新型抗耐药真菌化合物的作用靶点及分子机制研究

项目名称： 新型抗耐药真菌化合物的作用靶点及分子机制研究

项目编号： No.81330083

项目类型： 重点项目

立项/批准年度： 2014

项目学科： 医药、卫生

项目作者： 姜远英

作者单位： 中国人民解放军第二军医大学

项目金额： 290万元

中文摘要： 侵袭性真菌感染的发生率逐年上升，病死率居高不下，真菌天然耐药和获得性耐药为真菌感染治疗带来巨大困难，且近十多年来未发现基于新靶点的抗耐药真菌新药。本课题组从协同抗耐药真菌的新策略出发，发现了多个天然小分子化合物具有很强的抗耐药真菌活性，构效关系研究发现了它们共有的药效基团，但作用靶点和机制尚不明确。本项目拟对活性化合物深入进行构效关系及结构优化研究，以发现具有更高活性、特异性的化合物和能够固载于亲和凝胶上并具有靶点识别能力的配体；深入研究活性化合物抗耐药真菌的细胞和分子机制；采用基于SILAC的定量蛋白质组学技术结合亲和吸附技术研究活性化合物与真菌细胞内特异相互作用蛋白和作用靶点；采用基因敲除技术构建靶蛋白基因缺失菌，比较基因缺失菌和野生菌与活性化合物作用后的表型差异，关联分析靶蛋白的生物学功能及化合物作用机制，对其作用靶点进行确证，为研究新型高效的抗耐药真菌药物提供理论和实验依据。

中文关键词： 药物靶点;真菌感染;分子机制;耐药性;小分子化合物

英文摘要： The incidence of invasive fungal infections is increasing year by year, mortality remains high. Natural and acquired drug resistance of fungi brings enormous difficulties for the treatment of fungal infections. No new drug acting at novel target has been developed against drug-resistant fungi over the last decade. Based on the new strategy of synergistic effect, we have found multiple natural small molecules possess strong activity against drug-resistant fungi. The preliminary structure-activity relationship (SAR) study suggested that these compounds have similar pharmacophore. But until now, their target and molecular mechanisms have not been determined. In the present project, we first plan to continue the SAR and structural optimization study, for screening the compound with higher activity and specificity, and ensuring the ligand can be immobilized affinity gel with target recognition capability. Second, we want to investigate the cellular and molecular mechanisms of the active compounds against drug-resistant fungi. Third, we will screen the interaction protein in fungi with the above active compounds by SILAC-based quantitative proteomics technology and affinity adsorption technology, and then combine bioinformatics technologies to analyze the target of the active compounds against drug-resistant fungi. In the end, to validate the target of the novel compounds, we will knockout the target gene in fungi cell and compare the phenotype between gene deletion strain and wild type strain after incubation with active compound, combine association analysis between biological function of the target protein and mechanism of the active compounds against fungi. We aim to not only identify the target and clarify the antifungal mechanism of the active compound, but also provide theoretical and experimental basis for new effective antifungal agent development.

英文关键词： drug targets;fungal infection;molecular mechanisms ;drug resistance;small molecule compounds