The non-selective toxicity of colchicine remains a major barrier to its development as an anticancer agent. Here, we report a colchicine derivative, 8l, which exhibits potent and selective antiproliferative activity in prostate cancer cells. The present study investigates the impact of stereochemistry at the C10-substituted chiral amine fragment on the biological activity. Our findings reveal that the stereochemical configuration of 8l (7S,15R diastereomer) is critical for its efficacy, showing 12.5-fold greater antiproliferative activity than its counterpart, the 7S,15S diastereomer 8z. Additionally, 8l demonstrates superior α-tubulin polymerization inhibition compared to 8z, that were further corroborated by docking and simulation studies. Mechanistic insights indicate that 8l increases reactive oxygen species levels by modulating the NRF-2/KEAP-1 axis. In vivo, administration of 8l at doses of 0.3 and 0.6 mg/kg significantly suppresses tumor growth in a PC-3 xenograft mouse model. Collectively, this study highlights the therapeutic potential of 8l as a colchicine-based anticancer agent, effectively attenuating tumor progression through modulation of the NRF-2/KEAP-1 axis.