Abstract: Objective To investigate the chemical constituents and pharmacological activity of the ethyl acetate extract fromDisporposis pernyi. Methods Compounds in the ethyl acetate extract of Disporposis pernyiwere extracted and separated by systematic solvent extraction and column chromatography. The molecular structures were identified by nuclear magnetic resonance (NMR), mass spectrometry (MS) and thin layer chromatography (TLC). The inhibitory activity of extracts against human breast cancer cell line MCF-7 was determined by MTT method. Cell morphology was observed with AO-EB fluorescent staining . Results Three components were isolated and identified, which were betulin, β-daucosterol and luteolin. Betulin was first isolated from Disporposis pernyi. The extract has inhibitory effects on the proliferation of MCF-7 cells. Compared with the control group, the number of apoptotic cells of the drug group increased significantly. Conclusions The ethyl acetate extract from Disporposis pernyi has an inhibitory activity on human breast cancer cell MCF-7, among which betulin plays a certain role.