Novel Chlorambucil-Platinum(IV) (CP4) prodrugs are multi-mechanistic chemotherapeutics that exhibit promising anticancer potential. The platinum scaffold of the prodrugs has been previously found to induce changes in the microtubule cytoskeleton, specifically actin and tubulin of MDA-MB-231 cells, while chlorambucil alkylates DNA. These prodrugs demonstrated significant potency in 2D cell viability assays and in MDA-MB-231 cells as 3D printed spheroids. Significant production of reactive oxygen species was observed in MDA-MB-231 cells 72 h post treatment with the prodrugs. While the mitochondrial membrane potential was substantially reduced. Cellular uptake studies of CP4 prodrugs, assessed by ICP/MS established that active transport was the primary mechanism. Early apoptosis was induced at 48 h and continued to late apoptosis/necrosis at 72 h of treatment determined by an AnnexinV-FITC/PI assay using flow cytometry. Cell cycle analysis indicates cell cycle arrest at G0/G1 at 72 h. Immunofluorescence via confocal microscopy measured significant changes in the expression intensity and cellular distribution between actin and tubulin at 72 h of treatment. Drug effects on intrinsic and extrinsic pathway apoptotic markers, along with MAPK and PI3K pathway and autophagy markers were studied by Western blot analysis. In conclusion, this study validates that Chlorambucil-Platinum(IV) prodrugs act as multi-mechanistic chemotherapeutics that exhibit promising anticancer potential.