靶向血小板源性生长因子-BB的抗肝纤维化药物筛选研究  

Screening study of antifibrotic drug targeting platelet derived growth factor-BB

作　　者：雷蕾 李健蕊[1] 孙韩 王学凯 李虎[1] 彭宗根[1] LEI Lei;LI Jian-rui;SUN Han;WANG Xue-kai;LI Hu;PENG Zong-gen(CAMS Key Laboratory of Antiviral Drug Research,Institute of Medicinal Biotechnology,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100050,China)

机构地区：[1]中国医学科学院北京协和医学院医药生物技术研究所抗病毒药物研究重点实验室,北京100050

出　　处：《中国医药生物技术》2024年第6期533-540,共8页Chinese Medicinal Biotechnology

基　　金：“重大新药创制”国家科技重大专项(2018ZX09711001-003-010);中国医学科学院医学与健康科技创新工程(2022-I2M-2-002)。

摘　　要：目的基于表面等离子体共振(SPR)技术寻找以血小板源性生长因子-BB(PDGF-BB)为直接靶点的候选药物,为抗肝纤维化药物的开发提供基础。方法利用SPR分子相互作用检测系统,使用His-tagged人源重组PDGF-BB蛋白建立SPR高通量筛选方法,通过对1760个FDA批准上市的药物及640个天然产物进行筛选及结合特异性验证,初步得到能够与PDGF-BB结合的化合物。进一步在PDGF-BB诱导人肝星状细胞(LX-2细胞)中对筛选得到的化合物的抗纤维化作用进行生物学验证。最后,利用分子对接技术预测化合物与PDGF-BB的可能结合位点及模式。结果建立了良好的靶向PDGF-BB的SPR高通量筛选模型,发现洛美沙星、茶碱乙酸、洛伐他汀、丹酚酸C、硝酸毛果芸香碱和达比加群酯都能与PDGF-BB结合形成复合体。体外实验结果证实以上6种化合物可不同程度降低LX-2细胞中纤维化标志基因COL1A1 mRNA水平,其中洛美沙星可显著降低PDGF-BB诱导的纤维化因子COL1A1、α-SMA(ACTA2)及PDGF-BB受体PDGFRβ的mRNA及蛋白水平。分子对接结果表明洛美沙星与PDGF-BB蛋白的Asn34形成氢键,进而促进两者形成复合体。结论建立了一种靶向PDGF-BB的SPR药物高通量筛选模型,并首次发现洛美沙星是一种以PDGF-BB为直接靶点的潜在抗肝纤维化药物,为其新用途的开发提供了参考。Objective Platelet derived growth factor-BB(PDGF-BB)plays a crucial role in the occurrence and development of liver fibrosis.Surface plasmon resonance(SPR)technology was utilized to identify potential drugs that directly target PDGF-BB,providing a foundation for the development of antifibrotic drugs.Methods Using the molecular interaction detection system based on SPR technology,a high-throughput SPR screening method was established by utilizing the human recombinant PDGF-BB protein with histidine tag(His-tag).A total of 1760 FDA-approved drugs and 640 natural products were screened and compounds capable of binding to PDGF-BB were preliminarily obtained.Further biological verification of the anti-fibrotic effect of the screened compounds was further investigated in human hepatic stellate cells(LX-2 cells)activated by PDGF-BB.Finally,the molecular docking technology was utilized to predict the possible binding sites and patterns of the compounds with PDGF-BB.Results We established a well SPR high-throughput screening model targeting PDGF-BB and lomefloxacin,acefylline,lovastatin,salvianolic acid C,pilocarpine nitrate,and dabigatran etexilate were found to bind directly to PDGF-BB.In vitro results confirmed that the above six compounds reduced the mRNA level of fibrosis marker gene COL1A1 in the LX-2 cells activated by PDGF-BB,and among them lomefloxacin was verified to significantly reduce the fibrosis factor COL1A1 andα-SMA(ACTA2)and PDGF-BB receptor PDGFRβat mRNA and protein levels.Molecular docking results indicated that lomefloxacin formed hydrogen bound with Asn34 of PDGF-BB protein to promote the formation of a complex.

关 键 词：肝纤维化 血小板源性生长因子-BB 表面等离子体共振 靶向药物 洛美沙星 

分 类 号：R575.2[医药卫生—消化系统]