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Cô lập của SESQUITERPENOIDS từ thân rễ của CURCUMA AROMATICA SALISB Việt Nam.
96Journal of Chemistry, Vol. 38, No. 4, P. 96 - 99, 2000 ISOLATION OF SESQUITERPENOIDS FROM THE RHIROMES OF VIETNAMESE CURCUMA AROMATICA SALISB. Received 5-6-2000 Phan Minh Giang, Phan Tong Son College of Natural Science, Vietnam National University, Hanoi Summary The petroleum ether extract from the rhizomes of Curcuma aromatica Salisb. growing in Vietnam was subjected to repeated flash chromatography on silica gel to give six oxygenated sesquiterpenes furanodiene (1), furanodienone (2), curzerenone (3), germacrone (4), curcumenone (5) and zederone (6) together with three sesquiterpene hydrocarbons -humulene, -selinene, -selinene. INTRODUCTION The rhizomes of C. aromatica Salisb., Zingiberaceae, have been used medicinally in China, Japan and Southeast Asia [1]. As it contains bioactive principles, the chemical composition of C. aromatica have been investigated extensively and it is recognized to be a rich source of terpenoids. However, these investigations were focused on the plant growing in India, China and Japan [2-12], little is known about that growing in Vietnam. Hitherto, only the essential oil from C. aromatica rhizomes collected in Northern Vietnam has been analysed [13, 14]. Curzerenone, germacrone as the major constituents together with minor sesquiterpene hydrocarbons and monoterpenoids were identified [14]. In a previous paper [15], we reported the results of our investigation on the composition of the extracts from the rhizomes of C. aromatica Salisb. growing in Vietnam by means of GC, GC-MS and NMR methods. This paper goes into details of the isolation and identification of the sesquiterpenoid constituents. RESULTS AND DISCUSSION Repeated chromatography on silica gel of the petroleum ether soluble-fraction of an EtOH extract of C. aromatica rhizomes gave three sesquiterpene hydrocarbons (-humulene, -selinene and -selinene) and six oxygenated sesquiterpenes (furanodiene (1), furanodienone (2), curzerenone (3), germacrone (4), curcumenone (5) and zederone (6)). Of these, the hydrocarbons were identified by comparison of their MS and 1H-NMR spectra with those of authentic samples together with the relative retention indices (RRI) measured on GC capillary column CP Sil5 CB. Furanodiene (1) [16], furanodienone (2)[17] and curzerenone (3) [18] were first found in C. zedoaria, Zingiberaceae. We also isolated the interesting sesquiterpenoid germacrone (4)which also was found in many Curcuma species. The spectral data (MS, 1H- and 13C-NMR) of the compounds 1 - 4 are in full agreement with those isolated by us from C. aff. aeruginosa Roxb., Zingiberaceae [19]. Of the two main constituents 3 and 4 of this extract (GC: together 30.7%), curzerenone (3) has been 97found to be active against Mycobacterium tuberculosis H37RV [14]. Curcumenone (5) was isolated as colorless oil from a more polar fraction; 5 has also been isolated from C. zedoaria [20], C. longa [21] and C. aromatica of Japan [3]. Biogenetically, this seco-guaiane-type dicetone may be formed from germacrone via a guaiane-type cationic intermediate [20]. The IR spectrum of compound 6 (C15H18O3,from HR-MS) exhibited absorption bands due to an ,-unsaturated cetone (1674 cm-1), and an epoxide ring (846 cm-1), no hydroxyl or other carbonyl absorption bands were observed. The 13C-NMR and DEPT spectra of 6 indicated the presence of 15 C, including a cetonic carbonyl [ 192.1 ppm (s)], two oxygenated carbons [ 63.9 (s) and 66.5 ppm (d)], 6 olefinic carbons (4s and 2d), three methylene and three methyl groups. The 1H-NMR spectrum showed characteristic signals for a trisubstituted furan ring [ 7.06 (s), -H; 2.09 ppm (s), -CH3], and a vinylic proton [5.46 ppm (dd, J = 4 Hz, 12 Hz)]. The chemical shift and coupling constants of this vinylic proton suggested that the cetonic carbonyl could only be in conjugation with the furan ring. The presence of an epoxy function was supported by the signals at 1.32 (s, 4-Me) and 3.79 (s, H-5) in 1H-NMR spectrum as well as two carbon signals at 63.9 (s) and 66.5 (d) in 13C-NMR spectrum. The other major feature of the 1H-NMR spectrum of 6 was an AB system (2H) with doublets centered at 3.66 and 3.73 ppm (J = 16 Hz) (2 H-9). Thus on the basis of spectral data, a furanogermacrane skeleton for 6 was suggested. The complete 1H-1H COSY and HMQC spectral data confirmed the structure of zederone (6). Zederone was reported to be found in C. zedoaria [22, 23]. The relative stereochemistry of 6 was established by a NOESY experiment to be identical with the revised stereochemical structure proposed by I. Kitagawa et al. [24]. Figure 1: Important NOESY interactions of 6EXPERIMENTAL General procedures All melting points are uncorrected. Optical rotation was obtained on a Polartronic D CH3HCH3OOHHHHHHHO106987111213151454321OOO6513121114432110987153OO21OOOOOOH4 5 6 98(Schmidt + Haensch). 1H-NMR (CDCl3): Bruker AM 400. 13C-NMR (CDCl3): Bruker AC 200 with DEPT programme. For zederone (6)all NMR spectra were taken on a Bruker AMX 600. IR (CCl4): Perkin-Elmer 881. HR-MS:Finigan MAT 95 SQ. GC-EIMS: Hewlett-Packard 5890 II with a HP-1 column (25 m x 0.2 mm i.d., film thickness 0.5 µm) combined with MSD HP 5971A, carrier gas He. GC:Packard 439 with a CP Sil 5 CB column (25 m x 0.25 mm i.d., film thickness 0.39 µm); carrier gas: N2; detector: FID; plotter: Shimadzu C-R6A Chromatopack; temperature programme: 60 - 220oC, 5oC/min; injector temperature: 270oC; detector temperature: 300oC. FC: silica gel ICN Biomedicals (32-63 µm). TLC: DC-Alufolien 60 F254 No. (Merck 5554). Visualisation: UV light 254 nm, anisalde-hyde/H2SO4.Extraction and Isolation The dried rhizomes of C. aromatica (1kg) collected in Socson, Hanoi (Vietnam) were powdered and extracted with EtOH 96% by percolation at room temperature. The combined extract was partially concentrated under reduced pressure, then dissolved in distilled water, and partitioned with petroleum ether (PE, b.p. 30 - 60oC) and EtOAc successively to afford PE (15.9 g) and EtOAc (17.9 g) fractions. The PE fraction (3 g) was subjected to FC (applied pressure ~ 0.2 mbar) eluted with pentane-EtOAc (95 : 5) to give 6 fractions. Fraction 1 contained 33mg -selinene and -selinene. 171 mg fraction 2 was rechromato-graphed on AgNO3-impregnated silica gel using pentane as eluent to give 34 mg -humulene. Repeated FC of 515 mg fraction 3 eluted with PE-diethyl ether (4:1) afforded furanodiene (1) (52 mg), curzerenone (3) (43 mg) and germacrone (4) (63 mg). Fractions 4 and 5 were concentrated under reduced pressure and further purified by solid phase extraction using silica gel Waters cartridges to give furanodienone (2) in the yields of 298 mg and 424 mg, respectively. Fraction 6 was repeatedly chromatographed using pentane-diethyl ether (7 : 3) to yield 83 mg curcumenone (5) (GC: 83%) and 100mg crystalline zederone (6) (GC: 100%). Furanodiene (1), furanodienone (2), curzerenone (3) and germacrone (4) were obtained as colorless oils. For RRISil5, MS, 1H- and 13C-NMR, see reference 19. Curcumenone (3): Colorless oil. RRISil5: 1802. HR-MS: 234.1617 (C15H22O2). 1H-NMR: 0.40 (1H, ddd, J = 4.4 Hz, 7.3 Hz, 7.3 Hz, H-1), 0.64 (1H, m, H-5), 1.08 (3H, s, 10-CH3), 1.54 - 1.62 (2H, m, 2 H-2), 1.75 (3H, s, 11-CH3), 2.06 (3H, s, 11-CH3), 2.10 (3H, s, 4-CH3), 2.44 (2H, dd, J = 7 Hz, 7 Hz, 2 H-3), 2.52 (1H) and 2.98 (1H), AB system, J = 16 Hz (2 H-9), 2.70 - 2.80 (2H, m, 2 H-6). 13C-NMR: 24.0 (d, C-1), 23.3 (t, C-2), 43.7 (t, C-3), 208.8 (s, C=O, C-4), 23.3 (d, C-5), 28.1 (t, C-6), 127.9 (s, C-7), 201.7 (s, C=O, C-8), 48.7 (t, C-9), 19.9 (s, C-10), 147.5 (s, C-11), 23.3 (q, C-12), 23.3 (q, C-13), 29.9 (q, C-14), 18.9 (q, C-15). Zederone (6): Colorless needles. mp 149-150oC. RRISil5:1961. []D31 +290o(c 1.14, CHCl3). IR: max cm-1 1674, 1558, 1428, 1235, 1108, 1022, 931, 846. HR-MS: 246.1256 (C15H18O3). GC-MS: m/z (%) 246 (35), 231 (4), 213 (5), 188 (29), 175 (100), 161 (38), 147 (22), 133 (15), 122 (25), 119 (74), 105 (24), 91 (29), 77 (23), 65 (16), 53 (12). 1H-NMR (with COSY): 1.27 (1H, ddd, J = 4 Hz, 13.0 Hz, 13.5 Hz, H-3), 1.32 (3H, s, 4-CH3), 1.58 (3H, s, 10-CH3), 2.09 (3H, s br, 11-CH3), 2.21 (1H, d br, J = 13.5 Hz, H-2), 2.28 (1H, ddd, J = 3.5 Hz, 3.5 Hz, 13.0 Hz, H-3), 2.50 (1H, dddd, J = 3,5 Hz, 12.0 Hz, 13.5 Hz, 13.5 Hz, H-2), 3.66 (1H) and 3.73 (1H), AB system, J = 16 Hz (2 H-9), 3.79 (1H, s, H-5), 5.46 (1H, dd br, J = 4 Hz, 12 Hz, H-1), 7.06 (1H, s br, H-12). 13C-NMR (with HMQC): 131.1 (d, C-1), 24.6 (t, C-2), 37.9 (t, C-3), 63.9 (s, C-4), 66.5 99(d, C-5), 192.1 (s, C=O, C-6), 122.1 (s); 131.0 (s) (C-7; C-10), 157.0 (s, C-8), 41.8 (t, C-9), 123.2 (s, C-11), 138.0 (d, C-12), 10.2 (q, C-13), 15.7 (q, C-14), 15.1 (q, C-15). Acknowlegements The authors are grateful to the International Foundation for Science (IFS,Grant No. F/2841-1) for the financial support. 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