Phytochemical and pharmacological activities of Annona reticulata components suggest a wide range of clinical application in lieu of cancer therapy. Present study includes investigation of bioactive constituents from roots of Annona reticulata for cytotoxic properties against different cancer cell lines. Three aporphine alkaloids liriodenine (AAR-01), norushinsunine (AAR-02), reticuline (AAR-03) and one acetogenin neoannonin (TAR -01) isolated from the roots of Annona reticulata. The structures of the compounds were achieved by1HNMR, 13CNMR and Mass spectroscopic methods. All the isolated compounds subjected for cytotoxicity evaluation against A-549, K- 562, HeLa, MDA-MB cancer cell lines and normal cell lines (Vero cells) by MTT assay. All the isolated compounds exhibited prominent dose-dependent cytotoxicity against all the cancer cell lines at dilutions 5, 10 and 20 μg/ml, whereas TAR -01 showed strong cytotoxicity against cancer cell lines with IC50 values ranging from5.8 – 6.9μg/ml. Simultaneously, the effect of all the isolated compounds against Vero cell lines was lower in comparison with the cancer cell lines. The better cytotoxicity of all the isolated compounds the appearance of hydroxyl group at C-7 in AAR-02 could be favourable for increased cytotoxicity against cancer cell lines among aporphine alkaloids and the presence of two hydroxyl groups adjacent to the tetrahydrofuran ring in TAR-01 may be responsible for enhanced activity. The lower cytotoxicity against Vero cell line seems that the isolated constituents AAR-01, AAR-02, AAR-03 and TAR -01 may be used as chemopreventive agents in cancer therapy.