Six polymethoxylated flavonoids were isolated from ethyl acetate fraction of ethanol extracts from Vitex rotundifolia Linn. f. fruit, and their structures were identified by NMR and MS methods, and cytotoxic activity of six compounds are analyzed by MTT method. The six compounds are oroxylin A (1), chrysosplenol-D (2), casticin (3),artemetin (4), persicogenin (5) and 5,3’-dihydroxy-6,7,4’-trimethoxyflavanone (6). In which, persicogenin is isolated from V. rotundifolia for the first time. Oroxylin A, casticin, artemetin and 5,3’- dihydroxy- 6,7,4’ –trimethoxyflavanone can inhibit proliferation of human liver cancer cell line HepG2 in vitro, and their IC50 is 11. 90, 12. 54, 13. 69 and 11. 76 μmol·L^-1, respectively, showing that the four compounds have a certain cytotoxic activity.

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