Microenvironments of various solid tumors are characterized by hypoxia. Herein, we report a novel nanoparticle that can selectively release loaded drugs in hypoxic environments. The nanoparticle was prepared using a hypoxia-responsive amphiphilic polymer in aqueous media. The polymer was synthesized by conjugating a hydrophobic small molecule, 4-nitrobenzyl (3-azidopropyl) carbamate, to the side chains of an mPEG-PPLG copolymer. Doxorubicin (DOX) could be loaded into the nanoparticles with a high efficiency of 97.8%. The generated drug-loaded micellar nanoparticles (PPGN@DOX) presented hypoxia-sensitive drug release behavior in vitro. Meanwhile, PPGN@DOX could be effectively internalized by 4T1 cells and could release DOX into the cell nuclei under hypoxic conditions. The in vitro anticancer results suggested that PPGN@DOX presented superior tumor cell-killing ability compared with free DOX in hypoxic environments. Furthermore, PPGN@DOX prolonged the blood circulation time and improved the biological distribution of DOX, resulting in increased antitumor outcomes and reduced side effects in vivo. Overall, the present work demonstrates that hypoxia-responsive nanoparticles have great application potential in the treatment of hypoxic tumors.