CCR5受体抑制剂的虚拟筛选及体外活性评价

Virtual screening and evaluation of CCR5 receptor inhibitors in vitro

目的趋化因子C-C-基元受体5(CCR5受体)是1型艾滋病病毒(HIV-1)入侵靶细胞的关键辅助受体之一,利用计算机辅助药物设计的方法进行虚拟筛选,并进行体外活性测定,以期获得新的CCR5受体抑制剂的先导化合物。方法利用Discovery Studio 2.0软件构建HypoGen药效团模型;利用Sybyl-X 2.0软件构建Surflexdock分子对接模型;采用多级筛选的策略,运用"类药五原则"、药效团模型和分子对接模型对ZINC数据库进行逐级筛选;通过检测TZM-bl细胞中荧光素酶报告基因的表达情况来判断化合物的抗HIV-1活性,并通过MTT实验考查化合物的细胞毒性。结果成功构建了药效团和分子对接模型,并对ZINC数据库进行了多级虚拟筛选,得到6个待测化合物(B01-06);活性检测结果证实,B04的IC50为(29.52±3.73)μmol/L,CC50为(181.4±14.22)μmol/L;B06的IC50为(42.13±5.06)μmol/L,CC50为(354.9±28.82)μmol/L。结论化合物B04和B06具有一定的抗HIV-1活性,可作为潜在的先导化合物进行进一步的结构改造。

Objective To have virtual screening and evaluation of bio-active lead compounds of CCR5 receptor inhibitors in vitro by computer-aided drug design. Methods The HypoGen pharmacophore model was constructed by Discovery Studio 2.0. The Surflex-dock molecular docking model was constructed by Sybyl-X 2.0 multi-stage screening. The "Rule of Five", pharmacophore model and molecular docking model were used for screening step by step. The anti-HIV-1 activities of the compounds were tested by detecting the expression of luciferase reporter gene in TZM-bl cells. MTT assay examined the cytotoxicity of the compounds. Results The pharmacophore and molecular docking models were successfully built and six compounds(B01-06) were screened from the ZINC database by multi-level virtual screening. The median inhibition concentration(IC50) of B04 was 29.52±3.73 μmol/L, the cytotoxicity concentration(CC50 ) was 181.4±14.22 μmol/L;IC50 of B06 was 42.13±5.06 μmol/L, CC50 was 354.9±28.82 μmol/L. Conclusion B04 and B06 can be used as potential lead compounds for further structural modification.