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Dynamic Drug Design Targeting Botulinum Neurotoxins

针对肉毒杆菌神经毒素的动态药物设计

基本信息

批准号：
6787146
负责人：
JAMES M. BRIGGS
金额：
$ 25.99万
依托单位：
UNIVERSITY OF HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-08-15 至 2006-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6787146
关键词：
Clostridium botulinum active sites botulinum toxins computer simulation conformation drug design /synthesis /production drug receptors drug screening /evaluation electrophysiology endopeptidases inhibitor /antagonist molecular dynamics neurotoxins zinc

项目摘要

DESCRIPTION (provided by applicant): Botulinum neurotoxins (BoNTs) are a dangerous bioterrorism threat due to their extreme potency and lethality, as well as their ease of production and transport. If untreated, poisoning by the BoNTs can progress to flaccid paralysis and death due to respiratory failure. However, timely post-exposure intervention can limit the effects of the circulating toxin. Our overall, long-term research objective is to generate a novel class of therapeutics that can be administered to individuals who have been poisoned by BoNT. Each BoNT is composed of a catalytic light chain whose entry into neurons is mediated by the heavy chain. Our strategy is based on the model that botulism-related flaccid paralysis is a downstream consequence of the zinc-dependent endopeptidase activity elaborated by the BoNT light chain. One of the most powerful approaches to inactivate the endopeptidase function of the BoNT light chains is rational design of inhibitors targeting the active site. To achieve this, we wilt combine computational and experimental approaches to develop lead inhibitor templates. In Specific Aim 1, we will use a powerful computational approach called dynamic pharmacophore modeling to identify computational leads to block the endopeptidase activities of the BoNTs. In this approach, the conformational flexibility of the protein and active site are taken into account through molecular dynamics simulations and the generation of a consensus, or dynamic, pharmacophore model using an ensemble of molecular dynamics-generated protein conformations. The dynamic pharmacophore model is then used to search databases of commercially available small molecules to generate computational lead compounds. In Specific Aim 2, we will test each computational lead for inhibitory activity using enzyme assays and in vitro cellular assays. A milestone of this work will be the identification of one or more lead inhibitor templates that block the action of wild type toxin using in vitro model systems. The results from this research will establish the groundwork and justification for future development and in vivo testing of these novel inhibitors using established animal models.

描述（由申请人提供）：肉毒神经毒素（BoNT）是一种危险的生物恐怖主义威胁，因为它们具有极强的效力和致命性，而且易于生产和运输。如果不及时治疗，肉毒杆菌毒素中毒可能会发展为弛缓性麻痹，并因呼吸衰竭而死亡。然而，及时的暴露后干预可以限制循环毒素的影响。我们的总体长期研究目标是产生一类新的治疗方法，可以用于被BoNT中毒的个体。每个BoNT由催化轻链组成，其进入神经元由重链介导。我们的策略是基于这样的模型，即肉毒杆菌相关的弛缓性麻痹是由BoNT轻链阐述的锌依赖性内肽酶活性的下游后果。抑制BoNT轻链内肽酶功能的最有效方法之一是合理设计靶向活性位点的抑制剂。为了实现这一目标，我们将结合联合收割机的计算和实验方法，开发铅抑制剂模板。在具体目标1中，我们将使用一种强大的计算方法，称为动态药效团建模，以确定计算导致阻止BoNT的内肽酶活性。在这种方法中，蛋白质和活性位点的构象灵活性考虑到通过分子动力学模拟和生成的共识，或动态，药效团模型使用的分子动力学生成的蛋白质构象的合奏。然后使用动态药效团模型搜索市售小分子的数据库以生成计算先导化合物。在具体目标2中，我们将使用酶测定和体外细胞测定来测试每个计算引线的抑制活性。这项工作的一个里程碑将是使用体外模型系统鉴定一种或多种阻断野生型毒素作用的先导抑制剂模板。这项研究的结果将为未来的开发和使用已建立的动物模型进行这些新型抑制剂的体内测试奠定基础和理由。