Seven new oleanane-type triterpenoids (1-7), named fatsicarpains A-G, and the known compounds 3α-hydroxyolean-11,13(18)-dien-28-oic acid (8) and 3α-hydroxyolean-11-en-28,13β-olide (9) were isolated from the leaves and twigs of Fatsia polycarpa on the basis of bioassay-guided fractionation. The structures of compounds 1-7 were elucidated through spectroscopic analyses and single-crystal X-ray crystallography of 1, 8, and 9. Cytotoxicity against HepG2 2.2.15 and AGS cells and antihepatitis B virus (HBV) and antibacterial activities of 1-9 were also evaluated in vitro.

Chemical investigations of the Formosan soft coral Lemnalia flava have obtained a new ylangene-type sesquiterpenoid, (1S,2S,4R,6S,7R,8S)-4a a-formyloxy-b b-ylangene (1), along with two known sesquiterpenoids, lemnalol (2) and isolemnalol (3). Three new nardosinane-type sesquiterpenoids, designated as paralemnolins J-L (4-6), and five known sesquiterpenoids (7-11), were isolated from the other soft coral Paralemnalia thyrsoides. The structures of metabolites 1 and 4-6 were elucidated through extensive spectroscopic analysis and chemical methods. Moreover, the anti-inflammatory activity of metabolites 1-7 and 11 was evaluated in vitro.Key words Lemnalia flava; ylangene-type sesquiterpenoid; nardosinane-type sesquiterpenoid; Paralemnalia thyrsoides; antiinflammatory activity Chem. Pharm. Bull. 58(3) 381-385 (2010) © 2010 Pharmaceutical Society of Japan * To whom correspondence should be addressed. e-mail: yihduh@mail.nsysu.edu.tw . Moreover, the 1 H-1 H correlation spectroscopy (COSY) correlations (Fig. 2) between H-4 and H-16, and the heteronuclear multiple bond correlation (HMBC) correlations from H-16 to C-4 confirmed the location of the formyloxy functionality at C-4. Therefore, the planar structure of 1 was proposed decidedly.The computer modeled 3D structure of 1 was generated by using MM2 force field calculations for energy minimization with the molecular modeling program Chem3D Ultra 9.0. The relative stereochemistry of 1 assigned by the nuclear Overhauser effect spectroscopy (NOESY) spectrum was compatible with those of 1 offered by computer modeling, in which the close contacts of atoms calculated in space were consistent with the NOESY correlations (Fig. 3). A large long-range coupling constant ( 4 J 2,6 ϭ5.9 Hz) due to a W-type coupling between H-2 and H-6 further indicated the presence of a bridged cyclobutane system in 1. In addition, no coupling was observed between H-7 and the adjacent protons (H-2, H-6, and H-8), suggesting a dihedral angle of approximately 90°between these protons. The NOESY correlations between H 3 -11 with both H-4 and H-5b positioned the b-orientation of the aforementioned protons. Furthermore, H-7 382Vol. 58, No. 3

Eleven compounds were isolated from the methanolic extract of the leaves of Solanum erianthum D. Don, including five -linolenic acid analogs, -linolenic acid (1), 13S-hydroxy-9(Z),11(E)-octadecadienoic acid (2), 9S-hydroxy-10(E),12(Z), 15(Z)-octadectrienoic acid (3), 9(Z),11(E)-octadecadienoic acid (4), and octadecanoic acid (5); two benzofuran-type lactones, loliolide (6) and dihydroactinidiolide (7); two steroidal alkaloids, solasonine (8) and solamargine (9); a flavonol glycoside, camelliaside C (10); and a flavone, 5-methoxy-(3'',4''-dihydro-3'',4''-diacetoxy)-2'',2''-dimethylpyrano-(7,8:5'',6'')-flavone (11). Among these isolated compounds, 9 showed the most potent activity against HBsAg, with an IC 50 of 1.57 M, followed by 8 (IC 50 is 5.89 M). In the testing against HBeAg, 11 was the only active compound with an IC 50 of 36.11 M. Compound 9 also revealed strong inhibition of DNA replication towards HBV and its IC 50 was 2.17 M. However, -linolenic acid (1) showed a prominent selected index (SI), both in anti-HBsAg and inhibition of DNA replication with SI values of 7.75 and 7.18, respectively. This is the first report that unsaturated fatty acid 1, steroidal alkaloid glycoside 9 and flavone 11, all showed excellent activity against HBV. These results provide lead candidates in the development of anti-HBV drugs from natural sources.

Earlier studies of the genus Cespitularia (Xeniidae) led to the isolation of a diverse array of diterpenoids including alcyonolides, caryophyllanoids, cembranolides, cespitularanoids, dolabellanoids, norverticillanoids, verticillanoids, and xenicanoids. [1][2][3][4][5][6][7][8][9] Previous bioassay results of some verticillane-type, norverticillane-type, and cespitularane-type diterpenoids have demonstrated remarkable pharmacological activity such as in vitro cytotoxicity against various cancer cell lines.1,6-9) The ongoing researches for bioactive constituents prompted us to investigate the Formosan soft coral Cespitularia hypotentaculata ROXAS. We have previously obtained verticillanes, norverticillanes, and cespitularanes, cespitularins A-Q, 6,7) from the organic-soluble of the organism.Our continuing chemical examinations on the secondary metabolites of this soft coral have resulted in the purification of two new verticillane diterpenoids, cespitularins R and S (1, 2), and seven known compounds (3-9) [6][7][8] (Fig. 1). The structures of 1 and 2 were determined by detailed 1D and 2D NMR techniques, especially employing correlation spectroscopy (COSY), distortionless enhancement by polarization transfer (DEPT), heteronuclear multiple bond correlation (HMBC), heteronuclear single quantum correlation (HSQC), and nuclear Overhauser effect spectroscopy (NOESY) experiments. Furthermore, compound 2 (10 mM) was evaluated for the ability to inhibit the expression of the pro-inflammatory inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins. Compounds 7 and 8 did not inhibit the COX-2 protein expression, but significantly inhibited iNOS protein expression. With the exception of the above findings, the obtained negative results indicated that 1, 3-6, and 9 exhibited insignificantly anti-inflammatory activity against lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Results and DiscussionSpecimens of C. hypotentaculata were frozen immediately after collection. Conventional extraction procedures were used, and the acetone extract was exhaustively partitioned between EtOAc and H 2 O to afford the EtOAc-soluble fraction, which was evaporated under vacuum to yield a dark brown gum (15 g). Then, the concentrated residue was successively subjected to column chromatography and high-performance liquid chromatography (HPLC), leading to the isolation of two new verticillane diterpenoids, cespitularins R and S (1, 2), and seven previously characterized compounds 3-9 (see Experimental).Cespitularin R (1) was isolated as a colorless, viscous oil. High resolution electrospray ionization mass spectrometry (HR-ESI-MS) of 1 exhibited a pseudomolecular ion peak at m/z 365.2095 [MϩNa] (Table 1) coupled with COSY, HSQC, and HMBC correlations proved that the structure of 1 was identical to that of cespitularin A 6) except for the presence of a hydroxyl instead of an acetoxy group. The carbonyl signal at d C 170.1 was attributed to the acetate moiety linked to C-6, as further confirmed through the crucial...

The following paragraph should be included in the Results and Discussion section. "Compound 6 was isolated as a white, amorphous powder, and its molecular formula was found to be C 31 H 50 O 4 , as deduced from its HRESIMS. By comparison of the NMR spectroscopic data (Tables 1 and 3) of 6 with those of 4, it was found that an olefinic group at C-9/C-11 in 4 was converted to an additional methoxy group at C-11 [δ H 3.23 (3H, s) and δ C 54.2 (CH)] in 6. It was also confirmed by the 1 HÀ 1 H COSY correlations between H-12/H-11 and H-9/H-11 and the crucial HMBC correlations from H-9/11-OMe to C-11. The NMR spectroscopic data reported 20 and coupling constants of H-11 for 3α,21β-dihydroxy-11α-methoxyolean-12-ene [δ H 3.88 (1H, dd, J = 9.0, 3.5 Hz) and δ C 75.9 (CH)] are similar to those of 6 [δ H 3.80 (1H, dd, J = 8.4, 3.0 Hz) and δ C 75.6 (CH)], indicating an α-orientation of the methoxy group. Therefore, the structure of fatsicarpain F (6) was identified as 3α-hydroxy-11α-methoxyolean-12-en-28-oic acid." The authors apologize for any inconvenience.