Two chromone-secoiridoid glycosides and three indole alkaloid glycosides from Neonauclea sessilifolia

Article abstract of DOI:10.1016/S0031-9422(02)00541-1

From the dried roots of Neonauclea sessilifolia, two new chromone-secoiridoid glycosides, sessilifoside and 7″-O-β-D-glucopyranosylsessilifoside, and three novel indole alkaloid glycosides, neonaucleosides A, B, and C, were isolated along with the main known glycosides, 5-hydroxy-2-methylchromone-7-O-β-D-apiofuranosyl-(1→6)- β-D-glucopyranoside, sweroside, loganin, grandifloroside, and quinovic acid 3β-O-β-D-quinovopyranoside-28-O-β-D-glucopyranoside. The structures of these new glycosides were determined by spectroscopic and chemical means. Neonaucleoside A and its C-3 epimer were prepared from secologanin and tryptamine.

Full text of DOI:10.1016/S0031-9422(02)00541-1

Phytochemistry 62(2003) 359–369
www.elsevier.com/locate/phytochem
Two chromone-secoiridoid glycosides and three indole alkaloid
glycosides from Neonauclea sessilifolia
Atsuko Itoha, Takao Tanahashia,*, Naotaka Nagakuraa, Toyoyuki Nishib
aKobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, Japan
bThe Nippon Shinyaku Institute for Botanical Research, Yamashina-ku, Kyoto 607-8182, Japan
Received 19 August 2002; received in revised form 23 September 2002
Dedicated to Professor Meinhart H. Zenk on the occasion of his 70th birthday
Abstract
From the dried roots of Neonauclea sessilifolia, two new chromone-secoiridoid glycosides, sessilifoside and 700-O-b-d-glucopyr-
anosylsessilifoside, and three novel indole alkaloid glycosides, neonaucleosides A, B, and C, were isolated along with the main known
glycosides, 5-hydroxy-2-methylchromone-7-O-b-d-apiofuranosyl-(1!6)-b-d-glucopyranoside, sweroside, loganin, grandifloroside,
and quinovic acid 3b-O-b-d-quinovopyranoside-28-O-b-d-glucopyranoside. The structures of these new glycosides were determined
by spectroscopic and chemical means. Neonaucleoside A and its C-3 epimer were prepared from secologanin and tryptamine.
# 2003 Elsevier Science Ltd. All rights reserved.
Keywords: Neonauclea sessilifolia; Rubiaceae; Roots; Structure elucidation; Secoiridoid glycosides; Indole alkaloid glycosides; Sessilifoside;
Neonaucleosides A–C
1. Introduction
graphic procedures to afford five new glycosides 1–5,
along with the known compounds, loganin (Kawai et
al., 1988), sweroside (Cambie et al., 1990), epi-vogelo-
side (Recio-Iglesias et al., 1992), secologanin dimethyl-
acetal (Kawai et al., 1988), grandifloroside (Chapelle,
1976), methyl grandifloroside (Chapelle, 1976), loganetin
(Jensen et al., 1979), strictosamide (Erdelmeier et al.,
1991), desoxycordifoline (Brown et al., 1978), 5,7-dihy-
droxy-2-methylchromone (Brown et al., 1975), 5-hydroxy-
2-methylchromone-7-O-b-d-apiofuranosyl-(1!6)-b-d-
glucopyranoside (Abe and Yamauchi, 1993), scopolin
(Tsukamoto et al., 1985), hymexelsin (Rao et al., 1988),
3,4-di-O-caffeoylquinic acid (Nishizawa et al., 1988),
quinovic acid 3b-O-b-d-glucopyranoside-28-O-b-d-glu-
copyranoside (Aquino et al., 1988), quinovic acid
3b-O-b-d-quinovopyranoside-28-O-b-d-glucopyranoside
(Arriaga et al., 1990), and quinovic acid 3b-O-b-d-
glucopyranosyl-(1!4)-a-l-rhamnopyranoside-28-O-b-d-
glucopyranoside (Fan and He, 1997).
Neonauclea sessilifolia (Roxb.) Merr. (=Nauclea ses-
silifolia Roxb., Adina sessilifolia (Roxb.) Hook. f., Nau-
clea sericea Wall. ex G. Don), belonging to Rubiaceae,
grows in Southeast Asia 7–30 m in height (Lo et al.,
1999). Previous phytochemical investigations of some
Neonauclea plants resulted in the isolation of some
alkaloids and anthraquinones (Johns et al., 1970; Atta-
ur-Rahman et al., 1988; Tosa et al., 1998), but no phy-
tochemical study on N. sessilifolia has been reported.
From our interests in chemical constituents of Rubia-
ceae plants, especially alkaloidal glycosides in polar
fraction (Itoh et al., 2002), we examined the n-BuOH-
soluble fraction of the roots of N. sessilifolia to isolate
five new glycosides. The structure elucidation of these
glycosides is reported here.
2. Results and discussion
Sessilifoside (1) was isolated as an amorphous pow-
der. It showed UV maxima at 232 sh, 251, 259 sh, 294,
and 327 sh nm and IR bands at 3421, 1697, 1655, 1618,
and 1508 cmÀ1. The 1H NMR spectrum of 1 exhibited a
doublet for an olefinic proton at ꢀ 7.68 (J=2.5 Hz),
signals for terminal vinyl group at ꢀ 5.29 (dd, J=10.0,
1.5 Hz), 5.31 (dd, J=17.0, 1.5 Hz), and 5.61 (dt,
The n-BuOH fraction of the dried roots of N. sessili-
folia was separated by a combination of chromato-
* Corresponding author. Fax: +81-78-441-7545.
E-mail address: tanahash@kobepharma-u.ac.jp (T. Tanahashi).
0031-9422/03/$ - see front matter # 2003 Elsevier Science Ltd. All rights reserved.
PII: S0031-9422(02)00541-1

360
A. Itoh et al. / Phytochemistry 62 (2003) 359–369
J=17.0, 10.0 Hz), a doublet for an acetal proton at ꢀ
5.59 (J=1.5 Hz), and a doublet for an anomeric proton
at ꢀ 4.71 (J=8.0 Hz), indicating the presence of secolo-
ganin-type secoiridoid glucoside moiety in the molecule.
Furthermore a double doublet at ꢀ 5.84 (J=12.0, 2.5
Hz) was assigned to H-7 in the secoiridoid unit by
COSY correlations of H-5- H2-6- H-7, suggesting its
structural similarity to vogeloside (6) or epi-vogeloside
(7) (Recio-Iglesias et al., 1992). A singlet for vinyl
methyl group at ꢀ 2.45 and a pair of doublets for meta-
coupled aromatic protons at ꢀ 6.17 and 6.28 (J=2.0
Hz), together with its 13C NMR spectrum (Table 1),
suggested that compound 1 possessed the carbon fra-
mework of 5,7-dihydroxy-2-methylchromone, which has
been isolated in this study. The linkage of C-7 of the
secoiridoid glucoside moiety and C-300 of the chromone
moiety was demonstrated by the HMBC correlations
between H-7 of secoiridoid glucoside moiety and C-200,
300, and 400 of chromone moiety. The R-configuration of
C-7 was confirmed by the coupling constants between
H-7 and H2-6 and the NOESY correlation between H-5
and H-7 (Recio-Iglesias et al., 1992). Thus, sessilifoside
(1) was deduced as a shown structure.
The second new glycoside 2 was obtained as an amor-
phous powder. The NMR spectral features of 2 were clo-
sely similar to those of 1 except for the presence of signals
arising from an additional b-d-glucopyranosyl unit,
which was identified as being the d-form by acid hydro-
lysis of 2 (Hara et al., 1986). The attachment of the sec-
ond glucosyl unit at the C-700 hydroxyl group of the
chromone moiety was confirmed by the HMBC corre-
lation between H-1000 and C-700 and the NOESY correlation
between H-1000 and H-800. Accordingly, the glycoside 2 was
assigned as 700-O-b-d-glucopyranosylsessilifoside.
Sessilifoside (1) and 700-O-b-d-glucopyranosylsessilifo-
side (2) represent the first chromone linked secoiridoid

A. Itoh et al. / Phytochemistry 62 (2003) 359–369
361
glucosides. The compounds could be biosynthesized
through an aldol type condensation of secologanin with
the C10 unit by acetate-malonate pathway (Scheme 1),
as hydrangenosides had been assumed to be formed
from secologanin and the C15 unit by shikimate-mal-
onate route (Inouye et al., 1980).
The stereochemistry at C-3 of 3 was confirmed by the
synthesis of 3 and its C-3 epimer 10. Condensation of
tryptamine (11) and secologanin (12) at pH 4 for 3 days
followed by treatment with additional 12 and sodium
cyanoborohydride at pH 6 afforded 3, 8, 10, vincosa-
mide (13) (Erdelmeier et al., 1991), and 14. Compound
10 had the same molecular formula as 3 and its spectral
data were quite similar to those of 3 except for the
NMR signals assigned to C-3, 5, 14, 15, 500, 600 and H-3,
5, 500, 600. These findings revealed that 10 must be C-3
epimer of 3. The NMR spectral data of 14 showed an
indole unit and a secoiridoid glucoside unit. A singlet at
ꢀ 7.19 was assigned to H-2in indole unit and a two-
protons multiplet at ꢀ 3.04 was assigned to H2-7 in the
secoiridoid glucoside unit, i. e. H2-3 in 14. These spec-
tral data as well as detailed 2D NMR experiments and
SI mass spectrum demonstrated that 14 was a reduced
compound of the imine derived from tryptamine (11)
and secologanin (12). Compound 8 was confirmed to be
strictosidine with C-3S configuration by lactamization
to strictosamide (15), followed by acetylation to stricto-
samide tetraacetate (16) (Blackstock et al., 1972; Stock-
igt and Zenk, 1977). Condensation of strictosidine (8)
with 12 in the presence of sodium cyanoborohydride
yielded 3, but not 10. Thus, neonaucleoside A (3) was
formulated as shown.
Neonaucleoside A (3) was also obtained as an amor-
phous powder. Its HR-SIMS measurement showed a
pseudomolecular ion [M–H]À at 901.3605, consistent
1
with a molecular formula C44H58N2O18. Its H NMR
spectrum exhibited two doublets for two olefinic pro-
tons at ꢀ 7.42( d, J=1.0 Hz) and 7.57 (brs), signals for
two terminal vinyl groups at ꢀ 5.23 (dd, J=10.5, 1.5
Hz), 5.36 (brd, J=17.0 Hz), 5.75 (ddd, J=17.0, 10.5, 9.0
Hz), 5.29 (dt, J=10.5, 1.5 Hz), 5.41 (dt, J=17.5, 1.5
Hz), and 5.84 (ddd, J=17.5, 10.5, 7.0 Hz), two doublets
for acetal protons at ꢀ 5.46 (d, J=5.0 Hz) and 5.58 (d,
J=7.0 Hz), two doublets for anomeric protons at ꢀ 4.65
and 4.73 (each d, J=8.0 Hz), and two singlets for two
carbomethoxyl groups at ꢀ 3.66 and 3.71, indicating two
secoiridoid glucoside units. Further signals for four
aromatic protons at ꢀ 6.96 (td, J=7.5, 1.0 Hz), 7.04 (td,
J=7.5, 1.0 Hz), 7.27 (brd, J=7.5 Hz), 7.38 (brd, J=7.5
Hz) and eight sp2 carbon signals at ꢀ 107.0, 111.9, 118.6,
119.7, 122.1, 128.2, 137.9 (Â 2) and two sp3 methylene
carbon signals at ꢀ 17.4 and 43.1 demonstrated the pre-
sence of a tetrahydro-b-carboline nucleus. These spec-
tral features suggested that compound 3 consisted of an
indole alkaloid glucoside such as strictosidine (8)
(Smith, 1968; Nagakura et al., 1979; Patthy-Lukats et
al., 1997) or vincoside (9) (Nagakura et al., 1979) bound
to an additional secoiridoid glucoside moiety. The link-
age of C-700 of the second secoiridoid glucoside unit at
N-4 in the indole alkaloid glucoside was determined by
the 13C NMR spectrum of 3, which showed a signal for
C-700 at ꢀ 51.3 as a methylene carbon and by a HMBC
correlation between H-700 and C-5.
Neonaucleoside B (4), C45H54N2O20, was also isolated
as an amorphous powder. Its spectral features were
closely related to those of desoxycordifoline (17)
(Brown et al., 1978), which is also a constituent of the
plant material, but with an additional secoiridoid unit.
Glycoside 4 was treated with (trimethylsilyl)diazo-
1
methane to give 18. In the H NMR spectrum of 4, sig-
nals of two carbomethoxyl groups appeared at ꢀ 3.41
and 3.69, which were correlated to C-22 and C-1100 in its
HMBC spectrum, respectively, while in the spectrum of
18, an additional carbomethoxyl signal was observed at
ꢀ 3.98, which was correlated to C-23 in its HMBC
spectrum. Accordingly, the newly formed carbo-
methoxyl group in 18 originated from a carboxyl group
at C-5. When 13C NMR spectral data of 4 and 18 were
compared with those of 17, significant differences were
observed in the signals around C-6. While a methine
carbon signal at ꢀ 116.4 was assigned to C-6 in 17, a
quaternary carbon signal at ꢀ 135.1 in 4 was assigned to
C-6, suggesting the attachment of the second secoiridoid
glucoside at C-6 of desoxycordifoline (17) moiety. The
site of linkage in the second secoiridoid glucoside was
revealed to be C-7 by the correlations between H2-700
and C-5, 6, and 7 in the HMBC spectra of 4 and 18 and
the interactions between H-9 and H2-700, H2-600, and H-
500 in the ROESY experiments of 4 and 18. Accordingly,
the structure of neonaucleoside B was represented by 4.
Neonaucleoside C (5) was isolated as a colorless crys-
talline solid, mp >300 ꢀC and the molecular formula of
1
Scheme 1. Proposed biosynthetic sequence for sessilifosides.
C44H54N2O20 was deduced from its HR-SIMS. The H

362
A. Itoh et al. / Phytochemistry 62 (2003) 359–369
and 13C NMR spectral features of 5 were similar to
those of 3 and showed the presence of a tetrahydro-b-
carboline unit and two secoiridoid units in the molecule.
However, the 13C NMR spectrum of 5 differed remark-
ably in that one methoxyl and three carbonyl carbons
were observed and that C-5 and C-6 appeared as
methine carbons. Methylation of 5 with (trimethylsi-
lyl)diazomethane afforded two methylated compounds
19 and 20. The 1H NMR spectra of the two compounds
demonstrated two methoxyl groups in 19 and 20, but
with an additional N-methyl group in 20, suggesting
that a carbomethoxyl, a carboxyl group and a second-
ary amino group were present in 5. HMBC correlation
between OMe (ꢀ 3.66) and C-22 (ꢀ 167.5) in 5 as well as
HMBC correlations between OMe (19: ꢀ 3.71; 20: ꢀ
3.57) and C-22 (19: ꢀ 169.8; 20: ꢀ 169.3), and between
OMe (19: ꢀ 3.84; 20: ꢀ 3.78) and C-23 (19: ꢀ 174.8; 20: ꢀ
174.0) in 19 and 20 demonstrated the presence of a car-
bomethoxyl group at C-16 and a carboxyl group at C-5
in 5. A remaining carbonyl carbon was ascribed to a
lactone ring of the second secoiridoid glucose unit as in
sured on a Jasco DIP-370 digital polarimeter and CD
spectra on a Shimadzu-AVIV 62A DS circular dichro-
1
ism spectrometer. H (500 and 300 MHz) and 13C (125
and 75 MHz) NMR spectra were recorded on a Varian
VXR-500 and Varian Gemini-300 spectrometers with
TMS as an internal standard. MS and HR-MS were
obtained with a Hitachi M-4100 mass spectrometer.
Glycerol was used for SIMS and HR-SIMS as the
matrix. MPLC was carried out with Wakosil 40C18.
TLC was performed on precoated Kieselgel 60F254
plates (Merck).
3.2. Plant material
The roots of Neonauclea sessilifolia were collected at
Surat Thani, Thailand and identified by Dr. Tem Smi-
tinand, Forest Herbarium, Royal Forest Department,
Bangkok, Thailand. A voucher specimen (NST-592) is
deposited in the laboratory of the Nippon Shinyaku
Institute for Botanical Research.
1
1 by H and 13C NMR data and 2D-NMR experiments
3.3. Extraction and isolation
of 5, 19 and 20. The linkage of C-700 of the secoiridoid
glucoside unit to C-6 of tetrahydro-b-carboline unit was
revealed by COSY correlation between H-6 and H-700,
HMBC correlation between H-5 and C-700 and ROESY
correlations between H-9 and H-700 and between H-9
and H2-600 in 5, 19 and 20.
The stereochemistry was determined by the biogenetic
consideration as well as the following spectral data. The
ROESY correlation between H-3 and H-5 suggested that
both H-3 and H-5 adopted axial orientations. The cou-
pling constants around 3.5 Hz between H-5 and H-6 in
5, 19, and 20 therefore showed that H-6 take up an equa-
torial orientation. Assuming that the tetrahydro-b-car-
boline moiety is derived from l-tryptophane, the absolute
configuration at C-5 would be S. And following this, C-
6 was assigned to have the R-configuration. Similarly, a
ROESY correlation between H-500 and H-700 together
with obvious trans-diaxial couplings to one of the pro-
tons at C-600, allowed to assign the 700(R)-configuration
for 5 (Recio-Iglesias et al., 1992). Accordingly, neonau-
cleoside C (5) was deduced as shown structure.
Dried roots (1.58 kg) of N. sessilifolia were extracted
with hot MeOH and the MeOH extracts were evapo-
rated in vacuo. A part (21.4 g) of the residue (173 g) was
resuspended in H2O and extracted successively with
CHCl3 and n-BuOH. The residue from n-BuOH layers
(10.0 g) was fractionated on reversed-phase MPLC.
Elution with H2O–MeOH mixtures of indicated MeOH
content gave 9 fractions, 1 (20%, 1.69 g), 2 (20%, 556
mg), 3 (25%, 162mg), 4 (30%, 472mg), 5 (35–40%, 607
mg), 6 (50%, 366 mg), 7 (50%, 720 mg), 8 (60%, 2.18
g), and 9 (80%, 1.11 g). Fr. 1 was submitted to reversed-
phase MPLC eluted with MeOH–H2O (3:17, 1:4, 3:7),
and was further purified by prep. HPLC (mBondasphere
5m C18-100A, MeOH–H2O, 7:13) and prep. TLC
(CHCl3–MeOH, 4:1) to afford 5-hydroxy-2-methyl-
chromone-7-O-b-d-apiofuranosyl-(1!6)-b-d-glucopyr-
anoside (312mg), sweroside (520 mg), loganin (160 mg),
scopolin (11.7 mg), hymexelsin (34.0 mg), 7 (10.0 mg),
and loganetin (5.2mg). Fr. 2was purified by prep.
HPLC (MeOH-H2O, 7:13) to give 5-hydroxy-2-methyl-
chromone-7-O-b-d-apiofuranosyl-(1!6)-b-d-glucopyr-
anoside (153 mg), loganin (116 mg), and 7 (41.6 mg).
Frs. 3–9 were further purified by a combination of silica
gel CC (CHCl3–MeOH, 93:7-3:2), prep. HPLC
(MeOH–H2O, 7:13, 9:11, 1:1, 11:9, 3:2, 3:1, or 4:1) and
prep. TLC (CHCl3–MeOH, 7:3 or 4:1; CHCl3–MeOH-
H2O, 70:30:3; AcOEt–C6H6–EtOH, 4:1:2), respectively.
Fr. 3 yielded 3,4-di-O-caffeoylquinic acid (48.5 mg); fr.
4: 5,7-dihydroxy-2-methylchromone (8.8 mg), 17 (145
mg), 2 (9.5 mg), and secologanin dimethylacetal (3.6
mg); fr. 5: grandifloroside (206 mg); fr. 6: 5 (82.0 mg), 1
(5.7 mg), and methyl grandifloroside (11.0 mg); fr. 7: 4
(57.9 mg), quinovic acid 3b-O-b-d-glucopyranoside-28-
Neonaucleosides A-C (3–5) were quite unique glyco-
sides. These compounds are the first examples of indole
alkaloid glycosides linked with a second secoiridoid
glucoside unit.
3. Experimental
3.1. General
UV spectra were recorded on a Shimadzu UV-2500PC
spectrophotometer and IR spectra on a Shimadzu FTIR-
8200 spectrophotometer. Optical rotations were mea-

A. Itoh et al. / Phytochemistry 62 (2003) 359–369
363
O-b-d-glucopyranoside (73.6 mg), and strictosamide
(13.1 mg); fr. 8. quinovic acid 3b-O-b-d-quinovopyr-
anoside-28-O-b-d-glucopyranoside (1.09 g), 3 (19.6 mg),
and quinovic acid 3b-O-b-d-glucopyranosyl-(1!4)-a-l-
rhamnopyranoside-28-O-b-d-glucopyranoside (37.6 mg).
HMBC: H-10 to C-1; H-7 to C-200, 300, 400; Me-200 to C-200,
300.
3.5. 700-O-ꢃ-d-Glucopyranosylsessilifoside (2)
22
D
3.4. Sessilifoside (1)
Amorphous powder. ½ꢁŠ À165ꢀ (c 0.7, MeOH). UV
max
MeOH
l
nm ðlog "Þ: 245 (4.39), 251 (4.40), 261 sh (4.35),
26
D
Amorphous powder. ½ꢁŠ À151ꢀ (c 0.3, MeOH). UV
max
291 sh (3.77), 313 (3.64). IR ꢂ
cmÀ1: 3405, 1694,
KBr
max
MeOH
l
nm ðlog "Þ: 232 sh (4.25), 251 (4.33), 259 sh
1655, 1616, 1505. 1H NMR: Table 1. 13C NMR: Table 2.
Negative ion SIMS m/z: 709 [M–H]À, 547. HR-SIMS
Found 709.1988 [M–H]À; C32H37O18 requires 709.1981.
NOESY: H-5/H-7; H-1/H-10; H-800/H-1000. HMBC: H-1
to C-10; H-10 to C-1; H-6 (ꢀ 2.17) to H-300; H-7 to H-200,
KBr
max
(4.30), 294 (3.75), 327 sh (3.58). IR ꢂ
cmÀ1: 342 1,
1697, 1655, 1618, 1508. H NMR: Table 1. 13C NMR:
Table 1. HR-SIMS Found 547.1457 [M–H]À; C26H27O13
requires 547.1452. NOESY: H-5/H-7; H-1/H-10.
1
Table 1
13C (125 MHz) and 1H (500 MHz) NMR spectral data of 1 and 2 in CD3OD
1
2
Position
ꢀC
98.1
ꢀH
ꢀC
98.1
ꢀH
1
3
4
5
6
5.59 d (1.5)
7.68 d (2.5)
5.60 d (1.5)
7.69 d (2.5)
154.5
105.6
28.7
154.5
105.6
28.6
3.34 m
3.27–3.53 m
30.0
1.87 ddd (13.5, 4.5, 2.5)
2.18 td (13.5, 12.0)
5.84 dd (12.0, 2.5)
5.61 dt (17.0, 10.0)
2.74 ddd (9.5, 5.5, 1.5)
5.29 dd (10.0, 1.5)
5.31 dd (17.0, 1.5)
29.9
1.88 ddd (13.5, 4.5, 2.5)
2.17 td (13.5, 12.0)
5.85 dd (12.0, 2.5)
5.61 dt (17.0, 10.0)
2.75 ddd (10.0, 5.5, 1.5)
5.29 dd (10.0, 2.0)
5.31 dd (17.0, 2.0)
7
75.1
133.3
43.9
75.0
133.3
43.8
8
9
10
121.1
121.1
11
10
20
30
40
50
60
168.6
99.8
74.8
77.9
71.6
78.5
62.7
168.5a
99.7
74.7b
77.8c
71.2d
78.4e
62.4f
4.71 d (8.0)
3.23 dd (9.0, 8.0)
3.39 t (9.0)
4.71 d (8.0)
3.23 dd (9.5, 8.0)
3.27–3.53 m
3.27–3.53 m
3.29 dd (10.0, 9.0)
3.34 m
3.27–3.53 m
3.67g dd (12.0, 5.5)
3.90 dd (12.0, 2.0)
3.67 dd (12.0, 6.0)
3.90 dd (12.0, 2.0)
200
167.5
168.1a
300
118.2118.6
181.6
163.5
400
181.8
163.1
101.1
500
600
100.26.17
166.2164.9
94.7
d (2.0)
6.47 d (2.0)
6.64 d (2.0)
700
800
6.28 d (2.0)
95.7
900
159.2158.7
104.9
18.9
1000
Me
1000
2000
3000
4000
5000
6000
106.6
19.0
2.45 s
2.48 s
5.03 d (7.5)
101.6
74.8b
77.9c
71.6d
78.5e
62.7f
3.27–3.53 m
3.27–3.53 m
3.27–3.53 m
3.27–3.53 m
3.71g dd (12.0, 5.5)
3.90 dd (12.0, 2.0)
aÀg
Values with the same superscript are interchangeable.

364
A. Itoh et al. / Phytochemistry 62 (2003) 359–369
300, 400; Me-200 to C-200, 300; H-1000 to C-700; H-800 to C-600,
700, 900, 1000.
OMe (ꢀ 3.71) to C-22; H-21 to C-10; H-10 to C-21; H-300
to C-1100 H-700(ꢀ 2.80- 2.90) to C-5; H-500 to C-700, 1100;
OMe (ꢀ 3.66) to C-1100; H-100 to C-1000; H-1000 to C-100.
3.6. Neonaucleoside A (3)
3.7. Neonaucleoside B (4)
28
D
Amorphous powder. ½ꢁŠ À133ꢀ (c 1.0, MeOH). UV
max
28
D
MeOH
l
nm ðlog "Þ: 227 (4.54), 273 (3.73), 283 sh (3.72),
Amorphous powder. ½ꢁŠ À212ꢀ (c 1.0, MeOH). UV
max
KBr
MeOH
290 sh (3.64). IR ꢂ
cmÀ1: 3400, 1697, 1636. 1H
l
304 (3.73), 335 sh (3.56), 349 (3.65), 387 sh (3.21). IR
ꢂ
nm ðlog "Þ: 238 (4.58), 263 (4.43), 290 sh (3.89),
max
NMR: Table 2. 13C NMR: Table 5. Negative ion SIMS
m/z: 901 [M–H]À, 739. HR-SIMS Found 901.3605 [M–
H]À; C44H57N2O18 requires 901.3608. ROESY: H-21/H-
10; H-100/H-1000. HMBC: H-15 to C-3, 22; H-17 to C-22;
KBr
max
1
cmÀ1: 3384, 1697, 1624. H NMR: Table 3. 13C
NMR: Table 5. HR-SIMS Found 941.3177 [M-H]À;
C45H53N2O20 requires 941.3193. ROESY: H-9/H2-700;
Table 2
1H NMR spectral data of 3, 10 and 14 in CD3OD at 500 MHz
H
3
10
14
2
3
7.19 s
3.04 m
4.08 m
3.91 m
3
5
5
6
6
9
3.04 m
3.29 t (7.5)
3.29 t (7.5)
3.16 t (7.5)
3.16 t (7.5)
3.20–3.39 m
3.20–3.39 m
2.55 brd (15.0)
2.80–2.90 m
7.38 brd (7.5)
6.96 td (7.5, 1.0)
7.04 td (7.5, 1.0)
brd (7.5)
1.93 m
2.07 m
3.20–3.39 m
7.57 brs
5.29 dt (10.5, 1.5)
5.41 dt (17.5, 1.5)
5.84 ddd (17.5, 10.5, 7.0)
2.80–2.90 m
5.58 d (7.0)
3.08 dd (13.0, 6.0)
3.26–3.35 m
2.58 m
2.90 ddd (16.0, 11.0, 6.0)
7.40 brd (8.0)
6.98 td (8.0, 1.0)
7.06 td (8.0, 1.0)
7.32 brd (8.0)
1.95 m
2.13 m
3.23 m
7.55 brs
5.21 brd (10.5)
5.22 brd (17.0)
5.80 ddd (17.0, 10.5, 8.5)
2.67–2.74 m
5.63 d (6.5)
3.73 s
4.71 d (8.0)
3.22 dd (9.0, 8.0)
3.38 t (9.0)
3.26–3.35 m
3.26–3.35 m
3.65d dd (12.0, 5.0)
3.86e dd (12.0, 2.0)
5.45 d (5.5)
7.42 d (1.0)
2.77 m
1.67 m
2.04 m
2.67–2.74 m
2.67–2.74 m
5.71 ddd (17.0, 10.0, 9.0)
2.58 m
5.11 brd (17.0)
5.17 dd (10.5, 1.5)
3.67 s
7.58 brd (8.0)
7.05 td (8.0, 1.0)
7.13 td (8.0, 1.0)
7.37 brd (8.0)
1.87 ddd (14.0, 8.5, 7.0)
1.97 dt (14.0, 7.0)
2.80 brtd (7.0, 5.5)
7.54 d (0.5)
5.28 brd (10.5)
5.32 brd (17.0)
5.75 ddd (17.0, 10.5, 8.5)
2.63 ddd (8.5, 6.5, 5.5)
5.54 d (6.5)
3.69 s
4.69 d (8.0)
3.18 dd (9.0, 8.0)
3.37 t (9.0)
3.25 t (9.0)
3.32 ddd (9.0, 6.5, 2.0)
3.65 dd (12.0, 6.5)
3.91 dd (12.0, 2.0)
10
11
127.2
14
14
15
17
18
18
19
20
21
22-OMe
10
3.71 s
4.73 d (8.0)
20
3.19a dd (9.0, 8.0)
3.20–3.39 m
3.20–3.39 m
3.20–3.39 m
3.62b dd (12.0, 6.0)
3.87c dd (12.0, 2.0)
5.46 d (5.0)
30
40
50
600
600
100
300
7.42 d (1.0)
500
2.95 quint (5.0)
1.62 m
2.31 m
600
6
700
2.75 m
700
2.80–2.90 m
5.75 ddd (17.0, 10.5, 9.0)
2.80–2.90 m
5.23 dd (10.5, 1.5)
5.36 brd (17.0)
3.66 s
800
900
1000
1000
1010000-OMe
1000
2
4.65 d (8.0)
4.65 d (8.0)
3.22a dd (9.0, 8.0)
3.20–3.39 m
3.20–3.39 m
3.20–3.39 m
3.64b dd (12.0, 6.0)
3.87c dd (12.0, 2.0)
3.17 dd (9.0, 8.0)
3.26–3.35 m
3.26–3.35 m
3.26–3.35 m
3000
4000
5000
6000
6000
3.66d
3.91e dd (11.5, 2.0)
m
aÀe
Values with the same superscript are interchangeable.

A. Itoh et al. / Phytochemistry 62 (2003) 359–369
365
H-9/H2-600; H-9/H-500; H-21/H-10; H-100/H-1000. HMBC:
H2-14 to C-2, 3; H-15 to C-3, 14; H-17 to C-22; OMe (ꢀ
3.41) to C-22; H-300 to C-1100; OMe (ꢀ 3.69) to C-1100;
H2-700 to C-5, 6, 7; H-21 to C-10; H-10 to C-21; H-100 to
C-1000; H-1000 to C-100.
requires 929.3193. COSY: H-6/H-700. ROESY: H-3/H-5;
H-3/H-15; H-21/H-10; H-9/H-6; H-9/H2-600; H-9/H-700;
H-500/H-700; H-100/H-1000; H-300/H-1000. HMBC: H-17 to C-
1, 16, 22; OMe to C-22; H-5 to C-6, 23, 700; H-21 to C-10;
H-10 to C-21; H-300 to C-100, 400, 500, 1100; H-100 to C-1000; H-
1000 to C-100.
3.8. Neonaucleoside C (5)
3.9. Acid hydrolysis of 7-O-ꢃ-d-glucopyranosylsessilifoside
(2)
28
D
ꢀ
Colorless crystalline solid. mp >300 C (H2O). ½ꢁŠ
max
MeOH
À326ꢀ (c 0.2, Py). UV l
nm ðlog "Þ: 220 (4.61),
max
KBr
ꢀ
243 (4.34), 282 sh (3.84), 288 (3.74). IR ꢂ
1
cmÀ1
:
Glycoside 2 (1 mg) was heated at 95 C with dioxane
(0.5 ml) and 5% H2SO4 (0.5 ml) for 1 h. After neu-
tralization with Amberlite IRA-400 (OHÀ form), the
3421, 1690, 1626, 1508. H NMR: Table 4. 13C NMR:
Table 5. HR-SIMS Found 929.3180 [M-H]À; C44H53N2O20
Table 3
1H NMR spectral data of 4, 17 and 18 in CD3OD at 500 MHz
H
4
17
18
6
8.81 s
9
10
8.23 brd (8.0)
7.35 brt (8.0)
7.65 brt (8.0)
brd (8.0)
8.29 brd (8.0)
7.38 m
7.67 m
8.19 brd (8.0)
7.29 td (8.0, 1.0)
7.57 td (8.0, 1.0)
7.64 brd (8.0)
3.31 m
11
127.68
7.67 m
3.56 m
14
14
3.44 dd (14.0, 8.0)
3.52 dd (14.0, 6.0)
3.58 ddd (8.0, 6.0, 4.5)
7.56 s
3.56 m
3.61 m
7.55 s
3.47 dd (14.0, 6.5)
3.55 brq (6.5)
7.51 d (0.5)
15
17
18
18
5.05–5.09 m
5.05–5.09 m
5.08–5.12 m
5.08–5.12 m
5.90 ddd (17.0, 11.0, 8.5)
2.69 td (7.5, 5.0)
5.84 d (7.5)
3.32 s
5.00 brd (17.0)
5.03 dd (10.0, 1.0)
5.88 ddd (17.0, 10.0, 6.0)
2.66 ddd (7.5, 7.0, 6.0)
5.77 d (7.0)
19
20
5.84 ddd (17.0, 10.5, 8.5)
2.67 ddd (8.5, 7.0, 4.5)
5.77 d (7.0)
21
22-OMe
3.41 s
3.35 s
3.98 s
4.76 d (8.0)
23-OMe
10
4.76 d (8.0)
3.22a dd (9.0, 8.0)
3.32–3.42 m
4.79 d (8.0)
3.23 dd (9.0, 8.0)
3.41 t (9.0)
20
3.22 dd (9.0, 8.0)
3.24–3.41 m
3.24–3.41 m
30
40
3.27b t (9.5)
3.28 dd (10.0, 9.0)
3.38 ddd (10.0, 6.5, 2.0)
3.69 dd (12.0, 6.5)
3.96 dd (12.0, 2.0)
50
3.32–3.42 m
3.24–3.41 m
600
3.65c dd (12.0, 6.0)
3.83d dd (12.0, 1.5)
5.70 d (8.0)
3.70 dd (12.0, 6.0)
3.91 dd (12.0, 2.0)
5.61 d (7.5)
600
100
300
7.63 s
7.62 s
500
3.10 brq (6.0)
2.02 m
2.02 m
3.10 brq (6.0)
1.99 m
2.07 m
600
6
700
3.66 m
3.74 m
3.44 m
3.44 m
700
800
5.99 ddd (17.0, 10.5, 8.5)
2.75 ddd (8.5, 8.0, 5.0)
5.24 brd (10.5)
5.34 brd (17.0)
3.69 s
5.87 ddd (17.0, 10.5, 5.5)
2.75 td (7.5, 5.5)
5.26 brd (10.5)
5.37 brd (17.0)
3.69 s
900
1000
1000
1010000-OMe
1000
2
4.73 d (8.0)
3.22a dd (9.0, 8.0)
3.32–3.42 m
3.27b t (9.5)
3.32–3.42 m
3.67c dd (12.0, 6.0)
3.94d dd (12.0, 1.5)
4.71 d (8.0)
3.20 dd (9.0, 8.0)
3.24–3.41 m
3.24–3.41 m
3000
4000
5000
6000
6000
3.24–3.41 m
3.59 dd (11.5, 5.5)
3.79 dd (11.5, 1.0)
aÀd
Values with the same superscript are interchangeable.

366
A. Itoh et al. / Phytochemistry 62 (2003) 359–369
reaction mixture was concentrated and the residue was
passed through a Sep-Pak C18 cartridge with H2O. The
eluate was concentrated and the residue was treated
with l-cysteine methyl ester hydrochloride (1 mg) in
3.10. Condensation of tryptamine (11) with secologanin
(12)
A solution of secologanin (12) (50 mg, 0.13 mmol)
and tryptamine–HCl (11) (50 mg, 0.25 mmol) in citrate-
phosphate buffer (1.0 ml, pH 4.0) was incubated for 3
days at room temperature. To the reaction mixture
adjusted to pH 5–6 with 5% aq NaHCO3 12 (50 mg)
and NaBH3CN (40 mg) were added. After stirring for 2
h at room temperature, the reaction mixture was sub-
jected to a Sephadex LH-20 CC with MeOH as eluent
ꢀ
pyridine (0.125 ml) at 60 C for 1 h. The solution was
then treated with N,O-bis(trimethylsilyl)trifluoroacetamide
ꢀ
(0.05 ml) at 60 C for 1 h. The supernatant was applied
TM
to GLC; GLC conditions: column, SupelcoꢀSPB -1, 30
m  0.25 mm; column temperature, 230 C; N2 flow
rate, 0.8 ml/min; tR of derivatives, d-glucose 13.1 min,
l-glucose 13.6 min. d-Glucose was detected from 2.
Table 4
1H NMR spectral data of 5, 19 and 20 at 500 MHz
H
5a
19b
20b
1
3
5
11.00 brs
4.21 brd (11.0)
3.65-3.70 m
4.10 brd (10.5)
3.81 d (3.5)
3.68 td (3.5, 1.5)
7.44 brd (8.0)
6.98 td (8.0, 1.0)
7.05 td (8.0, 1.0)
7.29 brd (8.0)
1.98 ddd (13.5, 10.5, 3.5)
2.15 ddd (13.5, 10.0, 2.5)
3.14 ddd (10.0, 5.0, 3.5)
7.62 s
3.58 brd (8.0)
3.47 d (4.0)
3.79 m
7.47 brd (8.0)
7.03 td (8.0, 1.0)
7.10 td (8.0, 1.0)
7.37 brd (8.0)
1.97 ddd (14.5, 10.5, 2.5)
2.46 m
6
9
3.57 brt (3.0)
7.45 brd (8.0)
6.95 td (8.0, 1.0)
7.04 td (8.0, 1.0)
brd (8.0)
1.87 brt (11.0)
2.18 brt (11.0)
2.90-3.10 m
10
11
127.2
14
14
15
17
18
18
19
20
21
22-OMe
23-OMe
NMe
10
3.22-3.40 m
7.46 d (1.5)
7.62 s
5.23 brd (10.0)
5.35 brd (17.0)
5.81 ddd (17.0, 10.0, 7.5)
2.63 ddd (8.0, 7.5, 5.0)
5.59 d (8.0)
5.23 brd (10.5)
5.35 brd (17.0)
5.91 ddd (17.0, 10.5, 8.0)
2.68 td (8.0, 5.0)
5.74 d (8.0)
5.31-5.34 m
5.31-5.34 m
5.82 dt (18.0, 9.5)
2.86 ddd (9.5, 5.0, 4.0)
5.49 d (4.0)
3.66 s
3.71 s
3.84 s
3.57 s
3.78 s
2.44 s
4.69 d (8.0)
3.22-3.40 m
3.22-3.40 m
3.22-3.40 m
4.60 d (8.0)
2.90-3.10 m
3.12-3.20 m
2.90-3.10 m
4.76 d (8.0)
3.22 dd (9.0, 8.0)
3.39 t (9.0)
20
30
40
3.23d t (9.0)
50
3.12-3.20 m
3.29e ddd (9.0, 5.5, 2.0)
3.63f dd (12.0, 5.5)
3.86g dd (12.0, 2.0)
5.48 d (1.5)
3.22-3.40 m
600
3.41c dd (11.0, 6.0)
3.65-3.70 m
5.40 d (1.5)
3.64h dd (12.0, 5.5)
3.85i dd (12.0, 2.0)
5.51 d (1.5)
600
100
300
7.42 d (2.5)
2.90-3.10 m
7.52 d (2.5)
7.54 d (2.5)
500
2.89 dddd (13.0, 5.5, 4.0, 2.5)
1.64 ddd (13.5, 4.0, 1.5)
2.10 brq (13.0)
2.85 dddd (14.0, 5.5, 4.5, 2.5)
1.61 ddd (14.0, 4.5, 2.5)
2.49 td (14.0, 12.0)
4.89 m
600
1.69 brq (13.0)
1.91 brd (12.0)
4.79 brd (12.0)
5.20 ddd (17.0, 10.0, 8.5)
2.58 ddd (8.5, 5.5, 1.5)
5.07 dd (10.0, 2.5)
5.26 dd (17.0, 2.5)
4.46 d (8.0)
6
700
*
800
5.41 dt (17.0, 10.0)
2.54 ddd (9.0, 5.5, 1.5)
5.17 dd (10.0, 1.5)
5.24 dd (17.0, 1.5)
4.62 d (8.0)
5.57 ddd (16.5, 10.5, 9.0)
2.60 ddd (9.0, 5.5, 1.5)
5.21 dd (16.5, 1.5)
5.23 dd (10.5, 1.5)
4.63 d (8.0)
900
1000
1000000
1000
2000
3
2.90-3.10 m
3.12-3.20 m
3.12 dd (9.0, 8.0)
3.34 t (9.0)
3.11 dd (9.0, 8.0)
3.22-3.40 m
3.22-3.40 m
3.22-3.40 m
3.66h dd (12.0, 5.5)
3.89i dd (12.0, 2.0)
4000
5000
6000
6000
2.90-3.10 m
3.12-3.20 m
3.46c dd (11.0, 5.5)
3.65-3.70 m
3.25dt (9.0)
3.34e ddd (9.0, 6.5, 2.0)
3.65f dd (12.0, 6.5)
3.93g dd (12.0, 2.0)
a
Measured in DMSO-d6.
Measured in CD3OD.
Values with the same superscript are interchangeable.
b
c
* Overlapped with HOD signal.

A. Itoh et al. / Phytochemistry 62 (2003) 359–369
367
1
and the products were further purified by prep. TLC
(CHCl3–MeOH–H2O, 70:30:1.5; CHCl3–MeOH, 4:1;
AcOEt–C6H6–EtOH, 4:1:2) to afford 3 (2.2 mg), stric-
tosidine (8) (21.7 mg), 10 (5.4 mg), vincosamide (13) (5.9
mg), 14 (16.9 mg). Vincosamide (13) was identified with
ion SIMS m/z: 529 [M–H]À, 497. 3: H NMR spectral
data were identical with those of 3 from natural source.
22
D
227 (4.62), 272 sh (3.81), 280 (3.82), 290 sh (3.72). IR
10: ½ꢁŠ À157ꢀ (c 0.2, MeOH). UV l
nm ðlog "Þ:
MeOH
max
KBr
max
1
ꢂ
cmÀ1: 3400, 1697, 1636, 1437. H NMR: Table 2.
authentic sample (1H NMR, UV, SIMS and CD
13C NMR: Table 5. Negative ion SIMS m/z: 901 [M–
MeOH
max
l
nm ðꢀ "Þ: 217 (+17.8), 232 (À15.2), 267
H]À, 739. HR-SIMS Found 901.3607 [M–H]À;
C44H57N2O18 requires 901.3608. NOESY: H-3/H-20;
26
D
(À5.9)). 8: ½ꢁŠ
max
À206ꢀ (c 0.8, MeOH). UV
21
D
MeOH
l
289 (3.79). IR ꢂ
nm ðlog "Þ: 224 (4.53), 274 (3.85), 281 (3.86),
H-100/H-1000. 14: ½ꢁŠ
max
À84ꢀ (c 1.0, MeOH). UV
KBr
max
MeOH
cmÀ1: 3368, 1684, 1632. Negative
l
nm ðlog "Þ: 222 (4.09), 241 sh (4.57), 273 (3.77),
Table 5
13C NMR spectral data of 3-5, 10, 14, 17-20 at 125 MHz
C
3a
10a
14a
4a
17a
18a
5b
19a
20a
2137.9
3
5
6
7
137.74.3
59.1
43.1
12136.8
137.3
47.5
13173.26.7
13193.27.9
56.3
45.249.3
18.22 135.1
106.8
128.4
118.7
119.8
122.2
112.0
137.7
35.3
140.4
143.0
137.8
143.3
137.5c
51.0
59.8
38.1
52.4
60.1
40.8
60.3
67.5
41.9
i
n.d.
116.4
c
17.4
3.5
133.7
107.0
128.2
118.6
119.7
122.1
111.9
137.9
36.7
110.1
128.1
118.9
120.1
122.8
112.6
138.3
28.0
31.9
109.9
154.6
120.2
130.7
122.4
125.3
122.2
130.3
113.5
143.7
33.5
132.2
122.6
123.4
122.4
131.3
113.5
144.0
34.0
128.1c
122.6
124.7
121.4
129.0
113.2
142.5
35.2
106.0
126.6
118.6
118.6
120.8
111.1
136.1
36.5
106.9
127.9
119.0
120.1
122.2
112.1
138.1
38.6
106.2
127.1
118.5
120.4
122.6
112.4
138.4
35.7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
OMe
OMe
10
31.5
30.6
34.3
110.3
34.9
110.1
34.4
111.0
30.3
110.0
32.4
111.8
29.5
112.0
112.5
154.2154.4
119.7
136.1
45.5
112.0
154.7
119.8
135.8
45.4
154.8
154.2153.0
154.7
153.6
120.0
135.1
45.5
119.9
135.2135.6
45.7
119.4
119.2
119.6
120.8
135.0
45.1
97.5
134.7
136.1
135.9
45.7
97.4
169.5
169.5
51.8d
52.8
44.1
95.9
167.5
171.3
51.4
46.3
97.7
169.8
174.8
52.0
45.2
99.0
169.3
174.0
51.8
98.1
169.8
97.9
170.1
97.5
97.4
169.6
169.3
n.d.i
51.9
169.3
167.4
51.9
51.9
52.1
52.0
52.5
100.8
52.6
100.3c
74.6d
78.0e
71.6f
78.4g
62.8h
100.1
74.7c
78.1
71.6d
78.4e
62.7f
100.2100.3
74.7
78.0
71.6
78.5
100.4
100.2 98.9
100.4
20
74.6
78.0
71.6
78.4c
62.8d
74.7
78.1
71.7
78.7
63.0
74.7
78.0
71.6
78.6
62.8
73.0
74.6c
77.8d
71.5e
78.3f
62.6g
74.6
30
76.2c
69.9d
77.2e
61.0
77.8c
71.5d
78.4e
62.6f
42.9
40
50
60
62.8
NMe
100
98.4
153.2153.2
111.9
30.4
27.8
51.3
135.6
44.8
119.9
169.4
51.8
100.4c
74.7d
78.1e
71.7f
78.5g
62.9h
97.9
98.0
98.0
95.3
97.8
97.8
300
154.3
154.0
151.5
153.7
153.6
400
111.9
31.7
28.8
52.1
135.5
45.4
119.8
169.4
51.9
100.2100.3
74.8c
78.1
111.3
36.0
31.4
28.4
136.4
46.0
119.1
169.5
51.9
111.8
35.7
31.9
29.0
136.3
45.7
119.2
169.5
51.9d
104.3
26.9
28.4
80.1
131.9
41.3
106.1
28.8
29.0
106.4
28.6
27.1
500
600
700
82.6
82.6
800
133.3
43.7
120.6
133.7
43.5
120.4
900
1000
1100
OMe
1000
2000
3000
4000
5000
6000
120.1
164.4
168.2168.6
100.3
97.9
99.5
99.5
74.6
78.0
71.6
78.6c
62.9d
74.7
78.0
71.6
78.3
62.7
73.0
74.7c
78.0d
71.7e
78.6f
62.9g
74.6
76.6c
70.0d
77.3e
61.0
78.2c
71.7d
78.6e
62.8f
71.7d
78.5e
63.0f
a
Measured in CD3OD.
Measured in DMSO-d6.
Values with the same superscript in the same column are interchangeable.
Signals were not detected.
b
cÀh
i

368
A. Itoh et al. / Phytochemistry 62 (2003) 359–369
KBr
max
281 (3.77), 290 (3.70). IR ꢂ
cmÀ1: 3400, 1684, 1636,
3.14. Methylation of neonaucleoside C (5)
1458, 1437. 1H NMR: Table 2. 13C NMR: Table 5.
Negative ion SIMS m/z: 531 [M–H]À, 369. HR-SIMS
Found 531.2351 [M–H]À; C27H35N2O9 requires
531.2344. NOESY: H2-3/H2-5; H2-5/H2-14; H2-3/H-15;
H2-3/H-10; H-10/H-18 (ꢀ 5.32); H-11/H-18 (ꢀ 5.28); H-
12/H-19. HMBC: H2-5 to C-3.
A methanolic solution of 5 (9.8 mg) was treated with
2.0 M TMSCHN2 in n-hexane (0.1 ml) to afford 19 (4.7
27
mg) and 20 (4.0 mg). 19: ½ꢁŠD À187ꢀ (c 0.4, MeOH). UV
max
MeOH
l
nm ðlog "Þ: 225 (4.58), 246 sh (4.26), 281 sh
KBr
max
(3.84), 289 (3.75). IR ꢂ
cmÀ1: 3394, 1732, 1693,
1
1620. H NMR: Table 4. 13C NMR: Table 5. Negative
ion SIMS m/z: 943 [M–H]À, 782, 764. HR-SIMS Found
943.3330 [M–H]À; C45H55N2O20 requires 943.3350.
COSY: H-6/H-700. ROESY: H-3/H-5; H-21/H-10; H-9/
H-6; H-9/H2-600; H-6/H2-600; H-100/H-1000. HMBC: H-14
(ꢀ 1.98) to C-3, 20; H-14 (ꢀ 2.15) to C-15; H2-14 to C-16;
H-15 to C-3, 14, 16, 20, 22; H-17 to C-15, 16, 21, 22;
OMe (ꢀ 3.71) to C-22; H-21 to C-10; H-10 to C-21; H-5
to C-3, 6, 700; OMe (ꢀ 3.84) to C-23; H-300 to C-100, 400, 500,
3.11. Preparation of strictosamide tetraacetate (16)
A solution of strictosidine (8) (4.5 mg) in 10% aq
Na2CO3 was warmed at 70 ꢀC for 1 h. The reaction
mixture was purified by Sephadex LH-20 CC (MeOH)
and prep. TLC (CHCl3–MeOH, 4:1) to give strictosa-
mide (15) (1.5 mg). Spectral data of 15 were identical
with those of an authentic sample (1H NMR, UV,
26
MeOH
max
SIMS, CD l
nm ðꢀ "Þ: 2 2 6À( 11.4), 270 (+13.5)).
1100; H-100 to C-1000; H-1000 to C-100. 20: ½ꢁŠD À137ꢀ (c 0.2,
MeOH
max
Strictosamide (15) (1.5 mg) was treated with Ac2O-pyr-
idine. The crude residue (1.7 mg) was purified by prep.
TLC (CHCl3–MeOH, 49:1) to give strictosamide tetra-
MeOH). UV l
nm ðlog "Þ: 224 (4.58), 243 sh
KBr
max
(4.27), 283 sh (3.81), 289 (3.73). IR ꢂ
cmÀ1: 3405,
1734, 1695, 1622. 1H NMR: Table 4. 13C NMR: Table 5.
Negative ion SIMS m/z: 957 [M–H]À, 795, 778. HR-
SIMS Found 957.3526 [M–H]À; C46H57N2O20 requires
957.3507. COSY: H-6/H-700. ROESY: H-3/H-5; H-3/H-
20; H-5/NMe; H-5/OMe (ꢀ 3.78); H-21/H-10; H-6/H-9;
H-9/H-700; H-500/H-700; H-100/H-1000. HMBC: H-17 to C-
15, 16, 21, 22; OMe (ꢀ 3.57) to C-22; H-21 to C-10; H-10
to C-21; NMe to C-3, 5; H-5 to C-6, 23, 700; OMe (ꢀ
3.78) to C-23; H-300 to C-400, 1100; H-100 to C-1000 H-1000 to
C-100.
1
acetate (16) (1.1 mg). H NMR (CDCl3): ꢀ 1.23, 1.88,
nm ðꢀ "Þ:
MeOH
max
1.99, 2.07 (12H each s, 4 Â Ac). CD l
217 (À8.9), 240 (À3.2), 269 (+14.2).
3.12. Preparation of neonaucleoside A (3) from
strictosidine (8)
A solution of strictosidine (8) (15.3 mg, 0.029 mmol)
and secologanin (12) (11.2mg, 0.029 mmol) in MeOH
(0.5 ml) was adjusted to pH 5-6 with 5% HCl and trea-
ted with NaBH3CN (5.5 mg). The reaction mixture was
stirred for 3 h at room temperature and purified by
Sephadex LH-20 CC and prep. TLC (CHCl3–MeOH, 4:1),
giving rise to 3 (11.9 mg, 45.8%) with 8 (2.7 mg, 17.6%).
Compound 3 was identified with 3 from natural source (1H
Acknowledgements
Our thanks go to Dr. M. Sugiura (Kobe Pharmaceu-
tical University) for NMR spectra and to Dr. K. Saiki
(Kobe Pharmaceutical University) for MS measure-
ments. This work was supported, in part, by the Science
Research Promotion Fund from the Promotion and
Mutual Aid Corporation for Private Schools of Japan.
20
NMR, UV, IR, SIMS, and ½ꢁŠD À154ꢀ (c 0.7, MeOH)).
3.13. Methylation of neonaucleoside B (4)
A solution of 4 (10.0 mg) in MeOH (1.0 ml) was
treated with 2.0 M TMSCHN2 in n-hexane (0.1 ml).
Concentrated reaction mixture was purified by prep.
TLC (CHCl3–MeOH–H2O, 70:30:3) to give 18 (7.4 mg).
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27
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