Published: July 19, 2011 Copyright r 2011 American Chemical Society and American Society of Pharmacognosy 1744 dx.doi.org/10.1021/np2002435 | J. Nat. Prod. 2011, 74, 1744–1750 ARTICLE pubs.acs.org/jnp Oleanane-Type Triterpenoids from the Leaves and Twigs of Fatsia polycarpa Shi-Yie Cheng, † Chao-Min Wang, † Yuan-Man Hsu, ‡ Tsurng-Juhn Huang, § Shen-Chieh Chou, § En-Hung Lin, † and Chang-Hung Chou* , †, § † Research Center for Biodiversity, ‡ Department of Biological Science and Technology, and § Graduate Institute of Ecology and Evolutionary Biology, China Medical University, Taichung 40402, Taiwan, Republic of China b S Supporting Information P lants of the genus Fatsia (Araliaceae) are endemic to the Archipelago of the western Pacific rim and Bonin Islands. 1 There are three known Fatsia species: F. japonica, F. oligocar- pella, and F. polycarpa. 1 Flowers, mature fruits, and leaves of F. japonica are rich in triterpeniod saponins. 2À11 Saponins isolated from F. japonica have been recommended for treatment of ankylosing spondyloarthritis, osteoarthritis, rheumatism, rheu- matoid arthritis with accompanying reactive gout, osteochon- drosis, synovitis, and tendinitis. 10 They may also be helpful as a cofactor in antiaging cosmetics and in protein therapy for disorders related to the antioxidant enzymes Cu, Zn-SOD or reactive oxygen species. 11 Fatsia polycarpa Hayata (Araliaceae) is an evergreen shrub found in moist shady forests at middle elevations (2000À2800 m) in Taiwan. 12 Our search for bioactive constituents prompted us to investigate secondary metabolites of this plant that not been reported previously. Dichloromethane, EtOAc, and n-BuOH extracts of F. polycarpa were assayed for cytotoxicity against HepG2 2.2.15 (human hepatocellular carcinoma) cells. The active compounds isolated from the CH 2 Cl 2 extract were purified and structurally identified. Seven new oleanane-type triterpenoids (1À7) and two known analogues, 3R-hydroxyolean-11,13(18)-dien-28-oic acid (8) 13 and 3R-hydroxyolean-11-en-28,13β-olide (9), 13 were obtained. The structures of 1À7 were elucidated using extensive spectro- scopic analyses and by comparison with data reported in the literature. The structures of 1, 8, and 9 were confirmed by X-ray diffraction analysis. 14 Some oleanane-type compounds are of interest due to their antitumor, anti-HIV, anti-inflammation, antioxidation, antihy- perglycemia, and cardiovascular properties. 15 Of four million patients worldwide infected with hepatitis B virus (HBV), about 20% are expected to develop chronic hepatitis, liver cirrhosis, or hepatocarcinoma. 16 Helicobacter pylori infection is associated with an increased risk for development of duodenal ulcers, gastric ulcers, gastric adenocarcinomas, and gastric lymphomas. How- ever, antibiotic resistance to H. pylori and other bacterial pathogens is an increasing problem for eradicating infection. 17 Therefore, safe and efficient treatments to decrease the need for or to replace antibiotics for eradicating H. pylori and other infections are needed. Compounds 1À9 were evaluated in vitro for cytotoxicity against cancer cell lines HepG2 2.2.15 and AGS (human gastric cancer epithelial cells). Bioassays were also conducted for anti- hepatitis B virus and for antibacterial activity against H. pylori, Bacillus cereus, Enterococcus faecalis, Escherichia coli, Listeria monocytogenes, Salmonella enterica, Staphylococcus aureus, and Pseudomonas aeruginosa. ’ RESULTS AND DISCUSSION Conventional extraction procedures were used, and a MeOH extract of the leaves and twigs of F. polycarpa was suspended in H 2 O and then partitioned successively with CH 2 Cl 2 , EtOAc, and n-BuOH. Chromatographic separation of the CH 2 Cl 2 -soluble fraction using normal-phase Si-60, reversed-phase C 18 gel, and Sephadex LH-20 columns in combination with semipreparative reversed-phase C 18 HPLC led to the purification of oleanane- type triterpenoids 1À9. Compound 8 has been previously isolated from the aerial parts of the Argentinean shrub Junellia tridens. 13 Definitive support for Received: March 21, 2011 ABSTRACT: Seven new oleanane-type triterpenoids (1À7), named fatsicarpains AÀG, and the known compounds 3R- hydroxyolean-11,13(18)-dien-28-oic acid (8) and 3R-hydro- xyolean-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 1À7 were eluci- dated 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 1À9 were also evaluated in vitro.