高尿酸血症主要由肾尿酸盐排泄不足引起。尿酸转运蛋白1(URAT1),一种有机阴离子转运蛋白,是引起尿酸盐再吸收的主要蛋白质。在这项研究中,利用基于人工智能的AlphaFold2程序构建URAT1结构模型。经过分子对接和构象评价,基于丙磺舒-URAT1,苯溴马隆-URAT1,lesinurad-URAT1和verinurad-URAT1的复杂结构,构建了四个电子药效模型。结合药效基团建模,分子对接,MM/GBSA计算和ADME预测,从含有10,968个分子的天然产物数据库中选择25种类黄酮。然后,构建了过表达URAT1的HEK-293T细胞模型,并通过测量其对6-羧基荧光素(6-CFL)细胞摄取的影响来评估25种类黄酮对URAT1的抑制活性。Fisetin,黄芩素,刺槐素显示出最佳活性,IC50值为12.77、26.71和57.30µM,分别。最后,通过分子对接和分子动力学模拟分析了这三种黄酮类化合物的构效关系。结果表明,C-4上的羰基和C-7上的羟基,C-4',类黄酮中的C-5\'有利于URAT1抑制作用。这项研究促进了黄酮类化合物在URAT1抑制剂开发中的应用。由RamaswamyH.Sarma沟通。
Hyperuricemia is mainly caused by insufficient renal urate excretion. Urate transporter 1 (
URAT1), an organic anion transporter, is the main protein responsible for urate reabsorption. In this study, we utilized artificial intelligence-based AlphaFold2 program to construct
URAT1 structural model. After molecular docking and conformational evaluation, four e-pharmacophoric models were constructed based on the complex structures of probenecid-URAT1, benzbromarone-
URAT1, lesinurad-
URAT1, and verinurad-
URAT1. Combining pharmacophore modeling, molecular docking, MM/GBSA calculation and ADME prediction, 25 flavonoids were selected from the natural products database containing 10,968 molecules. Then, a model of HEK-293T cells overexpressing
URAT1 was constructed, and the inhibitory activity to
URAT1 of 25 flavonoids was evaluated by measuring their effect on cellular uptake of 6-carboxyfluorescein (6-CFL). Fisetin, baicalein, and acacetin showed the best activity with IC50 values of 12.77, 26.71, and 57.30 µM, respectively. Finally, the structure-activity relationship of these three flavonoids was analyzed by molecular docking and molecular dynamics simulations. The results showed that the carbonyl group on C-4 and hydroxyl group on C-7, C-4\', and C-5\' in flavonoids were conducive for URAT1 inhibitory effects. This study facilitates the application of flavonoids in the development of URAT1 inhibitors.Communicated by Ramaswamy H. Sarma.