Background The research and application of plants in food supplements and drugs have attracted great interest. This study aimed to examine the efficiency of several solvents for the extraction of the main compounds from Annona squamosa leaves and to evaluate the antioxidant, antibacterial, and anticancer activities of these extracts. Methods Gas chromatography-mass spectrometry was used to screen the bioactive compounds of A. squamosa methanolic extract. The free radical, hydrogen peroxide, and nitric oxide scavenging activities of the extracts were investigated. Furthermore, MTT, nuclear staining, LDH, and monolayer wound repair assays were performed to evaluate the potential anticancer activity of the extracts in colon cancer cells while the antibacterial activity was tested by using a well diffusion assay. Results A. squamosa leaves extracts were found to contain several bioactive compounds, of which the majority were sesquiterpenes (C15H24). These extracts exhibited strong antioxidant activity and antibacterial potency against both gram-positive and gram-negative bacteria. Different A. squamosa leaves extracts displayed remarkable antiproliferative, cytotoxic, antimigration, and apoptotic activities in colon cancer cells. Conclusions A. squamosa leaves contain major bioactive compounds that inhibit the growth of several types of bacteria and colon cancer cell lines, which demonstrated their efficacy as an alternative source of antibiotics and for the development of novel drugs for colon cancer therapy.

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: Annona squamosa L. is an important medicinal plant used in traditional medicine for the treatment of various diseases. Different parts of A. squamosa L. have various therapeutic effects; however, the anticancer activity of the leaves has not yet been identified. In vitro, MTT, nuclear staining, and LDH assays were used to evaluate cell survival and proliferation in cells exposed to the extracts. The effect of the extracts on cell migration was investigated using a monolayer wound repair assay, and the apoptotic effects were evaluated using flow cytometry. A breast cancer model was used to study the effect of the extract on the tumor size, and the expression of different proliferative and apoptotic markers was evaluated by immunohistochemical analysis. At a concentration of 100 µg/mL, A. squamosa leaf extracts exerted strong antiproliferative and cytotoxic effects against various cell lines. The extracts reduced wound closure and strongly induced apoptosis. In vivo study, rats were sacrificed 24 h after the last injection, and tumor size, as well as the expression of proliferative and apoptotic markers, were observed to be greatly affected by treatment with the extracts. Therefore, A. squamosa leaf extract may be developed as a potential novel drug to treat breast cancer in the future