Virtual screening of Kv1.3 inhibitors and investigation of their molecular mechanisms in alleviating alcoholic liver injury in mice.

Alcohol-related liver disease remains a global health challenge lacking effective pharmacotherapies. Given the role of the voltage-gated potassium channel Kv1.3 in immune regulation, this study aimed to identify potent Kv1.3 inhibitors for alcohol-related liver disease treatment. Guided by the GTEx database analysis linking Kv1.3 to hepatic inflammation, we conducted computer-aided virtual screening of 38,752 compounds. The lead compound, Z299576254, was selected and validated using membrane patch-clamp technology, demonstrating high affinity and significant inhibition of Kv1.3 outward currents. In a mouse model of alcohol-related liver disease, intraperitoneal administration of Z299576254 significantly alleviated hepatic injury, lipid deposition, and inflammatory responses without inducing hepatorenal toxicity. Mechanistic investigations using RNA-seq in RAW264.7 cells revealed that Z299576254 suppresses macrophage inflammatory responses by modulating the PI3K/AKT signaling pathway. Consequently, Z299576254 was identified as a safe and potent Kv1.3 inhibitor, offering a promising therapeutic strategy and molecular insight for managing alcohol-related liver disease.