We report an innovative reactive oxygen species (ROS)-based chemotherapy mechanism using doxorubicin (Dox)-loaded polyaspartamide (PA)-encapsulated superparamagnetic iron oxide nanoparticles (SPIONs). PA encapsulates SPIONs (PA-SPIONs) to create biocompatible nanocarriers with an average diameter of 152 nm. PA is conjugated with Dox, biotin, and 3-amino-1,2,4-triazole (3AT) to perform different functions in chemotherapy. As a result, 3AT inhibits endogenous catalase, biotin improves cancer cell targeting and uptake, and Dox, in conjunction with iron ions, stimulates exogenous ROS production above the lethal threshold of cancer cells. The fluorescence signal results from Dox confirm that PA-SPIONs have a great affinity for 4T1 cancer cells. Because of the pH-sensitive cleavage function of the hydrazone bond, approximately 80% of Dox is released in the cancer cell environment (pH ∼ 5). In in vitro cytotoxicity, PA-SPIONs are highly cytotoxic to 4T1 cells, with less than 50% cell viability. The 2′,7′-dichlorofluorescein diacetate test demonstrates the ROS production of the nanocarriers, while the cell apoptosis assay confirms the cell death mechanism. In an in vivo experiment, PA-SPIONs inhibited the tumor growth rate, with a tumor volume 4.5 times lower than that of the control after 28 days. The individual roles of Dox and 3AT in tumor volume growth inhibition are also demonstrated. PA-SPIONs are demonstrated for biosafety and blood compatibility. Because the nanocarriers demonstrate the high efficacy of 4T1 cancer cell treatment, it likely opens the door for future advancements in cancer-targeted chemotherapy.