This study aims to develop a novel formulation of chitosan-sorafenib-conjugated folic acid nanoparticles to overcome drug resistance and induce apoptosis in human breast cancer cell line. A drug release experiment was performed, and the loading capacity and entrapment efficiency were estimated. The combined nanoformula was tested as an anticancer treatment against breast cancer cells by a flowcytometry assay, cell cycle analysis, DNA fragmentation assay, quantitative real-time polymerase chain reaction, and Western blot. Furthermore, DNA fragmentation results indicate that this novel nanoformula caused higher apoptosis data. Inhibition of cell cycle progression was also higher with this combined nanoformula. Up-regulation of caspase 3, caspase 8 and caspase 9, cytochrome c was also observed; in contrast, the expression of the antiapoptotic marker B-cell lymphoma-2 was reduced in the combined nanoformula treated group compared to their individual treatments. Therefore, this combined nanoformula can improve the survival rate of patients with breast cancer.