Acidity-unlocked glucose oxidase as drug vector to boost intratumor copper homeostatic imbalance-enhanced cuproptosis for metastasis inhibition and anti-tumor immunity.

As one of the key tools of biocatalysis, natural enzymes have received extensive attention due to their unique activity. However, the non-selective catalysis and early leakage induced by delivery dependency of natural enzymes can cause side effects on normal tissues. Moreover, although cuproptosis is an emerging tumor-inhibiting programmed cell death, the occurrence of cuproptosis leads to high expression of Cu-dependent lysyl oxidase-like 2 (LOXL2), which promotes tumor metastasis. Herein, in order to intelligently regulate the "OFF-to-ON" catalytic activity of glucose oxidase (a natural enzyme called GOx) and simultaneously inhibit tumor metastasis caused by Cu imbalance, an acidity-unlocked GOx system drug carrier was constructed by co-assembling Cu ions and omeprazole (OPZ) on GOx exposing sulfhydryl and hydrophobic pockets. The GOx activity is significantly inhibited due to the coordination of Cu ions with sulfhydryl groups and the interaction of hydrophobic small molecule OPZ with hydrophobic bags, which results in specificity for tumor cells and ensures the safety of GOx in blood circulation. Meanwhile, dysregulation of intracellular Cu homeostasis that impairs the Cu-dependence of LOXL2 not only inhibits critical signaling during epithelial-mesenchymal transformation (EMT) and extracellular matrix (ECM) remodelling to prevent tumor metastasis, but also exacerbates enhanced cuproptosis induced by tumor metabolic stress, thereby reversing the immunosuppressive microenvironment. This strategy of acidity-unlocked the catalytic function of natural enzymes and LOXL2 activity inhibition provides a novel option for enhancing cuproptosis to inhibit tumor metastasis and anti-tumor immunity.