Bioactivity-guided isolation of neuritogenic triterpenoids from the leaves of Ilex latifolia Thunb.

Ilex latifolia Thunb is a traditional Chinese tea and herbal medicine. In this study, one new triterpene saponin (1) and six known triterpenoids (2-7) were isolated from the methanol extract of I. latifolia using a PC12 cell bioassay system. The structures and stereochemistry of these compounds were elucidated using spectroscopic methods and chemical derivatization. This new triterpene saponin (1) was characterized as an ursolic type acid with a 19 alpha-hydroxyl and a trisaccharide moiety at C-3. Compound 1 significantly promoted the neurite outgrowth in PC12 cells by 52% at 10 mu M, whereas compounds 2-7 showed less neuritogenic activity. Structure activity relationship studies indicated that introducing a trisaccharide moiety at C-3 is important for the neuritogenic activity, but the sugar group at C-28 decreased this activity. In addition, compound 1 increased the neurite outgrowth length in primary cortical neuron cells of mice and also exhibited a neuronal protection effect on H2O2-damaged PC12 cells at optimum concentrations.