Phlomisflavosides A and B, new flavonol bisglycosides from Phlomis spinidens

Article abstract of DOI:10.1248/cpb.49.1039

From the aerial parts of Phlomis spinidens, two new flavonol bisglycosides, phlomisflavosides A (1) and B (2), were isolated together with the known compounds, astragalin, isoquercitrin, lamiridoside, phlomoside A, shanzhiside methyl ester, 8-O-acetylshanzhiside methyl ester, phlorigidoside C, rodioloside (=salidroside), forsythoside B, citroside A and lariciresinol-4′-O-β-D-glucoside. The structures of the new compounds were elucidated based on spectral and chemical evidence.

Full text of DOI:10.1248/cpb.49.1039

August 2001
Notes
Chem. Pharm. Bull. 49(8) 1039—1041 (2001)
1039
Phlomisflavosides A and B, New Flavonol Bisglycosides from
Phlomis spinidens
,
a
a
a
b
c
Yoshio TAKEDA,* Natsuko ISAI, Toshiya MASUDA, Gisho HONDA, Yoshihisa TAKAISHI,
b
d
e
e
Michiho ITO, Hideaki OTSUKA, Ozodbek A. ASHURMETOV, and Olimjon K. KHODZHIMATOV
a
Faculty of Integrated Arts and Sciences, The University of Tokushima, Tokushima 770–8502, Japan, Graduate School of
b
Pharmaceutical Sciences, Kyoto University, Sakyo-ku, Kyoto 606–8501, Japan, Faculty of Pharmaceutical Sciences, The
c
University of Tokushima, Tokushima 770–8505, Japan, Institute of Pharmaceutical Sciences, Hiroshima University
d
Faculty of Medicine, Minami-ku, Hiroshima 734–8551, Japan, and Institute of Botany and Botanical Garden of Academy
e
of Sciences, Uzbek, Tashkent 700143, Uzbekistan. Received February 28, 2001; accepted April 21, 2001
From the aerial parts of Phlomis spinidens, two new flavonol bisglycosides, phlomisflavosides A (1) and B (2),
were isolated together with the known compounds, astragalin, isoquercitrin, lamiridoside, phlomoside A,
shanzhiside methyl ester, 8-O-acetylshanzhiside methyl ester, phlorigidoside C, rodioloside (؍salidroside),
forsythoside B, citroside A and lariciresinol-4؅-O-b-D-glucoside. The structures of the new compounds were eluci-
dated based on spectral and chemical evidence.
Key words Phlomis spinidens; Labiatae; phlomisflavoside A; phlomisflavoside B; flavonol bisglycoside
The plants classified to the genus Phlomis (Labiatae) have of b-glucopyranose and b-apiofuranose moieties which are
been known to contain several classes of compounds: iridoid linked to different oxygen atoms. GC analysis of the
1
)
2)
glucosides, phenyl-ethanoid glycosides, flavonoid glyco- methanolysis product of 1 revealed the presence of glucose
3
)
4)
5)
16)
13
sides, diterpene glycosyl ester, and nortriterpenes. Dur- and apiose moieties in the structure. The C-signals due to
ing the course of our studies on the constituents of traditional the aglycone part and b-glucopyranose unit are essentially
medicine and related plant material in the Uzbekistan Repub- the same as those of 4, whereas the signals due to ring B are
lic, we examined the glycosidic constituents of Phlomis slightly different from those of 4. On irradiation of the
spinidens NEVSKI (local name: Kuz kulok) which was used anomeric proton of apiofuranose moiety [d 5.58], a differ-
H
6
)
for treatment of allergy, and isolated two new flavonol bis- ential nuclear Overhauser enhancement (NOE) was observed
glycosides, termed phlomisflavosides A (1) and B (2) together for the proton at d 7.10 (H-5Ј) confirming that the apiofura-
7
)
with the known compounds, astragalin (3), isoquercitrin nose unit was connected to O-4Ј. A heteronuclear multiple
8
)
9)
1a)
(
4), lamiridoside, phlomoside A, shanzhiside methyl bond correlation (HMBC) experiment also supported these
1
0)
11)
ester, 8-O-acetylshanzhiside methyl ester, phlorigidoside results. Thus, phlomisflavoside A was elucidated as shown as
1
d)
12)
13)
C, rodioloside (ϭsalidroside), forsythoside B, citro- 1. In fact, partial hydrolysis of 1 with HCl-dioxane gave 4.
1
4)
15)
side A and lariciresinol-4Ј-O-b-D-glucoside. This paper
describes isolation and structure elucidation of the new com- amorphous powder, [a]D Ϫ93.5° (50% aq. pyridine). The
pounds.
molecular formula was assigned as C H O based on its
Compound 2 was also isolated as yellow colored
2
6
28 15
Compound 1 was isolated as yellow colored amorphous negative ion HR-FAB-MS, which is one oxygen less than
1
powder, [a] Ϫ140° (50% aq. pyridine) and gave a positive that of 1. The H-NMR spectrum of 2 exhibited A X type
D
2
2
coloration in the Mg–HCl test. The molecular formula was signals at d 7.10 and 8.08 (each 2H, d, Jϭ9.0 Hz) instead of
assigned as C H O based on its negative ion high resolu- ABX type signals observed in 1, and the other signals were
2
6
28 16
tion (HR)-FAB-MS. The UV spectrum showed maxima at very similar to those of 1. GC analysis of the methanolysis
57 and 351 nm, and the IR spectrum exhibited absorption product of 2 revealed the presence of glucose and apiose
2
Ϫ1
16)
13
bands ascribed to hydroxyls (3400 cm ), a,b-unsaturated moieties in the structure, and the C-NMR spectrum
Ϫ1
Ϫ1
ketone (1655 cm ) and aromatic rings (1606, 1502 cm ). (Table 1) also reflected the above mentioned differences.
Thus, 1 was suggested to be flavonol glycoside from its char- Acetylation of 2 gave nonaacetate (2b) [d 1.92, 2.00, 2.02,
acteristic color reaction and spectral properties. In systematic 2.08, 2.11, 2.15, 2.16, 2.34, 2.45 (each, 3H, s)]. On irradia-
investigation of the UV spectra, bathochromic shifts were ob-
served on addition of sodium acetate and aluminum chloride,
indicating that hydroxyl groups are located at C-5 and C-7
positions. Acetylation of 1 gave the decaacetate (1a) [d
H
1
3
.92, 2.00, 2.03, 2.07, 2.12, 2.14, 2.16, 2.34, 2.37, 2.45 (each
1
H, s)]. The H-NMR spectrum of 1 in DMSO-d showed an
6
ABX system due to 1,3,4-trisubstituted aromatic ring and
two meta-coupled doublets assigned to H-6 and H-8. Thus,
1
the aglycone part of 1 was analyzed as quercetin. The H-
NMR spectrum also showed signals due to anomeric protons
at d 5.37 (1H, d, Jϭ7.7 Hz, H-1Љ) and 5.58 (1H, d, Jϭ2.9 Hz,
1
3
H-1ٞ). The C-NMR spectrum (Table 1) showed, in addition
to the signals due to those of the aglycone part, the presence
∗
To whom correspondence should be addressed. e-mail: takeda@ias.tokushima-u.ac.jp
© 2001 Pharmaceutical Society of Japan

1
040
Vol. 49, No. 8
13
matographed over silica gel (400 g) with a mixture of CHCl and MeOH
Table 1.
C
C-NMR Data for 1, 2 and 4
3
with increasing amount of MeOH content. CHCl3 (2.4 l), CHCl –MeOH
3
a)
b)
b)
(19 : 1, 2 l), CHCl –MeOH (9 : 1, 2.4 l), CHCl –MeOH (4 : 1, 3.4 l), CHCl –
1
2
4
3
3
3
MeOH (3 : 1, 2.4 l) and CHCl –MeOH (7 : 3, 3.4 l) were eluted successively,
3
collecting 200 ml fractions. Fraction Nos. 34—39 gave a residue (489 mg),
2
3
4
5
6
7
8
9
0
1
2
3
4
5
6
1
2
3
4
5
6
1
2
3
4
5
155.6 (158.4)
134.2 (136.0)
177.3 (179.5)
161.1 (163.0)
99.3 (100.0)
166.0 (166.1)
94.0 (94.8)
158.4
135.9
179.4
162.9
100.1
166.2
94.9
158.4
105.7
125.1
132.0
116.9
160.6
116.9
132.0
104.2
75.7
78.0
71.3
78.3
62.6
108.4
78.4
80.4
75.8
64.8
158.2
135.6
179.3
162.8
99.9
an aliquot (101 mg) of which was separated by HPLC (solvent: MeOH–H O
2
1
0)
1d)
1
: 4) to give shanzhiside methyl ester (8.7 mg), phlorigidoside C (18.9
mg) and rodioloside (ϭsalidroside) (2.5 mg). Fraction Nos. 40—42 gave a
1
2)
residue (488 mg), an aliquot (100 mg) of which was separated by HPLC
1
2)
10)
as above to give phlomoside A (50.8 mg), shanzhiside methyl ester
(
165.9
94.8
1
4)
30.4 mg) and citroside A (3.7 mg). Fraction Nos. 43—47 gave a residue
418 mg), an aliquot (200 mg) of which was separated by HPLC as above
to give lamiridoside (59.0 mg). Fraction Nos. 21—32 in the HP-20 chro-
(
156.8 (158.4)
103.6 (105.8)
124.5 (125.8)
116.7 (116.7)
146.8 (147.7)
147.0 (148.6)
116.4 (118.0)
121.1 (122.7)
101.7 (104.2)
74.2 (75.7)
158.9
105.5
123.2
116.0
145.7
149.8
117.6
122.9
104.6
75.7
9
)
1
matography were combined and concentrated in vacuo to give a residue
Ј
Ј
Ј
Ј
Ј
Ј
Љ
Љ
Љ
Љ
Љ
Љ
ٞ
ٞ
ٞ
ٞ
ٞ
(
24.3 g) which was chromatographed over silica gel (1 kg) with a mixture of
CHCl and MeOH with increasing amounts of MeOH content. CHCl (7.5 l),
3
3
CHCl –MeOH (97 : 3, 7.5 l), CHCl –MeOH (19 : 1, 7.5 l), CHCl –MeOH
3
3
3
(
93 : 7, 5 l), CHCl –MeOH (9 : 1, 5 l), CHCl –MeOH (22 : 3, 7.5 l), CHCl –
3
3
3
MeOH (17 : 3, 7.5 l), CHCl –MeOH (4 : 1, 7.5 l), CHCl –MeOH (7 : 3, 7.5 l)
3
3
and MeOH (2.5 l) were eluted, collecting 500 ml fractions. Fraction Nos.
4—58 were combined and evaporated in vacuo to give a residue which was
separated by chromatography over silica gel (solvent: CHCl –MeOH with
3
c)
d)
76.6 (78.0)
78.0
71.1
78.2
62.5
3
11)
increasing amount of MeOH) to give 8-O-acetylshanzhiside methyl ester
70.2 (71.2)
77.3 (78.2)
c)
d)
(116 mg). Fraction Nos. 101—130 were combined and evaporated in vacuo
to give a residue (5.09 g) which was further separated by repeated chro-
61.2 (62.6)
matography over silica gel (solvent : CHCl –MeOH with increasing amounts
108.1 (109.4)
3
c)
d)
of MeOH) and Lobar RP-18 (solvent: MeOH–H O, 7 : 13) to give phlomis-
76.5 (78.3)
2
flavoside A (1) (398 mg) and phlomisflavoside B (2) (56.6 mg). Fraction
Nos. 33—35 in the HP-20 chromatography gave a residue (10.6 g) on evapo-
ration in vacuo which was separated by repeated silica gel chromatography
78.7 (80.4)
74.6 (75.9)
63.4 (64.9)
(
solvent: CHCl –MeOH with increasing amount of MeOH content) and
3
a) Measured in DMSO-d . The figures in parentheses are for CD OD solution.
HPLC (solvent: MeOH–H O, 2 : 3) to give additional 1 (31.3 mg) and 2
6
3
2
7
)
8)
b) Measured in CD OD. c—d) May be interchanged.
(48.9 mg) together with astragalin (3) (15.0 mg), isoquercitrin (4)
3
1
3)
(
74.3 mg), forsythoside B
(58.4 mg) and lariciresinol-4Ј-O-b-D-gluco-
1
5)
side (5.2 mg). Compounds 1 and 2 showed Rf values (0.16 and 0.24, re-
spectively) on silica gel TLC (solvent: CHCl –MeOH–H O, 15 : 6 : 1).
tion at d 5.60 due to the anomeric proton of apiofuranose
moiety, a differential NOE for the signal at d 7.10 was ob-
served. Thus, phlomisflavoside B was elucidated as shown as
3
2
Known compounds isolated were identified by comparisons of spectral data
with those reported.
21
Compound 1 Yellow colored amorphous powder, [a]D Ϫ140° (cϭ
2
. In fact, partial hydrolysis of 2 with 2% HCl-MeOH gave 3. 0.55, 50% aq. pyridine). UV l
(MeOH) nm (log e): 257 (4.39), 351
max
(
4.24); lmax (ϩAlCl ) nm: 263, 273, 355, 398; l
(ϩAlCl ϩHCl) nm:
max 3
3
Experimental
263, 273, 354, 398; lmax (ϩNaOAc) nm: 274, 320, 368; (ϩCH ONa) nm:
3
Ϫ1
Optical rotations were measured on a JASCO DIP-360 digital polarimeter. 272, 384; lmax (ϩH BO ) nm: 257, 351. IR n (KBr) cm : 3400, 1655,
3
3
max
1
IR spectra were measured on a Horiba Fourier transform infrared spectrome- 1606, 1502. H-NMR (DMSO-d , 90 °C) d: 3.16 (1H, m, H-4Љ), 3.26 (2H,
6
1
13
ter and UV spectra on a JASCO V-530 SR spectrophotometer. H- and C- H-2Љ, 3Љ), 3.40 (1H, dd, Jϭ11.8, 5.2 Hz, Ha-6Љ), 3.49 and 3.53 (each 1H, d,
NMR spectra were taken on a JEOL JNM EX-400 or a-400 spectrometer at Jϭ11.2 Hz, H-5ٞ), 3.59 (1H, dd, Jϭ11.8, 2.0 Hz, H -6Љ), 3.78 and 4.08
b
4
00 and 100 MHz with tetramethylsilane as an internal standard. HR-FAB- (each 1H, d, Jϭ9.4 Hz, H-4ٞ), 4.21 (1H, d, Jϭ2.9 Hz, H-2ٞ), 5.37 (1H, d,
MS were performed on a JEOL JMS SX-102 spectrometer with polyethyl-
Jϭ7.7 Hz, H-1Љ), 5.58 (1H, d, Jϭ2.9 Hz, H-1ٞ), 6.16 (1H, d, Jϭ2.0 Hz, H-
ene glycol-400 or 600 as calibration matrices. Column chromatography was 6), 6.34 (1H, d, Jϭ2.0 Hz, H-8), 7.10 (1H, d, Jϭ9.1 Hz, H-5Ј), 7.60 (1H, dd,
performed on Diaion HP-20 (Mitsubishi Chemical Co. Ltd., Tokyo), silica Jϭ9.1, 2.2 Hz, H-6Ј), 7.61 (1H, br s, H-2Ј), 12.37 (br s, 5-OH); (CD OD) d:
3
gel 60 (230—400 mesh, Merck) and Lober LiChroprep RP-18 column (40— 3.22 (1H, m, H-5Љ), 3.33 (1H, dd, Jϭ9.3, 9.3 Hz, H-4Љ), 3.42 (1H, dd, Jϭ9,
6
3 mm, fϭ25 mm, Lϭ310 mm, Merck), and TLC and preparative TLC were 9 Hz, H-3Љ), 3.47 (1H, dd, Jϭ9.3, 7.3 Hz, H-2Љ), 3.56 (1H, dd, Jϭ12.0,
performed on precoated silica gel plate 60 F (0.25 and 0.5 mm in thick-
5.4 Hz, H -6Љ), 3.68 (2H, s, H -5ٞ), 3.72 (1H, dd, Jϭ12.0, 2.0 Hz, H -6Љ),
2
54
a
2
b
ness, Merck). HPLC was performed on octadecylsilica gel (Cosmosil 3.91 and 4.15 (each 1H, d, Jϭ9.5 Hz, H-4ٞ), 4.35 (1H, br s, H-2ٞ), 5.29 (1H,
5
C18AR, Nacalai Tesque, Kyoto, fϭ20 mm, Lϭ250 mm) with a mixture of d, Jϭ7.3 Hz, H-1Љ), 5.66 (1H, br s, H-1ٞ), 6.20 (1H, br s, H-6), 6.39 (1H,
H O and MeOH at a flow rate of 6 ml/min, and the eluate was monitored by
br s, H-8), 7.18 (1H, d, Jϭ8.5 Hz, H-5Ј), 7.64 (1H, dd, Jϭ8.5, 1.5 Hz, H-6Ј),
2
1
3
UV (230 nm).
7.71 (1H, br s, H-2Ј), C-NMR: given in Table 1. HR-FAB-MS (negative)
Ϫ
Plant Material The aerial parts of Phlomis spinidens NEVSKI were har-
vested in June, 1997, in Uzbekistan Republic and identified by Dr. F. O.
m/z: 595.1340 [MϪH] (Calcd for C H O : 595.1299).
2
6
27 16
2
2
Compound 2 Yellow colored amorphous powder, [a]D Ϫ93.5° (cϭ
Khassanof of the Institute of Botany, Academy of Sciences, Uzbekistan Re- 0.64, 50% aq. pyridine). UV lmax (MeOH) nm (log e): 265 (4.21), 344
public. A voucher specimen (97B051) is deposited in the Herbarium of the (4.02); lmax (ϩAlCl ) nm: 275.5, 300, 341, 395; l (ϩAlCl ϩHCl): 276,
3
max
3
Graduate School of Pharmaceutical Sciences, Kyoto University.
300, 341, 395; lmax (ϩNaOAc) nm: 274, 357.5; lmax (ϩCH ONa) nm: 276,
3
Ϫ1
Extraction and Isolation The dried aerial parts (2.7 kg) of P. spinidens 372; l
were extracted with MeOH (27 lϫ2) for 3 weeks at room temperature. The 1502. H-NMR (DMSO-d , 90 °C): 3.13—3.28 (3H, H-2Љ, 3Љ, 4Љ), 3.38 (1H,
(ϩH BO ) nm: 265, 344. IR n
(KBr) cm : 3402, 1655, 1606,
max
max
3
3
1
6
MeOH extract was concentrated in vacuo to give a residue which was dis- dd, Jϭ11.7, 4.9 Hz, H -6Љ), 3.44, 3.49 (each 1H, d, Jϭ11.1 Hz, H-5ٞ), 3.58
a
solved in 90% MeOH (1 l). The 90% MeOH solution was washed with n- (1H, dd, Jϭ11.7, 2.2 Hz, H -6Љ), 3.78, 4.07 (each 1H, d, Jϭ9.3 Hz, H-4ٞ),
b
hexane (1 lϫ3) and then concentrated in vacuo. The residue was suspended 4.14 (1H, d, Jϭ3.2 Hz, H-2ٞ), 5.36 (1H, d, Jϭ7.1 Hz, H-1Љ), 5.60 (1H, d,
in H O (1 l) and the suspension was extracted with EtOAc (1 lϫ3) and n-
BuOH (1 lϫ3), successively. The n-BuOH extract was concentrated in vacuo
to give a residue (56.7 g) which was chromatographed over Diaion HP-20
Jϭ3.2 Hz, H-1ٞ), 6.12 (1H, d, Jϭ2.0 Hz, H-6), 6.31 (1H, d, Jϭ2.0 Hz, H-8),
7.10 (2H, d, Jϭ9.0 Hz, H-3Ј, 5Ј), 8.08 (2H, d, Jϭ9.0 Hz, H-2Ј, 6Ј), 12.34
(1H, br s, 5-OH); (CD OD) d: 3.21 (1H, m, H-5Љ), 3.39—3.46 (2H, H -2, 3),
2
3
2
(
fϭ11 cm, Lϭ35 cm) with a mixture of H O and MeOH with increasing 3.53 (1H, dd, Jϭ11.7, 5.4 Hz, H -6Љ), 3.65 (2H, s, H-5ٞ), 3.71 (1H, br d,
2
a
MeOH content. Six liters of 0, 10, 30, 40, 50, 70% aqueous MeOH and Jϭ11.7 Hz, H -6Љ), 3.91, 4.13 (each 1H, d, Jϭ9.8 Hz, H-4ٞ), 4.26 (1H, br s,
b
MeOH were eluted collecting 1 l fractions. Fraction Nos. 14—20 were com- H-2ٞ), 5.26 (1H, d, Jϭ7.3 Hz, H-1Љ), 5.68 (1H, br s, H-1ٞ), 6.20 (1H, br s,
bined and concentrated in vacuo to give a residue (5.95 g) which was chro- H-6), 6.40 (1H, br s, H-8), 7.13 (2H, d, Jϭ8.3 Hz, H-3Ј, 5Ј), 8.12 (2H, d,

August 2001
1041
13
Jϭ8.3 Hz, H-2Ј, 6Ј). C-NMR: given in Table 1. HR-FAB-MS (negative) part by a grants-in-aid from the Ministry of Education, Science, Sports, Cul-
Ϫ
m/z: 579.1346 [MϪH] (Calcd for C H O : 579.1350).
ture and Technology of Japan (No: 09041182 and 12576027).
2
6
27 15
Phlomisflavoside A Decaacetate (1b) Compound 1 (3.3 mg) was dis-
solved in pyridine (0.1 ml) and Ac O (0.1 ml) and the solution was kept References
2
overnight at 60 °C. Excess MeOH was added to the solution and the solvent
was removed in vacuo to give a residue which was purified by preparative
TLC (solvent: CHCl –Me CO, 9 : 1) to give the decaacetate (1b) (4.0 mg).
1) a) Muksudov M. S., Maksimov E. S., Umarova R. U., Saatov Z., Ab-
dullaev N. D., Khim. Prir. Soed., 1995, 243—246; b) Al-Hazinn H. M.
G., Alkhathlan H. Z., J. Chem. Soc. Pak., 18, 336—357 (1996); c)
Kasai R., Katagiri M., Ohtani K., Yamasaki K., Yang C.-R., Tanaka O.,
Phytochemistry, 36, 967—970 (1994); d) Takeda Y., Matsumura H.,
Masuda J., Honda G., Otsuka H., Takaishi Y., Sezik E., Yesilada E.,
ibid., 53, 931—935 (2000).
3
2
1
H-NMR (CDCl ) d: 1.92, 2.00, 2.03, 2.07, 2.12, 2.14, 2.16, 2.34, 2.37, 2.45
3
(
each 3H, s, 10ϫOAc), 3.60 (1H, m, H-5Љ), 3.91 (1H, dd, Jϭ12.5, 2.4 Hz,
H -6Љ), 4.01 (1H, dd, Jϭ12.5, 4.4 Hz, H -6Љ), 4.31, 4.39 (each 1H, d,
a
b
Jϭ10.7 Hz, H-4ٞ), 4.58 and 4.90 (each 1H, d, Jϭ12.5 Hz, H-5ٞ), 5.06 (1H,
dd, Jϭ9.8, 9.8 Hz, H-3Љ), 5.18 (1H, dd, Jϭ9.8, 7.8 Hz, H-2Љ), 5.29 (1H, dd,
Jϭ9.8, 9.8 Hz, H-4Љ), 5.58 (1H, d, Jϭ7.8 Hz, H-1Љ), 5.66 (1H, s, H-2ٞ), 5.71
2) a) Jimenez C., Riguera R., Nat. Prod. Rep., 11, 591—606 (1994); b)
Saracoglu I., Kojima K., Harput U. S., Ogihara Y., Chem. Pharm.
Bull., 46, 726—727 (1998); c) Takeda Y., Kinugawa M., Masuda T.,
Honda G., Otsuka H., Sezik E., Yesilada E., Phytochemistry, 51,
323—325 (1999).
(
1H, s, H-1ٞ), 6.83, 7.30 (each 1H, d, Jϭ2.0 Hz, H-6 and/or H-8), 7.30 (1H,
d, Jϭ7.3 Hz, H-5Ј), 7.77 (1H, d, Jϭ2.2 Hz, H-2Ј), 7.98 (1H, dd, Jϭ7.3, 2.2
ϩ
Hz, H-6Ј), HR-FAB-MS (positive) m/z: 1039.2377 [MϩNa] (ϩNaI) (Calcd
for C H O Na: 1039.2331).
Phlomisflavoside B Nonaacetate (2b) Compound 2 (2.7 mg) was
3) a) Harborne J. B., “The Flavonoid,” Chapman and Hall, London, 1988,
p. 317; b) Kumar R., Bhan S., Kallam A. K., Dhar K. L., Phytochem-
istry, 24, 1124—1125 (1985).
4) Katagiri M., Ohtani K., Kasai R., Yamasaki K., Yang C.-R., Tanaka O.,
Phytochemistry, 35, 439—442 (1994).
46
48 26
acetylated and the product purified as above to give the nonaacetate (2b)
1
(
2
2.8 mg). H-NMR (CDCl ) d: 1.92, 2.00, 2.02, 2.08, 2.11, 2.15, 2.16, 2.34,
3
.45 (each 3H, 9ϫOAc), 3.60 (1H, m, H-5Љ), 3.91 (1H, dd, Jϭ12.2, 2.4 Hz,
H -6Љ), 3.99 (1H, dd, Jϭ12.2, 4.2 Hz, H -6Љ), 4.29 and 4.42 (each 1H, d,
5) Kumar R., Bhan S., Kalla A. K., Dhar K. L., Phytochemistry, 31,
2797—2799 (1992).
6) “Opredelitel Rastenii Srednei Azii (Key for determining plants of
Middle Asia),” ed. by Adylov J. A., Vol. IX, Fan, Tashkent, 1987, p.
110.
a
b
Jϭ10.5 Hz, H-4ٞ), 4.68 and 4.87 (each 1H, d, Jϭ12.4 Hz, H-5ٞ), 5.06 (1H,
dd, Jϭ9.8, 9.8 Hz, H-3Љ), 5.17 (1H, dd, Jϭ8.1, 9.8 Hz, H-2Љ), 5.29 (1H, dd,
Jϭ9.8, 9.8 Hz, H-4Љ), 5.55 (1H, d, Jϭ8.1 Hz, H-1Љ), 5.67 (1H, s, H-2ٞ), 5.74
(
1H, s, H-1ٞ), 6.82, 7.29 (each 1H, d, Jϭ2.2 Hz, H-6 and/or H-8), 7.11 (2H,
d, Jϭ8.8 Hz, H-3Ј, 5Ј), 8.00 (2H, d, Jϭ8.8 Hz, H-2Ј, 6Ј). HR-FAB-MS (posi-
7) Nakabayashi T., J. Agr. Chem. Soc. Jpn., 26, 539—541 (1952).
8) a) Belley D. A., Justus Liebig’s Ann. Chem., 37, 101—113 (1841); b)
Hörhamner L., Wagner H., Arndt H., Discherl R., Farkas L., Chem.
Ber., 101, 450—453 (1968).
9) Eigtved P., Nielsen S. R., Nielsen B. J., Acta Chem. Scand., 28B, 85—
91 (1974).
ϩ
tive) m/z: 981.2286 [MϩNa] (ϩNaI) (Calcd for C H O Na: 981.2276).
4
4
46 24
Partial Hydrolysis of 1 Compound 1 (38.5 mg) was dissolved in a
mixture of 1 N aq. HCl (1.5 ml) and dioxane (1.5 ml) and the solution was
warmed at 60 °C for 15 min and then stirred at room temperature for 1 h.
The solution was neutralized by addition of Ag CO and the insoluble mate-
2
3
rial was filtered off. The filtrate was concentrated in vacuo. The residue was 10) Takeda Y., Nishimura H., Inouye H., Phytochemistry, 16, 1401—1404
purified by chromatography over silica gel (7 g, CHCl –MeOH with increas-
(1977).
ing amount of MeOH content) and HPLC (solvent: MeOH–H O, 9 : 11) to 11) Damtoft S., Jensen S. R., Nielsen B. J., Tetrahedron Lett., 23, 4155—
3
2
give isoquercitrin (4) (4.1 mg).
4156 (1982).
Partial Hydrolysis of 2 Compound 2 (37.3 mg) was dissolved in 2%
HCl–MeOH (5 ml) and the solution was kept at 40 °C for 1.5 h. After cool-
12) Shimomura H., Sashida Y., Adachi T., Phytochemistry, 26, 249—251
(1987).
ing, the solution was worked up as above to give a residue which was puri- 13) Endo K., Takahashi K., Abe T., Hikino H., Heterocycles, 19, 261—264
fied by chromatography over silica gel (as above) and finally HPLC (solvent:
(1982).
MeOH–H O, 2 : 3) to give astragalin (3) (9.0 mg).
14) Umehara K., Hattori I., Miyase T., Ueno A., Hara S., Kageyama C.,
Chem. Pharm. Bull., 36, 5004—5008 (1988).
2
Acknowledgements The authors thank Dr. F. O. Khassanof of the Insti- 15) Sugiyama M., Kikuchi M., Heterocycles, 36, 117—121 (1993).
tute of Botany, Uzbekistan Republic and the Cooperative Centre of the Uni- 16) Takeda Y., Zhang H., Matsumoto T., Otsuka H., Ooiso Y., Honda G.,
versity of Tokushima for identification of the plant material and for provid-
ing the facility for recording the NMR spectra. This work was supported in
Tabata M., Fujita T., Sun H., Sezik E., Yesilada E., Phytochemistry, 44,
117—120 (1997).