Anticancer agent-incorporating polymeric micelles accumulate effectively in tumors via the enhanced permeability and retention effect to exert potent antitumor effects. However, combined use of such micelles has not been elucidated. We compared the effect of combining the epirubicin-incorporating micelle NC-6300 and 1,2-diaminocyclohexane platinum (II) (oxaliplatin parent complex)-incorporating micelle NC-4016 (NCs) with that of epirubicin and oxaliplatin (E/O) in 44As3Luc cells using the combination index method. The in vivo antitumor activities of NCs and E/O were evaluated in mice bearing 44As3Luc xenografts. Pharmacokinetic analysis was performed by high-performance liquid chromatography and mass spectrometry. Cardiotoxicity of NC-6300 and epirubicin was assessed by echocardiography. Neurotoxicity of NC-4016 and oxaliplatin was evaluated by examining the paw withdrawal response to noxious mechanical stimuli. NCs showed a highly synergistic activity equivalent to E/O. In vivo, NCs exhibited higher antitumor activity in the subcutaneous tumor model and longer overall survival in the orthotopic tumor model than E/O (p < 0.001, p 5 0.015, respectively). The intratumor concentrations of epirubicin and platinum were significantly higher following NCs than following E/O administration. Moreover, the micelles showed lower cardiotoxicity and neurotoxicity than the corresponding conventional drugs. The combined use of the micelles was associated with remarkable efficacy and favorable toxicities in the human gastric cancer model, and warrants the conduct of clinical trials.The therapeutic window of cytotoxic anticancer agents (ACAs) is known to be narrow; at concentrations falling outside this window, these agents either fail to work or cause severe adverse events. To overcome this drawback, drug delivery systems have been developed. By utilizing the enhanced permeability and retention (EPR) effect, based on the leaky blood vessels and impaired lymphatic drainage in tumor tissues, drug delivery systems in nanoparticle-form enable ACAs to accumulate selectively in the tumors and exert prominent antitumor effect while decreasing the toxicity of the drug payload.1 Several preclinical studies have demonstrated the advantages of using ACA-incorporating polymeric micelles, and some are currently under clinical evaluation as single micelle agents alone 2-6 or single micelle agents in