Enzyme‐Responsive Metallopeptide Hydrogel Enables Cancer Cell‐Selective Prodrug Activation via Bioorthogonal Catalysis

Abstract Chemotherapy is often hindered by systemic toxicity and poor selectivity. To address these issues, we develop an enzyme‐responsive metallopeptide hydrogel (H₂Y_p‐Pd) that integrates enzyme‐instructed self‐assembly (EISA) and bioorthogonal catalysis for selective tumor‐targeted prodrug activation. Upon exposure to alkaline phosphatase (ALP), which is overexpressed in osteosarcoma cells (Saos‐2), H₂Y_p‐Pd selectively accumulates and self‐assembles into catalytic nanofibers. These assemblies function as both a reservoir of palladium catalyst and a “drug factory” to in situ activate a caged doxorubicin prodrug (Alloc‐Dox) via Pd‐mediated deallylation. This system achieves nearly 90% prodrug conversion in vitro, exhibiting potent cytotoxicity against cancer cells while sparing normal cell viability. Additionally, the H₂Y_p‐Pd/Alloc‐Dox combination significantly suppresses cancer cell migration and invasion, demonstrating therapeutic efficacy comparable to free doxorubicin. The hydrogel's injectability and biocompatibility further highlight its potential as an implantable catalytic depot for localized prodrug activation. By integrating enzyme targeting with bioorthogonal catalysis, this work offers an attractive paradigm for spatiotemporally controlled prodrug activation, and a promising solution to the efficacy versus safety dilemma of chemotherapy.