Corchorusin-D (COR-D), isolated from Corchorus acutangulus, was reported to induce apoptosis in leukemic cells. However, no studies concerning its activity on melanoma cells have been reported. We have evaluated its in vitro anti-cancer activity on melanoma cells (B16F10, SK-MEL-28, and A375). The results demonstrate that COR-D showed maximum inhibition of B16F10 cells in vitro. COR-D induced mitochondrial dysfunction and altered the Bax/Bcl-2 ratio with down regulation of pro-caspases 9 and activation of caspase 3 in B16F10 cells, triggering intrinsic pathway of apoptosis. Moreover, it inhibited the in vivo B16F10 tumor growth and increased the survival rate of mice. Greater number of Annexin V-FITC and propidium iodide (PI)-positive tumor cells signified that COR-D induced apoptosis in vivo also. The reduction in tumor growth is well correlated with decreased microvascular density of the tumor cells in treated mice. In conclusion, this study reveals that COR-D-induced mitochondrial dysfunction is responsible for the induction of apoptotic cell death.