摘 要 目的 基于分子对接和实验验证分析金银花不同溶剂提取物及主要成分抗RNA病毒蛋白酶的活性。方法 利用TCMSP（Traditional Chinese Medicine Systems Pharmacology）构建金银花小分子配体库，采用药物分子设计模拟 SYBYL 2.1.1 软件分析金银花与HIV-1蛋白酶和Cathepsin L蛋白酶结合情况。通过荧光法验证金银花不同溶剂提取物（水提物、30% 醇提物、60% 醇提物和85% ）抗HIV-1和Cathepsin L蛋白酶的活性。结果 收集金银花22个化学成分, 其中21个活性成分与HIV-1 蛋白酶有较好的结合，金银花抗HIV-1蛋白酶活性强于Cathepsin L蛋白酶。金银花水提物、30% 醇提物、60% 醇提物和85% 醇提物具有抗HIV-1的活性，IC50（half maximal inhibitory concentration）分别为0.14 ，0.015 ，0.05和0.121mg/mL，其中30% 醇提物具有较好的抗HIV-1的活性。金银花不同溶剂提取物未发现有强烈抑制组织蛋白酶的活性，实验结果和计算机筛选结果具有一致性。结论初步明确金银花抗HIV-1蛋白酶和Cathepsin L蛋白酶活性，为抗病毒药物的开发及应用提供理论依据。
ABSTRACT Objective Taking Lonicera japonica Thunb. as the research object, it has the effect of clearing away heat and detoxifying, the anti-RNA virus protease activity of L. japonica Thunb.was analyzed based on molecular docking and experimental verification. Methods Construction of small molecule ligand library of L. japonica Thunb. by TCMSP (Traditional Chinese Medicine Systems).The binding of L. japonica to HIV-1 protease and Cathepsin L protease was analyzed using drug molecular design simulation software SYBYL 2.1.1. The activities of different solvent extracts (water extract, 30% alcohol extract, 60% alcohol extract and 85% alcohol extract) of L. japonica were verified by fluorescence method against HIV-1 and Cathepsin L protease.Results Twenty-one active components were obtained from L. japonica Thunb. using TCMSP, and 20 of them had good binding with HIV-1 protease,the anti-HIV-1 protease activity of L. japonica Thunb. was stronger than Cathepsin L protease by molecular docking. By fluorescence analysis, it was found that the water extract, 30% alcohol extract, 60% alcohol extract and 85% alcohol extract had anti-HIV-1 activity with IC50 values of 0.14 mg/mL, 0.015 mg/mL. 0.05 mg/mL and 0.12 mg/mL. All extracts no inhibition against Cathepsin L protease. The experimental results are in good agreement with computer screening results.Conclusions This study preliminarily confirmed the activities of anti-HIV-1 protease and Cathepsin L protease in L. japonica Thunb., and the results provide theoretical basis for the development and application of antiviral drugs.
杨欣,危英,李星. 金银花对RNA病毒蛋白酶活性的影响[J]. 科学技术与工程, 2023, 23(30): 12827-12834.
Yangxin, Weiying, Lixing. The anti-RNA virus protease activity of Lonicera japonica Thunb. was analyzed based on molecular docking and experimental verification[J]. Science Technology and Engineering,2023,23(30):12827-12834.