基于分子对接及体外抑制实验预测芹菜素潜在药物相互作用

Prediction of potential drug interactions of apigenin based on molecular docking and in vitro inhibition experiments

该研究旨在考察芹菜素对UGT1A1酶活性的影响,从而预测其在临床用药时潜在的药物间相互作用。实验拟在前期实验基础上,首先采用计算机分子对接手段初步预测芹菜素与UGT1A1酶的结合靶点及结合强弱;其次采用体外人肝微粒体温孵体系,评价其对UGT1A1酶是否具有抑制作用。分子对接显示芹菜素对接进入UGT1A1酶蛋白F活性区,与内源性物质胆红素对接活性区重合,亲和作用中等;芹菜素黄酮母核主要与氨基酸残基ILE343和VAL345形成疏水结合Pi-Alkyl作用,同时,母核上羟基与氨基酸残基LYS346形成了额外的氢键作用,增加了分子与蛋白的结合。提示黄酮类母核结构具有与酶蛋白UGT1A1结合的特殊结构,同时母核引入羟基可增加结合能力。体外抑制实验提示芹菜素对UGT1A1酶具有中等抑制作用,抑制类型为竞争型抑制,与分子对接结果一致。综合两部分实验内容表明,芹菜素为UGT1A1酶底物,可竞争型抑制酶活性,临床用药中与UGT1A1酶底物具有药物间相互作用的风险。

The purpose of this study was to investigate the effect of apigenin on UGT1 A1 enzyme activity and to predict the potential drug-drug interaction of apigenin in clinical use.First,on the basis of previous experiments,the binding targets and binding strength of apigenin to UGT1 A1 enzyme were predicted by computer molecular docking method.Then the inhibitory effect of apigenin on UGT1 A1 enzyme was evaluated by in vitro human liver microsomal incubation system.Molecular docking results showed that apigenin was docked into the active region of UGT1 A1 enzyme protein F,consistent with the active region of bilirubin docking,with moderate affinity.Apigenin flavone mother nucleus mainly interacted with amino acid residues ILE343 and VAL345 to form hydrophobic binding Pi-Alkyl.At the same time,the hydroxyl group on the mother nucleus and the amino acid residue LYS346 formed an additional hydrogen bond,which increased the binding of the molecule to the protein.These results suggested that the flavonoid mother nucleus structure had a special structure binding to the enzyme protein UGT1 A1,and the introduction of hydroxyl groups into the mother nucleus can increase the binding ability.In vitro inhibition experiments showed that apigenin had a moderate inhibitory effect on UGT1 A1 enzyme in a way of competitive inhibition,which was consistent with the results of molecular docking.The results of two experiments showed that apigenin was the substrate of UGT1 A1 enzyme,which could inhibit the activity of UGT1 A1 enzyme competitively,and there was a risk of drug interaction between apigenin and UGT1 A1 enzyme substrate in clinical use.