In summary, this study introduces a new concept of regulating self-immolation and cargo release using the bioorthogonal cucurbit [7]uril-mediated supramolecular chemistry. Through a structure-activity relationship study of a subgroup of rationally designed self-immolative linkers, we identified the SIE2 scaffold featuring two CB-binding benzyl substitutes. When forming a dual-binding supramolecular complex with CB, the cyclization-elimination rate of SIE2 was downregulated up to 260-fold under our experimental conditions. Importantly, this rate can be reversibly restored through the addition of a competitive adamantane guest molecule, allowing for the triggered release of covalently bound cargo molecules from the CB/SIE2 supramolecular complex. Leveraging insights from the SIE2 design, we developed a SIE5 scaffold that enables the modular incorporation of different functional groups. An MB photosensitizer was incorporated into the SIE5 prodrug for a proof-ofconcept demonstration. Material characterizations and in-vitro cellular experiments confirmed the deactivation of MB release and PDT effect in the CB-SIE5 supramolecular complex, which could be reactivated by the introduction of the supramolecular trigger adamantane. This supramolecular strategy to regulate self-immolative chemistry provides a general bioorthogonal approach for prodrug activation and controlled release and finds promising applications in medicinal chemistry, drug delivery, materials science, and beyond.