<p>Triple-negative breast cancer (TNBC) is the most lethal and metastatic form of breast cancer (BC) that displays elevated levels of epithelial-mesenchymal transition (EMT). Standard chemotherapeutics are unable to reduce the metastatic rate; hence, anti-metastatic agents are instantly required. Zeylenone (ZEY) is an eminent anti-tumour and apoptotic agent that is naturally occurring as cyclohexene oxide, which is extracted from the <i>Uvaria grandiflora</i> leaves. However, the anti-proliferative, anti-metastatic, and apoptotic efficacy of ZEY in TNBC cells remains uncertain. Hence, our investigation proposed to study the antiproliferative, adhesive, EMT markers, and apoptotic activity of ZEY (10, and 15 µM/ml) on human TNBC cells MDA-MB-231 and to assess its latent mechanism. The cell toxicity exploration and cell apoptosis of ZEY action on MDA-MB-231 cells were evaluated by MTT, AO/EB, DAPI, PI, adherence assay, and mRNA expression levels. Data unveiled that ZEY (10 and 15 µM/ml) suppresses cell propagation and adhesion; however, it triggers apoptosis by the heightened mRNA intensities of Bax, caspase, while attenuating cyclin-D1, survivin, Bcl-2, and c-myc, in a dosage-relatedly. It generates a Bax/Bcl-2 ratio disparity, which activates the caspase cascade, Cyt-c, and leads to apoptosis. Furthermore, ZEY (10 and 15 µM/ml) inhibits relative gene levels of EMT markers as well as PI3K/AKT/mTOR pathways. According to MD modelling, ZEY docking investigation reveals that it has a low binding energy and a high binding affinity for the apoptotic and cell growth protein indicators mentioned above, as well as greater stability with Bax and mTOR. Our results emphasise that ZEY is beneficial as a protective remedy for BC. Molecular interaction studies of ZEY with apoptotic and cell proliferative proteins present findings that conclude that ZEY has customised the expression of proteins the breast cancer cells.</p>