Reactivation of Aged Acetylcholinesterase: Design and Development of Novel Therap

描述(申请人提供)：目前医学对策的主要缺陷之一是，目前的肟类药物不能重新激活已经老化的神经毒剂抑制的乙酰胆碱酯酶(AChE)。这项提议的重点是识别能够使老化的神经毒剂-AChE重新激活的有机分子。这项建议的目的是发现烷基化化合物，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乙酰胆碱酯酶加合物，可以用传统的肟类重新激活。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激活过程将作为独立的和顺序的问题进行调查。本建议的具体目的是：设计和合成新型烷基化化合物并测试其在体外对小分子甲基膦酸盐和模拟老化试剂-AChE的甲基膦酸肽的烷基化能力，设计并合成专用于AChE的新型烷基化化合物并测试其在体外对老化试剂-AChE的烷基化能力和恢复老化试剂-AChE的能力，以及选择将提供最大程度提高老化试剂-AChE的肟复活的烷基化化合物。烷基化化合物的设计将受到最先进的计算方法的指导，这些方法预测烷基化化合物与老化剂-AChE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。模型甲基膦酸类化合物、甲基膦酸类多肽和老化剂-乙酰胆碱酯酶的烷基化反应将通过核磁共振(核磁共振)和液质联用(LC-MS)以及新的固相方法进行测定。烷基化老化剂乙酰胆碱酯酶(AChE)的肟复活将用分光光度法测定。

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.