Abstract

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 α (PPARα) 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, PPARα 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 PPARα agonists. In current work, new 3H-benzo[b] [1,4] diazepine PPARα 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∼LY-19) were discovered to promote the expression of PPARα downstream gene, carnitine palmitoyl transterase-1 α (cpt1α). Among these active compounds, the EC₅₀ value of LY-2 against increasing cpt1α was 2.169 μΜ. Furthermore, the effect of LY-2 on cpt1α was weakened when PPARα knock down, which confirmed that it is a PPARα agonist again. Finally, the results from molecular dynamics simulations and binding free energy calculations showed that π-π stacking and hydrogen bonding interactions played key roles in the binding of LY-2 and PPARα protein and their complex maintained a stable structure to facilitate LY-2 to have a better binding affinity with PPARα protein. Taken together, compound LY-2 might be a novel lead compound for the development of potent PPARα agonists.

Communicated by Ramaswamy H. Sarma

 抽象

非酒精性脂肪性肝病 （NAFLD） 是世界上最常见的慢性肝病，其病理特征包括葡萄糖稳态失调和脂质蓄积。过氧化物酶体增殖物激活受体α （PPARα） 是脂肪酸代谢和生酮的关键调节因子，因为它的调节途径涉及激活脂肪酸摄取、加速脂肪酸氧化、抑制糖异生以及抑制炎症和纤维化。因此，PPARα 被认为是治疗 NAFLD 的潜在靶点，一些激动剂已进入临床试验，这促使我们发现更多新型 PPARα 激动剂。在目前的工作中，通过药效团建模、分子对接、衍生物结构搜索和生物测定，从 ChemDiv 数据库中鉴定出新的 3H-苯并[b] [1,4] 二氮卓类 PPARα 激动剂，其中发现化合物 LY-2 及其衍生物 （LY-10∼LY-19） 促进 PPARα 下游基因肉碱棕榈酰转α酶-1 （cpt1α） 的表达。在这些活性化合物中，LY-2 对增加 cpt1α 的 EC₅₀ 值为 2.169 μΜ。此外，当 PPARα 敲低时，LY-2 对 cpt1α 的作用减弱，这再次证实了它是 PPARα 激动剂。最后，分子动力学模拟和结合自由能计算结果表明，π-π 堆叠和氢键相互作用在 LY-2 和 PPARα 蛋白的结合中起关键作用，它们的复合物保持稳定的结构，促进 LY-2 与 PPARα 蛋白具有更好的结合亲和力。综上所述，化合物 LY-2 可能是开发强效 PPARα 激动剂的新型先导化合物。

通讯人：Ramaswamy H. Sarma