A Concerted Enzymatic and Bioorthogonal Approach for Extra‐ and Intracellular Activation of Environment‐Sensitive Ruthenium(II)‐Based Imaging Probes and Photosensitizers

Abstract In this article, we report a novel targeting strategy involving the combination of an enzyme‐instructed self‐assembly (EISA) moiety and a strained cycloalkyne to generate large accumulation of bioorthogonal sites in cancer cells. These bioorthogonal sites can serve as activation triggers in different regions for transition metal‐based probes, which are new ruthenium(II) complexes carrying a tetrazine unit for controllable phosphorescence and singlet oxygen generation. Importantly, the environment‐sensitive emission of the complexes can be further enhanced in the hydrophobic regions offered by the large supramolecular assemblies, which is highly advantageous to biological imaging. Additionally, the (photo)cytotoxicity of the large supramolecular assemblies containing the complexes was investigated, and the results illustrate that cellular localization (extracellular and intracellular) imposes a profound impact on the efficiencies of photosensitizers.