The present work focused on the synthesis of Vinca rosea leaf extract derived ZnO and vanadium-doped ZnO nanoparticles (V-ZnO NPs). The chemical composition, structural, and morphology of ZnO and vanadium-doped ZnO NPs were examined by FTIR, XRD, and SEM-EDX. The FTIR confirmed the presence of functional groups corresponding to ZnO and vanadium-doped ZnO NPs. SEM-EDX clearly indicated the morphology of synthesised NPs; the hexagonal crystal of NPs was confirmed from XRD. In addition, the cytotoxic effect of ZnO and V-ZnO NPs was estimated against the breast cancer (MCF-7) cell line. From the assay, Vinca rosea (V. rosea) capped ZnO NPs have shown improved cytotoxic activity than that of Vinca rosea capped V-ZnO NPs. ZnO and vanadium-doped ZnO NPs showed the strongest antibacterial activity against Enterococcus, Escherichia coli, Candida albicans, and Aspergillus niger. The alpha-amylase inhibition assays demonstrated antidiabetic activity of synthesised NPs. From the assay test, results obtained Vinca rosea capped ZnO nanoparticles prepared using the green approach showed high effective antioxidant, antidiabetic activity, and anticancer activity than vanadium-doped ZnO NPs.