Targeting PKLR and lipogenic enzymes through JNK inhibition to develop a therapeutic strategy for MASLD and MASH

Abstract Pyruvate kinase liver and red blood cells (PKLR) has emerged as a key gene associated with metabolic dysfunction-associated steatotic liver disease (MASLD). Through a computational drug repurposing approach, we identified JNK-IN-5A as a small molecule that significantly inhibits the c-Jun N-terminal kinase (JNK) family and suppresses PKL expression in HepG2 cells. In this study, we further evaluated JNK-IN-5A and its derivatives, including SET-151, SET-152, SET-162, and SET-130, as potential therapeutic candidates for MASLD. Building on our previously established HepG2 de novo lipogenesis (DNL) steatosis model, we demonstrated that JNK-IN-5A and its derivatives markedly reduced intracellular triacylglycerol (TAG) accumulation during DNL induction. These compounds also significantly inhibited the expression of key DNL pathway proteins, including PKL, FASN, ACACA, SCD1, SREBP1-c, and ChREBP. Global transcriptomic analyses revealed that SET-151, SET-152, and SET-162 exhibited superior anti-steatotic effects compared to SET-130 and JNK-IN-5A. These three derivatives uniquely downregulated genes involved in pyruvate metabolism, bile acid synthesis, fatty acid metabolism, and glycolysis pathways, effects not observed with JNK-IN-5A alone. Additionally, Compass analysis indicated that treatment with SET-151, SET-152, and SET-162 led to significant alterations in metabolic reactions related to lipid metabolism, whereas JNK-IN-5A showed minimal impact. Finally, we evaluated JNK-IN-5A and SET-152 in a high-sucrose, high-fat diet-induced in vivo rat model of MASLD. Both compounds significantly reduced hepatic lipid accumulation, liver stiffness, and key biochemical markers of MASLD. Collectively, our findings identified SET-152 as a promising drug candidate for the treatment of MASLD.