Glycosides of Atractylodes ovata.

Article abstract of DOI:10.1248/cpb.51.1106

A new coumarin glycoside and a new glycoside of an acetylene derivative were isolated from the water-soluble portion of the methanolic extract of Atractylodes ovata rhizome together with eight known compounds. Their structures were characterized as scopoletin beta-D-xylopyranosyl-(1-->6)-beta-D-glucopyranoside and (2E)-2-decene-4,6-diyne-1,8-diol 8-O-beta-D-apiofuranosyl-(1-->6)-beta-D-glucopyranoside, respectively, based on chemical and spectroscopic investigations. A comparison of the polar constituents among Atractylodes japonica, Atractylodes lancea, and A. ovata is led to the conclusion that A. ovata is distinguishable from A. lancea and A. japonica, as also shown by phylogenetic analysis.

Full text of DOI:10.1248/cpb.51.1106

1
106
Notes
Chem. Pharm. Bull. 51(9) 1106—1108 (2003)
Vol. 51, No. 9
Glycosides of Atractylodes ovata
,
a
a
a
a
b
Junichi KITAJIMA,* Akane KAMOSHITA, Toru ISHIKAWA, Akihito TAKANO, Tatsuo FUKUDA,
c
c
Susumu ISODA, and Yoshiteru IDA
a
b
Showa Pharmaceutical University; 3 Higashi-Tamagawagakuen, Machida, Tokyo 194–8543, Japan: Tokyo Metropolitan
c
Medical Plants Garden; 21–1 Nakajima-machi, Kodaira, Tokyo 187–0033, Japan: and Showa University School of
Pharmaceutical Sciences; Hatanodai, Shinagawa-ku, Tokyo 142–8555, Japan.
Received May 9, 2003; accepted June 20, 2003
A new coumarin glycoside and a new glycoside of an acetylene derivative were isolated from the water-solu-
ble portion of the methanolic extract of Atractylodes ovata rhizome together with eight known compounds. Their
structures were characterized as scopoletin b-D-xylopyranosyl-(1→6)-b-D-glucopyranoside and (2E)-2-decene-
4
,6-diyne-1,8-diol 8-O-b-D-apiofuranosyl-(1→6)-b-D-glucopyranoside, respectively, based on chemical and spec-
troscopic investigations. A comparison of the polar constituents among Atractylodes japonica, Atractylodes lancea,
and A. ovata is led to the conclusion that A. ovata is distinguishable from A. lancea and A. japonica, as also shown
by phylogenetic analysis.
Key words Atractylodes ovata; Atractylodes Rhizome; chemotaxonomy; scopoletin glycoside; 2-decene-4,6-diyne-1,8-diol gly-
coside
ϩ
In the previous paper, we reported the isolation and char- gested from the accurate mass number of the [MϩH] ion
acterization of eight sesquiterpenoid glycosides, including peak in the high-resolution positive FAB-MS.
atractyloside A—E, G, and a secoguaiane derivative, a
The major coumarin glycoside 1, C H O , showed
21 26 13
ϩ ϩ
monoterpenoid glucoside, seven aromatic compound glyco- [MϩK] and [MϩH] ion peaks at m/z 525 and 487, respec-
sides, and L-phenylalanine from the water-soluble portion of tively, in the positive FAB-MS. Enzymatic hydrolysis of 1
1
)
the rhizome of Atractylodes japonica. In addition, 16 gave an aglycone (1a), which was identified as scopoletin (7-
sesquiterpenoid glycosides, four monoterpenoid glucosides, hydroxy-6-methoxycoumarin), and D-glucose and D-xylose as
1
3
two hemiterpenoid glycosides, an alkyl glycoside, five aro- the sugar components. The C-NMR data of 1 were similar
6
)
matic compound glycosides, an acetylene derivative com- to those of scopoletin b-D-glucopyranoside (1b), except for
pound glucoside, two nucleosides, and L-tryptophan were the signals due to a b-D-xylopyranosyl group. Cross-peaks
isolated from the water-soluble portion of the rhizome of between the C-7/glucosyl H-1 and glucosyl C-6/xylosyl H-1
2
,3)
Atractylodes lancea. The rhizomes of Atractylodes plants were observed in the heteronuclear multiple bond connectiv-
are classified into two groups that contain b-eudesmol and ity (HMBC) spectrum of 1 (see Experimental), suggesting
hinesol as the main constituents of the essential oil (A. that the xylosyl group was located at C-6 of the glucose in
lancea and Atractylodes chinensis; so-jutsu), and atractylon 1b. This was also supported by the cross-peaks observed be-
as the main constituent of the essential oil (A. japonica and tween H-5/–OCH and between H-8/glucosyl H-1 in its nu-
3
Atractylodes ovata; byaku-jutsu) in the Japanese Pharma- clear Overhauser and exchange spectroscopy (NOESY) spec-
4
)
copoeia. Sometimes, however, the rhizomes are prescribed trum. Therefore 1 was characterized as scopoletin b-D-xy-
in traditional medicine indistinguishably. In addition, A. lopyranosyl-(1→6)-b-D-glucopyranoside.
ovata is distinguishable from other Atractylodes plants using
RAPD analysis, and the phylogenetic relationship between showed [MϩH] and [MϪC H O ϩH] ion peaks at m/z
Acetylene derivative compound glycoside 2, C H O ,
21 30 11
ϩ ϩ
5
)
1
1
20 10
A. japonica and A. lancea is suggested to be closer than that 459 and 183, respectively, in the positive FAB-MS, and
Ϫ
Ϫ
between A. japonica and A. ovata. We then undertook exami- [MϪH] and [MϪC H O ϪH] ion peaks at m/z 457 and
1
1
20 10
nation of the water-soluble portion of A. ovata to determine 181, respectively, in the negative FAB-MS. The NMR data
the chemotaxonomic relationships among A. ovata, A. japon- showed the presence of two triple bonds, one disubstituted
ica, and A. lancea.
double bond, one prim-methyl, one hydroxymethyl, one
The dried rhizome of A. ovata, which was cultivated in the methylene, and one oxygenated methine in addition to the b-
1
)
Tokyo Metropolitan Medical Plants Garden, was extracted D-apiofuranosyl-(1→6)-b-D-glucopyranosyl group. Com-
with 70% methanol, and the methanolic extract was sus- parison of its NMR data with those of (2E,8E)-decadiene-
pended in water and successively extracted with ether and 4,6-diyne-1,10-diol 1-O-b-D-glucopyranoside, which was
3
)
ethyl acetate. The aqueous layer was chromatographed on isolated from the rhizome of A. lancea, and the results of an
Amberlite XAD-II to give water and methanol eluate frac- HMBC experiment (Fig. 1) showed that the aglycone of 2
tions. The methanol eluate fraction was chromatographed on was 2-decene-4,6-diyne-1,8-diol, and the glycosyl group was
Sephadex LH-20 followed by silica gel, Lobar RP-8 column located at C-8. Therefore 2 was concluded to be (2E)-
chromatography, and HPLC, affording a coumarin glycoside decene-4,6-diyne-1,8-diol 8-O-b-D-apiofuranosyl-(1→6)-b-
(
1), an acetylene derivative compound glycoside (2), four D-glucopyranoside.
aromatic compound glycosides (3—6), three guaiane-type
Aromatic compound glycosides 3 to 6 and guaiane-type
sesquiterpenoid glucosides (7—9), and L-tryptophan (10). sesquiterpenoid glucosides 7 to 9 were identified as icariside
7
)
7)
7)
8)
Among them, 1 and 2 are new, and their structures were F , icariside D , syringin, dihydrosyrindine, atractylo-
2
1
2
)
3)
2)
characterized as follows. Their molecular formulae were sug- side A, 10-epi-atractyloside A, and atractyloside B, re-
∗
To whom correspondence should be addressed. e-mail: Kitajima@ac.shoyaku.ac.jp
© 2003 Pharmaceutical Society of Japan

September 2003
1107
spectively (Fig. 2).
(19 : 1)] to give 3 (65 mg), and fr. C6-10 was subjected to HPLC [ODS,
MeCN–H O (3 : 17)] to give 4 (20 mg). Fraction C (0.50 g) was passed
While the commom guaiane-type sesquiterpenoid gluco-
sides were found in the rhizomes of A. japonica, A. lancea,
and A. ovata, we were able to isolate the new characteristic
2
7
through a Lobar RP-8 column [MeCN–H O (3 : 17)] to give nine fractions
2
(
(
frs. C7-1—C7-9). Fraction C7-4 was recrystallized from methanol to give 1
200 mg), and fr. C7-8 was subjected to HPLC [ODS, MeCN–H O (3 : 17)
2
coumarin glycoside 1 as the main glycoside of the rhizome and CHA, MeCN–H O (24 : 1)] to give 2 (2 mg). Fraction C (0.33 g) was
2
9
passed through a Lobar RP-8 column [MeCN–H O (3 : 17)] to give seven
fractions (frs. C9-1—C9-7), and fr. C9-3 was subjected to HPLC [ODS,
of A. ovata. A comparison of the polar constituents among A.
japonica, A. lancea, and A. ovata (Table 1) showed that A.
ovata is distinguishable from A. lancea and A. japonica, as
did the phylogenetic analysis.
2
MeCN–H O (3 : 37)] to give 8 (7 mg) and 7 (40 mg). Fraction C (1.31 g)
2
11
was passed through a Lobar RP-8 column [MeCN–H O (3 : 17)] to give
2
seven fractions (frs. C11-1—C11-7). Fraction C11-3 was subjected to HPLC
[
CHA, MeCN–H O (14 : 1)] to give 9 (16 mg), and fr. C was subjected to
2
1
1
-
4
Experimental
Sephadex LH-20 (MeOH) to give 10 (15 mg).
The instruments used and the experimental conditions for spectral
mesurements and chromatography were the same as those reported in the
The following compounds were identified by comparison with authentic
compounds or published physical and spectral data: icariside F (3), icariside
D1 (4), syringin (5), dihydrosyrindine (6), atractyloside A (7), 10-epi-
atractyloside A (8), atractyloside B (9), and L-tryptophan (10).
2
1,3)
previous papers. Symmetryprep C18 7 mm (Waters; column size, 7.8ϫ
3
3
00 mm; ODS) and Carbohydrate analysis (Waters; column size, 3.9ϫ
00 mm; CHA) were used as columns for HPLC separations.
Scopoletin 7-O-b-D-Xylopyranosyl-(1→6)-b-D-glucopyranoside (1): Col-
2
3
Extraction and Separation The dried rhizome of A. ovata (1.5 kg),
orless needles (MeOH), mp 243—245 °C, [a] Ϫ148° (cϭ0.5, H O). Posi-
tive FAB-MS m/z: 525.1011 [MϩK] (Calcd for C H KO , 525.1011),
4
D
2
ϩ
which was cultivated in the Tokyo Metropolitan Medical Plant Garden (Ko-
daira, Tokyo, Japan), was extracted with 70% methanol (5 lϫ3) for 2 weeks,
and the extract (573.5 g) was partitioned between ether/water and then ethyl
acetate/water. The aqueous portion (555.3 g) was chromatographed over
Amberlite XAD-II (H O→MeOH) to give a water eluate (521.0 g) and a
methanol eluate (34.3 g).
2
1
26
13
ϩ
1
87.1465 [MϩH] (base, Calcd for C H O , 487.1452). H-NMR
21 27 13
(
DMSO-d , 500 MHz) d: 6.33 (1H, d, Jϭ9.5 Hz, H-3), 7.96 (1H, d,
6
Jϭ9.5 Hz, H-4), 7.30 (1H, s, H-5), 7.20 (1H. s. H-8), 3.82 (3H, s, 6-OCH ),
3
13
2
5.10 (1H, d, Jϭ7.5 Hz, Glc H-1), 4.12 (1H, d, Jϭ7.5 Hz, Xyl H-1). C-
NMR (DMSO-d , 125 MHz) d: 160.50 (C-2), 113.34 (C-3), 144.14 (C-4),
109.72 (C-5), 145.96 (C-6), 149.76 (C-7), 103.01 (C-8), 148.92 (C-9),
6
The methanol eluate was subjected to Sephadex LH-20 [MeOH–H O
2
(
9 : 1)] to give six fractions (frs. A—F). Fraction C (14.72 g) was chro-
1
12.31 (C-10), 56.01 (6-OCH ), 99.50 (Glc C-1), 72.99 (Glc C-2), 76.52
3
matographed on silica gel [CHCl –MeOH–H O (4 : 1 : 0.1→7 : 3 : 0.5→
3
2
(Glc C-3), 69.19 (Glc C-4), 75.33 (Glc C-5), 68.17 (Glc C-6), 104.02 (Xyl
C-1), 73.25 (Xyl C-2), 76.54 (Xyl C-3), 69.42 (Xyl C-4), 65.60 (Xyl C-5).
HMBC correlations: H-3/C-2, C-10; H-4/C-2, C-5, C-9, C-10; H-5/C-4, C-
6, C-7, C-9, C-10; H-8/C-6, C-7, C-9, C-10; –OCH /C-6; Glc H-1/C-7; Xyl
H-1/Glc C-6.
Enzymatic Hydrolysis of 1 A mixture of 1 (25 mg) and b-glucosidase
(5 mg; Toyobo Co. Ltd., Lot 93240) in water (5 ml) was shaken in a water
bath at 37 °C for 5 d. The mixture was concentrated in vacuo to dryness and
6
: 4 : 0.5)→MeOH] to give 15 fractions (frs. C —C ). Fraction C (0.13 g)
1 15 4
was passed through a Lobar RP-8 column [MeCN–H O (3 : 17)] to give 11
fractions (frs. C4-1—C4-11), and fr. C4-3 was subjected to HPLC [ODS,
2
3
MeCN–H O (3 : 17)] to give 5 (6 mg) and 6 (5 mg). Fraction C (0.13 g) was
2
6
passed through a Lobar RP-8 column [MeCN–H O (3 : 17)] to give 11 frac-
tions (frs. C6-1—C6-11), fr. C6-7 was subjected to HPLC [CHA, MeCN–H O
2
2
the residue was chromatographed over silica gel [CHCl –MeOH (4 : 1 to
3
1
: 1)] to afford an aglycone (1a; 10 mg) and a sugar fraction. The sugar frac-
tion was passed through Sephadex LH-20 (MeOH) to give a syrup, and
HPLC [carbohydrate analysis (Waters); detector, JASCO RI-930 detector
and JASCO OR-990 chiral detector; solv., MeCN–H O (9 : 1), 2 ml/min; tR
2
4
.20 min (same location as that of D-xylose) and tR 6.80 min (same location
as that of D-glucose)] showed the presence of D-glucose and D-xylose.
1
Scopoletin (1a): Colorless needles (EtOH), mp 205—207 °C, H-NMR
(
DMSO-d , 270 MHz) d: 6.21 (1H, d, Jϭ9.5 Hz, H-3), 7.91 (1H, d,
6
Fig. 1. Structure and HMBC Correlations of 2
Jϭ9.5 Hz, H-4), 7.21 (1H, s, H-5), 6.77 (1H. s. H-8), 3.81 (3H, s, 6-OCH ).
3
Fig. 2. Structures of 1—10

1
108
Vol. 51, No. 9
Table 1. Polar Constituents of A. japonica, A. lancea, and A. ovata Rhizome
Type
A. japonica
A. lancea
A. ovata
Sesquiterpenoid alcohol glycosides
Guaiane-type
7
8
9
1
2
4
5
ϩϩϩϩϩ
ϩϩ
ϩϩ
ϩϩϩ
ϩ
ϩϩϩϩϩ
ϩϩϩ
ϩϩ
ϩϩϩϩϩ
ϩϩϩ
ϩϩϩ
ϩϩ
ϩϩϩ
ϩ
ϩϩ
1
1
1
1
Secoatractylolactone-type
Eudesmane-type
13
35
ϩϩ
ϩϩϩ
ϩϩϩ
16
17
18
19
20
21
22
23
ϩϩϩϩ
ϩϩϩϩ
ϩ
ϩϩ
ϩϩ
ϩ
ϩϩϩ
ϩϩϩ
Hemiterpenoid alcohol glycosides
Monoterpenoid alcohol glucosides
2
2
8
9
ϩ
ϩ
1
,8-Cineole-type
24
ϩ
ϩ
ϩ
ϩ
ϩ
2
5
p-Menthane-type
26
2
7
Aromatic glycosides
C type
31
ϩ
ϩ
ϩ
ϩ
ϩϩ
ϩ
ϩ
ϩϩ
ϩϩ
6
3
3
3
2
3
6
3
4
7
5
1
6
C —C type
ϩϩ
ϩϩ
ϩϩϩϩ
ϩϩ
6
1
C —C type
6
2
3
ϩϩ
C —C type
ϩ
ϩϩϩϩϩϩ
ϩ
6
3
Acetylene glycoside
34
2
30
38
ϩ
ϩ
ϩ
Alkyl glycoside
Amino acids
ϩ
1
0
ϩϩϩϩ
ϩϩ
1
—9 mg, ϩ; 10—19 mg,ϩϩ; 20—39 mg, ϩϩϩ; 40—89 mg, ϩϩϩϩ; 90—199 mg, ϩϩϩϩϩ; м200 mg, ϩϩϩϩϩϩ (from 1.4 kg of A. japonica, 1.5 kg of A. lancea, and
.5 kg of A. ovata). 11: (1S,4S,5S,7R,10R)-10,11,14-trihydroxyguai-3-one 11-O-b-D-glucopyranoside, 12: (1S,4S,5R,7R,10R)-10,11,14-trihydroxyguai-3-one 11-O-b-D-glucopyra-
noside, 13: (1S,5R,7R,10R)-secoatractylolactone 11-O-b-D-glucopyranoside, 14: atractyloside A 14-O-b-D-fructofuranoside, 15: (1S,4S,5S,7R,10S)-10,11,14-trihydroxyguai-3-one
1
1
1-O-b-D-glucopyranoside, 16: (5R,7R,10S)-isopterocarpolone b-D-glucopyranoside, 17: atractyloside I, 18: cis-atractyloside I, 19: atractyloside C, 20: atractyloside D, 21: atracty-
loside E, 22: atractyloside G, 23: (2R,3R,5R,7R,10S)-atractyloside G 2-O-b-D-glucopyranoside, 24: (1R,2R,4S)-2-hydroxy-1,8-cineole b-D-glucopyranoside, 25: (1S,2S,4R)-2-hy-
droxy-1,8-cineole b-D-glucopyranoside, 26: (4S)-p-menth-1-ene-7,8-diol 8-O-b-D-glucopyranoside, 27: (1S,2R,4S)-p-menthane-1,2,8-triol 8-O-b-D-glucopyranoside, 28: 3-methyl-
3
-butenyl b-D-apiofuranosyl-(1→6)-b-D-glucopyranoside, 29: 3-methyl-2-butenyl b-D-apiofuranosyl-(1→6)-b-D-gluco-pyranoside, 30: isopropyl b-D-apiofuranosyl-(1→6)-b-D-
glucopyranoside, 31: 4-hydroxy-3-methoxyphenyl b-D-glucopyranoside, 32: 4-hydroxy-3-methoxyphenyl b-D-apiofuranosyl-(1→6)-b-D-glucopyranoside, 33: 4-hydroxy-3-
methoxyphenyl b-D-xylopyranosyl-(1→6)-b-D-glucopyranoside, 34: (2E,8E)-2,8-decadiene-4,6-diyne-1,10-diol 11-O-b-D-glucopyranoside, 35: (3R,5S,8S,10S)-3-hydroxya-
tractylerolide III 3-O-b-D-glucopyranoside, 36: seguinoside B, 37: phenethyl a-L-rhamnopyranosyl-(1→6)-b-D-glucopyranoside, 38: L-phenyalanine.
1
3
C-NMR (DMSO-d , 67.5 MHz) d: 160.72 (C-2), 111.51 (C-3), 144.52 (C- H2-9/C-7, C-8, C-10; H
-10/C-8, C-9; Glc H-1/C-8; Api H-1/Glc C-6.
6
3
4
1
), 109.56 (C-5), 145.36 (C-6), 151.49 (C-7), 102.77 (C-8), 149.61 (C-9),
10.40 (C-10), 56.01 (6-OCH3).
References
1
)
Kitajima J., Kamoshita A., Ishikawa T., Takano A., Fukuda T., Isoda
S., Ida Y., Chem. Pharm. Bull., 51, 152—157 (2003).
(
2E)-2-Decene-4,6-diyne-1,8-diol 8-O-b-D-Apiofuranosyl-(1→6)-b-D-
2
3
glucopyranoside (2): An amorphous powder, [a] Ϫ144° (cϭ0.1, MeOH).
Positive FAB-MS m/z: 459.1870 [MϩH] (Calcd for C H O , 459.1867),
1
[
br s, H -1), 6.67 (1H, td, Jϭ4.0, 16.0 Hz, H-2), 6.25 (1H, d, Jϭ16.0 Hz, H-
3
Jϭ7.5 Hz, H -10), 5.25 (1H, d, Jϭ7.5 Hz, Glc H-1), 5.81 (1H, d, Jϭ2.5 Hz,
Api H-1). C-NMR (pyridine-d , 125 MHz) d: 61.89 (C-1), 149.91 (C-2),
1
8
D
ϩ
2) Yahara S., Higashi T., Iwaki K., Nohara T., Marubayashi N., Ueda I.,
Kohoda H., Goto K., Izumi H., Nuno M., Katsuki S., Isoda S., Satake
M., Chem. Pharm. Bull., 37, 2995—3000 (1989).
2
1
31 11
ϩ
Ϫ
83 [MϪC H O ϩH] (base). Negative FAB-MS m/z: 457 [MϪH] , 181
11 20 10
Ϫ
1
MϪC H O ϪH] (base). H-NMR (pyridine-d , 500 MHz) d: 4.41 (2H,
11 20 10 5
3
)
Kitajima J., Kamoshita A., Ishikawa T., Takano A., Fukuda T., Isoda
S., Ida Y., Chem. Pharm. Bull., 51, 673—678 (2003).
2
), 5.11 (1H, dd, Jϭ6.5, 6.5 Hz, H-8), 1.84 (2H, m, H -9), 0.99 (3H, t,
2
4) “Japanese Pharmacopoeia,” 14th edition, Hirokawa Publishing Co.,
Tokyo, 2001, pp. 2708—2709, 2869—2871.
5) Kohjyouma M., Kohda H., Biol. Pharm. Bull., 20, 502—506 (1997).
3
13
5
06.92 (C-3), 77.92 (C-4), 74.33 (C-5), 71.20 (C-6), 82.26 (C-7), 69.14 (C-
), 29.34 (C-9), 9.76 (C-10), 101.91 (Glc C-1), 75.02 (Glc C-2), 78.53 (Glc
6
)
Tsukamoto H., Hisada S., Nishibe S., Chem. Pharm. Bull., 33, 396—
399 (1985).
C-3), 71.79 (Glc C-4), 77.34 (Glc C-5), 68.89 (Glc C-6), 111.31 (Api C-1),
7.80 (Api C-2), 80.45 (Api C-3), 75.03 (Api C-4), 65.57 (Api C-5). HMBC
correlations: H-2/C-1, C-3,C-4; H-3/C-5; H-8/C-6, C-7, C-9, C-10, Glc C-1;
7) Kitajima J., Ishikawa T., Tanaka Y., Ono M., Ito Y., Nohara T., Chem.
Pharm. Bull., 46, 1587—1590 (1998).
8) Kisiel W., Braszcz B., Fitoterapia, 71, 269—273 (2000).
7