Reactivation of Aged Acetylcholinesterase: Design and Development of Novel Therap

描述（由申请人提供）：目前医学对策的主要缺陷之一是目前的肟不能重新激活神经毒剂抑制的乙酰胆碱酯酶（AChE）。这一建议的重点是识别有机分子，使老化的神经毒剂乙酰胆碱酯酶被重新激活。本提案的目的是发现烷基化化合物选择性地与老化剂抑制乙酰胆碱酯反应，形成稳定的甲基烷基膦酸酯-乙酰胆碱酯加合物，可以与传统的肟重新激活。这项药物发现工作将作为一个体外研究项目进行，其中烷基化过程和肟再活化过程将作为单独和连续的问题进行研究。这项建议的具体目标是：设计合成新型烷基化化合物，并测试其体外烷基化小甲基膦酸盐和甲基膦酸肽的能力，模拟老化剂-AChE；设计合成针对AChE的新型烷基化化合物，并测试其体外烷基化老化剂-AChE和恢复老化剂-AChE再活化能力；烷基化化合物的设计将以最先进的计算方法为指导，这些计算方法可以预测烷基化化合物与衰老剂乙酰氨基甲酸酯的配体-受体相互作用，从而为新疗法的合成提供合理的指导。核磁共振（NMR）、液相色谱-质谱（LC-MS）以及新型固相方法将测定模型甲基膦酸盐、甲基膦酸肽和老化剂乙酰胆碱酯（ache）的烷基化反应。用分光光度法测定烷基化陈化剂乙酰胆碱酯的肟活化。

DESCRIPTION (provided by applicant): One of the major deficiencies of current medical countermeasures is that current oximes cannot reactivate nerve agent-inhibited acetylcholinesterase (AChE) that has aged. This proposal focuses on identifying organic molecules that will enable aged nerve agent-AChE to be reactivated. The objective of this proposal is to discover alkylating compounds that selectively react with aged agent-inhibited AChE to form stable methylalkylphosphonate-AChE adducts that can be reactivated with conventional oximes. This drug discovery effort will be conducted as an in vitro research project in which the alkylation process and the oxime reactivation process will be investigated as separate and sequential problems. The specific aims of this proposal are: to design and synthesize novel alkylating compounds and test their in vitro ability to alkylate small methylphosphonates and methylphosphonate-peptides that mimic aged agent-AChE, to design and synthesize novel alkylating compounds specific for AChE and test their in vitro ability to alkylate aged agent-AChE and restore the ability of aged agent-AChE to be reactivated, and to down-select for the alkylating compound that will provide the most improved oxime reactivation of aged agent-AChE. The design of alkylating compounds will be guided by state-of-the-art computational methods that predict the ligand-receptor interactions of alkylating compound with aged agent-AChE, thereby providing rational guidance for the synthesis of novel therapeutics. Alkylation of model methylphosphonates, methylphosphonate-peptides and aged agent-AChE will be determined by nuclear magnetic resonance (NMR) and liquid chromatography-mass spectrometry (LC-MS), along with novel solid-phase methods. Oxime reactivation of alkylated aged agent-AChE will be determined by spectrophotometric methods.