Targeting LXRα-R305 covalent activation for renal copper homeostasis restoration by natural pestaphilone A as an anti-AKI candidate

No first-in-class therapeutics for acute kidney injury (AKI) have gained global regulatory approval. Furthermore, the mechanism by which the copper modulation attenuates AKI remains elusive. Herein, natural pestaphilone A (PPA) was identified via high-content screening (HCS) of a structurally privileged natural products library and was found to be a potent anti-AKI lead targeting copper homeostasis. More importantly, PPA displayed robust drug development potential, underpinned by its desirable pharmacokinetics (43.9% oral bioavailability) and an acceptable safety margin (LD 50 70 mg/kg, i.v.). Combining DARTS analysis with probe (PPA-P) co-localization, we found that liver X receptor α (LXR α ), which was significantly downregulated in AKI patients, served as the direct binding target of PPA. A chemical proteomics approach was employed to verify that PPA directly induced the covalent modification of Arg305 on LXR α , an event that initiated a distinct signaling cascade. Mechanistically, this activated LXR α transcriptionally regulated the –1653/–1635 bp fragment of the SLC31A1 promoter, which elevated effector proteins CCS and SOD1, and thereby restored copper homeostasis in AKI. Our findings uncover an unreported LXR α –SCL31A1 axis governing copper homeostasis in AKI, and highlight PPA as a promising anti-AKI candidate.