The preparation technique of silver nanoparticles (AgNPs) is essential for the efficient drug delivery system. AgNPs coated PEG were prepared and investigated to control NPs physicochemical characteristics as well as a selective delivery of doxorubicin (DOX) to the affected site with lower side effects. In this context, AgNPs chemically synthesized and coated with PEG having different molecular weights via two different methods. The optimized AgNPs-PEG were further loaded with DOX. The prepared particles were characterized for their size, shape, and DOX in vitro release in phosphate buffer of two pH conditions 5.0 and 7.4. Moreover, the cytotoxicity of the prepared nanoformulations was investigated against MCF-7 and HepG2 cell lines, also the biodistribution of AgNPs was also investigated in rats. It was found that the investigated two methods for AgNPs-PEG preparation did not show a significant difference on the AgNPs characteristics. The prepared AgNPs were spherical in shape and showed mono-dispersibility distribution with encapsulation efficiency percent approximately 95%. DOX showed higher release in pH 5.0 than that observed in pH 7.4. Moreover, the prepared system showed sustained release behavior compared to the unloaded DOX. AgNPs loaded with DOX significantly reduced the IC_50% compared with AgNPs-PEGs and unloaded DOX against both MCF-7 and HepG2 cell lines. In addition, biodistribution study revealed the higher accumulation of AgNPs-PEG in liver compared to the other investigated organs. It could be concluded that the optimized AgNPs-PEG could be considered as an efficient carrier for DOX with high drug loading, sustained DOX delivery, higher synergistic cytotoxicity, and lower side effects on the other healthy cells.