Coumarin and steroid compound from stem bark of Kleinhovia ...

N.H. Soekamto et al. Proceeding of The International Seminar on Chemistry 2008 (pp. 231-234)Jatinangor, 30-31 October 2008J = 9.2 Hz, H-3), 7.60 (1H, d, J = 9.2 Hz, H-4), 6.84(1H, s, H-5), 6.92 (1H, s, H-8), 3.95 (3H, s, (OCH 3) );13 C NMR (Chloroform-D 6 , 500.2 MHz) δ: 161.67 (C-2), 150.41 (C-4’), 149.84 (C-6), 144.16 (C-7), 143.51(C-4), 113.58 (C-3), 111.66 (C-8’), 107.63 (C-5),103.35 (C-8), 56.57 (-OCH 3 ).Compound 2 was isolated as needles white crystal,with m.p. of 126 o C. The compound did not givephosphorescent under the UV lamp, The IR spectrumof 2 exhibited absorptions aliphatic (2956, 2918, 2848cm -1 ), -CH 2 (1462 cm -1 ) and -CH 3 (1377cm -1 )groups. The data indicated that compound 2 belong toterpenoid derivative. The sugestion was suported bynegatif result in the steroid test. 1 H NMR (CDCl 3 ,500.2 MHz) δ: 1.15 ppm (2H, dt, J =11.0; 3.65 Hz, H-1 eq ), 1.08, 1.84, 1.54, 3.51, 2.28, 2.22, 3.51, 1.98,1.94, 1.49, 0.91, 1.48, 2.00, 0.98, 1.58, 1.82, 1.07,1.34, 0.89, 1.65 ppm respectively (1H, m, for H-1 ax ,H-2eq, H-2 ax , H-3, H-4 eq , H-4 ax , H-5, 7eq and ax, 8, 9,11, 12eq, H-14, 15 eq , 16, 17, 20, 22 eq , 24, and 25 eq ),5.34 ppm (3H, d, J = 4.90 Hz, H-6), 0.67 and 1.00ppm (3H, s, H-18 and 19), 0.91 ppm (3H, d, J=6.1Hz), 1.14 ppm (2H, m, H-23), 0.82 and 0.80 ppmrespectively (3H, d, J = 6.7 Hz, for H-26 and 27),1.23 ppm (2H, m, H-28), 0.83 ppm (3H, t; J = 6.7 Hz,H-29); 13 C NMR (CDCl 3 , 500.2 MHz) δ: 140.8 (C-3),121.8 (C-6), 71.9 (C-2), 56.8 (C-15), 56.1 (C-17),50.2 (C-9), 45.9 (C-24), 42.4 (2C C-4 dan 14), 39.8(C-12), 37.3 (C-1), 36.6 (C-20), 36.2 (C-10),34.0 (C-22), 32.0 (2C, C-7 dan 8) 31.7 (C-2), 29.2 (C-25),28.4 (C-16), 26.1 (C-23), 24.4 (C-15), 23.1 (C-28),21.2 (C-11), 19.9 (C-26), 19.5 (C-27), 19.1 (C-19),18.9 (C-21), 12.1 (C-18), 12.0 (C-29).HOH 3 COOOHOH 3 CCH 3CH 3(1) (2)Figure 1 Scopoletin (1) and β-sitosterol (2)Results and DiscussionA coumarine derivative compound which hadbeen isolated for the first time From CHCl 3 fractionwas 7-hidroxy-6-methoxy coumarin (1). Thiscompound was purified by coulumn chromatographyand recrystallization and their structure wereH 3 CCH 3CH 3determined by spectral methods and comparison withthose reported in the literature.7-hidroxy-6-methoxy coumarin (1) was a whitepowder compound with m.p. of 182-184 o C. The UVspectrum of 1 showed absorption maxima at 228(3.98), 344 (4.40) nm, indicating the presence of anextended benzene chromophore. The IR spectrum of 1exhibited absorptions for hydroxyl (3338 cm -1 ),aliphatic (2924, and 2853 cm -1 ) and aromatic (, 1608,1566, 1508, 1464 cm -1 ) groups., -C=O (1701 cm -1 ),CH 3 (1376 cm -1 ), C-O (1291 cm -1 ).UV and IR data indicated that compound 1 iscoumarine derivative (Murray, et. al., 1982). The 1 HNMR spectrum of 1 showed two vinilic proton signalsat δ 6.27 ppm (1H, d, J = 9.2, H-3) and 7.60 ppm (1H,d, J = 9.2, H-4). Beside that 1 H NMR spectrum toshow two aromatic proton signals at δ 6.84 and 6.92ppm each 1H with multiplicity singlet assignable totetrasubstituted benzena moiety. Another informationin the 1 H NMR spectrum was the signal at δ 3.95 ppm(3H, s) for methyl –OCH 3 .The 13 C NMR spectrum of 1, showed that thereare ten signals including C carbonyl (C-2) at δ 161.67ppm, three oxycarbons at δ 144.16, 149.84 and 150.41ppm for C-6, C-7, and C-8a. And then other carbonatoms i.e. quarterner carbon C-4a, 6, 7, and 8a at δ111.66, 144.16, 149.84, and 150. 41 respectivelyassigned with the aid of HMBC measurements. It canbe proposed from these data that the structure of 1 is7-hidroxy-6-methoxy coumarin or scopoletin (Fig.2). The HMBC spectrum of 1, in particular, disclosedlong-range correlations between four the methynesignals at δ 6.27 ppm (H-3) with carbon signals at δ161.66 ppm (C-2), δ 7.60 ppm (H-4) with carbonsignals at δ 161.66 (C-2), 150.41 ppm (C-8a), δ 6.84ppm (H-5) with carbon signals at δ 149.84 (C-7),150.41 ppm (C-8a), δ 6.92 ppm (H-8) with carbonsignals at δ 144.16 (C-6), 149.84 (C-7), and δ 3.95ppm (H-9) with carbon signals at δ 144.16 (C-6) (Fig.3). Further support for structure 1 came fromcomparison of the NMR data with that of relatedcompound (Soekamto et.al., 2001), isolated fromMorus species (Table 1).Bioassay analysis to the compound 1 showmoderate activity against Artemia salina with LC 50 =569,64 µg/mL. Scopoletin was also isolated fromMorinda citrifolia L. and it can be used asantihypertensive, antiinflamatory and antiallergenic(Rosa et.al., 2003), and can also be used as inhibitorprostaglandin synthetase (MH Farah and Sauelsson G,1992).232