Abstract：
Acetylcholinesterase (AChE) stands as a primary target of commercial insecticides, notably organophosphates and carbamates. Despite their widespread use in agricultural and indoor pest control, concerns over their high toxicity and the emergence of resistance have restricted their efficacy. In this study, we conducted high-throughput virtual screening against both wild-type (WT) and resistant Cimex lectularius AChE utilizing a library encompassing 1 270 000 compounds. From this screening, we identified 100 candidate compounds and subsequently assessed their inhibitory effects on purified AChE enzymes. Among these candidates, AE027 emerged as a potent inhibitor against both WT and resistant AChE, exhibiting IC50 values of 10 and 43 μM, respectively. Moreover, the binding of AE027 significantly stabilized AChE, elevating its melting temperature by approximately 7 °C. Through molecular docking and molecular dynamics simulation, we delineated the binding mode of AE027, revealing its interaction with a site adjacent to the catalytic center, which is distinct from known inhibitors, with differing poses observed between WT and resistant AChE. Notably, the resistance mutation F348Y, positioned at a site directly interfacing with AE027, impedes ligand binding through steric hindrance. Furthermore, we evaluated the toxicity and pharmacokinetic properties of AE027 utilizing bioinformatics tools. These findings lay a crucial foundation for the development of a novel generation of insecticides that can combat both WT and resistant pest populations effectively and safely.
摘要：
乙酰胆碱酯酶（AChE）是商业杀虫剂的主要目标，尤其是有机磷酸酯和氨基甲酸酯。尽管它们广泛用于农业和室内虫害防治，对其高毒性和耐药性的担忧限制了其功效。在这项研究中,我们利用包含1270000种化合物的文库对野生型(WT)和抗性CimexlectulariusAChE进行了高通量虚拟筛选.从这次筛选来看,我们鉴定了100种候选化合物,随后评估了它们对纯化的AChE酶的抑制作用.在这些候选人中,AE027作为针对WT和抗性AChE的有效抑制剂出现，表现出10和43μM的IC50值，分别。此外，AE027的结合显着稳定了AChE，将其熔化温度提高约7°C。通过分子对接和分子动力学模拟,我们描绘了AE027的结合模式，揭示了它与催化中心附近位点的相互作用，这与已知的抑制剂不同，在WT和抗性AChE之间观察到不同的姿势。值得注意的是,抗性突变F348Y，位于与AE027直接连接的位点，通过位阻阻碍配体结合。此外，我们利用生物信息学工具评估了AE027的毒性和药代动力学特性.这些发现为开发新一代杀虫剂奠定了关键基础，该杀虫剂可以有效且安全地对抗WT和抗性害虫种群。
