摘要
[目的]获得能够治疗家蚕血液型脓病的小分子药物。[方法]基于主流的药物开发程序,采用计算机网络虚拟建模,以家蚕核型多角体病毒侵染细胞不可缺少的GP64蛋白为研究对象,构建其三维结构。紧接着虚拟筛选,选取其中前100个对接打分最高的分子,进行能量优化,依据CDOCKER Interaction Energy的函数得分值,筛选出前10个结果最好的活性分子。最后结合体外活性检测技术,对小分子化合物ZINC95401182进行抗病毒活性检测。[结果]得到一个与BmNPV GP64蛋白相似度高达92%的晶体结构,3 DUZ_A。以此为模型绘制出BmNPV GP64蛋白三维结构,拉式图结果显示核心区氨基酸总和大于90%,模型结构合理。在能量水平排名前10的配体中选出最适小分子化合物,ZINC95401182。在抗活性检测中,ZINC95401182设置0.1μg/mL、0.25μg/mL、1μg/mL、10μg/mL四个不同浓度处理家蚕细胞,结果显示在1μg/mL以上浓度时能够降低BmNPV的滴度,透射电子显微镜观察证实ZINC95401182抑制了BmNPV对细胞的入侵。[结论]小分子化合物ZINC95401182对BmNPV的侵染具有显著的抑制作用,可以作为一种潜在的家蚕血液型脓病治疗药物。
[Objective]To obtain small molecule drugs that can inhibit the infection of Bombyx mori nuclear polyhedrosis virus(BmNPV).[Method]Based on the mainstream drug development program and computer network virtual modeling,the three-dimensional structure of the essential GP64 protein of the cells infected by BmNPV was constructed.Then the first 100 molecules with the highest docking score were selected by virtual screening,and the first 10 molecules with the best docking score were selected according to the CDOCKER Interaction Energy score.Finally,the antiviral activity of small molecular compound ZINC95401182 was detected by in vitro assay.[Result]A crystal structure,3DUZ_A,was obtained,which was 92%similar to the GP64 protein of BmNPV.The three-dimensional structure of BmNPV GP64 protein was drawn by this model.The results showed that the sum of amino acids in the core region was more than 90%,and the model structure was reasonable.The most suitable small molecular compound,ZINC95401182,was selected from the top 10 ligands in energy level.In the test of anti-activity,ZINC95401182 treated Bombyx mori cells with different concentrations of 0.1μg/mL,0.25μg/mL,1μg/mL and 10μg/mL,the result showed that the titer of BmNPV could be reduced when the concentration of ZINC95401182 was above 1μg/mL,transmission electron microscope showed that ZINC95401182 inhibited the invasion of BmNPV.ZINC95401182 can reduce the titer of BmNPV when the concentration is above 1μg/mL in BmN cells.Transmission electron microscopy observations confirmed that ZINC95401182 inhibited the invasion of BmNPV into cells.[Conclusion]The small molecule compound ZINC95401182 has a significant inhibitory effect on BmNPV infection,and can be used as a potential drug for BmNPV treatment.
作者
刘梦瑾
浦仕奇
吕丁丁
陆浩杰
蒋师东
唐旭东
LIU Meng-jin;PU Shi-qi;LYU Ding-ding;LU Hao-jie;JIANG Shi-dong;TANG Xu-dong(Jiangsu University of Science and Technology,Zhenjiang 212003,China;The Sericultural Institute,Chinese Academy of Agricultural Sciences,Zhenjiang 212018,China;Zhenjiang College,Zhenjiang 212028,China)
出处
《生物技术》
CAS
2022年第3期303-308,共6页
Biotechnology
基金
江苏省自然科学基金青年项目(BK20190244)
镇江市科技创新资金项目(NY2021020)。
