July 2011
895
(MꢁH)ꢁ, 957 (MꢁC6H11O4)ꢁ, 941 (MꢁC6H11O5)ꢁ, 795 (MꢁC12H23O9)ꢁ,
633 (MꢁC18H31O14)ꢁ, 487 (MꢁC24H41O18)ꢁ.
gastric emptying (%)ꢂ(1ꢁamount of test sample/amount of standard)
ꢈ100
Desacyl-perennisoside X (3a): An amorphous powder, [a]D26 ꢀ3.8°
(cꢂ0.88, MeOH). High-resolution positive-ion FAB-MS: Calcd for
Statistics Values are expressed as meansꢆS.E.M. One-way analysis of
C60H98O29Na (MꢀNa)ꢀ: 1305.6091. Found: 1305.6086. IR (KBr): 3440, variance (ANOVA) followed by Dunnett’s test was used for statistical analy-
1736, 1655, 1230, 1075 cmꢁ1
.
1H-NMR (600 MHz, pyridine-d5) d: given
sis. Probability (p) values less than 0.05 were considered significant.
in Table 4. 13C-NMR data (150 MHz, pyridine-d5) dC: given in Table 5.
Positive-ion FAB-MS m/z: 1305 (MꢀNa)ꢀ. Negative-ion FAB-MS
m/z: 1281 (MꢁH)ꢁ, 1119 (MꢁC6H11O5)ꢁ, 957 (MꢁC12H21O10)ꢁ, 811
(MꢁC18H31O14)ꢁ, 649 (MꢁC24H41O19)ꢁ, 487 (MꢁC30H51O24)ꢁ.
Acknowledgements T. Morikawa, K. Ninomiya, and O. Muraoka were
supported by ‘High-tech Research Center’ Project for Private Universities:
matching fund subsidy from the Ministry of Education, Culture, Sports, Sci-
Desacyl-perennisoside XI (4a): An amorphous powder, [a]D25 ꢀ4.0° ence and Technology (MEXT) of Japan, 2007—2011 and by a Grant-in-Aid
(cꢂ0.29, MeOH). High-resolution positive-ion FAB-MS: Calcd for
for Scientific Research from MEXT. M. Yoshikawa, H. Matsuda, and S.
C60H98O29Na (MꢀNa)ꢀ: 1305.6091. Found: 1305.6085. IR (KBr): 3440, Nakamura were supported by the 21st COE Program, Academic Frontier
1736, 1655, 1260, 1051 cmꢁ1
.
1H-NMR (600 MHz, pyridine-d5) d: given
Project, and a Grant-in-Aid for Scientific Research from MEXT. H. Matsuda
was also supported by Hoh-ansha Foundation, Japan.
in Table 4. 13C-NMR data (150 MHz, pyridine-d5) dC: given in Table 5.
Positive-ion FAB-MS m/z: 1305 (MꢀNa)ꢀ. Negative-ion FAB-MS
m/z: 1281 (MꢁH)ꢁ, 1119 (MꢁC6H11O5)ꢁ, 957 (MꢁC12H21O10)ꢁ, 811
(MꢁC18H31O14)ꢁ, 649 (MꢁC24H41O19)ꢁ, 487 (MꢁC30H51O24)ꢁ.
References and Notes
1) Part XXXI: Matsuda H., Nakamura S., Fujimoto K., Moriuchi R.,
Kimura Y., Ikoma N., Hata Y., Muraoka O., Yoshikawa M., Chem.
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Thomson P. D. R., Montvale, 2004, pp. 877—878.
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Asao Y., Kumahara A., Matsuda H., Chem. Pharm. Bull., 55, 308—
316 (2007).
4) Yoshikawa M., Morikawa T., Asao Y., Fujiwara E., Nakamura S.,
Matsuda H., Chem. Pharm. Bull., 55, 606—612 (2007).
5) Morikawa T., Li X., Nishida E., Ito Y., Matsuda H., Nakamura S.,
Muraoka O., Yoshikawa M., J. Nat. Prod., 71, 828—835 (2008).
6) Yoshikawa M., Li X., Nishida E., Nakamura S., Matsuda H., Muraoka
O., Morikawa T., Chem. Pharm. Bull., 56, 559—568 (2008).
7) Xie Y., Morikawa T., Ninomiya K., Imura K., Muraoka O., Yuan D.,
Yoshikawa M., Chem. Pharm. Bull., 56, 1628—1631 (2008).
8) Nakamura S., Okazaki Y., Ninomiya K., Morikawa T., Matsuda H.,
Yoshikawa M., Chem. Pharm. Bull., 56, 1704—1709 (2008).
9) Morikawa T., Wang L.-B., Nakamura S., Ninomiya K., Yokoyama E.,
Matsuda H., Muraoka O., Wu L.-J., Yoshikawa M., Chem. Pharm.
Bull., 57, 361—367 (2009).
Acid Hydrolysis of 1a—4a and Perennisoside XII (5) Solutions of 1a
(5.2 mg), 2a (5.5 mg), 3a (4.0 mg), 4a (3.0 mg), and 5 (1.6 mg) in 5%
H2SO4–1,4-dioxane (1 : 1, v/v, 1.0 ml) were heated under reflux for 1 h. After
cooling, the reaction mixture was neutralized with Amberlite IRA-400 (OHꢁ
form), and the resin was removed by filtration. On removal of the solvent
from the filtrate under reduced pressure, the residue was partitioned in an
EtOAc–H2O (1 : 1, v/v) mixture, and the solvent was removed in vacuo from
the EtOAc-soluble fraction and as aqueous phase. The EtOAc-soluble frac-
tion was purified by HPLC [Cosmosil 5C18-MS-II, MeOH–H2O (80 : 20,
v/v)] to furnish bayogenin5,13,14) (1.2 mg, 52.2% from 1a, 1.2 mg, 50.0%
from 2a, 1.0 mg, 65.8% from 3a, 0.7 mg, 61.4%, from 4a, and 0.5 mg,
71.7% from 5), respectively. On the other hand, the aqueous layer was sub-
jected to HPLC analysis under the following conditions: HPLC column, Ka-
seisorb LC NH2-60-5, 4.6 mm i.d.ꢈ250 mm (Tokyo Kasei Co., Ltd., Tokyo,
Japan); detection, optical rotation [Shodex OR-2 (Showa Denko Co., Ltd.,
Tokyo, Japan); mobile phase, CH3CN–H2O (85 : 15, v/v); flow rate
0.5 ml/min]. Identification of L-rhamnose (i) from 1a—4a and 5, D-fucose
(ii) from 1a and 2a, D-glucose (iii) from 1a—4a and 5, and D-galactose (iv)
from 2a, 4a, and 5 present in the aqueous layer was carried out by compari-
son of their retention time and optical rotation with those of authentic sam-
ples. tR: (i) 12.0 min (negative optical rotation), (ii) 15.5 min (positive optical 10) Wang L.-B., Morikawa T., Nakamura S., Ninomiya K., Matsuda H.,
rotation, (iii) 20.7 min (positive optical rotation), and (iv) 22.2 min (positive
Muraoka O., Wu L.-J., Yoshikawa M., Heterocycles, 78, 1235—1242
optical rotation).
(2009).
Animals Male ddY mice were purchased from Kiwa Laboratory Ani- 11) Morikawa T., Wang L.-B., Ninomiya K., Nakamura S., Matsuda H.,
mal Co., Ltd. (Wakayama, Japan). The animals were housed at a constant
temperature of 23ꢆ2 °C and were fed a standard laboratory chow (MF, Ori-
ental Yeast Co., Ltd., Tokyo, Japan). The animals were fasted for 20—24 h
prior to the beginning of experiments, but were allowed free access to tap
water. All experiments were performed using conscious mice unless other-
Muraoka O., Wu L.-J., Yoshikawa M., Chem. Pharm. Bull., 57, 853—
859 (2009).
12) Morikawa T., Li X., Nishida E., Nakamura S., Ninomiya K., Matsuda
H., Oda Y., Muraoka O., Yoshikawa M., Helv. Chim. Acta, 93, 573—
586 (2010).
wise noted. The experimental protocol was approved by the Experimental 13) Eade R. A., Simes J. J. H., Stevenson B., Aust. J. Chem., 16, 900—905
Animal Research Committee of Kyoto Pharmaceutical University.
(1963).
Effect on Gastric Emptying in Olive Oil-Loaded Mice Gastric emp- 14) Kasai R., Miyakoshi M., Nie R.-L., Zhou J., Matsumoto K., Morita T.,
tying was determined by a modification of the phenol red method.16) Briefly,
each test sample suspended in 5% (w/v) acacia solution (10 ml/kg) was ad-
ministrated orally to fasted mice (ca. 30 g), and olive oil (0.15 ml/mouse)
containing 0.05% phenol red as a marker was given orally 30 min thereafter.
Two hours later, the mice were sacrificed by cervical dislocation under ether
anesthesia. The abdominal cavity was opened, and the gastroesophageal
junction and pylorus were clamped, then the stomach was removed,
weighted, and placed in 10 ml of 0.1 M NaOH and homogenized. The sus-
pension was allowed to settle for 1 h at room temperature, 1 ml of the super-
natant was added to 0.1 ml of 20% (w/v) trichloroacetic acid, and then the
mixture was centrifuged at 3000 rpm for 20 min. The supernatant (0.1 ml)
was mixed with 0.1 ml of 0.5 M NaOH, and the amount of phenol red was
determined from the optical density (OD) at 560 nm using a microplate
reader (SH-1000 Lab., Corona Electric Co., Ltd.). Escin IIa was used as a
reference compound.17—19) Gastric emptying (%) in the 30 min period was
calculated according to the following equation:
Nishi M., Miyahara K., Tanaka O., Phytochemistry, 27, 1439—1446
(1988).
15) The 1H- and 13C-NMR spectra of 1—5 and 1a—4a were assigned with
the aid of distortionless enhancement by polarization transfer (DEPT),
double quantum filter correlation spectroscopy (DQF COSY), hetero-
nuclear multiple quantum coherence (HMQC), heteronuclear multiple
bond correlation spectroscopy (HMBC), and total correlation spec-
troscopy (TOCSY) experiments.
16) Shimoda H., Ninomiya K., Nishida N., Yoshino T., Morikawa T.,
Matsuda H., Yoshikawa M., Bioorg. Med. Chem. Lett., 13, 223—228
(2003).
17) Matsuda H., Murakami T., Li Y., Yamahara J., Yoshikawa M., Bioorg.
Med. Chem., 6, 1019—1023 (1998).
18) Matsuda H., Li Y., Murakami T., Yamahara J., Yoshikawa M., Eur. J.
Pharmacol., 368, 237—243 (1999).
19) Matsuda H., Li Y., Yoshikawa M., Life Sci., 66, PL41—PL46 (2000).