Sequential nanocatalytic therapy and lysosomal dysfunction for overcoming castration-resistant prostate cancer.

Castration-resistant prostate cancer (CRPC) remains lethal due to adaptive resistance mechanisms such as stress-induced autophagy and NF-κB survival signaling. Here, an injectable fiber-in-hydrogel depot is developed for sequential delivery of a multi-enzyme nanozyme (cobalt-epigallocatechin gallate coordination nanozyme, CoNZ) and the lysosomal inhibitor chloroquine (CQ). The preferentially released CoNZ catalytically generates reactive oxygen species, including hydroxyl radicals, and oxygen in situ, inflicting oxidative damage while relieving tumor hypoxia. Simultaneously, it depletes antioxidants (glutathione, NADPH) and impedes NF-κB nuclear translocation, priming CRPC cells for apoptosis. The subsequently released CQ impedes enzymatic degradation or cleavage in endolysosomes and autolysosomes and blocks TLR9/NF-κB signaling, preventing tumor cells from repairing damage or activating pro-survival pathways. In vitro and in vivo, this two-pronged approach synergistically overcomes CRPC's defenses, achieving markedly enhanced cancer cell apoptosis and ∼80% tumor suppression (with occasional complete regression), resulting in an approximately 10-fold reduction in final tumor volume compared to the untreated control, without systemic toxicity, far surpassing single or co-administered treatments. This work demonstrates a spatiotemporally orchestrated combination of nanocatalytic therapy and lysosomal inhibition that dismantles CRPC's resistance mechanisms, highlighting a broadly applicable paradigm for overcoming therapeutic resistance in aggressive cancers.