Unraveling the structure-activity relationship of alloying in nanozyme design for synergistic tumor metabolic-immunotherapy

Catalyzing the glucose cascade reaction to impair tumor cell energy metabolism represents a promising strategy for tumor therapy. However, the application of natural enzymes as therapeutic agents remains limited by various challenges. Although nanozymes with multi-enzyme activities, including glucose oxidase-like (GOx-like) activity, have been reported, their development often involves complex material combinations and cumbersome synthesis processes. Here, we develop a nanozyme with GOx-, peroxidase (POD)-, superoxide dismutase (SOD)-, and NADH oxidase (NOX)-mimic activities by simply controlling the AuPt alloy ratio. The optimal cascade activity was observed for the nanozyme at an Au: Pt proportion of 13:7. Density functional theory (DFT) calculations revealed that Au sites drive glucose dehydrogenation (GOx-like), while Pt sites facilitate hydroxyl radical (•OH) generation (POD-like). Both in vitro and in vivo data indicated that Au 13 Pt 7 nanozymes disrupt tumor redox/metabolic homeostasis by depleting glucose and generating cytotoxic •OH, and impairing mitochondrial function via NOX-like, thereby inducing cell apoptosis. Notably, apoptotic immunogenic cell death (ICD) induces antitumor immunity and suppresses tumor metastasis. This study presents an innovative strategy for engineering nanozymes with multi-enzyme catalytic capabilities while demonstrating the promising application of alloy-based nanozymes in synergistic metabolic-immunotherapy.