Farnesoid X receptor (FXR) is a promising target for drug discovery against nonalcoholic fatty liver disease (NAFLD). However, no FXR agonist has been approved for NAFLD so far. The R & D of FXR agonists are somewhat hindered by the lack of effective and safe chemotypes. To this end, we developed a multi-stage computational workflow to screen the Specs and ChemDiv chemical library for FXR agonists, which consisted of machine learning (ML)-based classifiers, shape-based and electrostatic-based models, a FRED-based molecular docking protocol, an ADMET prediction protocol and substructure search. As a result, we identified a novel chemotype that has never been reported before, with compound XJ02862 (ChemDiv ID: Y020-6413) as the representative. By designing an asymmetric synthesis strategy, we were able to prepare four isomers of compound XJ02862. Interestingly, one of the isomers, 2-((S)-1-((2S,4R)-2-methyl-4-(phenylamino)-3,4-dihydroquinolin-1(2H)-yl)-1-oxopropan-2-yl)hexahydro-1H-isoindole-1,3(2H)-dione (XJ02862-S2), showed potent FXR agonistic activity in HEK293T cells. The molecular docking, molecular dynamics simulations and site-directed mutagenesis suggested the hydrogen bond between compound XJ02862-S2 and HIS294 of FXR is essential for ligand binding. We further demonstrated that compound XJ02862-S2 had no agonistic effect on TGR5. Further biological experiments have shown that compound XJ02862-S2 could ameliorate hypercholesterolemia, hepatic steatosis, hyperglycemia, insulin resistance (IR) in high-fat-diet induced obese (DIO) mice. In term of molecular mechanism, compound XJ02862-S2 regulates the expression of FXR downstream genes involved in lipogenesis, cholesterol transport and bile acid biosynthesis and transport. Taken together, we have discovered a novel chemotype as potent FXR agonists for NAFLD by computational modeling, chemical synthesis and biological evaluation.

法尼醇 X 受体 (FXR) 是针对非酒精性脂肪性肝病 (NAFLD) 的药物发现的有前途的目标。然而，到目前为止，还没有 FXR 激动剂被批准用于 NAFLD。由于缺乏有效和安全的化学型，FXR激动剂的研发在一定程度上受到阻碍。为此，我们开发了一个多阶段计算工作流程来筛选 FXR 激动剂的 Specs 和 ChemDiv 化学库，其中包括基于机器学习 (ML) 的分类器、基于形状和静电的模型、基于 FRED 的分子对接协议、ADMET 预测协议和子结构搜索。结果，我们发现了一种以前从未报道过的新型化学型，化合物XJ02862（ChemDiv ID：Y020-6413) 为代表。通过设计不对称合成策略，我们能够制备化合物XJ02862的四种异构体。有趣的是，其中一种异构体 2-((S)-1-((2S,4R)-2-methyl-4-(phenylamino)-3,4-dihydroquinolin-1(2H)-yl)-1-oxopropan -2-yl)hexahydro-1H-isoindole-1,3(2H)-dione ( XJ02862-S2 ) 在 HEK293T 细胞中表现出有效的 FXR 激动活性。分子对接、分子动力学模拟和定点诱变表明化合物XJ02862-S2和 FXR 的 HIS294之间的氢键对于配体结合是必不可少的。我们进一步证明化合物XJ02862-S2对 TGR5 没有激动作用。进一步的生物学实验表明，化合物XJ02862-S2可以改善高脂肪饮食诱导的肥胖 (DIO) 小鼠的高胆固醇血症、肝脂肪变性、高血糖症、胰岛素抵抗 (IR)。在分子机制方面，化合物XJ02862-S2调节参与脂肪生成、胆固醇转运和胆汁酸生物合成和转运的 FXR 下游基因的表达。总之，我们通过计算建模、化学合成和生物学评估发现了一种新型化学型作为 NAFLD 的有效 FXR 激动剂。