Novel Protein-Based Therapeutics for Nerve Agent Detoxification

用于神经毒剂解毒的新型蛋白质疗法

基本信息

批准号：
7292643
负责人：
Matthew R Redinbo
金额：
$ 47.5万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-27 至 2011-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7292643
关键词：
Acetylcholinesterase Active Sites Adverse effects Aging Amino Acids Animal Model Animals Architecture Binding Binding Sites Biochemical Biological Models Carboxylic Ester Hydrolases Chemical Weapons Chemicals Cholinesterases Complex Data Drug Metabolic Detoxication Engineering Enzymes Exposure to Goals Human Human Resources Hydrolase Hydrolysis In Vitro Investigation Life Medical Methods Military Personnel Molecular Mus Mutation Neurotransmitters North Carolina Organophosphates Pharmaceutical Preparations Process Proteins Purpose Range Reaction Research Institute Research Personnel Resistance Risk Saint Jude Children&apos s Research Hospital Sarin Serine Hydrolase Serum Site Soman Structure Surface Testing Therapeutic Toxic effect Universities analog base bioscavenger carboxylesterase catalyst cyclosarin design drug metabolism emergency service responder esterase gallium alloy GF human carboxylesterase 1 improved in vivo methylphosphonate mutant nerve agent novel programs prophylactic tabun

项目摘要

DESCRIPTION (provided by applicant): The organophosphate nerve agents tabun (GA), sarin (GB), soman (GD), cyclosarin (GF), VX, and Russian VX (R-VX) are among the deadliest compounds known. Current treatments for chemical weapons exposure offer limited protection, must be administered immediately, and can generate long-term toxic side effects. Human carboxylesterase 1 (hCE1), a promiscuous drug metabolism enzyme, offers promise as a broad- spectrum protein-based therapeutic for nerve agent detoxification. We have determined several crystal structures of hCE1 in complex with live nerve agents. These preliminary results have indicated that the enzyme is both stereoselective and resistant to aging, and provides two putative nerve agent binding sites: the active site and a promiscuous surface binding site. The focus of this application is to convert hCE1 into a highly efficient nerve agent hydrolase using a range of targeted structural, biochemical and in vivo studies. A team of investigators at the University of North Carolina at Chapel Hill, St. Jude Children's Research Hospital, and the US Army Medical Research Institute of Chemical Defense has been assembled, and four specific aims will be pursued: 1. Elucidate crystal structures of hCE1 in complex with a wide variety of live nerve agents. 2. Introduce targeted mutations designed to improve nerve agent binding and hydrolysis. 3. Examine the impact site-directed mutants have on hCETs nerve agent hydrolysis activity in vitro. 4. Assess the in vivo ability of engineered forms of hCE1 to protect serum esterase-deficient mice from exposure to nerve agents. The overall goal of these on-going investigations is to develop a novel, efficient, broad-spectrum protein- based therapeutic for prophylactic protection against nerve agent exposure. It is clear that effective catalytic countermeasures are urgently needed to safely protect military personnel and civilian first-responders from attacks involving chemical weapons.

描述（由申请人提供）：有机磷神经毒剂塔崩 (GA)、沙林 (GB)、索曼 (GD)、环沙林 (GF)、VX 和俄罗斯 VX (R-VX) 是已知最致命的化合物。目前针对化学武器暴露的治疗提供的保护有限，必须立即进行，并且可能产生长期毒副作用。人羧酸酯酶 1 (hCE1) 是一种混杂的药物代谢酶，有望作为一种广谱的基于蛋白质的神经毒剂解毒疗法。我们已经确定了 hCE1 与活神经毒剂复合物的几种晶体结构。这些初步结果表明，该酶既具有立体选择性又具有抗老化性，并提供两个假定的神经毒剂结合位点：活性位点和混杂的表面结合位点。该应用的重点是通过一系列有针对性的结构、生化和体内研究将 hCE1 转化为高效的神经毒剂水解酶。北卡罗来纳大学教堂山分校、圣裘德儿童研究医院和美国陆军化学防御医学研究所的研究小组已经集结，将实现四个具体目标： 1. 阐明 hCE1 的晶体结构与多种活神经毒剂复合。 2.引入旨在改善神经毒剂结合和水解的靶向突变。 3. 检查定点突变体对 hCET 神经毒剂体外水解活性的影响。 4. 评估 hCE1 的工程形式在体内保护血清酯酶缺陷小鼠免受神经毒剂暴露的能力。这些正在进行的研究的总体目标是开发一种新型、高效、广谱的基于蛋白质的治疗方法，用于预防神经毒剂暴露。显然，迫切需要采取有效的催化对策，以安全地保护军事人员和民事急救人员免受涉及化学武器的袭击。