Background: Pyruvate kinase liver and red blood cells (PKLR) is linked to metabolicdysfunction-associated steatotic liver disease (MASLD). Previous study, weidentified JNK-IN-5A, a c-Jun N-terminal kinase (JNK) inhibitor thatsuppresses PKL expression in HepG2 cells using computational drugrepurposing and screened out four hit JNK-IN-5A derivatives (SET-151, SET-152, SET-162, SET-130).
Materials and Methods: We validated therapeutic efficacy of JNK-IN-5A and fourderivative (SET-151, SET-152, SET-162, SET-130). HepG2 de novo lipogenesis(DNL) steatosis model was used in vitro validation. RNA sequencing data wereanalysed using systems biology approaches, including transcriptomic profilingand COMPASS analysis. GLP-like toxicity assessment in rat model shows in vivosafety and MASLD rat model revealed in vivo therapeutic effect to MASLDand MASH.
Results: In a HepG2 DNL steatosis model, all compounds reduced intracellulartriacylglycerol (TAG) and inhibited key DNL proteins (PKL, FASN, ACACA, SCD1,SREBP1-c, ChREBP). Transcriptomic profiling revealed stronger anti-steatoticeffects with SET-151, SET-152, and SET-162, which uniquely downregulatedgenes in pyruvate metabolism, bile acid synthesis, fatty acid metabolism, andglycolysis. Compass analysis showed these derivatives significantly altered lipidrelatedmetabolic reactions, unlike JNK-IN-5A. In a high-sucrose, high-fat dietinducedMASLD rat model, JNK-IN-5A and SET-152 reduced hepatic lipidaccumulation, liver stiffness, and MASLD biomarkers.
