靶向ERK2抑制剂的活性筛选与机理研究

Bioassay Screening and Binding Mode Study of Inhibitors Targeting at ERK2

关键词:  细胞外信号调节激酶2  分子对接  蛋白表达  活性筛选  结合模式

Key words:  extracellular signal-regulated kinase 2  molecular docking  protein expression  bioassay screening  binding mode

摘要:

细胞外信号调节激酶2(Extracellular Signal-Regulated Kinase 2，ERK2)是恶性肿瘤发生、发展过程中的关键激酶，目前还没有靶向ERK2的药物获批上市。本研究旨在获得结构新颖的靶向ERK2小分子抑制剂。通过质粒构建、蛋白表达与体外激酶活性测试，对本实验室构建的北部湾次级代谢产物及化学合成化合物库中的化合物开展生物活性筛选，利用分子对接阐明化合物与ERK2的结合模式及作用机理。试验最终筛选获得6-甲氧基红镰霉素B、异红镰霉素及嘧啶脲类等5个具有较高抑制活性的化合物(统一命名为化合物1-5)，其半抑制浓度(IC₅₀)分别为(4.17±1.82)、(13.39±0.93)、(27.85±2.55)、(19.14±1.02)和(8.55±3.72) μmol/L。化合物1-5对ERK2具有抑制活性，它们与ERK2中的LYS-54、MET-108及GLN-105残基形成的氢键是影响两者结合稳定性的关键因素。

Abstract:

Extracellular Signal-Regulated Kinase 2 (ERK2) is a key kinase in the occurrence and development of malignant tumors.At present,no drugs targeting ERK2 have been approved for marketing.The purpose of this study is to obtain novel small molecule inhibitors targeting ERK2.Through plasmid construction,protein expression and kinase activity test in vitro,biological activity screening was carried out on the secondary metabolites and compounds in the chemical synthesis compound library of Beibu Gulf constructed in our laboratory.Molecular docking was used to elucidate the binding mode and mechanism of the compounds with ERK2.Finally,5 compounds(named as compound 1-5) with high inhibitory activity of 6-methoxy-erythromycin B,isoerythromycin and pyrimidine urea were obtained by screening,and their half-maximal inhibitory concentrations (IC₅₀) were (4.17±1.82),(13.39±0.93),(27.85±2.55),(19.14±1.02) and (8.55±3.72) μmol/L,respectively.Compound 1-5 have inhibitory activity against ERK2 and the hydrogen bond formed between them and residues of LYS-54,MET-108 and GLN-105 in ERK2 are the key factors affecting the stability of the binding.