Abstract: OBJECTIVE To design a novel nanoparticle delivery system(HMME-PEI-HAuNS) based on hollow gold nanospheres(HAuNS), and to investigate the photothermal and photodynamic antitumor activity under the illumination of near-infrared light. METHODS HAuNS were prepared by a redox reaction using cobalt nanoparticles as a template. Hematopor phyrin monomethylether(HMME) was loaded on the surface of HAuNS via poly(ethylene imine)(PEI) as a linker to form a final nanoparticle delivery system(HMME-PEI-HAuNS). The structure of HMME-PEI-HAuNS was confirmed by NMR, IR and UV spectra. A tumor model of SKOV3 in mice was established and the biodistribution of HMME-PEI-HAuNS was investigated by fluorescence in vivo imager. Targeting peptide with specific affinity for tumor cell surface EphB4 receptors TNYL was further modified to enhance the tumor-targeting accumulation of the nanosystem. SKOV3 cells were stained with Hoechst, and the fluorescence intensity in the cells was observed using a laser confocal microscope. Cytotoxicity was evaluated by MTT colorimetry. RESULTS The HMME was successfully loaded in HAuNS with a rate of (63.4±5.2)%. Due to the enhanced permeability and retention effect of tumor, HMME-PEI-HAuNS had more accumulation and longer retention time in the tumor site than free HMME and HMME-PEI micelles, and the cumulative efficiency was about 1.6%. It was found that the targeting of nanosphere was significantly higher under the mediation of TNYL polypeptides. Under 808 nm laser irradiation, TNYL-HMME-PEI-HAuNS played a synergistic effect of photothermal and photodynamic therapy, inducing a powerful tumor cell killing, and the cell survival rate was less than 10% at high concentrations. CONCLUSION Active targeting nanosphere(TNYL-HMME-PEI-HAuNS) have strong photothermal and photodynamic antitumor effect under 808 nm NIR irradiation.