ATP‐Activated Tumor Targeting Therapy via Bioorthogonal Alkyne‐Calixarene Host–Guest Assembly

Bioorthogonal chemistry enables precise therapeutic activation trough targeted click reactions. However, the challenge of insufficient reaction kinetics in vivo and limited tumor penetration of reagents hurdles its clinical translation. We herein report a supramolecular strategy to achieve precise tumor‐specific chemoimmunotherapy through ATP‐responsive bioorthogonal chemistry. Pre‐loading of activated alkyne derivative within a calixarene macrocycle prevents off‐target reactions during circulation while enabling tumor‐specific activation via ATP‐mediated competitive displacement. This controlled release triggers an in situ amino‐yne click reaction, simultaneously initiating targeted chemotherapy and robust immunogenic cell death (ICD). Comprehensive biological evaluations demonstrate that this host–guest complex significantly reduces systemic toxicity and enhances cellular uptake efficiency compared to the free drug. The dual targeting mechanism of combining the enhanced permeability and retention (EPR) effect with ATP responsiveness, overcomes the selectivity limitation of conventional bioorthogonal systems. The successful merger of these two fields opens new avenues for developing next‐generation stimuli‐responsive therapeutics with spatiotemporal control.