Identification of 3H-benzo[b] [1,4] diazepine derivatives as PPARα agonists by in silico studies and biochemical evaluation

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease in the world, whose pathologic features include dysregulated glucose homeostasis and lipid accumulation. Peroxisome proliferators-activated receptor a (PPAR alpha) is a key regulator of fatty acid metabolism and ketogenesis due to its regulatory pathways involve activating fatty acid uptake, accelerating fatty acid oxidation, inhibiting gluconeogenesis, and suppressing inflammation and fibrosis. Therefore, PPAR alpha is considered as a potential target for the treatment of NAFLD and some agonists have entered clinical trials, which drove us to discover more novel PPAR alpha agonists. In current work, new 3H-benzo[b] [1,4] diazepine PPAR alpha agonists were identified from the ChemDiv database by pharmacophore modeling, molecular docking, derivative structure search, and bioassays, where compound LY-2 and its derivatives (LY-10 similar to LY-19) were discovered to promote the expression of PPAR alpha downstream gene, carnitine palmitoyl transterase-1 alpha (cpt1 alpha). Among these active compounds, the EC50 value of LY-2 against increasing cpt1 alpha was 2.169 mu M. Furthermore, the effect of LY-2 on cpt1a was weakened when PPAR alpha knock down, which confirmed that it is a PPAR alpha agonist again. Finally, the results from molecular dynamics simulations and binding free energy calculations showed that pi-pi stacking and hydrogen bonding interactions played key roles in the binding of LY-2 and PPAR alpha protein and their complex maintained a stable structure to facilitate LY-2 to have a better binding affinity with PPAR alpha protein. Taken together, compound LY-2 might be a novel lead compound for the development of potent PPAR alpha agonists. [GRAPHICS] .