Glochidionionosides A-D: Megastigmane glucosides from leaves of Glochidion zeylanicum (Gaertn.) A. Juss

Article abstract of DOI:10.1248/cpb.51.286

Five megastigmane glucosides were isolated from the leaves of Glochidion zeylanicum. One of them was a known compound, blumenol C O-β-D- glucopyranoside (1), and the structures of the four new compounds, glochidionionosides A-D (2-5), were mainly elucidated by spectroscopic methods, including a modified Mosher's method. The absolute configurations of the six-membered ring of glochidionionoside D (5) were deduced by β-D-glucopyranosylation-induced shift trends in the ₁₃C-NMR spectra and confirmed by X-ray analysis as its pbromobenzoate (5b), and the axis chirality of C-7 was determined to be R.

Full text of DOI:10.1248/cpb.51.286

286
Chem. Pharm. Bull. 51(3) 286—290 (2003)
Vol. 51, No. 3
Glochidionionosides A—D: Megastigmane Glucosides from Leaves of
Glochidion zeylanicum (GAERTN.) A. JUSS
Hideaki OTSUKA, ,a Hidehiko KIJIMA,a Eiji HIRATA,b Takakazu SHINZATO,c Anki TAKUSHI,d
*
f
Masahiko BANDO,e and Yoshio TAKEDA
a Department of Pharmacognosy, Graduate School of Biomedical Sciences, Hiroshima University; 1–2–3 Kasumi, Minami-
ku, Hiroshima 734–8551, Japan: b Faculty of Agriculture, University of the Ryukyus; 1 Senbaru, Nishihara-cho, Nakagami-
c
gun, Okinawa 903–0129, Japan: Yona Field, Subtropical Field Science Center, Faculty of Agriculture, University of the
Ryukyus; 635 Aza Yona, Kunigami-son, Kunigami-gun, Okinawa 905–1427, Japan: d Okinawa Prefectural Experimental
Station of Forestry; 134 Furugen, Yomitan-son, Nakagami-gun, Okinawa 904–0314, Japan: e Medicinal Chemistry
Research Institute, Otsuka Pharmaceutical Co., Ltd.; 463–10 Kagasuno, Kawauchi-cho, Tokushima 771–0192, Japan: and
f Faculty of Integrated Arts and Sciences, The University of Tokushima; 1–1 Minamijosanjima-cho, Tokushima 770–8502,
Japan. Received October 28, 2002; accepted December 6, 2002
Five megastigmane glucosides were isolated from the leaves of Glochidion zeylanicum. One of them was a
known compound, blumenol C O-b-D-glucopyranoside (1), and the structures of the four new compounds,
glochidionionosides A—D (2—5), were mainly elucidated by spectroscopic methods, including a modified Mosh-
er’s method. The absolute configurations of the six-membered ring of glochidionionoside D (5) were deduced by
b-D-glucopyranosylation-induced shift trends in the 13C-NMR spectra and confirmed by X-ray analysis as its p-
bromobenzoate (5b), and the axis chirality of C-7 was determined to be R.
Key words Glochidion zeylanicum; Euphorbiaceae; megastigmane glucoside; glochidionionoside; modified Mosher’s method;
X-ray analysis
In our continuing studies on plants collected in the Oki- spectrum showed the presence of six signals assignable to b-
nawan islands, we investigated the constituents of Glochidion glucopyranose. The remaining 13 signals comprised those of
zeylanicum (Euphorbiaceae). Plants belonging to the Euphor- one carbonyl carbon, di- and trisubstituted double bonds,
biaceae are of taxonomic interest, since the family contains three methyls, two methylenes, one of which was assigned to
various morphologically different plants.
be a primary carbinol from its chemical shift, one methine
The isolation of butenolide glucosides has been reported with an oxygen atom, and two quaternary carbons with and
from the leaves of Glochidion zeylanicum (GAERTN.) A. without an oxygen functional group, respectively. From the
JUSS.1) Further investigation afforded five megastigmane glu- above, together with the 1H-NMR data, the structure of 2 was
cosides, blumenol C glucoside (1), and glochidionionosides assumed to be a megastigmane glucoside with hydroxyl
A—D (2—5). Megastigmane glycoside was first isolated functional groups at the C-6, -9, and -13 positions, like
from Vinca rosea as roseoside.2) The glycosides are a rapidly roseoside, except for the methyl group on C-5 was oxidized
expanding family and have been isolated from a variety of to a primary alcohol. The position of the glucosidic linkage
plant sources since the aglycone of one, vomifoliol, was iso- was deduced to be on the hydroxyl group at C-13 from the
lated from Rauwolfia vomitoria in 1969.3,4) This paper deals 13C-NMR chemical shift of C-9 (dC 68.8),6) and this was fur-
with the spectroscopic structural elucidation of new ther confirmed by the heteronuclear multiple-bond correla-
megastigmane glucosides and the structure of a compound tion spectrum in which dH 4.67 (H-1Ј) crossed dC 68.1 (C-
analogous to grasshopper ketone was analyzed using an X- 13). The absolute configuration of the 6-position was de-
ray diffraction method.
duced to be S by comparison of the CD maxima with those
of a known compound.6) The absolute configuration of C-9
was determined to be S by a modified Mosher’s method, after
Results and Discussion
Compounds 1—5 were isolated from the n-BuOH-soluble hydrolysis of 2 and esterification with R- and S-a-methoxy-
fraction of a MeOH extract of the leaves of G. zeylanicum by a-trifluoromethylphenylacetic acids (MTPA) (Fig. 2, 2b, c).7)
a combination of Diaion HP-20 and normal- and reversed- Therefore the structure of glochidionionoside A (2) was
phase silica gel column chromatography (CC). Droplet elucidated to be (6S,7E,9S)-megastigman-3-one-4,7-diene-
counter-current chromatography (DCCC) and HPLC were 6,9,13-triol 13-O-b-D-glucopyranoside.
also used for purification of compounds (Fig. 1, Table 1).
The details and yields are given in Experimental.
Glochidionionosides B (3), C19H32O8, and C (4), C19H30O8,
were also isolated as amorphous powders. The NMR spectra
Blumenol C glucoside (1) was isolated as an amorphous showed that 3 and 4 were compounds analogous to 2, except
powder, and the NMR spectra indicated the presence of a for the absence of a hydroxyl group at the 6-position, and the
megastigmane skeleton and a glucopyranose unit. From the former does not have a double bond between C-7 and -8,
1
spectroscopic data including H- and 13C-NMR, and circular whereas the latter does. The absolute stereochemistries at the
dichroism (CD), the structure of 1 was identified as (6S,9R)- 6 and 9 positions were determined similarly to the previous
megastigman-3-on-4-en-9-ol 9-O-b-D-glucopyranoside.5)
compoyund, from their CD spectra6,8) and the results ob-
Glochidionionoside A (2) was isolated as an amorphous tained using the modified Mosher’s method, as shown in Fig.
powder, and the elemental composition was determined to be 2, respectively (Fig. 2, 3b, c, and 4b, c). Therefore the struc-
C19H29O9 by high-resolution (HR)-FAB-MS. The 13C-NMR tures of glochidionionosides B (3) and C (4) were elucidated
* To whom correspondence should be addressed. e-mail: hotsuka@hiroshima-u.ac.jp
© 2003 Pharmaceutical Society of Japan

March 2003
287
Fig. 1. Structures of Blumenol C Glucoside (1) and Glochidionionosides A (2)—D (5)
Table 1. 13C-NMR Data for Glochidionionosides A—D (2—5) and 5a
(100 MHz, CD3OD)
C
2
3
4
5
5a
1
2
3
4
5
6
7
8
9
10
11
12
13
42.8
50.8
201.3
124.8
165.0
79.2
130.2
137.2
68.8
23.9
23.5
24.2
68.1
37.8
48.6
202.2
123.2
167.9
47.8
27.8
39.8
68.9
23.6
27.6
28.9
70.8
103.5
75.1
78.2
71.7
78.1
62.8
37.1
49.0
202.0
124.1
164.0
52.0 123.8 (123.4)
127.3 202.0 (198.6)
140.5 102.8 (102.8)
68.9 211.5 (210.4)
23.8
27.6
28.9
70.1
32.7 (32.7)a)
35.8
45.2 (44.8) (Ϫ3.5)b) 48.7
73.4 (72.8) (ϩ7.4) 66.0
48.1 (48.3) (Ϫ1.5) 49.6
72.5 (71.5)
72.7
123.8
202.0
102.7
211.5
26.7
30.2
32.7
30.4
26.7 (26.6)
29.9 (29.7)
32.7 (32.7)
30.5 (30.5)
1Ј
103.9
75.2
78.2
71.8
78.1
103.5 102.8 (103.2)
2Ј
3Ј
4Ј
5Ј
6Ј
75.0
78.2
71.7
78.1
62.8
75.2 (75.3)
78.1 (78.7)
71.7 (71.7)
78.0 (78.6)
62.9 (62.8)
Fig. 2. Results Obtained with the Modified Mosher’s Method for Glochid-
ionionosides A—C (2—4)
62.9
a) Data for C5D5N. b) Dd5a—5 (CD3OD).
The Dd values are in Hz (dS–dR, 400 MHz).
to be (6R,9S)-megastigman-3-on-4-ene-9,13-diol and (6R, field shifts of the axial proton of H-3 (dH 4.95 in 6 and 4.14
7E,9S)-megastigman-3-one-4,7-diene-9,13-diol 13-O-b-D- in 5) and the axial methyl protons of C-12 (dH 1.51 in 6 and
glucopyranosides, respectively.
1.23 in 5) disappeared, due to 1,3-diaxial interaction of the
Glochidionionoside D (5) was isolated as an amorphous hydroxyl group on C-5. These data imply that one of the ab-
powder, with the elemental composition of C19H32O8 by HR- solute configurations at C-3 and -5 must be different from
FAB-MS, and the 13C-NMR spectrum showed the presence that of 6, 7, and 8. To confirm this, 5 was enzymatically hy-
of three characteristic signals for an allenic moiety [dC 123.8 drolyzed for determination of the absolute configuration of
(s), 202.0 (s) and 102.8 (d)], together with six for b-glucopy- C-3 using the b-D-glucopyranosylation-induced shift-trend
ranose and 10 other signals, one of which was highly rule for 13C-NMR spectrometry.12) When the 13C-NMR data
deshielded, appearing at dC 211.5. These results and the IR of 5 and 5a were compared, the absolute configuration of C-3
absorption band at 1941 cmϪ1 indicated that the aglycone was assigned to be R, which is opposite to that of 6, 7, and 8
must have the same planar structure as that of grasshopper (Table 1). However, the orientation of the hydroxyl sub-
ketone9) and its 3-O-b-D-glucopyranoside (6) (icariside B1). stituent was found to be in the equatorial position, which was
5-O-b-D-Glucopyranosides (7, 8) (citrosides A, B), which are the same as in other grasshopper ketone glucosides. In turn,
isomers at the 8 position, and previously isolated from the hydroxyl group at the 5-position must be oriented in the
Epimedium grandiflorum var. thunbergianum10) and Citrus equatorial position. These findings were confirmed by the
unshu,11) respectively. However, the spectroscopic data were H–H long-range correlation (H–H COSY) spectrum and the
not identical to those of 6 or 7 and 8. Furthermore, the down- results of difference NOE experiments. In the H–H COSY

288
Vol. 51, No. 3
spectrum, H-13 (dH 1.53) crossed H-4ax (dH 1.80) in a W- recrystallized from i-PrOH, and a suitable crystal was sub-
figure correlation, and on irradiation of H-3 (dH 4.14), signif- jected to X-ray analysis using the heavy atom method. The
icant enhancement was observed at dH 1.28 (H3-11ax), 1.53 absolute configuration was determined by Vijvoet’s anom-
(H3-13ax), 1.98 (H-2eq), 2.25 (H-4eq), and 4.22 (H-1Ј). Thus alous dispersion method (Table 2).13) The absolute structure
the methyl group (C-13) on C-5 must be in the axial orienta- of the ring system was the same as that determined by NMR
tion. Spectral determination of the axis chirality of the al- spectroscopy. The axis chirality of the side chain was ascer-
lenic bond was ambiguous. Thus the aglycone was treated tained by X-ray analysis to be R, as shown in Fig. 3.
with p-bromobenzoyl chloride, the resulting ester (5b) was
Compounds closely related to glochidionionosides A (2)
and C (4) were isolated from Apocynum venetum, apocyno-
sides II and I, respectively.14) On the C-13 methyl groups,
they similarly possessed hydroxyl groups to which sugar
moieties were attached. However, they are respective epimers
at the C-9 position. Therefore the stereochemical structures
at the 9-positions, which do not carry glucose moieties, have
been determined independently in this study.
Table 2. Determination of Absolute Stereochemistry of 5b
Obs. (Fo)
(ϩ) (Ϫ)
Calcd. (Fc)
(ϩ) (Ϫ)
h
k
l
2
1
1
1
1
0
3
3
3
3
3
1
4
2
3
0
1
3
1
4
1
4
2
4
1
2
Ϫ2
0
Ϫ2
Ϫ1
1
Ϫ1
0
Ϫ1
Ϫ2
Ϫ3
1
Ϫ1
Ϫ1
1
0
3
0
Ϫ2
2
Ϫ3
1
Ϫ3
Ϫ3
Ϫ1
1
61Ͻ132
193Ͼ145
149Ͻ201
202Ͻ254
220Ͼ172
120Ͻ163
292Ͼ239
235Ͼ188
204Ͼ162
118Ͻ156
142Ͻ176
49Ͻ104
111Ͻ151
244Ͻ287
229Ͻ271
64Ͼ30
268Ͼ230
151Ͼ64
64Ͻ132
193Ͼ142
152Ͻ204
198Ͻ249
220Ͼ172
116Ͻ167
280Ͼ227
235Ͼ191
206Ͼ167
112Ͻ154
139Ͻ178
68Ͻ110
103Ͻ141
241Ͻ283
228Ͻ268
62Ͼ22
264Ͼ229
150Ͼ62
2
Ϫ4
0
5
10
8
Ϫ5
Ϫ2
0
2
Ϫ10
3
3
5
2
3
Ϫ4
0
Ϫ5
4
1
1
Experimental
A highly porous synthetic resin (Diaion HP-20) was purchased from Mit-
subishi Chemical Co., Ltd. (Tokyo, Japan). Silica gel column chromatogra-
phy (CC) and reversed-phase [octadecyl silica gel (ODS)] open CC (RPCC)
were performed on silica gel 60 (E. Merck, Darmstadt, Germany) and Cos-
mosil 75C18-OPN (Nacalai Tesque, Kyoto, Japan) [Fϭ50 mm, Lϭ25 cm,
linear gradient: MeOH–H2O (1 : 9, 1 l)®(7 : 3, 1 l), fractions of 10 g being
collected], respectively. Droplet counter-current chromatography (DCCC)
(Tokyo Rikakikai, Tokyo, Japan) was equipped with 500 glass columns
(Fϭ2 mm, Lϭ40 cm), and the lower and upper layers of a solvent mixture
of CHCl3–MeOH–H2O–n-PrOH (9 : 12 : 8 : 2) were used for the stationary
and mobile phases, respectively. Five-gram fractions were collected and
numbered according to their order of elution with the mobile phase. HPLC
was performed on ODS (Inertsil; GL Science, Tokyo, Japan; Fϭ20 mm,
Lϭ250 mm), and the eluate was monitored with a UV detector. Hesperidi-
nase was purchased from Sigma Chemical Co. (St. Louis, MO, U.S.A.). (R)-
(ϩ)- and (S)-(Ϫ)-a-methoxy-a-trifluoromethylphenylacetic acids were from
Nacalai Tesque. (Kyoto, Japan).
A melting point was determined with a Yanagimoto micromelting point
apparatus and is uncorrected. Optical rotations were measured on a Union
Giken PM-101 digital polarimeter. IR spectra were measured on a Shimadzu
IR-408 spectrophotometer and UV spectra on a Shimadzu UV-160A spec-
trophotometer. 1H- and 13C-NMR spectra were taken on a JEOL JNM a-400
spectrometer at 400 MHz and 100 MHz, respectively, with tetramethylsilane
(TMS) as an internal standard. Negative-ion HR-FAB-MS were recorded on
a JEOL JMS SX-102 spectrometer. CD spectra were obtained on a JASCO
J-720 spectropolarimeter.
217Ͻ253
98Ͻ135
438Ͼ385
78Ͻ102
282Ͻ316
162Ͼ136
90Ͼ60
208Ͻ241
100Ͻ135
441Ͼ389
73Ͻ108
275Ͻ314
168Ͼ135
88Ͼ60
10
10
Plant Material Leaves of G. zeylanicum (GAERTN.) A. JUSS (Euphor-
biaceae) were collected in Okinawa, Japan, in August 1990, and a voucher
specimen was deposited in the Herbarium of the Institute of Pharmaceutical
Sciences, Hiroshima University Faculty of Medicine (90-GZ-Okinawa-
0822).
Extraction and Fractionation The air-dried leaves of G. zeylanicum
(4.72 kg) were extracted three times with MeOH. The MeOH extract was
concentrated to 1.5 l, and then 75 ml of H2O was added to form a 95% aque-
ous solution. This solution was washed with 1.5 l of n-hexane and then the
methanolic layer was concentrated to a viscous gum. The gummy residue
was suspended in 1.5 l of H2O, and then extracted with 1.5 l each of EtOAc
and n-BuOH successively to give 75.7 and 108 g of EtOAc- and n-BuOH-
soluble fractions, respectively. The n-BuOH extract (107 g) was subjected to
highly porous synthetic resin (Diaion HP-20) CC (Mitsubishi Chemical Co.,
Ltd.; Fϭ80 mm, Lϭ55 cm), using H2O–MeOH (4 : 1, 6 l), (2 : 3, 6 l), (3 : 2,
6 l), and (1 : 4, 6 l), and MeOH (6 l), with 2-l fractions collected. Fractions
4—6 were combined (28.7 g) and then subjected to silica gel (530 g) CC
with elution with CHCl3 (1.5 l), CHCl3–MeOH [(99 : 1, 3 l), (49 : 1, 6 l),
(24 : 1, 6 l), (47 : 3, 6 l), (23 : 2, 6 l), (9 : 1, 6 l), (7 : 1, 6 l), (17 : 3, 4.5 l), (4 : 1,
3 l), and (7 : 3, 3 l)], and CHCl3–MeOH–H2O (70 : 30 : 4, 3 l), with 500-ml
fractions collected. Combined fractions (45—64, 2.35 g) were then sepa-
rated by RPCC. The residue (480 mg) of fractions 74—90 was subjected to
DCCC, and the residue (229 mg) of fractions 35—49 was subjected to
HPLC [H2O–MeOH (7 : 3)], which afforded an allenic compound 5 in a
yield of 77 mg. Combined RPCC fractions (91—105, 311 mg) were sub-
jected to DCCC, and the residue (161 mg) of fractions 45—56 was subjected
to HPLC [H2O–MeOH (7 : 3)] to give 59 mg of 3 and 57 mg of 4. Combined
silica gel CC (65—87, 2.17 g) were subjected to RPCC. The residue
Fig. 3. Perspective ORTEP Drawing of 5b
There are two independent molecules in an asymmetric unit. Atom numberings were
omitted for clarification, except for noncarbon atoms.

March 2003
289
(485 mg) of fractions 60—75 was subjected to DCCC, and 2 (25 mg) was
purified from the residue (125 mg) of fractions 4—16 by HPLC [H2O–
MeOH (7 : 3)].
ϩ37.5° (cϭ0.40, H2O, 24 h after being dissolved in the solvent).
Preparation of (R)- and (S)-MTPA Esters (2b, c) from Glochidion-
ionol A (2a) A solution of 2a (3.4 mg) in 1 ml of dehydrated CH2Cl2 was
reacted with (R)-MTPA (50 mg) in the presence of N,NЈ-dicyclohexylcar-
bodiimide (38 mg) (DCC) and 4-dimethylaminopyridine (DMAP) (17 mg),
and the resulting mixture was occasionally stirred at 25 °C for 40 min. After
the addition of 1 ml each of H2O and CH2Cl2, the solution was washed suc-
cessively with 5% HCl, NaHCO3-saturated H2O and brine. The organic layer
was dried over Na2SO4 and then evaporated under reduced pressure. The
residue was purified by preparative TLC [silica gel (0.25 mm thickness,
spotted for 18 cm, and developed with CHCl3–(CH3)2CO (19 : 1) for 9 cm
and eluted with CHCl3–MeOH (9 : 1)] to furnish the ester, 2b (5.7 mg, 63%).
Using a similar procedure, 2c (5.4 mg, 59%) was prepared from 2a (3.4 mg)
with (S)-MTPA (46 mg), DCC (38 mg), and 4-DMAP (17 mg).
Glochidionionol A 9,13-Di-(R)-MTPA Ester (2b): Amorphous powder,
1H-NMR (CDCl3) d: 0.894 (3H, s, H3-12), 0.999 (3H, s, H3-11), 1.43 (3H, d,
Jϭ7 Hz, H3-10), 2.23 (2H, s, H2-2), 3.52—3.54 (6H, m, –OCH3ϫ2), 4.78
(1H, dd, Jϭ16, 2 Hz, H-13a), 4.93 (1H, dd, Jϭ16, 2 Hz, H-13b), 5.62 (1H,
quid, Jϭ7, 1 Hz, H-9), 5.71 (1H, dd, Jϭ16, 1 Hz, H-7), 5.82 (1H, dd, Jϭ16,
7 Hz, H-8), 5.93 (1H, t, Jϭ2 Hz, H-4), 7.37—7.52 (10H, m, aromatic pro-
tons); HR-FAB-MS (negative-ion mode) m/z 671.2103 [MϪH]Ϫ (Calcd for
C33H33O8F6: 671.2080).
From combined fractions (7—8, 25.9 g) obtained on Diaion HP-20 CC,
14 mg of 1 was isolated using similar chromatographic methods.
Blumenol C O-b-D-Glucopyranoside (1): [a]D28 ϩ43.9° (cϭ0.96, MeOH);
CD De (nm): ϩ2.15 (218), ϩ2.84 (239), ϩ0.63 (331) (cϭ1.05ϫ10Ϫ4 M,
MeOH).
Glochidionionoside A (2): Amorphous powder; [a]D28 ϩ29.1° (cϭ1.48,
MeOH); IR nmax (KBr) cmϪ1: 3407, 2971, 2930, 2882, 1659, 1413, 1372,
1317, 1163, 1076, 1042; UV lmax (MeOH) nm (log e): 237 (3.86); 1H-NMR
(CD3OD) d: 1.00 (3H, s, H3-12eq), 1.05 (3H, s, H3-11ax), 1.24 (3H, d,
Jϭ6 Hz, H3-10), 2.20 (1H, d, Jϭ17 Hz, H-2a), 2.51 (1H, d, Jϭ17 Hz, H-2b),
3.24 (1H, t, Jϭ8 Hz, H-2Ј), 3.66 (1H, dd, Jϭ12, 6 Hz, H-6Јa), 3.86 (1H, dd,
Jϭ12, 2 Hz, H-6Јb), 4.27 (1H, d, Jϭ8 Hz, H-1Ј), 4.29 (1H, dd, Jϭ14, 2 Hz,
H-13a), 4.32 (1H, m, H-9), 4.67 (1H, dd, Jϭ14, 2 Hz, H-13b), 5.82 (1H, s,
H-7), 5.83 (1H, d, Jϭ4 Hz, H-8), 6.30 (1H, br s, H-4); 13C-NMR (CD3OD):
Table 1; CD De (nm): ϩ8.18 (240), Ϫ1.20 (331) (cϭ1.11ϫ10Ϫ4 M, MeOH);
HR-FAB-MS (negative-ion mode) m/z: 401.1829 [MϪH]Ϫ (Calcd for
C19H29O9: 401.1812).
Glochidionionoside B (3): Amorphous powder, [a]D28 ϩ7.4° (cϭ1.22,
MeOH); IR nmax (KBr) cmϪ1: 3409, 2965, 2930, 2878, 1651, 1371, 1163,
1078, 1045; UV lmax (MeOH) nm (log e): 239 (3.93); 1H-NMR (CD3OD) d:
1.03 (3H, s, H3-12), 1.11 (3H, s, H3-11), 1.16 (3H, d, Jϭ6 Hz, H3-10), 2.02
(1H, t, Jϭ5 Hz, H-6), 2.03 (1H, d, Jϭ18 Hz, H-2a), 2.54 (1H, d, Jϭ18 Hz,
H-2b), 3.25 (1H, dd, Jϭ12, 6 Hz, H-6Јa), 3.68 (1H, m, H-9), 3.88 (1H, dd,
Jϭ12, 2 Hz, H-6Јb), 4.35 (1H, d, Jϭ8 Hz, H-1Ј), 4.38 (1H, dd, Jϭ17, 2 Hz,
H-13a), 4.52 (1H, dd, Jϭ17, 2 Hz, H-13b), 6.17 (1H, br s, H-4); 13C-NMR
(CD3OD): Table 1; CD De (nm): ϩ1.47 (217), ϩ1.34 (239), ϩ0.42 (332)
(cϭ9.43ϫ10Ϫ5 M, MeOH); HR-FAB-MS (negative-ion mode) m/z: 387.2016
[MϪH]Ϫ (Calcd for C19H31O8: 387.2019).
Glochidionionol A 9,13-Di-(S)-MTPA Ester (2c): Amorphous powder,
1H-NMR (CDCl3) d: 0.947 (3H, s, H3-12), 1.03 (3H, s, H3-11), 1.39 (3H, d,
Jϭ7 Hz, H3-10), 2.30 (2H, s, H2-2), 3.52—3.54 (6H, m, –OCH3ϫ2), 4.67
(1H, dd, Jϭ16, 2 Hz, H-13a), 5.03 (1H, dd, Jϭ16, 2 Hz, H-13b), 5.59 (1H,
qui, Jϭ7 Hz, H-9), 5.85 (1H, d, Jϭ16 Hz, H-7), 5.91 (1H, t, Jϭ2 Hz, H-4),
5.93 (1H, dd, Jϭ16, 7 Hz, H-8), 7.37—7.52 (10H, m, aromatic protons);
HR-FAB-MS (negative-ion mode) m/z 671.2052 [MϪH]Ϫ (Calcd for
C33H33O8F6: 671.2080).
Enzymatic Hydrolysis of Glochidionoionosides B (3) and C (4) to
Glochidionionols B and C (3a and 4a, Respectively) Glochidionoiono-
sides B (3) (21 mg) and C (4) (25 mg) were hydrolyzed with emulsin (25,
18 mg, respectively) in 2 ml of H2O at 37 °C for 15 h. The reaction mixtures
were concentrated, and then the aglycones and D-glucose were purified in a
similar manner to that for 2a. The aglycones (3a, 4a) were recovered in frac-
tions 25—29 (5.7 mg, 47%) and 24—28 (10.2 mg, 70%), and D-glucose in
fractions 44—50 (5.7 mg, 59%) and 44—51 (9.0 mg, 68%), respectively.
Glochidionionol B (3a): Colorless syrup; [a]D28 ϩ89.5° (cϭ0.38, MeOH);
1H-NMR (CD3OD) d: 1.02 (3H, s, H3-12), 1.11 (3H, s, H3-11), 1.16 (3H, d,
Jϭ6 Hz, H3-10), 1.46—1.64 (3H, m, H-7a, 7b, 8a), 1.73—1.83 (1H, m, H-
8b), 2.04 1H, d, Jϭ18 Hz, H-2a), 2.55 (1H, d, Jϭ18 Hz, H-2b), 3.69 (1H,
sextet, Jϭ6 Hz, H-8), 4.15 (1H, dd, Jϭ18, 2 Hz, H-13a), 4.30 (1H, dd,
Jϭ18, 2 Hz, H-13b), 6.65 (1H, br s, H-4); 13C-NMR (CD3OD) d: 23.5 (C-
10), 27.6 (C-11), 28.0 (C-7), 28.8 (C-12), 37.4 (C-1), 39.8 (C-8), 48.0 (C-6),
48.7 (C-2), 65.1 (C-13), 68.8 (C-9), 121.6 (C-4), 172.3 (C-5), 202.3 (C-3);
HR-FAB-MS (negative-ion mode) m/z: 225.1506 [MϪH]Ϫ (Calcd for
C13H21O3: 225.1491). D-Glucose: [a]D25 ϩ44.7° (cϭ0.38, H2O, 24 h after
being dissolved in the solvent).
Glochidionionoside C (4): Amorphous powder; [a]D28 ϩ112.1° (cϭ1.42,
MeOH); IR nmax (KBr) cmϪ1: 3395, 2966, 2930, 2878, 1657, 1413, 1370,
1298, 1163, 1076; UV lmax (MeOH) nm (log e): 236 (4.06); 1H-NMR
(CD3OD) d: 0.98 (3H, s, H3-12), 1.04 (3H, s, H3-11), 1.23 (3H, d, Jϭ6 Hz,
H3-10), 2.09 (1H, d, Jϭ17 Hz, H-2a), 2.49 (1H, d, Jϭ17 Hz, H-2b), 2.75
(1H, d, Jϭ9 Hz, H-6), 3.24 (1H, dd, Jϭ9, 8 Hz, H-2Ј), 3.68 (1H, dd, Jϭ12,
6 Hz, H-6Јa), 3.68 (1H, m, H-9), 3.87 (1H, dd, Jϭ12, 2 Hz, H-6Јb), 4.27
(1H, d, Jϭ8 Hz, H-1Ј), 4.27 (1H, dd, Jϭ17, 2 Hz, H-13a), 4.48 (1H, dd,
Jϭ17, 2 Hz, H-13b), 5.61 (1H, ddd, Jϭ15, 9, 1 Hz, H-7), 5.72 (1H, dd,
Jϭ15, 6 Hz, H-8), 6.25 (1H, br s, H-4); 13C-NMR (CD3OD): Table 1, CD De
(nm): ϩ20.8 (244), Ϫ1.06 (322) (cϭ1.09ϫ10Ϫ4 M, MeOH), HR-FAB-MS
(negative-ion mode) m/z: 385.1843 [MϪH]Ϫ (Calcd for C19H29O8:
385.1862).
Glochidionionoside D (5): Amorphous powder; [a]D28 Ϫ47.8° (cϭ1.15,
MeOH); IR nmax (KBr) cmϪ1: 3409, 2967, 1941, 1669, 1366, 1246, 1076
1028; UV lmax (MeOH) nm (log e): 229 (4.14), 279 (3.19); 1H-NMR
(CD3OD) d: 1.23 (3H, s, H3-12eq), 1.28 (3H, s, H3-11ax), 1.53 (3H, s, H3-
13), 1.56 (1H, dd, Jϭ10, 13 Hz, H-2ax), 1.80 (1H, dd, Jϭ13, 10 Hz, H-4ax),
1.98 (1H, ddd, Jϭ13, 4, 1 Hz, H-2eq), 2.22 (3H, s, H3-10), 2.25 (1H, ddd,
Jϭ13, 4, 1 Hz, H-4eq), 3.16 (1H, dd, Jϭ9, 8 Hz, H-2Ј), 3.28 (1H, t, Jϭ9 Hz,
H-4Ј), 3.37 (1H, t, Jϭ9 Hz, H-3Ј), 3.68 (1H, dd, Jϭ12, 6 Hz, H-6Јa), 3.88
(1H, dd, Jϭ12, 2 Hz, H-6Јb), 4.14 (1H, tt, Jϭ10, 4 Hz, H-3), 4.42 (1H, d,
Jϭ8 Hz, H-1Ј), 5.93 (1H, s, H-8); 13C-NMR (CD3OD and C5D5N): Table 1;
CD De (nm): Ϫ1.75 (210), ϩ0.29 (242), Ϫ0.25 (278), Ϫ0.27 (323)
(cϭ8.94ϫ10Ϫ5 M, MeOH); HR-FAB-MS (negative-ion mode) m/z: 385.1841
[MϪH]Ϫ (Calcd for C19H31O8: 385.1862).
Enzymatic Hydrolysis of Glochidionoionoside A (2) Glochidiono-
ionoside A (2) (19 mg) was hydrolyzed with emulsin (20 mg) in 2 ml of H2O
at 37 °C for 18 h. The reaction mixture was concentrated and then subjected
to silica gel CC (Fϭ15 mm, Lϭ20 cm) with C6H6 (40 ml), C6H6–CHCl3
(1 : 1, 40 ml), CHCl3 (100 ml), and CHCl3–MeOH (19 : 1, 100 ml, 9 : 1,
100 ml, 17 : 3, 100 ml, 7 : 3, 300 ml), with 10-ml fractions collected. Glo-
chidionionol A (2a) and D-glucose were recovered in fractions 28—32
(6.9 mg, 61%) and 44—50 (6.0 mg, 71%), respectively.
Glochidionionol
C
(4a): Colorless syrup; [a]D26 ϩ230.9° (cϭ0.68,
MeOH); 1H-NMR (CD3OD) d: 0.991 (3H, s, H3-12), 1.03 (3H, s, H3-11),
1.23 (3H, d, Jϭ6 Hz, H3-10), 2.09 (1H, d, Jϭ17 Hz, H-2a), 2.48 (1H, d,
Jϭ17 Hz, H-2b), 2.57 (1H, d, Jϭ8 Hz, H-6), 4.11 (1H, dd, Jϭ12, 2 Hz, H-
13a), 4.20 (1H, dd, Jϭ12, 2 Hz, H-13b), 4.26 (1H, qui, Jϭ6 Hz, H-9), 5.60
(1H, dd, Jϭ16, 8 Hz, H-7), 5.67 (1H, dd, Jϭ15, 6 Hz, H-8), 6.14 (1H, t,
Jϭ2 Hz, H-4); 13C-NMR (CD3OD) d: 23.8 (C-10), 27.4 (C-11), 27.9 (C-12),
37.3 (C-1), 49.2 (C-2), 52.3 (C-6), 64.2 (C-13), 68.8 (C-9), 122.5 (C-4),
127.5 (C-7), 140.2 (C-8), 168.3 (C-5), 202.1 (C-3); HR-FAB-MS (negative-
ion mode) m/z: 223.1338 [MϪH]Ϫ (Calcd for C13H19O3: 223.1334). D-Glu-
cose: [a]D26 ϩ43.1° (cϭ0.53, H2O, 24 h after being dissolved in the solvent).
Preparation of (R)- and (S)-MTPA Esters (3b, c, and 4b, c) from 3a
and 4a, Respectively Using a similar procedure to that used for the prepa-
ration of 2b and 2c from 2a, 3b (6.3 mg, 77%) and 3c (5.3 mg, 65%) were
prepared from 3a (2.8 mg each) using the respective amounts of (R)- and
(S)-MTPA (44, 43 mg), DCC (33, 34 mg), and 4-DMAP (16, 16 mg). The
developing solvent for preparative TLC was CHCl3–(CH3)2CO, 19 : 1.
Glochidionionol B 9,13-Di-(R)-MTPA Ester (3b): Amorphous powder;
1H-NMR (CDCl3) d: 0.917 (3H, s, H3-12), 0.936 (3H, s, H3-11), 1.24—1.30
(1H, m, H-7a), 1.33 (3H, d, Jϭ7 Hz, H3-10), 1.53—1.64 (3H, m, H-7b, 8a,
8b), 1.77 (1H, t, Jϭ5 Hz, H-6), 2.03 (1H, d, Jϭ18 Hz, H-2a), 2.27 (1H, d,
Jϭ18 Hz, H-2b), 3.53—3.55 (6H, m, –OCH3ϫ2), 4.68 (1H, dd, Jϭ16, 2 Hz,
H-13a), 4.78 (1H, dd, Jϭ16, 2 Hz, H-13b), 5.08 (1H, m, H-9), 5.85 (1H, br s,
H-4), 7.37—7.52 (10H, m, aromatic protons); HR-FAB-MS (positive-ion
mode) (ϩNaI) m/z: 681.2228 [MϩNa]ϩ (Calcd for C33H36O7F6Na:
Glochionionol A (2a): Colorless syrup, [a]D28 ϩ156.7° (cϭ0.46, MeOH),
1H-NMR (CD3OD) d: 0.996 (3H, s, H3-12eq), 1.04 (3H, s, H3-11ax), 1.23
(3H, d, Jϭ6 Hz, H3-10), 2.17 (1H, d, Jϭ17 Hz, H-2a), 2.49 (1H, d, Jϭ17 Hz,
H-2b), 4.18 (1H, dd, Jϭ19, 2 Hz, H-13a), 4.30 (1H, qd, Jϭ6, 2 Hz, H-9),
4.38 (1H, dd, Jϭ19, 2 Hz, H-13b), 5.80 (2H, m, H-7, 8), 6.16 (1H, br s, H-
4); 13C-NMR (CD3OD) d: 23.3, 23.7, 24.0 (C-10, 11, 12), 42.7 (C-1), 50.7
(C-2), 61.3 (C-13), 68.6 (C-9), 79.3 (C-6), 123.1 (C-4), 130.2 (C-7), 136.9
(C-8), 169.4 (C-5), 201.3 (C-3); HR-FAB-MS (negative-ion mode): m/z:
239.1288 [MϪH]Ϫ (Calcd for C13H19O4: 239.1283). D-Glucose: [a]D28

290
Vol. 51, No. 3
a Rigaku AFC-5S automated four-circle diffractometer with graphite-
monochromated MoKa radiation. Crystal data: C20H23O4Br, Mrϭ430.30,
triclinic, space group P1, aϭ7.977(1) Å, bϭ21.323(2) Å, cϭ6.406(1) Å,
aϭ91.48(1)°, bϭ112.56(1)°, gϭ82.911(9)°, Vϭ998.3(3) Å3, Zϭ2, Dxϭ
1.352 Mg mϪ3, F(000)ϭ418, and m(MoKa)ϭ20.842 cmϪ1. The intensities
were measured in the w-scan mode, 2qϽ55°, and measurements were con-
ducted on one component of Bijvoet pairs. Three standard reflections were
monitored every 150 measurements. The data were corrected for Lorentz
and polarization factors. The absorption correction was applied (transmis-
sion factorϭ0.620—0.998).15) Of 8867 independent reflections which were
collected, 4579 reflections with IϾ2.00s(I) were used for the structural de-
termination and refinement. The structure was solved by the direct method
using the TEXSAN crystallographic software package.16) This compound
has two independent molecules in the asymmetric unit. All nonhydrogen
atoms were found in the Fourier map. All H atoms were found in the differ-
ence Fourier map and refined isotropically. The refinement of atomic para-
meters was carried out using the full-matrix least-squares with anisotropic
temperature factors for all non-H atoms. The final refinement converged
with Rϭ0.043 and Rwϭ0.048 for 452 parameters. Then 25 Bijvoet pairs
with high intensity and high measurement accuracy were selected (Table 2).
The absolute configuration of 5b was determined as shown in Fig. 3 using
Bijvoet’s anomalous-dispersion method.13) Atomic scattering factors were
taken from the “International Tables for X-ray Crystallography.”17)
681.2263).
Glochidionionol B 9,13-Di-(S)-MTPA Ester (3c): Amorphous powder;
1H-NMR (CDCl3) d: 0.926 (3H, s, H3-12), 0.993 (3H, s, H3-11), 1.27 (3H, d,
Jϭ6 Hz, H3-10), 1.42—1.48 (1H, m, H-7a), 1.59—1.74 (3H, m, H-7b, 8a,
8b), 1.84 (1H, t, Jϭ5 Hz, H-6), 2.07 (1H, d, Jϭ18 Hz, H-2a), 2.33 (1H, d,
Jϭ18 Hz, H-2b), 3.48—3.54 (6H, m, –OCH3ϫ2), 4.74 (1H, dd, Jϭ16, 2 Hz,
H-13a), 4.88 (1H, dd, Jϭ16, 2 Hz, H-13b), 5.07 (1H, m, H-9), 5.89 (1H, br s,
H-4), 7.36—7.51 (10H, m, aromatic protons); HR-FAB-MS (positive-ion
mode) (ϩNaI) m/z: 681.2224 [MϩNa]ϩ (Calcd for C33H36O7F6Na:
681.2263).
Using a similar procedure to that used for the preparation of 3b and 3c
from 3a, 4b (11.1 mg, 74%) and 4c (10.2 mg, 68%) were prepared from 4a
(5.3 mg each) using of the respective amounts of (R)- and (S)-MTPA (46,
49 mg), DCC (32, 34 mg), and 4-DMAP (18, 22 mg). The developing sol-
vent for preparative TLC was CHCl3–(CH3)2CO, 19 : 1.
Glochidionionol C 9,13-Di-(R)-MTPA Ester (4b): Amorphous powder;
1H-NMR (CDCl3) d: 0.888 (3H, s, H3-12), 0.954 (3H, s, H3-11), 1.42 (3H, d,
Jϭ6 Hz, H3-10), 2.08 (1H, d, Jϭ17 Hz, H-2a), 2.23 (1H, d, Jϭ17 Hz, H-2b),
2.56 (1H, d, Jϭ8 Hz, H-6), 3.51—3.55 (6H, m, –OCH3ϫ2), 4.67 (1H, dd,
Jϭ16, 1 Hz, H-13a), 4.77 (1H, dd, Jϭ16, 1 Hz, H-13b), 5.57 (1H, qui,
Jϭ6 Hz, H-9), 5.55 (1H, dd, Jϭ15, 6 Hz, H-8), 5.59 (1H, dd, Jϭ15, 8 Hz, H-
7), 5.98 (1H, s, H-4), 7.37—7.52 (10H, m, aromatic protons); HR-FAB-MS
(positive-ion mode) (ϩNaI) m/z: 679.2111 [MϩNa]ϩ (Calcd for
C33H34O7F6Na: 679.2106).
Glochidionionol C 9,13-Di-(S)-MTPA Ester (4b): Amorphous powder;
1H-NMR (CDCl3) d: 0.918 (3H, s, H3-12), 0.961 (3H, s, H3-11), 1.38 (3H, d,
Jϭ6 Hz, H3-10), 2.11 (1H, d, Jϭ17 Hz, H-2a), 2.28 (1H, d, Jϭ17 Hz, H-2b),
2.59 (1H, d, Jϭ9 Hz, H-6), 3.50—3.53 (6H, m, –OCH3ϫ2), 4.66 (1H, dd,
Jϭ15, 1 Hz, H-13a), 4.83 (1H, dd, Jϭ15, 1 Hz, H-13b), 5.52 (1H, qui,
Jϭ6 Hz, H-9), 5.61 (1H, dd, Jϭ15, 6 Hz, H-7), 5.70 (1H, dd, Jϭ15, 9 Hz, H-
8), 5.98 (1H, s, H-4), 7.36—7.52 (10H, m, aromatic protons); HR-FAB-MS
(positive-ion mode) (ϩNaI) m/z: 679.2110 [MϩNa]ϩ (Calcd for
C33H34O7F6Na: 679.2106).
The final atom coordinates and a list of the temperature factors and final
structure factors have been deposited at the Cambridge Crystallographic
Centre, University Chemical Laboratory, Lensfield Road, Cambridge CB12
1EW, U.K.
Acknowledgments The authors are grateful for access to the supercon-
ducting NMR instrument at the Analytical Center of Molecular Medicine of
Hiroshima University Faculty of Medicine. This work was supported by a
grant-in-aid from the Ministry of Education, Science, Sports, Culture and
Technology of Japan (No. 13672216). Thanks are also due to the Okinawa
Foundation for financial support with an Okinawa Research Promotion
Award (H.O.).
Enzymatic Hydrolysis of Glochidionionoside D (5) Glochidioniono-
side D (5, 23 mg) in 2 ml of H2O was hydrolyzed with emulsin (20 mg) at
37 °C for 18 h. The reaction mixture was dried and then subjected to silica
gel (Fϭ15 mm, Lϭ20 cm) CC with CHCl3 (150 ml), and CHCl3–MeOH
(9 : 1, 100 ml) and (7 : 3, 300 ml), with 10-ml fractions collected. An agly-
cone (5a) and D-glucose were recovered in fractions 18—21 (9.8 mg) and
31—37 (8.0 mg), respectively.
References and Notes
1) Otsuka H., Hirata E., Takushi A., Shinzato T., Takeda Y., Bando M.,
Kido M., Chem. Pharm. Bull., 48, 547—551 (2000).
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2541—2543 (1974).
3) Pouset J. L., Poison J., Tetrahedron Lett., 1969, 1173—1174 (1969)
4) Galbraith M. N., Horn D. H. S., J. Chem. Soc. Chem. Commun., 1972,
113—114 (1972).
5) Miyase T., Ueno A., Takizawa H., Kobayashi H., Oguchi H., Chem.
Pharm. Bull., 36, 2475—2484 (1988).
6) Otsuka H., Yao M., Kamada K., Takeda Y., Chem. Pharm. Bull., 43,
754—759 (1995).
7) Ohtani I., Kusumi T., Kashman Y., Kakisawa H., J. Am. Chem. Soc.,
113, 4092—4096 (1991).
8) Weiss G., Koreeda M., Nakanishi K., J. Chem. Soc. Chem. Commun.,
1972, 565—566 (1972).
9) Meinwald J., Erickson K., Hartshorn M., Meinwald Y. C., Eisner T.,
Tetrahedron Lett., 1968, 2959—2962 (1968).
10) Miyase T., Ueno A., Takizawa N., Kobayashi H., Karasawa H., Chem.
Pharm. Bull., 35, 1109—1117 (1987).
11) Umehara K., Hattori I., Miyase T., Ueno A., Hara S., Kageyama C.,
Chem. Pharm. Bull., 36, 5004—5008 (1988).
12) Kasai R., Suzuno M., Asakawa I., Tanaka O., Tetrahedron Lett., 1977,
175—178 (1977).
13) Bijvoet J. M., Peerdeman A. F., van Bommel A. J., Nature (London),
168, 271—272 (1951).
14) Murakami T., Kishi A., Matsuda H., Hattori M., Yoshikawa M., Chem.
Pharm. Bull., 49, 845—848 (2001).
Aglycone (5a): Colorless syrup; [a]D25 Ϫ37.9° (cϭ0.66, MeOH); UV lmax
(MeOH) nm (log e): 229 (4.07); H-NMR (CD3OD) d: 1.24 (3H, s, H3-12),
1
1.27 (3H, s, H3-11), 1.51 (3H, d, Jϭ1 Hz, H3-13), 1.50 (1H, dd, Jϭ12,
10 Hz, H-2ax), 1.69 (1H, ddq, Jϭ12, 10, 1 Hz, H-4ax), 1.84 (1H, ddd, Jϭ12,
4, 2 Hz, H-2eq), 2.08 (1H, ddd, Jϭ12, 4, 2 Hz, H-4eq), 2.22 (3H, d,
Jϭ0.5 Hz, H3-10), 3.99 (1H, tt, Jϭ10, 4 Hz, H-3), 5.93 (1H, d, Jϭ0.5 Hz, H-
8); 13C-NMR (CD3OD): Table 1; HR-FAB-MS (negative-ion mode) m/z:
223.1357 [MϪH]Ϫ (Calcd for C13H19O3: 223.1334). D-Glucose: [a]D24
ϩ41.5° (cϭ0.53, H2O, 24 h after being dissolved in the solvent).
Synthesis of p-Bromobenzoyl Ester (5b) of 5a The aglycone (5a,
9 mg) was treated with 18 mg of p-bromobenzoyl chloride and 15 ml of pyri-
dine in 1 ml of CDCl3 at 25 °C. Equal amounts of the reagents were added at
12 and 24 h. Six hours after the final addition of the reagents, 2 ml of H2O
and then 1 ml of CHCl3 were added to the reaction mixture. The organic
layer was washed successively with 2 N HCl, NaHCO3-saturated H2O and
brine. The residue, taken up in the organic layer, was subjected to prepara-
tive TLC (silica gel, 0.25 mm thickness, 18 cm width, developed for 9 cm
with C6H6–(CH3)2CO, 4 : 1, and eluted with CHCl3–MeOH, 9 : 1) to give
10 mg of the ester.
p-Bromobenzoyl Ester (5b): Colorless plates (EtOH); mp 112—114 °C;
[a]D29 Ϫ21.0° (cϭ0.67, CHCl3), 1H-NMR (CDCl3) d: 1.31 and 1.34 (each
3H, each s, H3-11, 12), 1.58 (3H, s, H3-13), 1.89 (1H, dd, Jϭ14, 5 Hz, H-
2ax), 1.98 (1H, ddd, Jϭ14, 4, 1 Hz, H-2eq), 2.05 (1H, dd, Jϭ14, 4 Hz, H-
4ax), 2.23 (3H, d, Jϭ0.5 Hz, H3-10), 2.25 (1H, ddd, Jϭ14, 4, 1 Hz, H-4eq),
5.37 (1H, septet, Jϭ4 Hz, H-3), 6.01 (1H, d, Jϭ0.5 Hz, H-8), 7.89 and 7.58
(each 2H, each d, each Jϭ9 Hz, H-2Љ, 3Љ, 5Љ, 6Љ); 13C-NMR (CDCl3) d: 26.6
(C-10), 30.2, 30.6, 31.7 (C-11, 12, 13), 34.8 (C-1), 43.3 (C-2), 44.6 (C-4),
69.1 (C-3), 71.2 (C-5), 102.3 (C-8), 121.3 (C-6), 128.3, 129.2 (C-1Љ, 4Љ),
131.1ϫ2, 131.9ϫ2 (C-2Љ, 3Љ, 5Љ, 6Љ), 165.3 (C-7Љ), 198.5 (C-7), 209.0 (C-9);
HR-FAB-MS (positive-ion mode) (ϩNaI) m/z: 429.0660 and 431.0660
[MϩNa]ϩ (Calcd for C20H23O479BrNa and C20H23O481BrNa: 429.0677 and
431.0657, respectively).
15) North A. C., Phillips D. C., Mathews F. S., Acta Crystallogr., 1968,
A24, 351—359 (1968).
16) Molecular Structure Corporation and Rigaku Corporation (2000).
teXsan. Single Crystal Structure Analysis Software. Version 1.11.
32000 Research Forest Drive, The Woodlands, TX 77381, U.S.A., and
3–9–12 Matsubara-cho, Akishima-shi 196–8666, Tokyo, Japan, re-
spectively.
17) “International Tables for X-ray Crystallography,” Vol. C, Kynoch
Press, Birmingham, England, 1992.
X-Ray Structure Determination of 5b A suitable crystal (0.50 mmϫ
0.50 mmϫ0.10 mm) was used for analysis. All data were obtained with