中文摘要
农作物真菌细胞色素P450羊毛甾醇14α脱甲基化酶(P450-14DM，CYP51)是杀菌剂类农药作用的重要靶标酶之一。研究真菌CYP51活性中心作用位点与底物、配体相互作用的机理对研发新一代超高效、低抗性脱甲基化抑制剂类（DMI）杀菌剂而言是一个非常关键的科学问题。本项目旨在综合运用分子模拟、基因克隆、定点突变、同位素标记、光谱表征和有机合成等技术，系统地研究农作物真菌CYP51活性中心结构特征及其作用位点（特别是氨基酸残基的作用位点）与底物、配体相互作用的详细机理。在此基础上，进行农作物真菌CYP51新型抑制剂的生物合理设计、合成与筛选研究。以期得到具有进一步结构优化和开发前景的农作物真菌CYP51超高效、低抗性抑制剂的全新先导结构，为我国农作物真菌DMI新型杀菌剂的创制工作提供合理的设计指导，并打下一定的前期研发基础。
英文摘要
Cytochrome P450 14DM (CYP51) of crop is being one of the most important enzyme target for the research and development of novel potent and environment-friendly inhibitors of germicide. However, the mechanism of inhibitors interacting with active site of CYP51 is little known, except for the obvious active point of hemachrome porphyrin. Therefore, the investigation of the detailed interactions between ligands and active site (residual amino acids) is a crucial scientific research issue for designing and developing next generation of green germicide. In the present research project, a sensible mix of molecular modeling, gene clone, mutation, isotope-labled, spectroscopic and synthesis techniques has been employed to explore the detailed interacting information of ligands interacting with the active site. Furthermore, based on the research results foresaid we do structure-based inhibitors design and attempt to find novel potent hits and lead compounds which are could be further optimized and refined for germicide commercial use in China.
结题摘要
农作物真菌细胞色素P450羊毛甾醇14α脱甲基化酶(P450-14DM，CYP51)是杀菌剂类农药作用的重要靶标酶之一。研究真菌CYP51活性中心作用位点与底物、配体相互作用的机理对研发新一代超高效、低抗性脱甲基化抑制剂类（DMI）杀菌剂而言是一个非常关键的科学问题。本项目旨在综合运用分子模拟、基因克隆、定点突变、同位素标记、光谱表征和有机合成等技术，系统地研究农作物真菌CYP51活性中心结构特征及其作用位点（特别是氨基酸残基的作用位点）与底物、配体相互作用的详细机理。在此基础上，进行农作物真菌CYP51新型抑制剂的生物合理设计、合成与筛选研究。以期得到具有进一步结构优化和开发前景的农作物真菌CYP51超高效、低抗性抑制剂的全新先导结构，为我国农作物真菌DMI新型杀菌剂的创制工作提供合理的设计指导，并打下一定的前期研发基础。