Induction of cuproptosis via targeting mitochondrial metabolism using dPTBD with potent therapeutic activity against KRAS-driven cancer.

Cuproptosis is a type of recently reported cell death characterized by aberrant accumulation of copper ions within cells, leading to mitochondrial stress and protein aggregation. Recent studies suggest that certain cancer cells are particularly susceptible to cuproptosis-inducing agents. However, the genetic determinants of cellular sensitivity to cuproptosis and the therapeutic potential of cuproptosis inducers in cancer treatment remain unclear. Here, we report the discovery of a small molecule, N1,N1-dimethyl-N4-(4-(pyridin-2-yl)thiazol-2-yl)benzene-1,4-diamine (dPTBD), that targets KRAS-driven cancer via a tetracycline-inducible cell-based drug screening. dPTBD exhibited significant antitumor efficacy in KRAS-driven cancers both in vitro and in vivo. Mechanistic studies revealed that dPTBD acted as a copper ionophore, promoting intracellular copper accumulation particularly in the mitochondria, leading to metabolic disruption and cuproptotic cell death. Adding trace amount of copper massively enhanced the cytotoxic effect of dPTBD, resulting in an immediate mitochondrial dysfunction and cuproptosis. In preclinical models, dPTBD, either alone or combined with physiologically tolerable amount of copper, significantly suppressed tumor growth in KRAS-mutant pancreatic and colon cancer xenografts. Taken together, our study reveals that induction of cuproptosis is a new therapeutic strategy for KRAS-driven cancer and identifies dPTBD as a lead compound for future development.