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The purpose of this study is to prepare nanoencapsulation synthesized with a new formulation of Fe@Au-CS-CU-FA nanoparticle (NPs) by pulsed laser ablation in liquid (PLAL) method as drug delivery to treat breast cancer (T-47D) and (MCF12A) as a normal cell line. The method synthesized Fe@Au NPs using the PLAL working at wavelength 532 nm with different laser fluence (1.9, 2.2, and 2.5) J/cm2. These Fe@Au NPs were characterized by atomic force microscope (AFM), field emission scanning electron microscopy (FESEM), and high-resolution transmission electron microscopy (HRTEM). The obtained mean sizes of Fe@Au NPs were 63.65, 32.47, and 31.18 nm at 1.9, 2.2, and 2.5 J/cm2, respectively. Results of the MTT assay of CU loaded Fe@Au-CS-FA NPs on human breast cancer cell line (T-47D) confirmed that cytotoxicity of CU can enhance when they are loaded on Fe@Au-CS-FA NPs in comparison with free CU. While results of flow cytometry showed that this combination can increase the therapeutic effects of CU by apoptosis induction in the T-47D cell line. Conclusion of Fe@Au-CS-CU-FA NPs causes a decrease in T-47D cell viability and caused induces 85% apoptosis. The in vivo study of Fe@Au-CS-CU-FA nanoformulation confirmed that the mean tumor size decreases in time. [ABSTRACT FROM AUTHOR]