Terpenoids và coumarin từ Atractylodes lancea phát triển tại Việt Nam.
499Journal of Chemistry, Vol. 42 (4), P. 499 - 502, 2004 Terpenoids and coumarin from Atractylodes lancea growing in Vietnam Received 29th-Sept.-2003 Chau Van Minh1, Phan Van Kiem1, Hoang Thanh Huong1, Jung Joon Lee2and Young Ho Kim31Institute of Natural Products Chemistry, VAST 2Korea Research Institute of Bioscience and Biotechnology, Korea 3College 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, 1H-, 13C-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, acetylatractylodinol, (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),11-eudesmadiene, -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 13C-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 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 500was deposited at the herbarium of the Institute of Natural Product Chemistry, VAST, Vietnam. H1234657891011121314151OHH1234657891011121314152OOH3CO23456789101'2'3'4'5' 3H3C-C-O1317293010O4Fig. 1: Structurtes of 1, 2, 3 and 4Extraction 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, FAB-MS m/z: 205 [M+H]+(positive); 1H-NMR (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). 13C-NMR (CDCl3) (75 MHz) : 152.2 (s, C-11), 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, C-3), 37.3 (t, C-9), 36.6 (s, C-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 m/z: 223 [M+H]+(positive); 1H-NMR (CDCl3)(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 - 83oC; FAB-MS m/z: 241 [M+H]+(positive); 1H-NMR (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 - 501242oC; FAB-MS m/z: 469 [M+H]+(positive); 1H-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, C-21), 33.7 (q, C-29), 33.0 (t, C-7), 32.9 (s, C-17), 31.5 (s, C-20), 28.8 (q, C-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=CH2groups {H4.75 (2H, br s, H2-12), 4.74 (1H, s, Ha-15), 4.55 (1H, s, Hb-15); C152.2 (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=CH2group {H4.75 (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 {H3.93 (3H, s, 7-OCH3); C56.4 (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); C33.7, 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 {H2.07 (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. 502References 1. U. V. Ahmad and A. Rahman. Handbook of Natural Products data, 2, 163-1164, Amster-dam-London-New York-Tokyo, Academic Press (1994). 2. V. V. Chi (ed.). Vietnamese Medical Plant Dictionary, Hanoi Medicine Pub. (1997). 3. K. Bruns, H. Dolhaine and U. Weber, World crops: Productions, Utilization and Description, 7, 207 (1982). 4. N. X. Dung, T. D. Chinh, D. D. Rang, P. A. Lectercq. J. of Essential Oil Research, Vol. 6, No. 6, 637 - 638 (1994). 5. T. Fujioka, K. Furumi, H. Fujii, H. Okabe, K. Mihashi, Y. Nakano, H. Matsunaga, M. Katano, and M. Mori, Chem. Pharm. Bull., Vol. 47, No. 1, P. 96 - 100 (1999). 6. M. S. Lehner, A. Steigel, and R. Bauer, Phytochemistry, 46, P. 1023 - 1028 (1997). 7. Y. Naya et al. Tetrahedron letters, 23, 3047 (1982). 8. Y. Nakai, T. Kido, K. Hashimoto, Y. Kase, I. Sakakibara, M. Higuchi, and H. Sasaki. J. of Ethnopharmacology, 84, 51 - 55 (2003). 9. Y. Nishikawa, I. Yasuda, Y. Watanabe, and T. Seto, Yagugaku Zasshi, 96, 1322 (1976). 10. Y. Nishikawa, I. Yasuda, Y. Watanabe, and T. Seto, Shoyakugaku Zasshi, 30, 132 (1976). 11. R. Phila, J. P. Bianchni, A. R. P. Ramanoelina, J. R. E. Rasoarahona, R. Faure, A. Cambon, Phytochemistry, Vol. 47, No. 6, P. 1085 - 1088 (1998). 12. M. Resch, A. Steigel, Z. Chen, and R. Bauer, J. Nat. Prod. 61, 347-350 (1998). 13. M. Shunyo, T. Reiko, and A. Masao, Phytochemistry, 27, 535-537 (1988). 14. S. Yahara, T. Higashi, K. Iwaki, T. Nohara, N. Marubayashi et al, Chem. Pharm. Bull, 37, 2995-3000 (1989). 15. Yosioka, T. Nishino, and I. Kitagawa, Yagugaku Zasshi, 96, 1229 (1976). . from Atractylodes lancea. Keywords: Atractylodes lancea, 4(15),11-eudesmadiene, -eudesmol, osthol, 3-acetyl-amyrin. I - Introduction Atractylodes lancea. Chemistry, Vol. 42 (4), P. 499 - 502, 2004 Terpenoids and coumarin from Atractylodes lancea growing in Vietnam Received 29th-Sept.-2003 Chau Van Minh1,