Terpenoids và coumarin từ Atractylodes lancea phát triển tại Việt Nam

Terpenoids và coumarin từ Atractylodes lancea phát triển tại Việt Nam.

Journal of Chemistry, Vol 42 (4), P 499 - 502, 2004

Terpenoids and coumarin from Atractylodes lancea

growing in Vietnam

Received 29 th -Sept.-2003

Chau Van Minh1, Phan Van Kiem1, Hoang Thanh Huong1, Jung Joon Lee2

and Young Ho Kim3

1 Institute of Natural Products Chemistry, VAST

2 Korea Research Institute of Bioscience and Biotechnology, Korea

3 College of Pharmacy, Chungnam National University Korea

Summary

Two sesquiterpenes 4(15),11-eudesmadiene and -eudesmol, one coumarin osthol, and one pentacyclic triterpene 3-acetyl- -amyrin have been isolated from the dichloromethane extract of the roots of Atractylodes lancea growing in Vietnam, by various chromatography methods Their structures were determined by spectroscopic means such as FAB-MS, 1 H-, 13 C-NMR, DEPT 135 and DEPT 90 Of which, 4(15),11-eudesmadiene is the main compound of the constituents, and

3 -acetyl-amyrin was isolated for the first time from Atractylodes lancea

Keywords: Atractylodes lancea, 4(15),11-eudesmadiene, -eudesmol, osthol, 3 -acetyl-amyrin

I - Introduction

Atractylodes lancea (Thunb.) DC

(Vietnamese name is Thuong truat) has been

used to treat digestive disorders, mild

diarrhorea, rheumatic diseases, and influenza

[2] Previous phytochemical investigations of A

lancea, which growing in China, showed the

presence of polyacetylenes, atractylodin,

atract-ylodinol, acetylatractatract-ylodinol,

(4E,6E,12E)-tetradecatriene-8,10-diyne-1,3,14-triol [6, 8 -

10]; sesquiterpene-glycosides [14];

sesquiterpe-ne atractylon, the coumarin osthol,

atractyleno-lides I, II and III [12]; hinesol, -eudesmol [8]

Several reports deal with the composition of

essential oil [3, 15] Herein, we report the

isolation and structural elucidation of two

sesquiterpenes 1-2, the cumarin 3 and one

triterpene 4, which were obtained from the

dichloromethane extract of the roots of A

lancea growing in Vietnam Their structures

were determined as 4(15),11eudesmadiene, -eudesmol, osthol and 3-acetyl- -amyrin, respec-tively, by spectroscopic means

II- MATERIALS AND METHODS

General experimental procedures

FAB-MS was obtained using a JMS-SX 102 spectrometer 1H-NMR (300 MHz) and 13 C-NMR (75 MHz) were recorded on a Bruker DRX300 spectrometer and TMS was used as an internal standard Column chromatography (CC) was performed on silica gel (Kieselgel 60,

70-230 mesh and 70-230-400 mesh, Merck)

Plant material

The roots of A lancea was collected in

Langson province, Vietnam in January 2003 and identified by Prof Vu Van Chuyen, Hanoi University of Pharmacy A voucher specimen

was deposited at the herbarium of the Institute of Natural Product Chemistry, VAST, Vietnam

H

1 2

3

4 5 6

7 8 9 10

11 12

13 14

15

1

OH H

1 2 3

4 5 6

7 8 9 10

11 12

13

14

15

2

H3CO

2 3 4 5 6 7

8 9 10

1' 2' 3' 4' 5'

3

H3C-C-O

1 3

17

29 30

10

O

4

Fig 1: Structurtes of 1, 2, 3 and 4

Extraction and isolation

The dried and powdered roots of A lancea

(680 g) were extracted repeatedly with hot

MeOH three times The combined solutions

were evaporated under reduced pressure to give

MeOH extract (45.0 g), which was suspended in

water and then partitioned with

dichlorome-thane The dichloromethane fraction (32.0 g)

was then chromatographed on a silica gel

column, using hexane-acetone (from 100 : 1 to

5 : 1) as the eluent yielded five fractions

(AL.1-AL.5) Fraction AL.1 was then

chromatogra-phed on a silica gel column, using

hexane-ethylacetate (100:1) as the eluent yielded 1 (1.2

g) and 2 (28.0 mg) as white oils Fraction AL.3

was then chromatographed on a silica gel

column, using hexane-ethylacetate (100 : 2) as

the eluent yielded 3 (12.0 mg) and 4 (15.0 mg)

as white crystals

4(15),11-Eudesmadien (1): A white oil,

(CDCl3) (300 MHz) : 4.75 (2H, br s, H2-12),

4.74 (1H, s, Ha-15), 4.55 (1H, s, Hb-15), 1.82

(3H, s, H3-13) and 0.78 (3H, s, H3-14). 13

151.4 (s, C-4), 108.6 (t, C-12), 105.7 (t, C-15),

50.3 (d, C-5), 46.3 (d, C-7), 42.4 (t, C-1), 41.6

(t, 3), 37.3 (t, 9), 36.6 (s, 10), 29.9 (t, C-6), 27.2 (t, C-2), 23.9 (t, C-8), 21.4 (q, C-13) and 16.7 (q, C-14)

-Eudesmen-11-ol (2): A white oil, FAB-MS

(300 MHz) : 4.75 (1H, s, Ha-15), 4.55 (1H, s,

Hb-15), 1.25 (6H, s, H3-12, 13) and 0.71 (3H, s,

H3-14). 13C-NMR (CDCl3) (75 MHz) : 151.5 (s, C-4), 105.7 (t, C-15), 74.0 (s, C-11), 50.2 (d, C-5), 49.9 (d, C-7), 42.2 (t, C-1), 41.6 (t, C-3), 36.7 (t, C-9), 36.6 (s, C-10), 27.5 (q, C-12, 13), 25.4 (t, C-2), 23.9 (t, C-6), 22.7 (t, C-8) and 16.7 (q, C-14)

Osthol (3): White crystals, m.p 82 - 83o

C;

(CDCl3) (300 MHz) : 7.61 (1H, d, 9.6 Hz, H-4), 7.29 (1H, d, 8.4 Hz, H-5), 6.84 (1H, d, 8.4

Hz, H-6), 6.23 (1H, d, 9.6 Hz, H-3), 5.24 (t, 7.2,

Hz, H-2’), 3.93 (3H, s, C7-OCH3); 3.54 (2H, d, 7.2 Hz, H-1’) 1.85 (3H, s, H-4’) and 1.68 (3H,

s, H-5’) 13C-NMR (CDCl3) (75 MHz) : 161.7

(s, C-2), 160.6 (s, C-7), 153.2 (s, C-9), 144.1

(d, C-4), 133.0 (s, C-3’), 126.6 (d, C-5), 121.6 (d, C-2’), 118.4 (s, C-8), 113.4 (d, C-3), 113.3 (s, C-10), 107.8 (d, C-6), 56.4 (q, C7-OCH3), 26.2 (q, C-5’), 22.3 (t, C-1’) and 13.8 (q, C-4’)

3 -acetyl-amyrin (4): White crystals, m.p 241 -

242C; FAB-MS m/z: 469 [M+H] (positive);

1

H-NMR (300 MHz) : 5.21 (1H, t, 3.5, H-12),

4.54 (1H, dd, 6.0, 2.1, H-3), 2.07 (3H, s,

COCH3), 1.14 (3H, s, H3-27), 0.98 (6H, s, H3

-26, 25), 0.88 (12H, s, H3-30, 29, 24, 23) and

0.84 (3H, s, H3-28) 13C-NMR (CDCl3) (75

MHz) : 171.4 (s, CH3CO), 145.6 (s, C-13),

122.1 (d, C-12), 81.3 (d, C-3), 55.7 (d, C-5),

48.0 (d, C-18), 47.7 (d, C-9), 47.2 (t, C-19),

42.1 (s, C-14), 40.2 (s, C-8), 38.7 (t, C-1), 38.1

(s, C-4), 37.5 (t, C-22), 37.3 (s, C-10), 35.1 (t,

21), 33.7 (q, 29), 33.0 (t, 7), 32.9 (s,

17), 31.5 (s, 20), 28.8 (q, 23), 28.4 (q,

C-28), 27.3 (t, C-2), 26.6 (t, C-15), 26.3 (q, C-27),

24.1 (q, C-30), 24.0 (t, C-16), 23.9 (t, C-11),

21.7 (q, CH3CO), 18.7 (t, C-6), 17.2 (q, C-26),

17.1 (q, C-25) and 15.9 (q, C-24)

Iii - Results and discussion

Repeated column chromatography on silica

gel of the dichloromethane extract of the dried

roots of A lancea yielded 1-4 (see experimental

part) Compounds 1 and 2 were yielded as white

oils Compound 3 and 4 formed as white

crystals

The positive FAB-MS spectrum of 1 showed

the molecular ion at m/z 205 [M+H]+,

corresponded to the formula of C15H24 The 1H-

and 13C-NMR spectra of 1 confirmed the

presence of 15 carbons, including two >C=CH2

groups { H 4.75 (2H, br s, H2-12), 4.74 (1H, s,

Ha-15), 4.55 (1H, s, Hb-15); C 152.2 (s, C-11),

108.6 (t, C-12), 151.4 (s, C-4), 105.7 (t, C-15)};

Two tertiary methyl groups { H1.82 (3H, s,

H3-13) and 0.78 (3H, s, H3-14); C21.4 (q, C-13),

16.7 (q, C-14)} The spectral data of 1 were in

good agreement with those of reported data of

4(15),11-eudesmadiene [11], thus 1 was

identified to be 4(15),11-eudesmadiene, a

sesquiterpene

Compound 2 showed the molecular ion at

m/z 223 [M+H]+, in the positive FAB-MS

spectrum, which corresponded to the formula of

C15H26O The 1H- and 13C-NMR spectra of 2

also revealed 15 carbons of a sesquiterpene,

including one >C=CH2 group { H 4.75 (1H, s,

Ha-15) and 4.55 (1H, s, Hb-15); C151.5 (s, C-4) and 105.7 (t, C-15)}; three tertiary methyl groups { H1.25 (6H, s, H3-12, 13), 0.71 (3H, s,

H3-14); C27.5 (q, C-12, 13), 16.7 (q, C-14)}, a oxygene bearing tertiary carbon (74.0, s, C-11)

The structure of 2 was determined to be 4(15),

11-eudesmen-11-ol ( -eudesmol) by comparison

the spectral data of 2 with those of 1 and of the

reported data [7, 11]

Compound 3 showed the molecular ion at

m/z 245 [M+H]+ in the positive FAB-MS spectrum, which corresponded to the formula of C15H16O3 The 1H- and 13C-NMR spectral signals

of 2 almost assignable to those of a coumarin

including one methoxy group { H 3.93 (3H, s, 7-OCH3); C 56.4 (q, 7-OCH3)}; two tertiary methyl groups { H1.85 (3H, s, H-4’) and 1.68 (3H, s, H-5’); C13.8 (q, C-4’), 26.2 (q, C-5’)}

The spectral data of 3 matched well with those

of osthol in the reported data [5], thus 3 was

identified as osthol

The positive FAB-MS spectrum of 4 showed

the molecular ion at m/z 469 [M+H]+, corresponded to the formula of C32H52O2 The

MS and NMR spectra confirmed that 4 was a

pentacyclic-triterpene, which had eight methyl groups { H1.14 (3H, s, H3-27), 0.98 (6H, s, H3

-26, 25), 0.88 (12H, s, H3-30, 29, 24, 23), and 0.84 (3H, s, H3-28); C 33.7, 28.8, 28.4, 26.3, 24.1,17.2, 17.1, 15.9}; a trisubstituted double bond { H5.21 (1H, t, 3.5, H-12); C145.6 (s, C-13), 122.1 (d, C-12)}, and an acetoxy group { H 2.07 (3H, s, COCH3); C21.7 (q, CH3CO), 171.4 (s, CH3CO)} at axial configuration {4.54 (1H,

dd, J = 6.0, 2.1, H-3 )} [1,13] Based on above

data and comparison with the reported data

[1,13], compound 4 was identified to be 3

-acetyl-amyrin

This is first report of 4 from Atractylodes

lancea

Acknowledgements: This study was supported

by a grant from the Korea-Vietnam cooperation project We are grateful to the KBSI for measuring the MS and NMR spectra We thank

to Prof Vu Van Chuyen, Hanoi University of Pharmacy for the plant identification

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