Phenolic and iridoid glycosides from Strychnos axillaris

Article abstract of DOI:10.1016/j.phytochem.2007.12.002

Five phenolic glycosides 1-5 and an iridoid glucoside 6 were isolated, together with 22 known compounds, from the dried barks and woods of Strychnos axillaris. Their structures were determined by application of spectroscopic (NMR, MS) and chemical methodo

Full text of DOI:10.1016/j.phytochem.2007.12.002

Available online at www.sciencedirect.com
PHYTOCHEMISTRY
Phytochemistry 69 (2008) 1208–1214
www.elsevier.com/locate/phytochem
Phenolic and iridoid glycosides from Strychnos axillaris
Atsuko Itoh a, Yasuhiro Tanaka a, Naotaka Nagakura a, Toru Akita b,
Toyoyuki Nishi b, Takao Tanahashi a,*
a Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan
b The Nippon Shinyaku Institute for Botanical Research, Yamashina-ku, Kyoto 607-8182, Japan
Received 30 August 2007; received in revised form 13 November 2007
Available online 4 February 2008
Abstract
Five phenolic glycosides 1–5 and an iridoid glucoside 6 were isolated, together with 22 known compounds, from the dried barks and
woods of Strychnos axillaris. Their structures were determined by application of spectroscopic (NMR, MS) and chemical methodologies.
Ó 2007 Elsevier Ltd. All rights reserved.
Keywords: Strychnos axillaris; Loganiaceae; Phenolic glycosides; Iridoid glucoside; Calleryanin; Vanilloloside; Cantleyoside
1. Introduction
extract was successively partitioned between H2O and
CHCl3 and between H2O and n-BuOH. The n-BuOH-sol-
uble fraction was separated by a combination of chro-
matographic procedures to afford six new compounds
1–6 together with 22 known compounds: tachioside
(Zhong et al., 1999), isotachioside (Zhong et al., 1999),
3,4,5-trimethoxyphenol 1-O-b-D-apiofuranosyl-(1?6)-b-
D-glucopyranoside (Duynstee et al., 1999), calleryanin
(7) (Challice et al., 1980), vanilloloside (8) (Ida et al.,
1994), 3,5-dimethoxy-4-hydroxybenzyl alcohol 4-O-b-D-
glucopyranoside (Kitajima et al., 1998), caffeoylcallerya-
nin (9) (Challice and Williams, 1968), (7S, 8R)-balano-
phonin 4-O-b-D-glucopyranoside (Warashina et al.,
2005), (+)-syringaresinol O-b-D-glucoside (Lami et al.,
1991), scorzonoside (Tolstikhina and Semenov, 1998), lir-
iodendrin (Kobayashi et al., 1985), (ꢀ)-pinoresinol 4-O-
b-D-glucopyranoside (Sugiyama and Kikuchi, 1991),
(+)-lariciresinol 40-O-b-D-glucoside (Sugiyama and Kiku-
chi, 1993), loganic acid (Nakamoto et al., 1988), loganin
(Nakamoto et al., 1988), sweroside (Jensen et al., 1981),
picconioside I (Damtoft et al., 1997), cantleyoside (10)
(Kocsis et al., 1993), triplostoside A (Ma et al., 1992),
strictosidinic acid (Arbain et al., 1993), 3,4-di-O-caffeoyl-
quinic acid (Wang et al., 1992), and 3,5-di-O-caffeoylqui-
nic acid (Wald et al., 1989). The structures of the new
compounds were determined as follows.
The genus Strychnos (Loganiaceae) is well-known as a
rich source of various bioactive indole alkaloids repre-
sented by strychnine (Bisset, 1980). Several b-carboline glu-
coalkaloids closely related to indole alkaloids have been
isolated from Strychnos mellodora (Brandt et al., 1999).
In continuation of our phytochemical studies on constitu-
ents of the genus Strychnos (Itoh et al., 2005, 2006), we
examined S. axillaris Colebr. (=S. pubescens C.B. Clarke)
collected in Thailand. This species has been traditionally
used in East Asia as an arrow poison (Perry, 1980), but
no phytochemical study of this plant has been reported.
From our interests in glucoalkaloids in the polar fraction
(Itoh et al., 2003), we investigated the n-BuOH-soluble
fraction of this plant. In this paper, we report the isolation
and structure elucidation of six new glycosides along with
22 known compounds from S. axillaris.
2. Results and discussion
The dried barks and woods of S. axillaris were
extracted with MeOH under conditions of reflux. The
*
Corresponding author. Tel./fax: +81 78 441 7545.
E-mail address: tanahash@kobepharma-u.ac.jp (T. Tanahashi).
0031-9422/$ - see front matter Ó 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.phytochem.2007.12.002

A. Itoh et al. / Phytochemistry 69 (2008) 1208–1214
1209
7
6
O
O
1"
HO
HO
5"
R2O
6
7"
2"
5"
7
6'
5
O
OH
OH
5
O
O
9"
2
8"
O
6"
2
OR2
GlcO
HO
OH
1'
OR1
OH
OR1
4: R1=Me, R2=H
5: R1=H, R2=Glc
1: R1=R2=H
2: R1=H, R2=trans-feruloyl
3: R1=Me, R2=trans-feruloyl
OR2
O
7"'
6"'
2"'
5"'
GlcO
OMe
OH
trans-feruloyl =
OR1
9"'
8"'
7: R1=R2=H
8: R1=Me, R2=H
9: R1=H, R2=trans-caffeoyl
11
COOMe
H
6
O
R
11"
3
COOH
HOOC
COOMe
5
7
8
COO
H
9
H
H
7"
6"
1
O
H
H
3"
HO
OGlc
O
O
10
1"
O
H
8"
H
OGlc
OGlc
OGlc
10"
11
12
6: R=OH
10: R=H
Compound 1 was obtained as an amorphous powder,
[a]D ꢀ81. Its HR-SIMS showed a pseudomolecular ion
[M–H]ꢀ at m/z 433.1354, indicating a molecular formula
of C18H26O12. It also had UV maxima at 202 and 278 nm
group [d 3.86 (s)]. These resonances were assignable to a
trans-feruloyl group by its NOESY correlation between a
methoxy singlet and an aromatic proton at d 7.16 (d,
J = 2.0 Hz) and HMBC interactions between aromatic
protons at dH 7.05 and 7.16 and an olefinic carbon at dC
147.4. The ester linkage of the trans-feruloyl group to the
hydroxyl group at C-500 of apiose moiety was demonstrated
by the downfield shift of C-500 relative to those in 1 and the
HMBC cross peaks between H2-500 [dH 4.27, 4.30 (each d,
J = 11.5 Hz)] and C-9000 of trans-feruloyl group (dC 169.0).
Thus, compound 2 was deduced to be 500-O-trans-feru-
loyl-60-O-b-D-apiofuranosylcalleryanin.
The spectroscopic properties of compound 3 were quite
similar to those of 2 except for additional signals for a
methoxy group at dH 3.84 and dC 56.7 (Table 1). The loca-
tion of the methoxy group was determined to be at C-3 by
comparison of the 1H and 13C NMR spectroscopic data for
the benzyl alcohol moiety with those of vanilloloside (8)
(Ida et al., 1994). Further evidence was obtained from
the NOESY correlation between H-2 [dH 6.99 (d,
J = 2.0 Hz)] and the methoxy singlet. Accordingly, com-
pound 3 was established as 500-O-trans-feruloyl-60-O-b-D-
apiofuranosylvanilloloside.
1
and IR bands at 3386 and 1508 cmꢀ1. Its H NMR spec-
trum (Table 1) exhibited an AMX spin system at d 6.79
(dd, J = 8.5, 2.0 Hz), 6.87 (d, J = 2.0 Hz), and 7.15 (d,
J = 8.5 Hz), a signal for an oxygenated benzylic methylene
group at d 4.48 (s), as well as resonances for a b-glucopyr-
anosyl unit at d 3.35–4.70, implying the presence of a cal-
leryanin (7) moiety (Challice et al., 1980). This was
supported by the NOESY correlations between H-2 (d
6.87), H-6 (d 6.79) and H2-7 (d 4.48) and between H-5 (d
1
7.15) and H-10 of glucose unit (d 4.70). The H and 13C
NMR spectra of 1 showed additional signals for a b-apio-
furanosyl unit at dH 3.59 (2 H, s), 3.76 (1H, d, J = 9.5 Hz),
3.92 (1H, d, J = 2.5 Hz), 3.97 (1H, d, J = 9.5 Hz), 4.99 (1H,
d, J = 2.5 Hz) and dC 65.6, 75.0, 78.1, 80.5, 111.1, besides
the resonances for a calleryanin moiety. Acid hydrolysis
of 1 liberated D-glucose and D-apiose, which were identified
by GLC analysis of their thiazolidine derivatives (Hara
et al., 1986). On the basis of the HMBC correlation
between the H-100 of the apiose moiety and C-60 of the glu-
cose moiety, it was concluded that an apiose was linked to
C-60 of the calleryanin moiety, which was supported by the
downfield shift of C-60 and the upfield shift of C-50 by gly-
cosidation when compared with 7. Accordingly, compound
1 was characterized as 60-O-b-D-apiofuranosylcalleryanin.
The second new glycoside 2 was also obtained as an
Compound 4, C23H26O11, was also isolated as an amor-
1
phous powder. Its H and 13C NMR spectroscopic data
(Table 2) showed that 4 consisted of a vanilloloside (8)
moiety esterified with a trans-caffeoyl group. Acylation of
the hydroxyl group at C-7 was determined by the HMBC
correlation between H2-7 and C-900 of the caffeoyl group,
and by comparison of the 13C NMR spectroscopic data
of 4 with those of 8 and caffeoylcalleryanin (9) (Challice
and Williams, 1968). Accordingly, the structure of 4 was
determined to be 7-O-trans-caffeoylvanilloloside.
1
amorphous powder. The H NMR spectroscopic features
of 2 were closely similar to those of 1 except for the signals
for another AMX spin system [d 6.79 (d, J = 8.5 Hz), 7.05
(dd, J = 8.5, 2.0 Hz), 7.16 (d, J = 2.0 Hz)], trans-olefinic
protons [d 6.38, 7.65 (each d, J = 16.0 Hz)], and a methoxy

1210
A. Itoh et al. / Phytochemistry 69 (2008) 1208–1214
Table 1
1H and 13C NMR spectral data of compounds 1–3 in CD3OD
1
2
3
dC
dH
dC
dH
dC
dH
1
2
3
4
5
6
7
138.5
116.0
148.4
146.0
118.9
119.7
64.7
138.6
116.1
148.4
146.1
118.9
119.7
64.9
137.8
112.7
150.8a
147.3
118.1
120.9
65.0
6.87
d
(2.0)
6.84
d
(2.0)
6.99
d
(2.0)
7.15
6.79
4.48
d
dd
s
(8.5)
(8.5, 2.0)
7.15
6.76
4.42
d
dd
s
(8.5)
(8.5, 2.0)
7.12
6.86
4.49
3.84
d
dd
s
(8.0)
(8.0, 2.0)
3-OMe
glc
10
56.7
s
104.5
74.9
77.6
71.6
77.2
68.8
4.70
3.48
3.44
3.35
3.55
3.63
4.02
d
brt
t
brt
ddd
dd
dd
(8.0)
(8.5)
(9.0)
(9.0)
(9.5, 6.5, 2.0)
(11.0, 6.5)
(11.0, 2.0)
104.5
74.9
77.6
71.7
77.1
68.7
4.67
3.48
3.42
3.34
3.57
3.63
4.05
d
dd
t
brt
ddd
dd
dd
(8.0)
(9.0, 8.0)
(9.0)
103.0
75.0
77.9
71.8
77.1
68.8
4.81
3.49
3.43
3.33
3.56
3.61
4.00–4.04
d
dd
t
brt
ddd
dd
m
(7.5)
(9.0, 7.5)
(9.0)
(9.5)
(9.5, 6.5, 2.0)
(11.0, 6.5)
20
30
40
(9.0)
50
(9.0, 6.5, 2.0)
(11.0, 6.5)
(11.0, 2.0)
60
60
a0p0 i
1
111.1
78.1
80.5
75.0
4.99
3.92
d
d
(2.5)
(2.5)
110.8
78.6
79.0
75.1
5.02
3.98
d
d
(2.0)
(2.0)
110.8
78.6
79.0
75.1
4.99
3.94
d
d
(2.0)
(2.0)
200
300
400
3.76
3.97
3.59
d
d
s
(9.5)
(9.5)
3.84
4.05
4.27
4.30
d
d
d
d
(9.5)
(9.5)
(11.5)
(11.5)
3.82
4.02
4.24
4.28
d
d
d
d
(9.5)
(9.5)
(11.0)
(11.0)
400
500
65.6
67.4
67.4
500
a0c0y0 l
1000
2000
3000
4
5000
6000
7000
8000
9000
3000-OMe
a
127.6
111.8
149.5
150.9
116.6
124.3
147.4
115.1
169.0
56.5
127.7
111.8
149.5
150.9a
116.6
124.3
147.3
115.1
168.9
56.5
7.16
d
(2.0)
7.17
d
(2.0)
6.79
7.05
7.65
6.38
d
dd
d
(8.5)
6.80
7.06
7.64
6.38
d
dd
d
(8.5)
(8.5, 2.0)
(16.0)
(16.0)
(8.5, 2.0)
(16.0)
(16.0)
d
d
3.86
s
3.87
s
Assignments with the same superscript may be interchanged.
Compound 5, C28H34O16, was obtained as an amor-
phous powder and its 1H and 13C NMR spectroscopic fea-
tures were closely similar to those of 9, except for
additional signals arising from a b-glucopyranosyl unit.
The attachment of the second glucopyranosyl unit at C-
400 of caffeoylcalleryanin moiety was confirmed by the
HMBC correlation between H-1000 of glucose unit and C-
400 and by the NOESY interaction between H-500 and H-
1000. Accordingly, compound 5 was assigned to 400-O-b-D-
glucopyranosyl-caffeoylcalleryanin.
coside unit in the molecule. These spectroscopic features
were similar to those of cantleyoside (10), which was also
isolated in this study, except for the absence of the signal
1
for an aldehyde proton in its H NMR spectrum (Kocsis
et al., 1993). Its 13C NMR spectrum exhibited three car-
bonyl carbon resonances at d 168.0, 169.5, and 176.1.
These observations suggested that glucoside 6 was com-
posed of loganin (11) unit and secologanoside (12) units.
The presence of a secologanoside unit was further sup-
ported by a fragment ion peak at m/z 389.
Compound 6, named axillaroside, was assigned the
molecular formula C33H46O20 from its HR-SIMS. Its H
The site of ester linkage was confirmed by HMBC exper-
iments. The signal at dc 169.5 was assigned to C-11 of the log-
anin moiety by HMBC correlations from H-3, H-5 and the
methoxyl resonance at d 3.69, and the signal at dc 176.1
was assigned to C-700 ofthe secologanoside moiety by HMBC
correlations from H-500 and H2-600. The remaining carbonyl
carbon resonance at dc 168.0, which was assigned to C-1100
of the secologanoside moiety by HMBC interactions with
H-300 and H-500, was correlated with H-7 [d 5.20 (br t,
J = 5.5 Hz)] of the loganin unit, indicating that the C-11 car-
boxyl group of the secologanoside moiety was linked to the
1
NMR spectrum showed signals for two olefinic protons
at d 7.24 (d, J = 1.5 Hz) and 7.49 (d, J = 2.0 Hz), two acetal
proton resonances at d 5.28 (d, J = 5.0 Hz) and 5.50 (d,
J = 4.5 Hz), two sets of signals for b-glucopyranosyl units
at d 3.20–4.68, a doublet for a secondary methyl group at
d 1.07 (J = 6.5 Hz), and resonances for a terminal vinyl
group at d 5.23 (dd, J = 10.0, 1.5 Hz), 5.27 (dd, J = 17.0,
1.0 Hz), and 5.66 (br dt, J = 17.0, 10.0 Hz), indicating the
presence of an iridoid glucoside unit and a secoiridoid glu-

A. Itoh et al. / Phytochemistry 69 (2008) 1208–1214
1211
Table 2
1H and 13C NMR spectral data of compounds 4 and 5 in CD3OD
4
5
dC
dH
dC
dH
1
2
3
4
5
6
7
132.7
114.0
150.9
148.0
117.9
122.5
67.1
133.3
117.2
148.4a
146.7
118.7
121.0
67.0
7.06
d
(2.0)
6.91
d
(2.0)
(8.0)
7.16
6.96
5.15
3.87
d
dd
s
(8.0)
(8.0, 2.0)
7.18
6.84
5.11
d
dd
s
(8.0, 2.0)
3-OMe
glc
10
56.8
s
102.8
75.0
77.9
71.4
78.3
62.5
4.91
3.50
3.46
3.36–3.46
3.36–3.46
3.69
d
(7.0)
(8.0)
(9.0)
104.2
74.8b
77.5c
71.3
78.3d
62.4
4.77
d
(7.5)
20
brt
brt
m
m
dd
dd
3.51
dd
m
m
m
dd
dd
(9.0, 7.5)
30
3.38–3.50
3.38–3.50
3.38–3.50
3.71
40
50
60
(12.0, 5.0)
(12.0, 1.5)
(12.0, 5.0)
(12.0, 2.0)
60
3.87
3.90
a0c0yl
100
200
3
400
500
600
700
800
900
g0l0c0
1000
2
127.7
115.2
146.9
149.7
116.5
123.1
147.2
115.0
169.1
131.3
116.0
148.6a
148.9
118.1
122.3
146.3
117.2
168.6
7.03
d
(2.0)
7.11
d
(2.0)
6.77
6.94
7.56
6.28
d
dd
d
(8.0)
7.19
7.04
7.59
6.38
d
dd
d
(8.5)
(8.0, 2.0)
(16.0)
(16.0)
(8.5, 2.0)
(16.0)
(16.0)
d
d
103.5
74.9b
77.6c
71.3
78.4d
62.4
4.85
3.51
3.38–3.50
3.38–3.50
3.38–3.50
3.71
d
dd
m
m
m
(7.5)
(9.0, 7.5)
3000
4000
5000
6000
6000
a–d
dd
dd
(12.0, 5.0)
(12.0, 2.0)
3.90
Assignments with the same superscript may be interchanged.
C-7 hydroxy group of loganin component. Accordingly,
axillaroside was formulated as shown in formula 6.
8200 spectrophotometer. 1H (500 MHz) and 13C
(125 MHz) NMR spectra were recorded on a Varian
VXR-500 spectrometer with TMS as an internal standard.
MS and HRMS were obtained with a Hitachi M-4100 mass
spectrometer. Glycerol or 3-nitrobenzyl alcohol was used
as the matrix for SIMS and HRSIMS. MPLC was carried
out with Wakogel FC-40 or Wakosil 40C18. TLC was
performed on precoated Kieselgel 60F254 plates (Merck).
3. Concluding remarks
In the present study, five new phenolic glycosides 1–5
with a benzyl alcohol moiety and a new iridoid glucoside
6 were isolated along with a glucoalkaloid strictosidic acid.
Iridoid glycosides and their related indole alkaloids have
previously been reported from the genus Strychnos. How-
ever, this is the first instance of the isolation of benzyl alco-
hol glycosides such as calleryanin and vanilloloside from
the genus.
4.2. Plant material
The bark and wood of Strychnos axillaris Colebr. were
collected at Sakae Rat, Nakhon Ratchasima, Thailand in
March 1986 and identified by Dr. T. Smitinand, The Forest
Herbarium, Royal Forest Department, Bangkok, Thailand.
4. Experimental
4.3. Extraction and isolation
4.1. General
Dried and powdered mixture (2000 g) of bark and wood
of S. axillaris was extracted with MeOH under conditions
of reflux for 1 h, with the extracts concentrated in vacuo
and a part (39 g) of the resulting residue (130 g) resuspended
UV spectra were recorded on a Shimadzu UV-2500PC
spectrophotometer and IR spectra on a Shimadzu FTIR-

1212
A. Itoh et al. / Phytochemistry 69 (2008) 1208–1214
in H2O and extracted successively with CHCl3 and n-BuOH.
The residue (23.6 g) from the n-BuOH layer was fractionated
using reversed-phase MPLC. Elution with H2O-MeOH mix-
tures of the indicated MeOH content (%v/v) gave nine frs, 1
(5%, 286 mg), 2 (10%, 160 mg), 3 (15%, 414 mg), 4 (20%,
197 mg), 5 (20%, 797 mg), 6 (25–30%, 1.11 g), 7 (30–35%,
457 mg), 8 (35%, 4.59 g), and 9 (40–45%, 1.54 g). Fr. 1 was
purified by preparative HPLC (lBondasphere 5l C18-
H-7 and H-3, 5; H-6 and H-10; H-60 and H-100; H-2000, 6000
and H-7000; OMe and H-2000; HMBC: H-3 to C-1, 5; H-5
to C-1; H-6 to C-1, 2; H-7 to C-3, 4, 5; H- 10 to C-1; H-
60 to C-100; H- 500 to CO; H-2000 to C-3000, 4000, 7000; H-5000 to
C-3000, 4000; H-6000 to C-2000, 4000; H-7000 to C-2000, 6000, CO; H-8000
to C-1000, CO; OMe to C-3000; negative ion SIMS m/z 609
[MꢀH]ꢀ, 139; negative ion HRSIMS m/z 609.1823 (calcd
for C28H33O15, 609.1820).
˚
100 A, MeOH–H2O, 1:4, MeOH–MeCN–H2O, 1:2:97,
v/v/v) and preparative TLC (EtOAc–C6H6–EtOH, 4:1:3,
CHCl3–MeOH, 7:3, CHCl3–MeOH–H2O, 70:30:3, v/v/v)
to afford 7 (68.7 mg), 8 (20.7 mg), loganic acid (18.8 mg),
tachioside (3.4 mg), and isotachioside (3.3 mg), respectively.
In the same way, frs. 2–9 were purified by a combination of
reversed-phase MPLC with MeOH–H2O, normal-phase
MPLC with CHCl3–MeOH, preparative HPLC (lBonda-
4.6. 500-O-trans-Feruloyl-60-O-b-D-apiofuranosylvanilloloside
(3)
26
MeOH
max
White powder; ½aꢁD ꢀ 45 (c 0.3, MeOH); UV k
nm
(log e): 220sh (4.26), 230sh (4.19), 285 (3.92), 297sh
(3.91), 327 (4.09); IR m cmꢀ1 : 3421, 1716, 1595, 1516;
KBr
max
For H and 13C NMR spectroscopic data, see: Table 1;
1
˚
sphere 5l C18-100 A, MeOH–H2O, 2:8–11:9 (by vol.),
NOESY: H-3 and H-7, OMe; H-5 and H-7; H-6 and H-
10; H-60 and H-100; H-6000 and H-7000; OMe and H-2000;
HMBC: H-3 to C- 5; H-5 to C-3, 7; H-6 to C-4; H-7 to
C-3, 4, 5; H-10 to C-1; H-100 to C-60; H- 500 to CO; H-2000
to C-3000, 6000; H-5000 to C-1000, 3000; H-6000 to C-2000; H-7000 to
C-1000, 2000, 6000, 8000, CO; H-8000 to C-1000, CO; OMe to C-3000;
negative ion SIMS m/z 623 [M-H]ꢀ, 153; negative ion
HRSIMS m/z 623.1979 (calcd for C29H35O15, 623.1977).
MeOH–H2O–AcOH, 40:60:1, MeCN–H2O, 3:17, MeCN–
H2O–AcOH, 20:85:1, 15:85:1, v/v/v) and preparative TLC
(CHCl3–MeOH, 7:3; CHCl3–MeOH–H2O, 70:30:3, 14:6:1,
v/v/v). Fr. 2 yielded 1 (15.2 mg), 8 (19.6 mg), loganic acid
(20.9 mg), and 3,5-dimethoxy-4-hydroxybenzyl alcohol 4-
O-b-D-glucopyranoside (7.6 mg); fr. 3: loganic acid
(196 mg); fr. 4: loganin (4.2 mg), sweroside (24.6 mg), and
3,4,5-trimethoxyphenol 1-O-b-D-apiofuranosyl-(1 ? 6)-b-
D-glucopyranoside (20.1 mg); fr. 5: sweroside (46.8 mg),
3,4-di-O-caffeoylquinic acid (17.0 mg), liriodendrin
(158 mg), and 3,5-di-O-caffeoylquinic acid (15.7 mg); fr. 6:
3,4-di-O-caffeoylquinic acid (40.2 mg), scorzonoside
(9.7 mg), liriodendrin (588 mg), and (+)-lariciresinol 4-O-
b-D-glucoside (21.1 mg); fr. 7: 2 (2.9 mg), 3 (2.8 mg), 5
(57.8 mg), (7S, 8R)-balanophonin 4-O-b-D-glucopyranoside
(3.7 mg), 10 (4.9 mg), (+)-syringaresinol O-b-D-glucoside
(10.8 mg), 3,4-di-O-caffeoylquinic acid (28.4 mg), scorzono-
side (8.8 mg), (ꢀ)-pinoresinol 4-O-b-D-glucopyranoside
(5.7 mg), and strictosidinic acid (8.7 mg); fr. 8: 4 (13.4 mg),
5 (7.5 mg), 6 (5.6 mg), 9 (535 mg), 10 (768 mg), and 3,4-di-
O-caffeoylquinic acid (78.4 mg); fr. 9: 10 (25.7 mg), picconio-
side I (11.9 mg), and triplostoside A (160 mg).
4.7. 7-O-trans-Caffeoylvanilloloside (4)
25
MeOH
max
White powder; ½aꢁD ꢀ 39 (c 0.7, MeOH); UV k
nm
(loge): 249sh (3.98), 285sh (4.02), 300sh (4.07), 329 (4.16);
IR m cmꢀ1 : 3421, 1697, 1601, 1516; For 1H and 13C
KBr
max
NMR spectroscopic data, see: Table 2; NOESY: H-3, 5
and H-7; OMe and H-3; H-6 and H-10; HMBC: H-3 to
C- 1, 2, 7; H-5 to C-1, 3, 7; H-6 to C-1, 2; H-7 to C-3, 4,
5, 900; H-200 to C-400, 700; H-500 to C-300, 400; H- 600 to C-200,
400, 700; H-700 to C-200, 600, 800, 900; H-800 to C-100, 900; OMe to
C-2, H-10 to C-1; negative ion SIMS m/z 477 [M-H]ꢀ,
179; negative ion HRSIMS m/z 477.1387 (calcd for
C23H25O11, 477.1398).
4.8. 400-O-b-D-Glucopyranosyl-caffeoylcalleryanin (5)
4.4. 60-O-b-D-Apiofuranosylcalleryanin (1)
24
MeOH
max
White powder; ½aꢁD ꢀ 83 (c 1.0, MeOH); UV k
nm
23
MeOH
max
White powder; ½aꢁD ꢀ 81 (c 1.5, MeOH); UV k
nm
(log e): 244sh (4.04), 286 (4.20), 320 (4.11); IR
1
(log e): 202 (3.82), 278 (3.39); IR m
cmꢀ1 : 3386, 1508;
m
cmꢀ1 : 3389, 1697, 1635, 1508; For H and 13C NMR
KBr
max
KBr
max
For H and 13C NMR spectroscopic data, see: Table 1;
NOESY: H-7 and H-3, 5; H-6 and H-10; HMBC: H-3 to
C-1, 2, 5, 7; H-5 to C-1, 3, 7; H-6 to C-1, 2, 4; H-7 to C-
3, 4, 5; H-10 to C-1; H-100 to C-60; negative ion HRSIMS
m/z 433.1354 (calcd for C18H25O12, 433.1347).
spectroscopic data, see: Table 2; NOESY: H-3, 5 and H-
7; H-6 and H-10; H-500 and H-1000; HMBC: H-3 to C- 1, 5,
7; H-5 to C-1, 3, 6, 7; H-6 to C-1, 4; H-7 to C-3, 4, 5, 900;
H-200 to C-400, 700; H-500 to C-100, 600; H- 600 to C-200, 400, 500,
700; H-700 to C-100, 200, 600, 800, 900; H-800 to C-100, 900; H-10 to
C-1; H-1000 to C-400; negative ion SIMS m/z 625 [M-H]ꢀ,
463; negative ion HRSIMS m/z 625.1777 (calcd for
C28H33O16, 625.1770).
1
4.5. 500-O-trans-Feruloyl-60-O-b-D-apiofuranosylcalleryanin
(2)
25
MeOH
max
White powder; ½aꢁD ꢀ 44 (c 0.3, MeOH); UV k
nm
4.9. Axillaroside (6)
(log e): 219sh (4.26), 231sh (4.14), 286 (3.98), 298sh (3.99),
25
1
MeOH
nm
max
327 (4.14); IR m cmꢀ1 : 3426, 1717, 1597, 1516; For H
White powder; ½aꢁD ꢀ 60 (c 0.6, MeOH); UV k
KBr
max
and 13C NMR spectroscopic data, see: Table 1; NOESY:
(loge): 234 (4.24); IR mKBrcmꢀ1 : 3421, 1697, 1636; 1H
max

A. Itoh et al. / Phytochemistry 69 (2008) 1208–1214
1213
NMR (CD3OD) d 1.07 (1H, d, J = 6.5 Hz, H-10), 1.74 (1H,
ddd, J = 14.5, 8.0, 5.0 Hz, H-6), 2.07 (1H, td, J = 9.0,
5.0 Hz, H-9), 2.10–2.18 (1H, m, H-8), 2.29 (1H, ddd,
J = 14.5, 7.5, 1.5 Hz, H-6), 2.30 (1H, dd, J = 16.5,
9.0 Hz, H-600), 2.81 (1H, br dt, J = 9.0, 5.0 Hz, H-900),
2.91 (1H, dd, J = 16.5, 4.0 Hz, H-600), 3.11 (1H, br q,
J = 8.0 Hz, H-5), 3.20, 3.22 (2H, each dd, J = 9.0, 8.0 Hz,
H-20, 2000), 3.26-3.38 (2H, m, H-50, 5000), 3.27, 3.28 (2 H, each
br t, J = 9.0 Hz, H-40, 4000), 3.28-3.38 (1 H, m, H-500), 3.36,
3.37 (2H, each t, J = 9.0 Hz, H-30, 3000), 3.66, 3.67 (2H, each
dd, J = 12.0, 6.0 Hz, H-60, 6000), 3.69 (3H, s, OMe), 3.89,
3.90 (2H, each dd, J = 12.0, 2.0 Hz, H-60, 6000), 4.66 (1H,
d, J = 8.0 Hz, H-10), 4.68 (1H, d, J = 8.0 Hz, H-1000), 5.20
(1H, br t, J = 5.5 Hz, H-7), 5.23 (1H, dd, J = 10.0,
1.5 Hz, H-1000), 5.27 (1H, dd, J = 17.0, 1.0 Hz, H-1000),
5.28 (1H, d, J = 5.0 Hz, H-1), 5.50 (1H, d, J = 4.5 Hz, H-
100), 5.66 (1H, br dt, J = 17.0, 10.0 Hz, H-800), 7.24 (1H, d,
J = 1.5 Hz, H-3), 7.49 (1H, d, J = 2.0 Hz, H-300); 13C
NMR (CD3OD) d 13.9 (C-10), 29.1 (C-500), 32.7 (C-5),
35.3 (C-600), 40.4 (C-6), 41.4 (C-8), 45.4 (C-900), 47.2 (C-9),
51.8 (OMe), 62.8 (C-60, 6000), 71.6, 71.7 (C-40, 4000), 74.7,
74.8 (C-20, 2000), 78.1 (C-30, 3000), 78.3 (C-7), 78.5 (C-50, 5000),
97.6 (C-1), 97.7 (C-100), 100.1 (C-1000), 100.3 (C-10), 110.5
(C-400), 113.3 (C-4), 120.6 (C-1000), 134.5 (C-800), 152.7 (C-
3), 153.7 (C-300), 168.0 (C-1100), 169.5 (C-11), 176.1 (C-700);
HMBC: H-1 to C-3, 5; H-3 to C-1, 4, 5, 11; H-5 to C-6,
9, 11; H-6 to C-5; H-7 to C-5, 9, 1100; H-8 to C-9; H-9 to
C-8, 10; H-10 to C-8, 9; H-100 to C-300, 500; H-300 to C-100,
400, 500, 1100; H-500 to C-700, 1100; H-600 to C-400, 500, 700, 900; H-
900 to C-400, 500, 800; H-1000 to C-900; OMe to C-11; H-10 to
C-1; H-1000 to C-100; negative ion SIMS m/z 761 [M–H]ꢀ,
599, 389; negative ion HRSIMS m/z 761.2503 (calcd for
C33H45O20, 761.2506).
Pharmaceutical University) for MS measurements. This re-
search was financially supported by Kobe Pharmaceutical
University Collaboration Fund.
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