Wang_2026_J.Am.Chem.Soc__

Photodynamic therapy (PDT) is effective for localized cancers, but its reliance on external light limits treatment depth and uniformity. Herein, we report a programmable supramolecular chemiluminescence (CL)-induced PDT platform that generates reactive oxygen species (ROS) in the absence of light irradiation and displays intrinsic cancer selectivity. This platform is constructed through the supramolecular interaction between enzyme-responsive chemiluminescent spiroadamantyl phenoxy-1,2-dioxetane probes (ADO(P), ADO(E), and ADO(G)) and luminescent iridium(III) complexes bearing a trimethyl-beta-cyclodextrin (TMCD) host [Ir(N()C)(2)(bpy-TMCD)](Cl) (HN()C = 2-phenylpyridine (Hppy) (1a), 2-phenylquinoline (Hpq) (2a), and 2-(1-naphthyl)benzothiazole (Hbsn) (3a)). The chemiluminescent probes exhibited CL upon reaction with alkaline phosphatase (ALP), porcine liver esterase (PLE), and beta-galactosidase (beta-gal); while the iridium(III) complexes showed intense phosphorescence with efficient singlet oxygen production, distinct intracellular localization, and tunable (photo)cytotoxicity. In aqueous solutions, mixing the chemiluminescent probes with TMCD-tagged iridium(III) complexes afforded supramolecular adducts that, upon enzyme activation, underwent chemiluminescence resonance energy transfer (CRET) from the dioxetane donors to the iridium(III) acceptors. Remarkably, the ALP-responsive conjugate formed from probe ADO(P) and complex 3a (adduct ADO(P)-3a) selectively generated ROS and elicited potent cytotoxicity in cancerous HeLa cells and spheroids under light-free conditions, with apoptosis as the predominant cell death pathway, while remaining noncytotoxic in normal HEK293 cells. This modular platform couples supramolecular host-guest assembly, enzyme-specific activation, and CRET-induced ROS generation to overcome the limitations of conventional PDT, enabling cancer-selective, light-free therapy.