光亲和标记串联点击化学研究啶虫丙醚作用靶标

Pyridalyl belongs to a recent generation of chemical insecticides that exhibit high insecticidal activity against Lepidoptera, and the toxicity of this compound on various beneficial insects is very low. Though the insecticidal mechanism studies indicated that the insecticidal activity of pyridalyl is closely related the functional inhibition of the cultured sf9 cells derived from ovarian of Spodoptera frugiperda, the interaction target and its exact mechanism of pyridalyl is unknown. Photoaffinity labeling is one of the key tools to investigate the interaction of ligand and its receptor at molecular level. However, the application of photoaffinity labeling always showed restricted limitation due to the bulky tag introduced to the probe in advance that may result in negative influence on the biological activity of the probe. Click chemistry provided an efficient method to overcome the inherent defect of the designed probe by using stepwise manner. In this methodology, the tag can be introduced after the probe photolabeled its biological target, thus avoid the negative influence of the bulky tag on their biological activity. In general, the cultured sf9 cells which exhibited high relationship with the insecticidal activity of pyridalyl were selected to investigated the interaction target and mechanism of pyridalyl by using tandem photoaffinity labeling-click chemistry methodology in this project.The elucidation of the interaction mechanism of pyridalyl can provide direction for rational design of novel insecticide.

啶虫丙醚是一种新型高效、低毒、环境友好的鳞翅目杀虫剂，研究表明其杀虫活性与昆虫卵巢细胞sf9功能抑制密切相关，但其作用靶标和作用机制并不清楚。光亲和标记技术是在分子水平上研究配体与受体相互作用的核心工具之一，决定光亲和标记成败的关键因素是光亲和探针分子的合理设计，传统的探针分子设计中必须引入大体积标签基团易对探针分子活性产生不利影响。"点击化学"可实现标签基团的分步连接：即探针分子标记靶蛋白后再连接标签基团，避免传统探针分子设计时的固有缺陷。因此，本项目拟以昆虫卵巢细胞sf9为研究对象，将光亲和标记和"点击化学"技术有效串联，鉴定啶虫丙醚的作用靶标，进一步阐释其作用机制，为新型杀虫剂的合理设计提供指导。

项目以啶虫丙醚为先导开展结构修饰，重点对其烷基链及末端吡啶环开展优化，通过引入不同杂环、氨基甲酸酯、双酰胺等结构合成了十类总计近200个不同结构类型衍生物，测试了所合成目标化合物的杀虫活性，总结了构效关系并在此基础上通过引入叠氮光敏基团合成了四类光敏探针分子，筛选到两个对小菜饿、甜菜夜娥活性优于啶虫丙醚本身的探针分子，其中光敏探针分子III-6在10 ppm浓度下对小菜蛾有100%的防效，杀虫谱测定表明其对多种鳞翅目类的昆虫均有较好的杀虫活性，对缨翅目的西花蓟马若虫也有一定的防效。室内毒力LD50值测定结果表明：III-6对小菜蛾室内毒力的LD50值为0.831 ppm，跟啶虫丙醚（2.021 ppm）相比降低了近2.5倍，两者差异显著，有进一步研究的价值；在国际期刊发表了6篇SCI论文，申请中国发明专利2项。