基于同源建模和虚拟筛选松材线虫海藻糖磷酸酶抑制剂的发现研究  

Discovery of Trehalose Phosphatase Inhibitors in Pine Wood Nematodes based on Homology Modeling and Molecular Docking Virtual Screening

作　　者：杨鑫 陈双扣 黄夏 陈秋月 朱万宏 王瑜 陈俊杰 YANG Xin;CHEN Shuangkou;HUANG Xia;CHEN Qiuyue;ZHU Wanhong;WANG Yu;CHEN Junjie(School of Chemistry and Chemical Engineering,Chongqing University of Science and Technology,Chongqing 401331,P.R.China;Chongqing Key Laboratory of Pharmaceutical Process Digitization,Chongqing University of Science and Technology,Chongqing 401331,P.R.China)

机构地区：[1]重庆科技大学化学化工学院,重庆401331 [2]重庆科技大学制药工程数字化重庆市重点实验室,重庆401331

出　　处：《西部林业科学》2025年第1期73-84,共12页Journal of West China Forestry Science

基　　金：重庆市科技兴林科学研究类项目(渝林科研YB2024-2);重庆科技大学研究生创新计划(YKJCX2420509)。

摘　　要：采用同源建模技术和虚拟筛选方法,从天然产物化合物库中筛选出环境友好型松材线虫海藻糖磷酸酶抑制剂。通过同源建模构建松材线虫TPP的三维蛋白结构,并结合AUC、EF值等参数评估对接模型的活性分子富集能力,筛选出与TPP结合能低于阈值的化合物并通过T.E.S.T毒性预测软件和类农药性评估,以确保其毒性水平在安全范围内;进行杀线虫实验,结合可视化分析,以验证其杀虫活性及作用机制。结果显示:同源建模得到的对接模型具有优异的活性分子富集能力,基于该模型筛选出天然产物芦竹碱,杀线虫活性测试表明,在1 mg·mL^(-1)浓度下,72 h对松材线虫的校正致死率为98.5%,LC50值为0.056 g·L^(-1),表明芦竹碱具有显著杀虫效果;通过分析杀虫作用机制发现,芦竹碱可与靶点蛋白通过氢键和疏水相互作用形成稳定的复合物,从而抑制松材线虫体内海藻糖磷酸酶的活性,影响线虫的生命活动。研究结果表明,基于同源建模、虚拟筛选研究及活性测试的研究策略相结合可以有效地发现环境友好型松材线虫抑制剂,研究为从天然产物化合物库筛选出杀线虫剂的先导化合物提供了理论依据。To screen for environmentally friendly inhibitors of trehalose phosphatase in pine wood nematodes from a library of natural product compounds using homology modeling and virtual screening methods.The three-dimensional protein structure of TPP from pine wood nematodes was constructed through homology modeling,and the enrichment capacity of the docking model for active molecules was evaluated using parameters such as AUC and EF values.Compounds with binding energies below the threshold were selected and further assessed with T.E.S.T toxicity prediction software and pesticide-like property evaluation to ensure their toxicity levels were within a safe range.Nematicidal experiments were conducted,and visualization analysis was used to verify their nematicidal activity and mechanisms of action.The docking model obtained from homology modeling demonstrated excellent enrichment capacity for active molecules.Based on this model,the natural product gramine was selected.Nematode activity tests showed that at a concentration of 1mg·mL^(-1),the corrected mortality rate of pine wood nematodes after 72 hours was 98.5%,with an LC50 value of 0.056 g·L^(-1),indicating that gramine has significant nematicidal effects.Analysis of the nematicidal mechanism revealed that gramine can form stable complexes with the target protein through hydrogen bonds and hydrophobic interactions,thereby inhibiting the activity of trehalose phosphatase within the nematode and affecting its life activities.The combined research strategy of homology modeling,virtual screening,and activity testing can effectively discover environmentally friendly inhibitors of pine wood nematodes.This study provides a valuable exploration for screening nematicidal lead compounds from a library of natural product compounds.

关 键 词：松材线虫病 虚拟筛选 分子对接 天然产物 芦竹碱 

分 类 号：S763[农业科学—森林保护学]