Reactivation of Aged Acetylcholinesterase: Design and Development of Novel Therap

描述（由申请人提供）：当前医疗对策的主要缺陷之一是，当前肟类药物无法重新激活已老化的神经毒剂抑制的乙酰胆碱酯酶（AChE）。这项建议的重点是确定有机分子，将使老年神经代理-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，其选择性地与老化的试剂抑制的AChE反应，形成稳定的甲基烷基膦酸酯-AChE加合物，其可以用常规肟再活化。这一药物发现工作将作为体外研究项目进行，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：设计和合成新的烷基化化合物，并测试它们在体外烷基化模拟老化剂-AChE的小的甲基膦酸酯和甲基膦酸酯-肽的能力，设计和合成对AChE特异的新的烷基化化合物，并测试它们在体外烷基化老化剂-AChE的能力和恢复老化剂-AChE再活化的能力，并向下选择将提供老化试剂-AChE的最大改进的肟再活化的烷基化化合物。预测烷基化化合物与老化剂-AChE的配体-受体相互作用的最先进的计算方法将指导烷基化化合物的设计，从而为新型治疗剂的合成提供合理的指导。将通过核磁共振（NMR）和液相色谱-质谱（LC-MS）以及新型固相方法沿着来确定模型甲基膦酸酯、甲基膦酸酯-肽和老化剂-AChE的烷基化。用分光光度法测定烷基化老化剂AChE的肟再活化。

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.