Self-Boosting Programmable Release of Multiple Therapeutic Agents by Activatable Heterodimeric Prodrug-Enzyme Assembly for Antitumor Therapy

Chemotherapy suffers from systemic toxicity, and tumor microenvironment (TME)-responsive prodrugs often fail to release sufficient drugs under low reactive oxygen species (ROS) levels. This study develops a self-boosting, ROS-activatable prodrug-enzyme assembly for multimodal antitumor therapy. A heterodimeric prodrug (NTP) comprising paclitaxel (PTX), a Type-I photosensitizer (NBS), and a thioketal (TK) linker was synthesized and assembled with glucose oxidase (GOx) via hydrophobic interactions, enabling therapeutic activation through acidic TME-triggered dissociation and hydrogen peroxide (H2O2)-responsive drug release. NTP@GOx released 83.57% PTX and 94.84% NBS in acidic TME, generating superoxide radical (O2-•) for Type-I photodynamic therapy (PDT). This resulted in 98% tumor inhibition in mice, with complete tumor eradication in 60% of cases. Dual-modality imaging confirmed enhanced tumor accumulation and hypoxia induction. NTP@GOx synergizes starvation therapy (GOx), chemotherapy (PTX), and O2-independent PDT (NBS) through self-boosting ROS generation and pH feedback, offering a promising strategy for multimodal cancer treatment.