Nanoscale particles, as drug carriers, have a potential to improve drug therapeutic efficiency. However, the feasible design of the nanostructure containing chemotherapeutic agents and the behavior of the delivery to tumor tissues and cells has not been adequately investigated. In this study, we developed a novel nanoparticle, consisting of a gelatinase-cleavage peptide with poly(ethylene glycol) (PEG) and poly(epsilon-caprolactone) (PCL)-based structure for tumor-targeted EVO (an alkaloid isolated from Evodia rutaecarpa) delivery. We found that EVO-NPs were transformed by gelatinases, which could significantly promote drug release and enhance the cellular uptake of EVO (P < 0.01). In vivo biodistribution study demonstrated that targeted EVO-NPs could accumulate and remain in the tumor regions. Moreover, EVO-NPs exhibited higher tumor growth suppression than EVO on hepatic H22 tumor model via intravenous administration (P < 0.01). Both in vitro and in vivo experiments suggested that the gelatinase-mediated nanoscale delivery system was promising for improvement of antitumor efficacy in various over-expressed gelatinase cancers.