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    Area of Science:

    • Immunology and Pharmacology
    • Polymer Chemistry and Drug Delivery

    Background:

    • Stimulator of interferon genes (STING) pathway activation is crucial for cancer and viral infection treatments.
    • Dimeric-amidobenzimidazoles (diABZI) are promising small-molecule STING agonists, often conjugated to carriers.
    • Previous conjugates utilized enzyme-cleavable linkers, but stable conjugation strategies were unexplored.

    Purpose of the Study:

    • To evaluate the STING-activating potential of macromolecular diABZI conjugates with stable linkers.
    • To develop novel polymer-drug conjugates for enhanced STING agonist delivery.
    • To investigate the in vivo efficacy and cellular uptake mechanisms of these new conjugates.

    Main Methods:

    • Covalent ligation of diABZI to polyethylene glycol (mPEG) chains via amide bonds.
    • Synthesis of large dimethylacrylamide (DMA) polymers directly from a diABZI-functionalized RAFT agent.
    • In vitro STING activation assays, in vivo tumor growth inhibition studies, and flow cytometry/microscopy for cellular uptake analysis.

    Main Results:

    • Stable macromolecular diABZI conjugates activated STING in vitro with kinetics comparable to small molecules.
    • These conjugates demonstrated significant in vivo tumor growth inhibition.
    • Cellular uptake via endocytosis was observed, with colocalization to the ER suggesting effective STING delivery.

    Conclusions:

    • Stable macromolecular diABZI conjugates possess high immunostimulatory potency.
    • These findings support the development of polymer-drug conjugates for STING agonist delivery.
    • Endosomal and intracellular trafficking pathways offer alternative routes for STING activation.