-6′), 3.36 (1H, dd, J = 9.0, 8.8 Hz, H-5′), 3.28 (1H, dd,
J = 9.0, 8.7 Hz, H-4′), 3.24 (1H, m, H-3′), 3.14 (1H, dd, J = 8.8, 7.8 Hz,
You M et al. Glycosides from the… Planta Med 2011; 77: 1939–1943

Letters 1943
[M + Na]+, 1163 [2M + Na]+, 569 [M − H]−, 1139 [2M − H]−; HR-
ESIMS: m/z 593.1821 [M
593.1841, Δ = + 3.3 ppm).
16 Liu R, Sun QH, Sun AL, Cui JC. Isolation and purification of coumarin
compounds from Cortex Fraxinus by high-speed counter-current chro-
matography. J Chromatogr A 2005; 1072: 195–199
17 Angyal SJ, Bethell GS. Conformational analysis in carbohydrate chemis-
try. III. The 13C N.M.R. spectra of the hexuloses. Austrian J Chem 1976;
29: 1249–1265
18 Prawat H, Mahidol C, Ruchirawat S, Prawat U, Tuntiwachwuttikul P,
Tooptakong U, Taylor WC, Pakawatchai C, Skelton BW, White AH. Cyano-
genic and non-cyanogenic glycosides from Manihot esculenta. Phyto-
chemistry 1995; 40: 1167–1173
+
Na]+ (calcd. for C26H34O14Na:
Pregn-5-en-3β-ol-20-one-3-O-bis-β-D-glucopyranosyl-(1 → 2,1 →
1
6)-β-D-glucopyranoside (6): White amorphous powder; H NMR
+
and 13C NMR data, see l Table 1. LR-ESIMS: m/z = 825 [M + Na] ;
"
HR-ESIMS: m/z 825.3831 [M + Na]+ (calcd. for C39H62O17Na:
825.3879, Δ = + 5.8 ppm).
19 Itokawa H, Xu JP, Takeya K. Studies on chemical constituents of antitu-
mor fraction from Periploca sepium Bge. I. Chem Pharm Bull 1987; 35:
4524–4529
20 Yoshimura T, Kisyuku H, Kamei T, Takabatake K, Shindo M, Shishido K.
Enantiocontrolled synthesis of (+)-curcuquinone and (−)-curcuhydro-
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21 Klyne W. The configuration of the anomeric carbon atoms in some car-
diac glycosides. Biochem J 1950; 47: xli–xlii
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23 Matano Y, Okuyama T, Shibata S, Hoson M, Kawada T, Osada H, Noguchi
T. Studies on coumarins of a Chinese drug “Qian-Hu”; VII. Structures of
new coumarin-glycosides of Zi-Hua Qian-Hu and effect of coumarin-
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Supporting information
The extraction and isolation of compounds 1–18, details on the
acid hydrolysis of compounds 1, 2, 5, 9, 10, sugar analysis, acety-
lation of 3, full assignment of NMR data for compounds 3a, 7, 11,
and the NMR and MS spectra of compounds 1–7 and 11 are avail-
able as Supporting Information.
Acknowledgements
!
The work was supported by an NSFC grant (No. 90713040).
Conflict of Interest
!
24 Talapatra SK, Chaudhuri MK, Talapatra B. Coumarins of the root bark of
Feronia elephantum. Phytochemistry 1973; 12: 236–237
We declare that we have no conflict of interest.
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Bibliography
DOI http://dx.doi.org/10.1055/s-0031-1279995
Published online July 15, 2011
Planta Med 2011; 77: 1939–1943
© Georg Thieme Verlag KG Stuttgart · New York ·
ISSN 0032‑0943
Correspondence
Prof. Dr. Jin-Feng Hu
Department of Natural Products for Chemical Genetic Research
Key Laboratory of Brain Functional Genomics
Ministry of Education, East China Normal University
No. 3663 Zhongshan Road N
Shanghai 200062
P.R. China
Phone: + 862162607510
Fax: + 862162606791
jfhu@brain.ecnu.edu.cn
jfhu@fudan.edu.cn
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