基于药效团模型虚拟筛选活络丸中治疗骨关节炎的COX-2抑制成分

Virtual Screening of COX-2 Inhibitory Components in Huoluowan(HLW)for Treatment of Osteoarthritis Based on Pharmacophore Model

目的采用药效团模型-分子对接-结合自由计算的分层虚拟筛选策略结合酶活性抑制实验,挖掘活络丸中对环氧合酶2(cyclooxygenase-2,COX-2)有抑制作用的成分。方法使用Schrödinger 2020-4软件中PHASE模块进行COX-2小分子抑制剂药效团模型的构建,应用筛选出来的最佳药效团模型对活络丸中药化合物库进行虚拟筛选,对筛选到符合药效团模型的成分进行基于靶点结构的分子对接、结合自由能计算,选择潜在抑制成分进行酶活性测定实验,并对活性较好的小分子进行药代动力学、毒理学性质预测。结果构建的最佳药效团模型具有两个氢键受体和两个芳环中心;体外酶活性抑制实验结果显示,筛选出的11个潜在靶向抑制成分均对COX-2具有不同程度的抑制作用,其中抑制活性较强的成分为表儿茶素IC_(50)为(0.93±0.15)μmol·L^(-1)、木犀草素IC_(50)为(1.96±0.19)μmol·L^(-1)及槲皮素IC_(50)为(2.09±0.28)μmol·L^(-1),ADMET计算发现木犀草素、表儿茶素、槲皮素成药性较好。结论采用药效团模型、分子对接、结合自由能计算及体外酶活性抑制实验,挖掘活络丸中对COX-2有抑制作用的成分,为骨关节炎中药来源的单体成分的现代化研究提供线索。

OBJECTIVE To explore the components in Huoluowan(HLW)that inhibit cyclooxygenase-2(COX-2)by using the hierarchical virtual screening strategy of pharmacophore,molecular docking,free energy calculation combined enzyme activity inhibition experiment.METHODS The phase module in Schrödinger 2020-4 software was used to construct the pharmacophore model of COX-2 small molecule inhibitor,the selected optimal pharmacophore model was used to screen virtually the traditional Chinese medicine compound library of HLW,and the screened components that meet the pharmacophore model are subjected to molecular docking and binding free energy calculation based on target structure.The potential inhibitory components were selected for enzyme activity determination,and the pharmacokinetic and toxicological properties of small molecules with good activity were predicted.RESULTS The best pharmacophore model has two hydrogen bond receptors and two aromatic ring centers.The enzyme activity assay results showed that all the small molecules had different degrees of inhibition on COX-2 and the component with stronger inhibition activity was epicatechin with IC_(50) of(0.93±0.15)μmol·L^(-1),the IC_(50) of luteolin was(1.96±0.19)μmol·L^(-1),the IC_(50) of quercetin was(2.09±0.28)μmol·L^(-1),ADMET calculation results showed that luteolin,epicatechin and quercetin had good drug properties.CONCLUSION In this study,pharmacophore model,molecular docking,combined free energy calculation and in vitro enzyme activity inhibition experiment are used to excavate the components in HLW that inhibit COX-2.At the same time,it provides clues for the modern research of the monomer components of traditional Chinese medicine for osteoarthritis.