Reactivation of Aged Acetylcholinesterase: Design and Development of Novel Therap

老化乙酰胆碱酯酶的重新激活：新型疗法的设计和开发

• 批准号: 8735550
• 负责人: Christopher M. Hadad
• 金额: $ 37.7万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8735550
• 关键词: Acetylcholine; Acetylcholinesterase; Aging-Related Process; Alkylation; Ammonium; Binding; Biochemical; Chemical Agents; Chemical Warfare Agents; Chemicals; Cleaved cell; Computer Simulation; Computing Methodologies; Country; Development; Docking; Enzymes; Exposure to; Health; Human; Human body; Hydrolysis; In Vitro; Lead; Libraries; Ligands; Mechanics; Medical; Methods; Military Personnel; Modeling; Molecular; Neurotransmitters; Nuclear Magnetic Resonance; Organic Synthesis; Organophosphorus Compounds; Oximes; Peptides; Pesticides; Pharmaceutical Preparations; Phase; Population; Process; Proteomics; Reaction; Reagent; Research Project Grants; Serine; Societies; Solid; Structure; Syria; Testing; adduct; aged; cholinergic; design; drug discovery; improved; liquid chromatography mass spectrometry; methylphosphonate; molecular dynamics; nerve agent; novel; novel therapeutics; phosphonate; quantum; quinone methide; receptor; scaffold; toxicant

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.

描述（由申请人提供）：当前医疗对策的主要缺陷之一是当前的肟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）。该提案的重点是识别有机分子，使老化的神经毒剂 - 乙酰胆碱酯酶 (AChE) 重新激活。该提案的目的是发现烷基化化合物，选择性地与老化剂抑制的乙酰胆碱酯酶反应，形成稳定的甲基烷基膦酸酯-乙酰胆碱酯酶加合物，该加合物可以用常规肟重新激活。这项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。该提案的具体目标是：设计和合成新型烷基化化合物，并测试其体外烷基化小甲基膦酸酯和模仿老化剂-AChE的甲基膦酸酯肽的能力；设计和合成针对AChE的新型烷基化化合物，并测试其在体外的能力。体外烷基化老化剂-AChE并恢复老化剂-AChE重新激活的能力，以及向下选择将提供最改进的肟的烷基化化合物的能力老化剂-AChE的重新激活。烷基化化合物的设计将以最先进的计算方法为指导，预测烷基化化合物与老化药物-AChE的配体-受体相互作用，从而为新型治疗药物的合成提供合理指导。模型甲基膦酸酯、甲基膦酸酯肽和老化剂-AChE 的烷基化将通过核磁共振 (NMR) 和液相色谱-质谱 (LC-MS) 以及新型固相方法进行测定。烷基化老化剂-AChE的肟再活化将通过分光光度法测定。