Three acylated glycosidic acid methyl esters and two acylated methyl glycosides generated from the convolvulin fraction of seeds of Quamoclit pennata by treatment with indium(III) chloride in methanol

Article abstract of DOI:10.1248/cpb.c13-00355

Treatment of the ether-insoluble resin glycoside (convolvulin) fraction from seedsof Quamoclit pennata (Convolvulaceae) with indium(III) chloride in methanol provided three oligoglycosides of hydroxy fatty acid (glycosidic acid) methyl esters and two methyl glycosides, which were partially acylated by a glycosidic acid, 75-hydroxydecanoic acid 7-0-β-d-quinovopyranoside (quamoclinic acid B) and/or two organic acids, (E)-2-methylbut-2-enoic (tiglic) acid and/or 3R-hydroxy-2R-methylbutyric (nilic) acid. Theirstructures were elucidated on the basis of spectroscopic data and chemical conversions.

Full text of DOI:10.1248/cpb.c13-00355

952
Regular Article
Chem. Pharm. Bull. 61(9) 952–961 (2013)
Vol. 61, No. 9
Three Acylated Glycosidic Acid Methyl Esters and Two Acylated
Methyl Glycosides Generated from the Convolvulin Fraction of Seeds of
Quamoclit pennata by Treatment with Indium(III) Chloride in Methanol
Kousuke Akiyama,a Tomoko Mineno,b Masafumi Okawa,c Junei Kinjo,c Hiroyuki Miyashita,d
,a
Hitoshi Yoshimitsu,d Toshihiro Nohara,d and Masateru Ono*
a School of Agriculture, Tokai University; 5435 Minamiaso, Aso, Kumamoto 869–1404, Japan: b Faculty of
Pharmacy, Takasaki University of Health and Welfare; 60 Nakaorui, Takasaki, Gunma 370–0033, Japan: Faculty
c
of Pharmaceutical Sciences, Fukuoka University; 8–19–1 Nanakuma, Jonan-ku, Fukuoka 814–0180, Japan: and
d Faculty of Pharmaceutical Sciences, Sojo University; 4–22–2 Ikeda, Nishi-ku, Kumamoto 860–0082, Japan.
Received May 8, 2013; accepted June 8, 2013
Treatment of the ether-insoluble resin glycoside (convolvulin) fraction from seeds of Quamoclit pen-
nata (Convolvulaceae) with indium(III) chloride in methanol provided three oligoglycosides of hydroxy fatty
acid (glycosidic acid) methyl esters and two methyl glycosides, which were partially acylated by a glycosidic
acid, 7S-hydroxydecanoic acid 7-O-β-D-quinovopyranoside (quamoclinic acid B) and/or two organic acids,
(E)-2-methylbut-2-enoic (tiglic) acid and/or 3R-hydroxy-2R-methylbutyric (nilic) acid. Their structures were
elucidated on the basis of spectroscopic data and chemical conversions.
Key words resin glycoside; convolvulin; Quamoclit pennata; acylated glycosidic acid methyl ester; acylated
methyl glycoside; indium(III) chloride
The so-called resin glycosides, which are commonly found acids, upon alkaline hydrolysis of the crude jalapin fraction
in plants of the Convolvulaceae family, are well known as pur- of the plant seeds.18) We also isolated six genuine jalapins,
gatives, and are characteristic ingredients of crude drugs such quamoclins I, II, III, IV, V, and VI from the fraction.18,19)
as Pharbitis Semen (the seed of Pharbitis nil CHOISY), Mexi- In the case of convolvulins from the seeds, we recently re-
can Scammoniae Radix (the root of Convolvulus scammonia ported the structures of seven glycosidic acids, quamoclinic
L.), Orizabae Tuber (the tuber of Ipomoea orizabensis (PEL- acids B, C, D, E, F, G, and H, and six organic acids, isobu-
LET) LEDANOIS), Jalapae Tuber (the tuber of I. purga HAYNE), tyric, 2S-methylbutyric, (E)-2-methylbut-2-enoic (tiglic), 3R-
and Rhizoma Jalapae Braziliensis (the root of I. operculata hydroxy-2R-methylbutyric (2R,3R-nilic), 7S-hydroxydecanoic,
(GOMES) MART.).1) They can be roughly divided into an ether- and 7S-hydroxydodecanoic acids, which were obtained by
soluble resin glycoside called jalapin, and an ether-insoluble alkaline hydrolysis of the crude convolvulin fraction of the
one called convolvulin.2) In 1987, Noda et al. succeeded in the seeds.14,15) As part of an ongoing study of the resin glycosides
isolation and structural elucidation of four jalapins.3) Almost from these seeds, this paper reports the isolation and structur-
all jalapins hitherto isolated and characterized had common al elucidation of three acylated glycosidic acid methyl esters
intramolecular macrocyclic ester structures composed of an and two acylated methyl glycosides, which were generated by
oligoglycoside of a hydroxy fatty acid partially acylated by treatment of the crude convolvulin fraction with indium(III)
some organic acids at the sugar moiety (acylated glycosidic chloride in MeOH.
acid); some examples were ester-type dimers.4–9) On the other
hand, convolvulin is regarded as an oligomer of various of
Results and Discussion
Despite numerous attempts, the isolation of pure resin gly-
acylated glycosidic acids.10) However, no pure convolvulin has
thus far been isolated, and chemical studies had been confined cosides from the crude convolvulin fraction19) of the seeds of
only to characterization of the component organic acids and Q. pennata has been unsuccessful. This fraction was there-
glycosidic acids afforded by alkaline hydrolysis of mixtures fore treated with indium(III) chloride in MeOH, as in the
of convolvulins.11–15) We recently reported the isolation and case of pharbitin.16) The treated fraction exhibited a number
structural elucidation of seven acylated glycosidic acid methyl of separate spots in TLC on silica gel. This treated fraction
ester monomers that were generated by treatment of the crude was successively subjected to Diaion HP20, Sephadex LH-20,
convolvulin fraction (named pharbitin) from Pharbitis Semen silica gel, and Chromatorex ODS column chromatography, and
using indium(III) chloride in methanol (MeOH),16) which was HPLC on octadecyl silica (ODS), to afford five compounds,
reported by Mineno and Kansui17) to be a catalyst for mild temporarily referred to as QM-1 (1)–QM-5 (5).
methyl esterification of carboxylic acids.
QM-1 (1) was obtained as an amorphous powder and
Quamoclit pennata BOJER is a Convolvulaceae plant na- exhibited a [M−H]− ion peak at m/z 1509 in negative-ion
tive to tropical regions of South America, and is primarily FAB-MS and a [M+Na]+ ion peak at m/z 1533 in positive-
cultivated as an ornamental plant. We previously reported the ion FAB-MS. High-resolution (HR)-positive-ion FAB-MS
isolation and structural elucidation of a glycosidic acid called indicated the molecular formula of 1 to be C67H114O37. On
quamoclinic acid A, which was obtained along with three or- alkaline hydrolysis, 1 furnished an organic acid fraction and
ganic acids, 2S-methylbutyric, n-decanoic, and n-dodecanoic a glycosidic acid. GC of the former revealed the presence of
nilic acid and tiglic acid. The latter was identified as qua-
1
moclinic acid F (6) by comparison of the H-NMR spectrum
The authors declare no conflict of interest.
© 2013 The Pharmaceutical Society of Japan
*To whom correspondence should be addressed. e-mail: mono@agri.u-tokai.ac.jp

September 2013
953
1
with that of an authentic sample.14) The H-NMR spectrum of OH-2 of Rha of 1. In the HMBC spectrum of 2, cross-peaks
1 showed signals due to H-2 [δ 2.90 (1H, dq, J=7.0, 7.0Hz)] were observed between H-4 of Qui or H-4 of Qui′ and C-1 of
of the first niloyl unit (Nla), H-3 [δ 7.04 (1H, brq, J=7.0Hz)] Nla, H-4 of Qui′ or H-4 of Qui and C-1 of Tig, and methoxy
of the tigloyl unit (Tig), one methoxy group [δ 3.63 (3H, s)], protons and C-1 of Agl (Fig. 1). However, the counterparts of
one primary methyl group [δ 0.90 (3H, t, J=7.0Hz)], two non- C-1 of Nla and C-1 of Tig could not be identified because the
equivalent methylene protons [δ 2.72 (1H, dd, J=6.5, 15.0Hz), 1H-NMR signals due to H-4 of Qui and H-4 of Qui′ appeared
2.69 (1H, dd, J=4.5, 15.0Hz)] adjacent to a carbonyl group, at almost the same chemical shifts. Although no cross-peak
seven anomeric protons [δ 6.26 (1H, s), 6.20 (1H, d, J=7.5Hz), between H-2 of Rha and C-1 of QaB was observed in the
6.03 (1H, s), 5.92 (1H, d, J=8.0Hz), 5.61 (1H, d, J=7.5Hz), HMBC spectrum of 2, the above data suggested that QaB was
5.06 (1H, d, J=8.0Hz), 4.80 (1H, d, J=7.0Hz)], eight sec- attached to OH-2 of Rha.
ondary methyl groups [δ 1.88 (3H, d, J=6.5Hz), 1.73 (3H,
To determine the sites of each ester linkage of Nla and
d, J=7.0Hz), 1.54 (3H, d, J=6.0Hz), 1.53 (3H, d, J=6.0Hz), Tig, partial deacylation of 2 was conducted. Compound 2
1.45 (3H, d, J=6.0Hz), 1.42 (3H, d, J=6.5Hz), 1.39 (3H, d, was refluxed with 5% triethylamine–MeOH for 1h, and the
J=6.5Hz), 1.33 (3H, d, J=7.0Hz)], and one tertiary methyl products were purified by HPLC to give 8. The positive-ion
group [δ 1.97 (3H, br s)] (Table 1). The 13C-NMR spectrum of FAB-MS of 8 showed a [M+Na]+ ion peak at m/z 1749, which
1 yielded signals due to 67 carbons, including seven anomeric was 100 mass units (niloyl unit) smaller than that of 2. In the
carbons (δ 106.1, 102.5, 102.3, 102.2, 102.1, 101.6, 101.0), two 1H-NMR spectrum of 8, the signal due to H-4 of Qui was
olefinic carbons (δ 138.0, 128.9), and three carboxyl carbons (δ shifted upfield (1.66ppm) compared with that of 2, and the
1
175.4, 172.9, 167.1) (Table 2). These H- and 13C-NMR signals signals due to the nilic acid unit disappeared; however, the
1
were assigned on the basis of H–1H correlation spectroscopy signals due to H-4 of Qui′ and H-2 of Rha were observed at
(COSY), heteronuclear multiple-quantum coherence (HMQC), quite similar chemical shifts to those of 2 (Table 1). Also, a
heteronuclear multiple-bond correlation (HMBC), and 1D- key correlation was observed between H-4 of Qui′ and C-1
total correlation spectroscopy (TOCSY) spectra, and indicated of Tig in the HMBC spectrum of 8. These data suggested
that 1 contains 1mol each of nilic acid, tiglic acid, and 6. A that Nla and Tig of 2 were located at OH-4 of Qui and OH-4
1
comparison of the chemical shifts of the H-NMR signals14) of Qui′, respectively. Accordingly, the structures of 2 and 8
due to the sugar moieties between 1 and 6 showed that the were respectively defined as methyl 3S,11S-dihydroxytetra-
signals due to H-4 of the first quinovosyl unit (Qui) and H-4 decanoate 11-O-α-L-rhamnopyranosyl-(1→3)-O-(4-O-tigloyl)-
of the second quinovosyl unit (Qui′) of 1 showed significant β-D-quinovopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→
downfield shifts of 1.58ppm and 1.67ppm, respectively, as a 3)-[O-(4-O-2R,3R-niloyl)-β-D-quinovopyranosyl-(1→4)]-O-
result of acylation. These data indicated that the ester link- (2-O-7S-hydroxydecanoyl
7-O-β-D-quinovopyranoside)-α-L-
ages were located at OH-4 of Qui and OH-4 of Qui′. The sites rhamnopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→2)-β-D-
of each ester linkage of Nla and Tig were determined from fucopyranoside and its deacyl derivative, in which Nla of 2
the HMBC spectrum of 1, with key cross-peaks observed was cleaved (Fig. 2).
between H-4 of Qui and C-1 of Nla, H-4 of Qui′ and C-1
QM-3 (3) was obtained as an amorphous powder, and
of Tig, and methoxy protons and C-1 of the aglycone moi- HR-positive-ion FAB-MS showed that its molecular for-
ety (Agl) (Fig. 1). Therefore, Nla and Tig were attached to mula was found to be the same as that of 2. On alkaline
OH-4 of Qui and OH-4 of Qui′, respectively. The configura- hydrolysis, 3 furnished nilic acid, tiglic acid, 7, and quamo-
1
tion of the nilic acid unit of this crude convolvulin fraction clinic acid E (9).14) The H- and 13C-NMR spectra of 3 had
was previously determined to be 2R,3R.14) Consequently, the signals due to one each of methoxy group, Nla, Tig, QaB,
structure of 1 was defined as methyl 3S,11S-dihydroxytetra- and quamoclinic acid E unit (Tables 1, 2). Comparison of
1
decanoate 11-O-α-L-rhamnopyranosyl-(1→3)-O-(4-O-tigloyl)- the H-NMR signals14) due to the sugar moiety in 3 and 9
β-D-quinovopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→ indicated acylation shifts [Δδ=δ(3)–δ(9)] of the signals due
3)-[O-(4-O-2R,3R-niloyl)-β-D-quinovopyranosyl-(1→4)]-O-α- to H-2 (Δδ=1.11) of Rha, H-4 (Δδ=1.64) of the second fu-
L-rhamnopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→2)-β-D- cosyl unit (Fuc′), and H-4 (Δδ=1.79) of Qui′. In addition,
fucopyranoside (Fig. 2).
the HMBC spectrum of 3 showed cross-peaks between H-4
QM-2 (2) was obtained as an amorphous powder, and af- of Fuc′ and C-1 of Nla, H-4 of Qui′ and C-1 of Tig, and the
forded tiglic acid, nilic acid, 6, and quamoclinic acid B (7)14) methoxy protons and C-1 of Agl (Fig. 1). Accordingly, the
on alkaline hydrolysis. Its positive-ion FAB-MS indicated a structure of 3 was assigned as methyl 3S,11S-dihydroxytetra-
[M+Na]+ ion peak at m/z 1849, which was 316 mass units decanoate 11-O-β-D-fucopyranosyl-(1→3)-O-(4-O-tigloyl)-β-
[quamoclinic acid B unit (QaB)] lager than that of 1. The D-quinovopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→3)-
molecular formula of 2 was found to be C83H142O43 by HR- [O-(4-O-2R,3R-niloyl)-β-D-fucopyranosyl-(1→4)]-O-(2-O-
positive-ion FAB-MS. The 1H- and 13C-NMR spectra of 2 7S-hydroxydecanoyl 7-O-β-D-quinovopyranoside)-α-L-rham-
were analogous to those of 1, except for the appearance of nopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→2)-β-D-fuco-
signals due to one QaB (Tables 1, 2). Thus, 2 is composed of pyranoside (Fig. 2).
1
1mol each of nilic acid, tiglic acid, 6, and 7. The H-NMR
QM-4 (4) was obtained as an amorphous powder. Nega-
signals due to the sugar moiety of 2 were compared with those tive-ion FAB-MS and positive-ion FAB-MS of 4 showed a
of 1, and the signal due to H-2 of the rhamnosyl unit (Rha) of [M−H]− ion peak at m/z 1139 and a [M+Na]+ ion peak at m/z
2 showed a downfield shift of 1.15ppm. On the other hand, the 1163, respectively, indicating its molecular weight to be 1140.
signals due to H-4 of Qui and H-4 of Qui′ of 2 were observed The molecular formula of 4 was shown by HR-positive-ion
1
at similar chemical shifts to those of 1. From these data, 2 was FAB-MS to be C47H80O31. The H-NMR spectrum of 4 showed
assumed to be a derivative of 1, in which 7 was attached to signals due to two H-2 of niloyl units, one methoxy group,

954
Vol. 61, No. 9
Table 1. 1H-NMR Spectral Data for 1–3 and 8 (in Pyridine-d5, 500MHz)
Position
1
2
8
3
Fuc-1
4.80 d (7.0)
4.47a)
4.82 d (7.0)
4.85 d (7.5)
4.79 d (7.5)
2
4.48 dd (7.0, 8.5)
4.51 dd (3.5, 8.5)
4.10 brd (3.5)
3.96a)
4.51 dd (7.5, 8.5)
4.58a)
4.14a)
4.04a)
4.47 dd (7.5, 9.0)
4.50 dd (3.0, 9.0)
4.08 dd (3.0)
3.95a)
3
4.48a)
4
4.06 brs
3.92a)
5
6
1.39 d (6.5)
1.37 d (6.5)
1.41 d (6.5)
1.44 d (6.5)
Fuc′-1
5.86 d (8.0)
2
4.20 dd (8.0, 9.0)
4.36 dd (3.0, 9.0)
5.62 d (3.0)
3
4
5
4.34a)
6
1.48 d (6.5)
Fuc″-1
4.93 d (7.5)
2
4.20 dd (7.5,9.0)
4.02 dd (3.0, 9.0)
3.95 d (3.0)
3
4
5
3.77a)
6
1.44 d (6.5)
Glc-1
5.61 d (7.5)
5.59 d (8.0)
5.67 d (7.5)
4.21 dd (7.5, 9.0)
4.11 dd (9.0, 9.0)
4.02a)
5.59 d (8.0)
2
4.18 dd (7.5, 9.0)
4.13 dd (9.0, 9.0)
4.04 dd (9.0, 9.0)
3.60 m
4.18 dd (8.0, 9.0)
4.10 dd (9.0, 9.0)
4.01 dd (9.0, 9.0)
3.55 ddd (3.0, 5.5, 9.0)
4.28 dd (3.0, 11.5)
4.16 dd (5.5, 11.5)
6.14 d (7.5)
4.18a)
3
4.09 dd (9.0, 9.0)
4.02 dd (9.0, 9.0)
3.56 ddd (3.0, 5.0, 9.0)
4.29 dd (3.0, 11.0)
4.18a)
4
5
3.58 ddd (3.0, 5.0, 9.0)
4.29 dd (3.0, 11.5)
4.20a)
6
4.28 dd (3.5, 11.5)
4.19 dd (5.0, 11.5)
6.20 d (7.5)
6
Glc′-1
6.26 d (8.0)
3.97a)
6.13 d (7.5)
3.95a)
2
3.94 dd (7.5, 9.0)
4.43 dd (9.0, 9.0)
4.10 dd (9.0, 9.0)
4.01 m
3.96 dd (7.5, 9.0)
4.38 dd (9.0, 9.0)
4.01 dd (9.0, 9.0)
3.96 m
3
4.43 dd (9.0, 9.0)
3.99a)
4.04a)
4.36 dd (9.0, 9.0)
4.01 dd (9.0, 9.0)
3.95a)
4
5
6
4.48 dd (2.5, 11.5)
4.17 dd (6.0, 11.5)
6.26 s
4.47 brd (11.0)
4.12 dd (5.5, 11.0)
6.29 d (1.5)
4.51 dd (1.5, 12.0)
4.14a)
4.44 dd (2.0, 12.0)
4.11 dd (5.5, 12.0)
6.27 d (1.0)
6
Rha-1
6.34 d (1.5)
6.00 dd (1.5, 3.5)
5.41 dd (3.5, 9.5)
4.59 dd (9.5, 9.5)
5.08 dq (9.5, 6.5)
1.94 d (6.5)
6.08 s
2
4.89 brs
6.04 dd (1.5, 3.0)
5.36 dd (3.0, 9.0)
4.53 dd (9.0, 9.0)
5.03 dq (9.0, 6.0)
1.88 d (6.0)
6.06 dd (1.0, 3.5)
5.36 dd (3.5, 9.5)
4.55 dd (9.5, 9.5)
5.03 dq (9.5, 6.0)
1.89 d (6.0)
3
5.18 dd (3.0, 9.0)
4.69 dd (9.0, 9.0)
4.97 dq (9.0, 6.5)
1.88 d (6.5)
4
5
6
Rha′-1
6.03 s
6.11 dd (1.5)
2
4.57 brs
4.58 dd (1.5, 3.0)
4.31 dd (3.0, 9.0)
4.18 dd (9.0, 9.0)
4.11 m
4.57a)
3
4.30 dd (2.5, 9.5)
4.18 dd (9.5, 9.5)
4.08 dq (9.5, 6.0)
1.54 d (6.0)
4.34 dd (3.0, 9.0)
4.19 dd (9.0, 9.0)
4.15a)
4
5
6
1.56 d (6.0)
1.58 d (6.0)
5.94 d (8.0)
3.98a)
Qui-1
5.92 d (8.0)
6.01 d (8.0)
2
3.98 dd (8.0, 9.5)
4.34 dd (9.5, 9.5)
5.34 dd (9.5, 9.5)
4.22 dq (9.5, 6.5)
1.53 d (6.5)
3.99 dd (8.0, 9.0)
4.40 dd (9.0, 9.0)
5.36 dd (9.0, 9.0)
4.35 m
3
4.25 dd (9.0, 9.0)
3.70 dd (9.0, 9.0)
4.15a)
4
5
6
1.63 d (6.0)
1.70 d (6.0)
5.12 d (8.0)
3.97a)
Qui′-1
5.06 d (8.0)
5.17 d (8.0)
5.22 d (8.0)
2
4.06 dd (8.0, 9.5)
4.16 dd (9.5, 9.5)
5.26 dd (9.5, 9.5)
3.85 dq (9.5, 6.0)
1.45 d (6.0)
4.15 dd (8.0, 9.5)
4.42 dd (9.5, 9.5)
5.36 dd (9.5, 9.5)
4.02 m
4.04 dd (8.0, 9.5)
4.30 dd (9.5, 9.5)
5.31 dd (9.5, 9.5)
4.02 dq (9.5, 6.0)
1.63 d (6.0)
3
4.39 dd (9.5, 9.5)
5.33 dd (9.5, 9.5)
4.04a)
4
5
6
Agl-2
2
1.64 d (6.5)
1.66 d (6.0)
2.73 dd (8.0, 14.5)
2.69 dd (5.0, 14.5)
4.41a)
2.72 dd (8.0, 14.5)
2.69 dd (4.5, 14.5)
4.41a)
2.73 dd (8.0, 15.0)
2.69 dd (5.0, 15.0)
4.40a)
2.73 dd (8.0, 15.0)
2.69 dd (5.0, 15.0)
4.40 m
3

September 2013
955
1
Table 11.. CHon-NtinMuRedSpectral Data for 1–3 and 8 (in Pyridine-d5, 500MHz)
Position
1
2
8
3
11
3.88a)
3.89a)
3.88 m
3.87a)
14
0.90 t (7.0)
3.63 s
0.90 t (7.5)
3.63 s
0.89 t (7.0)
3.63 s
0.93 t (7.0)
3.63 s
OCH3
Nla-2
2.90 dq (7.0, 7.0)
4.42a)
2.91 dq (7.0, 7.0)
4.35a)
2.79 dq (7.5, 7.5)
4.27a)
3
4
1.42 d (6.5)
1.33 d (7.0)
7.04 brq (7.0)
1.73 d (7.0)
1.97 brs
1.43 d (6.5)
1.39 d (6.5)
5
1.34 d (7.0)
1.30 d (7.5)
Tig-3
7.10 qq (1.5, 7.5)
1.76 d (7.5)
7.11 q (7.0)
1.77 d (7.0)
2.03 brs
7.13 qq (1.0, 7.5)
1.73 d (7.5)
4
5
2.01 brs
1.97 brs
QaB-2
2.50 ddd (8.0, 8.0, 16.0)
2.45 ddd (8.0, 8.0, 16.0)
3.90a)
2.52a)
2.51 ddd (7.5, 7.5, 16.0)
2.45 ddd (7.5, 7.5, 16.0)
3.89a)
2
2.49a)
7
3.91 m
10
0.96 t (7.5)
0.96 t (7.0)
4.80 d (7.5)
3.97a)
0.96 t (7.0)
Qui″-1
4.79 d (8.0)
4.79 d (7.5)
2
3
4
5
6
3.96 dd (8.0, 9.0)
4.14 dd (9.0, 9.0)
3.71 dd (9.0, 9.0)
3.78 dq (9.0, 6.0)
1.63 d (6.0)
3.95 dd (7.5, 9.0)
4.14 dd (9.0, 9.0)
3.71 dd (9.0, 9.0)
3.78 dq (9.0, 6.0)
1.63 d (6.0)
4.15a)
3.72 dd (9.0, 9.0)
3.79 dq (9.0, 6.5)
1.63 d (6.5)
δ in ppm from TMS. Coupling constants (J) in Hz are given in parentheses. a) Signals were overlapped with other signals.
1
six anomeric protons, and eight secondary methyl groups and
A comparison of the H-NMR spectrum of 4 with that of 10
its 13C-NMR spectrum exhibited signals due to six anomeric showed significant downfield shifts [Δδ=δ(4)–δ(10)] of the sig-
carbons and two carboxyl carbons (Tables 3, 4). However, nals due to H-4 (Δδ=1.66) of Qui and H-4 (Δδ=1.55) of Qui′
no signals attributable to a hydroxy fatty acid, which is the in 4. In addition, the HMBC spectrum of 4 showed key cross
aglycone unit of resin glycosides, were observed. From the peaks between H-4 of Qui and C-1 of Nla and H-4 of Qui′
assigned data, it was considered that 4 was a methyl hexagly- and C-1 of the second niloyl unit (Nla′) (Fig. 1). Accordingly,
coside esterified by two nilic acids. On alkaline hydrolysis, 4 the structure of 4 was assigned as methyl (4-O-2R,3R-niloyl)-
furnished nilic acid and a methyl glycoside (10), temporarily β-D-quinovopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→
referred to as methyl quamoside A. The negative-ion FAB-MS 3)-[O-(4-O-2R,3R-niloyl)-β-D-quinovopyranosyl-(1→4)]-O-α-
of 10 showed a [M−H]− ion peak at m/z 939 along with frag- L-rhamnopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→2)-α-D-
ment ion peaks at m/z 793 [939−146 (6-deoxyhexosyl unit)]−, fucopyranoside (Fig. 2).
631 [793−162 (hexosyl unit)]−, 485 [631−146]−, and 339
QM-5 (5) was obtained as an amorphous powder. Alkaline
[485−146]−. On acidic hydrolysis, 10 afforded monosaccharide hydrolysis of 5 gave nilic acid, 7, and 10. The positive-ion
fraction, which was converted into thiocarbamoyl-thiazolidine FAB-MS of 5 revealed a [M+Na]+ ion peak at m/z 1479,
derivatives and then analyzed using HPLC, according to the which was 316 mass units larger than that of 4. The molecular
process reported by Tanaka et al.20) Derivatives of D-glucose, formula of 5 was found to be C63H108O37 using HR-positive-
D-fucose, D-qinovose, and L-rhamnose were detected. The ion FAB-MS. The H- and 13C-NMR spectra of 5 were impos-
1
1H- and 13C-NMR signals of 10 were assigned with the aid able on those of 4, apart from the appearance of signals due
of 2D-NMR spectra (Tables 3, 4); the assigned data of the to one QaB (Tables 3, 4). A comparison of the chemical shifts
sugar moiety of 10 were quite similar to those of quamoclinic of the signals in 4 and 5 showed a significant downfield shift
acid C,14) except that the data for the β-D-fucopyranosyl unit (1.17ppm) of the signal assignable to H-2 of Rha of 5; in con-
attached to the aglycone of quamoclinic acid C were re- trast, the signals due to H-4 of Qui and H-4 of Qui′ resonated
placed by those for the α-D-fucopyranosyl unit. From these at similar chemical shifts to those of 4. These data suggest
data, the structure of 10 was considered to be methyl β-D- that 5 is a derivative of 4, in which 7 is attached to OH-2 of
quinovopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→3)-[O- Rha of 4. This inference was supported by the following evi-
β-D-quinovopyranosyl-(1→4)]-O-α-L-rhamnopyranosyl-(1→ dence. Partial deacylation of 5 in a similar manner to that per-
1
2)-O-β-D-glucopyranosyl-(1→2)-α-D-fucopyranoside (Fig. 2). formed on 2 gave 11. A comparison of the H-NMR signals of
This structure was confirmed by the following evidence. 5 and 11 showed two significant upfield shifts [Δδ=δ(5)–δ(11)]
The 13C-NMR spectrum of 10 showed, compared with those of the signals due to H-4 (Δδ=1.65) of Qui and H-4 (Δδ=
of methyl pyranosides in the literature,21,22) glycosylation 1.60) of Qui′ and the disappearance of the signals due to
shifts23,24) of the signals due to C-2 (+9.2ppm) of the first the two niloyl units in the spectrum of 11; the signal due to
fucosyl unit (Fuc), C-2 (+3.8ppm) of the first glucosyl unit H-2 of Rha in 11 resonated at a similar position to that in 5
(Glc), C-3 (+6.4ppm) and C-4 (+5.4ppm) of Rha, and C-2 (Tables 3, 4). The HMBC spectrum of 5 indicated key cor-
(+10.0ppm) of the second glucosyl unit (Glc′). In the HMBC relations between H-4 of Qui or H-4 of Qui′ and C-1 of Nla
spectrum of 10, key cross-peaks between H-1 of Glc′ and C-3 and H-4 of Qui′ or H-4 of Qui and C-1 of Nla′ (Fig. 1). Con-
of Rha, H-1 of Qui and C-4 of Rha, H-1 of Qui′ and C-2 of sequently, the structures of 5 and 11 were respectively identi-
Glc′, and methoxy protons and C-1 of Fuc, were observed.
fied as methyl (4-O-2R,3R-niloyl)-β-D-quinovopyranosyl-(1→

956
Vol. 61, No. 9
Table 2. 13C-NMR Spectral Data for 1–3 and 8 (in Pyridine-d5, 125MHz)
Position
1
2
8
3
1
2
8
3
Fuc-1
102.5
78.6
76.1
72.9
71.0
17.1
102.4
78.5
76.0
73.1
71.0
17.1
102.4
78.6
76.2
73.1
71.0
17.1
102.6
78.6
76.0
73.4
71.0
17.0
102.2
73.8
72.7
74.3
69.2
16.7
107.1
73.5
75.2
72.7
71.7
17.1
102.2
76.6
78.6
72.5
77.2
63.0
101.5
86.3
77.4
71.8
78.2
62.6
97.6
73.9
75.2
79.3
67.9
19.1
Qui-1
102.1
76.3
75.8
77.5
69.8
18.6
106.1
77.9
79.0
74.4
71.3
18.1
175.4
48.4
69.6
21.1
13.4
167.1
128.9
138.0
14.3
12.4
101.8
76.3
75.8
77.5
70.0
18.6
106.8
77.7
79.1
74.5
71.4
18.2
175.4
48.4
69.7
21.2
13.5
167.2
128.9
138.1
14.4
12.4
173.3
34.7
78.8
14.4
103.4
75.6
78.2
76.9
72.7
18.8
172.9
43.4
68.2
80.1
14.5
51.3
102.0
76.6
78.4
77.0
72.5
18.6
106.6
77.6
79.1
74.6
71.7
18.4
2
2
3
3
4
4
5
5
6
6
Fuc′-1
Qui′-1
106.1
77.8
84.8
74.6
71.5
18.3
174.9
50.3
69.7
20.8
14.2
167.3
129.0
137.6
14.2
12.5
173.4
34.7
78.6
14.4
103.4
75.6
78.2
76.9
72.7
18.7
172.9
43.4
68.2
80.2
14.5
51.2
2
2
3
3
4
4
5
5
6
6
Fuc″-1
Nla-1
2
2
3
3
4
4
5
5
6
Tig′-1
167.1
129.0
138.0
14.5
12.5
173.3
34.7
78.9
14.4
103.4
75.6
78.2
76.9
72.7
18.8
172.9
43.4
68.2
80.2
14.5
51.2
Glc-1
102.3
77.3
79.0
72.4
77.2
63.0
101.6
85.7
77.4
72.0
78.2
62.7
101.0
72.0
77.9
79.5
68.1
19.2
102.2
72.3
72.3
73.7
69.9
18.8
102.2
76.9
78.6
72.5
77.2
63.0
101.8
86.3
77.3
71.8
78.2
62.6
97.4
74.1
75.3
79.3
67.7
19.1
102.2
72.3
72.3
73.7
69.9
18.7
102.0
76.9
78.2
72.3
77.3
63.0
101.7
86.0
77.3
72.1
78.2
62.7
97.2
74.1
74.5
79.8
68.0
19.2
102.3
72.3
72.4
73.7
70.0
18.7
2
2
3
3
4
4
5
5
QaB-1
6
2
Glc′-1
7
2
10
3
Qui″-1
4
2
5
3
6
4
Rha-1
5
6
2
3
Agl-1
2
172.9
43.4
68.2
80.0
14.4
51.2
4
5
3
6
11
Rha′-1
14
2
3
4
5
6
OCH3
δ in ppm from TMS.
2)-O-β-D-glucopyranosyl-(1→3)-[O-(4-O-2R,3R-niloyl)-β-D- acylated saccharides with a reducing terminal during the
quinovopyranosyl-(1→4)]-O-(2-O-7S-hydroxydecanoyl 7-O- treatment with indium(III) chloride in MeOH. It is therefore
β-D-quinovopyranoside)-α-L-rhamnopyranosyl-(1→2)-O-β-D- presumed that one of the reasons for the difficulty in isolating
glucopyranosyl-(1→2)-α-D-fucopyranoside, and its deacyl de- convolvulin from the methanol extract of Q. pennata is the
rivative, in which two niloyl units of 5 were cleaved (Fig. 2).
co-existence of the acylated saccharides with a reducing ter-
Mannich and Schumann10) speculated that convolvulin minal, because α- and β-anomers easily reach equilibrium.25)
from I. purga was an oligomer of acylated glycosidic acid. Although QM-1–QM-5 were all considered to be artifacts
Although QM-2 and QM-3 were acylated by one quamoclinic formed during the treatment, they gave the new information of
acid B, QM-1, QM-2, and QM-3 were all regarded as methyl the structures of genuine convolvulins of Q. pennata.
ester monomers of acylated glycosidic acid. On the other
hand, both of QM-4 and QM-5 were acylated methyl glyco-
sides. Two acylated trisaccharides, which are closely related
Experimental
General Procedures Optical rotations were determined
resin glycosides, were previously reported as natural constitu- with a JASCO P-1020 polarimeter. The 1H- and 13C-NMR
ents of the seeds of Cuscuta chinensis.25) Therefore, QM-4 and spectra were recorded by using a JEOL ECA-500 spectrom-
QM-5 were considered to be formed from the corresponding eter, and chemical shifts are given on a δ (ppm) scale with

September 2013
957
Fig. 1. 1H–13C Long-Range Correlations Observed in the HMBC Spectra of 1–5 (in Pyridine-d5, 500MHz)
tetramethylsilane (TMS) as an internal standard. MS data chromatograph with
a
flame-ionization detector. Colum
were collected using a JEOL JMS-700 mass spectrometer. chromatography (CC) was carried out over Diaion HP20
Analytical GC was carried out with a Shimadzu GC-8A gas (Mitsubishi Chemical Industries, Japan), Sephadex LH-20

958
Vol. 61, No. 9
Fig. 2. Structures of 1–5
(Pharmacia Fine Chemicals), silica gel 60 (Merck, Art. No. of mixtures of MeOH–H2O (60% MeOH, 70% MeOH, 80%
1.09385), and Chromatorex ODS (Fuji Silysia Chemical, Ltd., MeOH, 85% MeOH, 90% MeOH, 100% MeOH) as eluents
Japan). HPLC separation was performed on a Shimadzu LC- to give fractions 2.10.1–2.10.18. HPLC (column 1) of fractions
10AS micro pump with a Shimadzu RID-10A RI detector. 2.10.11 (140mg) and 2.10.14 (355mg), using 85% MeOH as
For HPLC column chromatography, COSMOSIL 5C18-AR-II eluent, afforded 1 (25mg) from fraction 2.10.11 and 2 (131mg)
(Nacalai Tesque, Inc., Japan, 20mm i.d.×250mm, column and 3 (73mg) from fraction 2.10.14. Fraction 2.11 (2666mg)
1) and COSMOSIL 5C18-AR (Nacalai Tesque, Inc., 6.0mm was chromatographed on a Chromatorex ODS column using
i.d.×250mm, column 2) were used.
a gradient of mixtures of MeOH–H2O (60% MeOH, 65%
Plant Material The seeds of Quamoclit pennata BOJER MeOH, 70% MeOH, 75% MeOH, 80% MeOH, 85% MeOH,
were purchased from Heiwaen, a gardening shop in Nara 90% MeOH, 95% MeOH, 100% MeOH) as eluents to give
Prefecture, Japan, and were identified by Prof. Kazumoto fractions 2.11.1–2.11.38. Fractions 2.11.4 (40mg) and 2.11.16
Miyahara (Faculty of Pharmaceutical Sciences, Setsunan Uni- (108mg) were each subjected to HPLC (column 1) using 60%
versity). A voucher specimen (QP1990) has been deposited at MeOH for fraction 2.11.4 and 75% MeOH for fraction 2.11.16
the laboratory of Natural Products Chemistry, School of Agri- as eluents to give 4 (20mg) from fraction 2.11.4 and 5 (37mg)
culture, Tokai University.
from fraction 2.11.16.
Treatment of the Convolvulin Fraction with Indium(III)
QM-1 (1): Amorphous powder. [α]D26 −49.7° (c=0.8, MeOH).
Chloride in MeOH and Isolation of 1–5 The crude covol- Positive-ion FAB-MS m/z: 1533 [M+Na]+. HR-positive-ion
vulin fraction (15032mg) previously obtained18) from the seeds FAB-MS m/z: 1533.6923 (Calcd for C67H114O37Na+, 1533.6931).
of Quamoclit pennata was dissolved in MeOH (300mL), and Negative-ion FAB-MS m/z: 1509 [M−H]−. H-NMR spectral
1
indium(III) chloride (7500mg) was added to the solution at data: see Table 1. 13C-NMR spectral data: see Table 2.
the room temperature. The mixture was heated at reflux for
QM-2 (2): Amorphous powder. [α]D19 −48.1° (c=1.6, MeOH).
27d, while being monitored by TLC. The concentrated reac- Positive-ion FAB-MS m/z: 1849 [M+Na]+. HR-positive-ion
tion mixture was chromatographed on a Diaion HP20 column, FAB-MS m/z: 1849.8843 (Calcd for C83H142O43Na+, 1849.8817).
eluted with H2O and MeOH. The MeOH eluate (11162mg) was 1H-NMR spectral data: see Table 1. 13C-NMR spectral data:
subjected to Sephadex LH-20 CC eluted with MeOH to give see Table 2.
fractions 1 (1630mg) and 2 (8113mg). CC of fraction 2 on sil-
QM-3 (3): Amorphous powder. [α]D26 −34.1° (c=0.7, MeOH).
ica gel eluted with a gradient of mixtures of CHCl3–MeOH– Positive-ion FAB-MS m/z: 1849 [M+Na]+. HR-positive-ion
H2O (14:2:0.1, 10:2:0.1, 8:2:0.2, 7:3:0.5, 6:4:1, 0:1:0) FAB-MS m/z: 1849.8816 (Calcd for C83H142O43Na+, 1849.8817).
afforded fractions 2.1–2.15. Fraction 2.10 (2142mg) was chro- Negative-ion FAB-MS m/z: 1825 [M−H]−, 333 [quamoclinic
matographed on a Chromatorex ODS column using a gradient acid B−H]−. H-NMR spectral data: see Table 1. 13C-NMR
1

September 2013
959
Table 3. 1H-NMR Spectral Data for 4, 5, 10, and 11 (in Pyridine-d5, 500MHz)
Position
4
10
5.30 d (3.5)
5
11
5.30 d (3.5)
Fuc-1
5.31 d (3.0)
4.48 dd (3.0, 10.0)
4.56 dd (3.0, 10.0)
4.05 d (3.0)
4.02 m
5.30 d (3.0)
2
4.50 dd (3.5, 10.5)
4.57 dd (3.5, 10.5)
4.07a)
4.46 dd (3.0, 9,5)
4.59 dd (3.0, 9.5)
4.07 d (3.0)
4.48 dd (3.5, 10.5)
4.60 dd (3.5, 10.5)
4.09 d (3.5)
4.00a)
3
4
5
4.00a)
4.03a)
6
1.40 d (7.0)
4.96b)
1.40 d (6.5)
5.01 d (7.0)
4.11a)
1.36 d (7.0)
1.38 d (6.5)
4.96a)
Glc-1
4.93 d (7.5)
2
4.08a)
4.16 dd (7.5, 9.0)
4.04a)
4.04a)
4.18a)
3
4.08a)
4.08a)
4.08a)
4.06a)
4.06a)
4
4.08a)
5
3.61 m
3.62 m
3.56 m
3.59 m
6
4.35 dd (4.0, 12.0)
4.26 dd (5.0, 12.0)
5.97 d (8.0)
3.92 dd (8.0, 9.0)
4.38 dd (9.0, 9.0)
4.13 dd (9.0, 9.0)
3.87 m
4.36a)
4.33 dd (2.5, 11.5)
4.24 dd (5.0, 11.5)
5.89 d (8.0)
4.35a)
6
4.26 dd (5.0, 11.5)
5.90 d (8.0)
3.97 dd (8.0, 9.5)
4.37a)
4.26 dd (5.0, 11.5)
5.90 d (8.0)
3.97 dd (8.0, 9.5)
4.34a)
Glc′-1
2
3.97 dd (8.0, 8.5)
4.30a)
4.04a)
3
4
4.11a)
4.06a)
5
3.88 ddd (3.0, 5.5, 9.5)
4.39a)
3.80 m
3.86a)
6
4.36a)
4.13a)
4.32a)
4.37a)
6
4.14 dd (5.5, 12.0)
6.20 d (1.5)
4.94 dd (1.5, 3.0)
5.17 dd (3.0, 9.0)
4.70 dd (9.0, 9.0)
4.95 dq (9.0, 6.0)
1.86 d (6.0)
5.78 d (8.0)
3.95 dd (8.0, 9.0)
4.31 dd (9.0, 9.0)
3.68 dd (9.0, 9.0)
4.04a)
4.06a)
4.09a)
Rha-1
6.17 s
6.16 s
6.19 d (2.0)
6.09 dd (2.0, 3.0)
5.40 dd (3.0, 9.0)
4.59 dd (9.0, 9.0)
5.01 dq (9.0, 6.5)
1.88 d (6.5)
5.94 d (7.5)
4.01 dd (7.5, 9.0)
4.34a)
2
4.92 d (3.0)
5.16 dd (3.0, 9.0)
4.67 dd (9.0, 9.0)
4.90 dq (9.0, 6.0)
1.81 d (6.0)
5.92 d (7.5)
3.97 dd (7.5, 9.0)
4.45 dd (9.0, 9.0)
5.34 dd (9.0, 9.0)
4.29 m
6.11 d (3.5)
3
5.36 dd (3.5, 9.0)
4.51 dd (9.0, 9.0)
5.00 dq (9.0, 6.0)
1.82 d (6.0)
4
5
6
Qui-1
5.99 d (7.5)
2
3.97 dd (7.5, 9.0)
4.50 dd (9.0, 9.0)
5.37 dd (9.0, 9.0)
4.43 m
3
4
3.72 dd (9.0, 9.0)
4.18a)
5
6
1.53 d (6.5)
5.13 d (8.0)
4.19 dd (8.0, 9.0)
4.04 dd (9.0, 9.0)
5.29 dd (9.0, 9.0)
3.83 dq (9.0, 6.5)
1.54 d (6.5)
2.99 dq (7.0, 7.0)
4.36a)
1.56 d (6.5)
5.13 d (8.0)
4.17 dd (8.0, 9.0)
4.01a)
1.63 d (6.0)
1.64 d (6.5)
5.16 d (7.5)
4.17a)
Qui′-1
5.24 d (7.5)
2
4.26 dd (7.5, 9.0)
4.21 dd (9.0, 9.0)
5.37 dd (9.0, 9.0)
3.97 m
3
4.12 dd (8.5, 8.5)
3.77a)
3.85a)
4
3.74a)
5
3.74a)
1.67a)
6
1.66 d (6.5)
1.77 d (6.5)
Nla-2
3.02 dq (7.0, 7.0)
4.38 dq (7.0, 7.0)
1.38 d (7.0)
3
4
1.36 d (6.5)
1.31 d (7.0)
2.82 dq (7.0, 7.0)
4.30a)
5
1.32 d (7.0)
Nla′-2
2.87 dq (7.0, 7.0)
4.33a)
3
4
1.39 d (6.5)
1.28 d (7.0)
1.41 d (6.5)
5
1.30 d (7.0)
QaB-2
2.47 ddd (7.5, 7.5, 15.5)
2.38 ddd (7.5, 7.5, 15.5)
3.88 m
2.54 ddd (7.0, 8.5, 16.0)
2.40 ddd (6.5, 8.0, 16.0)
3.87a)
2
7
10
0.94 t (7.0)
0.93 t (7.0)
Qui″-1
4.79 d (8.0)
4.79 d (8.0)
2
3.96 dd (8.0, 9.0)
4.15 dd (9.0, 9.0)
3.71 dd (9.0, 9.0)
3.78 dq (9.0, 6.0)
1.62 d (6.0)
3.96 dd (8.0, 9.0)
4.15 dd (9.0, 9.0)
3.71 dd (9.0, 9.0)
3.78a)
3
4
5
6
1.62 d (6.0)
OCH3
3.29 s
3.28 s
3.29 s
3.28 s
δ in ppm from TMS. Coupling constants (J) in Hz are given in parentheses. a) Signals were overlapped with other signals. b) Signals were deformed by virtual coupling.

960
Vol. 61, No. 9
Table 4. 13C-NMR Spectral Data for 4, 5, 10, and 11 (in Pyridine-d5, 125MHz)
Position
4
10
5
11
4
10
5
11
Fuc-1
100.9
79.4
70.0
73.0
66.5
16.9
105.1
78.3
78.5
71.6
78.0
62.5
101.7
85.8
77.5
71.4
78.0
62.5
101.4
71.7
79.0
78.8
68.4
18.8
102.2
76.2
75.6
77.7
69.9
18.2
100.9
79.2
70.1
73.0
66.5
16.9
105.1
78.2
78.6
71.6
77.9
62.6
101.4
84.8
77.5
71.4
78.2
62.5
101.4
71.6
79.0
78.9
68.7
18.9
102.8
76.6
78.0
77.3
72.3
18.5
100.8
79.2
70.0
73.1
66.5
16.9
104.7
77.6
78.4
71.6
77.9
62.4
101.7
85.8
77.5
71.2
78.1
62.3
98.0
73.7
76.2
78.6
67.9
18.8
101.9
76.2
75.4
77.8
69.9
18.2
100.8
79.1
70.1
73.2
66.5
16.9
104.7
77.2
78.5
71.7
78.0
62.5
101.6
84.8
77.5
71.4
78.2
62.5
98.0
73.6
75.8
78.7
68.3
18.9
102.4
76.6
77.8
77.3
72.4
18.6
Qui′-1
106.2
76.3
75.3
76.6
71.2
18.2
175.4
48.4
69.7
20.9
13.2
175.0
48.5
69.5
21.0
13.3
105.5
75.9
78.0
76.5
73.8
18.6
106.4
76.2
75.3
76.4
71.0
18.3
175.4
48.3
69.6
20.8
13.2
175.1
48.4
69.4
20.9
13.3
173.5
34.7
78.4
14.3
103.3
75.5
78.1
76.9
72.7
18.6
55.1
105.8
76.0
77.9
76.5
74.0
18.7
2
2
3
3
4
4
5
5
6
6
Glc-1
Nla-1
2
2
3
3
4
4
5
5
6
Nla′-1
Glc′-1
2
2
3
3
4
4
5
5
QaB-1
173.7
34.7
78.8
14.4
103.4
75.6
78.2
76.9
72.7
18.7
55.1
6
2
Rha-1
7
2
10
3
Qui″-1
4
2
5
3
6
4
Qui-1
5
6
2
3
4
5
6
OCH3
55.1
55.1
δ in ppm from TMS.
spectral data: see Table 2.
CHP 20 column chromatography using H2O and acetone as
QM-4 (4): Amorphous powder. [α]D19 −26.5° (c=1.4, MeOH). eluents to give glycosidic acid (3mg) derived from 1 and gly-
Positive-ion FAB-MS m/z: 1163 [M+Na]+. HR-positive-ion cosidic acid fractions (7mg from 2, 7mg from 3) derived from
FAB-MS m/z: 1163.4586 (Calcd for C47H80O31Na+, 1163.4576). 2 and 3. The glycosidic acid derived from 1 was identical with
Negative-ion FAB-MS m/z: 1139 [M−H]−. H-NMR spectral 6 by comparison of H-NMR spectrum with that of authentic
1
1
data: see Table 3. 13C-NMR spectral data: see Table 4.
sample.14) The glycosidic acid fractions derived from 2 and
QM-5 (5): Amorphous powder. [α]D19 −25.6° (c=0.8, MeOH). 3 were each chromatographyied on silica gel column eluted
Positive-ion FAB-MS m/z: 1479 [M+Na]+. HR-positive-ion with a gradient of mixtures of CHCl3–MeOH–H2O (14:2:0.1,
FAB-MS m/z: 1479.6472 (Calcd for C63H108O37Na+, 1479.6462). 10:2:0.1, 8:2:0.2, 7:3:0.5, 6:4:1, 0:1:0) to afford 6 (4mg)
Negative-ion FAB-MS m/z: 1455 [M−H]−. H-NMR spectral and 7 (1mg) from the fraction derived from 2 and 7 (1mg)
1
data: see Table 3. 13C-NMR spectral data: see Table 4.
and 9 (4mg) from the fraction derived from 3, which were
1
Alkaline Hydrolysis of 1–5 Suspentions of 1 (4mg), 2 identified by comparison of H-NMR spectra with those of
(10mg), 3 (10mg), 4 (10mg), and 5 (5mg) in 1M KOH (1mL) authentic samples.14)
were heated at 95°C for 1h. The reaction mixture was adjust-
The aqueous layers of 4 and 5 were neutralized with 0.1M
ed to pH 4 with 1M HCl, then diluted with H2O (10mL), and NaOH. After removal of the solvent, the residues were sub-
extracted with ether (99.5%, Nacalai Tesque, Inc.; 3×5mL). jected to Sephadex LH-20 CC eluted with MeOH to give 10
The ether layer was dried over MgSO4 and evaporated to (7mg) from the residue derived from 4 and 7 (0.5mg) and 10
furnish an organic acid fraction, which was analyzed by GC (2mg) from the residue derived from 5. The generated com-
[Shimadzu GC-8A gas chromatograph with flame-ionaization pounds derived from 5 were each identified by comparison of
detector; column, Unisole 30T (5%), 3.2mm i.d.×2m glass 1H-NMR spectra with those of authentic samples.14)
column (column 3); carrier gas N2, 1.0kg/cm2; column tem-
Methyl Quamoside A (10): Amorphous powder. [α]D24 −18.5°
perature, 120°C; tR (min): 10.38 (tiglic acid) for 1–3]. A part of (c=1.3, MeOH). Positive-ion FAB-MS m/z: 963 [M+Na]+. HR-
the organic acid fraction was methylated with diazomethane- positive-ion FAB-MS m/z: 963.3569 (Calcd for C37H64O27Na+,
ether and then the reaction mixture was analyzed by GC [col- 963.3527). Negative-ion FAB-MS m/z: 939 [M−H]−, 793
umn, column 3; column temperature, 100°C; carrier gas, N2 [939−146]−, 631 [793−162]−, 485 [631−146]−, 339 [485−146]−.
1.2 kg/cm2; tR (min): 4.20 (methyl nilate) for 1–5].
1H-NMR spectral data: see Table 3. 13C-NMR spectral data:
The aqueous layers of 1–3 were each desalted over MCI gel see Table 4.

September 2013
961
Partial Deacylation of 2 and 5 Solutions of 2 (32mg)
and 5 (7mg) in 5% triethylamine–MeOH (1mL) were refluxed
for 1h and 2h, respectively. After removal of the solvent, the
residues were successively subjected to silica gel CC eluted
with a gradient of mixtures of CHCl3–MeOH–H2O (14:2:0.1,
10:2:0.1, 8:2:0.2, 7:3:0.5, 6:4:1, 0:1:0) and HPLC (col-
umn 2) using 85% MeOH for residue derived from 2 and 70%
MeOH for residue derived from 5 as eluents to give 8 (15mg)
from the reaction mixture of 2 and 11 (4mg) from the reaction
mixture of 5.
References
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Compound 8: Amorphous powder. [α]D25 −46.2° (c=1.2,
MeOH). Positive-ion FAB-MS m/z: 1749 [M+Na]+. HR-pos-
itive-ion FAB-MS m/z: 1749.8273 (Calcd for C78H134O41Na+,
1749.8293). Negative-ion FAB-MS m/z: 1725 [M−H]−, 333.
1H-NMR spectral data: see Table 1. 13C-NMR spectral data:
see Table 2.
Compound 11: Amorphous powder. [α]D25 −22.9° (c=0.5,
MeOH). Positive-ion FAB-MS m/z: 1279 [M+Na]+. HR-
positive-ion FAB-MS m/z: 1279.5419 (Calcd for C53H92O33Na+,
1279.5413). Negative-ion FAB-MS m/z: 1255 [M−H]−,
939 [1255−316]−, 793 [939−146]−, 631 [793−162]−, 485
1
[631−146]−, 339 [485−146]−, 333. H-NMR spectral data: see
Table 3. 13C-NMR spectral data: see Table 4.
Sugar Analysis of 7 Compound 7 (1.0mg) was heated in
1M HCl (0.2mL) at 95°C for 1h. The reaction mixture was
neutralized with Amberlite MB-3 (Organo Co.) and then evap-
orated under reduced pressure to give a monosaccharide frac-
tion. This fraction was dissolved in pyridine (0.2mL) contain-
ing L-cysteine methyl ester hydrochrolide (1.0mg) and heated
at 60°C for 1h. A solution (0.01mL) of o-tolylisothiocyanate
(0.1mL) in pyridine (1.0mL) was added to the mixture, which
was heated at 60°C for 1h. The reaction mixture was analyzed
by HPLC [detector, Shimadzu SPD-10A UV detector (250nm),
column, column 2; eluent 25% CH3CN in 50mM H3PO4; flow
ate 0.8mL/min; column temperature, 35°C; tR (min): 27.39
(D-glucose deriv.), 36.91 (D-fucose deriv.), 42.09 (D-quinovose
deriv.), 45.48 (L-rhamnose deriv.)].
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Acknowledgments We express our appreciation to Mr.
H. Harazono of Fukuoka University for the measurement of
the FAB-MS. The authors thank Prof. Kazumoto Miyahara
(Faculty of Pharmaceutical Sciences, Setsunan University) for
identifying the plant material.
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