Redox-responsive dendritic copolymer-drug conjugates enhance therapeutic mitophagy through coordinated microtubule destabilization for synergistic triple-negative breast cancer therapy

Application of microtubule-targeting agents (MTAs) for triple-negative breast cancer (TNBC) is hampered by their limited efficacy and strong systemic toxicity. Herein, we reported dendritic copolymer-drug conjugates to synergistically disrupt microtubule dynamics and induce therapeutic mitochondrial autophagy (mitophagy), thus enhancing therapeutic efficacy of MTAs. Paclitaxel (PTX) and 2-methoxyestradiol (2ME) were conjugated to glutathione-stimuli responsive dendritic copolymers, resulting in DDS-PTX and DDS-2ME, respectively. PTX and 2ME were tumor-specifically released from DDS-PTX and DDS-2ME, and simultaneously acted on microtubule polymerization and depolymerization, respectively. Dual perturbation of microtubules triggered catastrophic microtubule network collapse, prolonged mitotic arrest and amplified mitochondrial stress. Mechanistically, severe mitotic stress activated the PINK1/Parkin pathway, driving excessive mitophagy and caspase-dependent apoptosis. In a murine TNBC model, treatment with combined DDS-PTX and DDS-2ME resulted in a tumor inhibition rate of 95.01 %, and the median survival was significantly extended compared to monotherapies with DDS-PTX or DDS-2ME. This combined formulation also remarkably reduced side effects of free PTX and 2ME. Mitophagy-mediated apoptotic amplification was explored as a therapeutic paradigm in this study to bridge cytoskeletal disruption with organelle-level vulnerability for enhanced tumor therapy. STATEMENT OF SIGNIFICANCE: Distinct redox-responsive dendritic copolymer-drug conjugates (DDS-PTX and DDS-2ME) were constructed to deliver paclitaxel and 2-methoxyestradiol for synergistic triple-negative breast cancer therapy. Tumor-specific drug release enabled spatiotemporal coordination of microtubule stabilization and depolymerization, thus inducing catastrophic microtubule fragmentation, prolonged mitotic arrest, and amplified mitochondrial stress. These effects subsequently triggered PINK1/Parkin-mediated therapeutic mitophagy and caspase-dependent apoptosis, achieving a 95.01 % tumor suppression rate and extending median survival to 56 days in murine models. Notably, the conjugates significantly reduced systemic toxicity compared to free drugs while maintaining hemocompatibility and organ safety. By integrating molecular-scale tumor microenvironment responsiveness with cytoskeletal-organelle crosstalk, this work establishes a mechanistically driven paradigm to amplify subcellular stress responses, offering a transformative strategy for refractory cancers with enhanced efficacy and safety.