Enzyme‐Activatable Near‐Infrared Photosensitizer with High Enrichment in Tumor Cells Based on a Multi‐Effect Design

Abstract Enzyme‐activatable near‐infrared (NIR) fluorescent probes and photosensitizers (PSs) have emerged as promising tools for molecular imaging and photodynamic therapy (PDT). However, in living organisms selective retention or even enrichment of these reagents after enzymatic activation at or near sites of interest remains a challenging task. Herein, we integrate non‐covalent and covalent retention approaches to introduce a novel “1‐to‐3” multi‐effect strategy—one enzymatic stimulus leads to three types of effects—for the design of an enzyme‐activatable NIR probe or PS. Using this strategy, we have constructed an alkaline phosphatase (ALP)‐activatable NIR fluorogenic probe and a NIR PS, which proved to be selectively activated by ALP to switch on NIR fluorescence or photosensitizing ability, respectively. Additionally, these reagents showed significant enrichment (over 2000‐fold) in ALP‐overexpressed tumor cells compared to the culture medium, accompanied by massive depletion of intracellular thiols, the major antioxidants in cells. The investigation of this ALP‐activatable NIR PS in an in vivo PDT model resulted in complete suppression of HeLa tumors and full recovery of all tested mice. Encouragingly, even a single administration of this NIR PS was sufficient to completely suppress tumors in mice, demonstrating the high potential of this strategy in biomedical applications.