Studies On Chemical Constituents Of Beesia Calthifolia(Maxim.)Ulbr

Liver diseases have become a serious public health problem in China. Liver injury is the pathologic basis of many kinds of liver diseases. Our previous findings and literature survey has showed that cycloartane triterpenoids have potential therapeutic effects against liver injury. In this project, we focused on phytochemical studies of these cycloartane triterpenoids in the folkloric medicine, rhizomes of Beesia calthifolia (Maxim.)Ulbr.(Ranunculaceae), as well as their hepatoprotective effects.B. calthifolia is a perennial herb and a folkloric medicine widely used in southwest China. The rhizome or whole plant has been applied for the treatment of ostalgia, rheumatism, abscesses, toothache, as well as external application for boils and snakebites.In the current study, chemical constituents of B. calthifolia and their hepatoprotective activities were investigated. Twenty-seven compounds, including thirteen new and nine known cycloartane triterpenoids, were isolated from the 95% EtOH extract by using modern chromatography techniques, including silica gel, D-101 macroporous resin, Sephadex LH-20, and C18 reverse-phased silica gel chromatographic columns, as well as HPLC.MS, IR and NMR techniques were applied to elucidate the structures of the twenty-seven isolated compounds and their structures were identified as: (20S*,24R*)-20,24-epoxy-12α,15α,25-trihydroxy-3-(β-D-xylopyranosyloxy)-9,19-cyclolanostan-16-one (1); (20S*,24R*)-20,24-epoxy-3β,12α,15α,25-tetrahydroxy-9,19-cyclolanostan-16-one (2); (20S*,24R*)-15α-acetoxy-20,24-epoxy-9,19-cyclolanostane-3β,12a,25-triol-16-one (3); (20S*,24R*)-20,24-epoxy-3β,12β,15α,25-tetrahydroxy-9,19-cyclolanostan-16-one (4); (20S*,24R*)-20,24-epoxy-3β-)β-D-xylopyranosyloxy)-9,19-cyclolanostane-12α,15α,16β,18,25-pentol (5); (20S*,24R*)-20,24-epoxy-3β-(β-D-xylopyranosyloxy)-9,19-cyclolanostane-12β,15α, 16β,25-tetrol (6); (20S*,24R*)-15α,16β-diacetoxy-20,24-epoxy-9,19-cyclolanostane-3β,12β,25-triol (7); (20S*,24R*)-15α-acetoxy-20,24-epoxy-9,19-cyclolanostane-3β,12β,16β,25-tetrol (8); (20S,24S)-15α,16β-diacetoxy-20,24-epoxy-9,19-cyclolanostane-3β,12β,24,25-tetrol (9); (20S,24S)-15β,16β-diacetoxy-18,24;20,24-diepoxy-9,19-cyclolanostane-3β,12β,25-triol (10); (20S,24S)-15α-acetoxy-16β,24;20,24-diepoxy-3β-(β-D-xylopyranosyloxy)-9,19-cyclolanostane-18,25-diol (11); (20S,24S)-15α-acetoxy-16β,24;20,24-diepoxy-9,19-cyclolanostane-3β,18,25-triol (12); (20S,24S)-15α-acetoxy-16β,24;20,24-diepoxy-9,19-cyclolanostane-3β,12β,25-triol (13); beesioside Ⅲ (14); aglycone of beesioside Ⅲ (15); aglycone of beesioside Ⅱ (16); beesioside Ⅱ (17); beesisiode N (18); beesioside M (19); beesioside I (20); aglycone of beesioside IV (21); beesioside O (22); 2-methoxy-4-methylbenzene-1-O-β-D-glucopyranoside (23); β-rosasterol (24); fukinolic acid (25); dibutyl terephthalate (26); palmitic acid (27). Among them, compounds 1-13 were new compounds. Compounds 15,16,21 and 23 were isolated from nature for the first time. Compounds 24～27 were isolated from B. calthifolia for the first time.Hepatoprotective activities of some of the compounds were evaluated on a D-galactosamine induced cell damage model in human hepatic L02 cells.In conclusion, the results has provided a solid experimental basis for further development of B. calthifolia.