In this study, polyhydroxybutyrate (PHB)-coated magnetic nanoparticles (MNPs) were prepared by coprecipitation of iron salts (Fe and Fe) by ammonium hydroxide. Characterizations of PHB-coated MNPs were performed by Fourier transform infrared spectroscopy, x-ray diffraction, dynamic light scattering, thermal gravimetric analysis, vibrating sample magnetometry, and transmission electron microscopy analyses. Doxorubicin was loaded onto PHB-MNPs, and the release efficiencies at different pHs were studied under in vitro conditions. The most efficient drug loading concentration was found about 87% at room temperature in phosphate-buffered saline (pH 7.2). The drug-loaded MNPs were stable up to 2 months in neutral pH for mimicking physiological conditions. The drug release studies were performed with acetate buffer (pH 4.5) that mimics endosomal pH. Doxorubicin (60%) released from PHB-MNPs within 65 hours. Doxorubicin-loaded PHB-MNPs were about 2.5-fold more cytotoxic as compared with free drug on resistant Michigan Cancer Foundation-7 (human breast adenocarcinoma, MCF-7) cell line (1 μM doxorubicin) in vitro. Therefore, doxorubicin-loaded PHB-MNPs lead to overcome the drug resistance.