TARGET-GUIDED BOTULINUM NEUROTOXIN INHIBITOR DISCOVERY

靶向引导的肉毒杆菌神经毒素抑制剂的发现

• 批准号: 6963343
• 负责人: HARTMUTH C KOLB
• 金额: $ 18.39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6963343
• 关键词: Clostridium botulinum; X ray crystallography; biomimetics; bioterrorism /chemical warfare; botulinum toxins; chemical structure; chemical synthesis; combinatorial chemistry; inhibitor /antagonist; intermolecular interaction; metalloendopeptidases; molecular dynamics; neurotoxins

DESCRIPTION (provided by the applicant): Neurotoxins produced by Clostridium botulinum are among the most potent toxins known to man. They are zinc endopeptidases that cleave a group of proteins integral to neurotransmitter release, causing flaccid paralysis, which may lead to death. The organism, and the toxin it produces, poses a significant threat as an agent of biological warfare and terrorism. Current therapy of botulism is based on macromolecular antitoxins, which are only able to neutralize the neurotoxins before they enter nerve cells, requiring treatment to commence as soon as possible to avoid permanent nerve damage. Consequently, there is a great need for cell-permeable small molecule drugs that selectively inhibit the action of botulinum neurotoxins. In combination with antibiotic therapy, such inhibitors could be life-saving. Of all seven serotypes produced by C. botulinum, we have chosen botulinum neurotoxin B (BoNT B) as the initial target for inhibitor development, since it is one of the most common forms and since it has been well-characterized structurally. In situ click chemistry, an exploratory new technique in which the target protein assembles its own inhibitors from simple precursors, will be employed for lead discovery and optimization. The main goals of the project are to (a) investigate the use of in situ click chemistry for the discovery and optimization of BoNT B inhibitors; (b) develop a detailed understanding of the structural requirements for BoNT B inhibition through structure-activity profiling, molecular modeling and X-ray crystallography; (c) develop compounds through in situ click chemistry that inhibit BoNT B with high affinity (<1 mu/M). The specific aims are as follows: (1) Search for anchor molecules: Produce and screen several libraries of peptidomimetic compounds that are targeted against the BoNT B metalloproteinase; (2) Perform target-guided synthesis of BoNT B inhibitors through in situ click chemistry; (3) Investigate the structure-activity relationship through in situ click chemistry and perform structure based studies.

描述（由申请人提供）：肉毒梭菌产生的神经毒素是人类已知的最有效的毒素之一。它们是锌内肽酶，它裂解了一组与神经递质释放不可或缺的蛋白质，从而导致脆性麻痹，这可能导致死亡。作为生物战和恐怖主义的推动者，生物体及其产生的毒素构成了重大威胁。当前的肉毒杆菌疗法基于大分子抗毒素，该抗毒素只能在进入神经细胞之前对神经毒素进行中和，需要尽快开始治疗以避免永久性神经损伤。因此，非常需要细胞可渗透的小分子药物，这些药物有选择地抑制肉毒神经毒素的作用。结合抗生素疗法，这种抑制剂可能会挽救生命。在肉毒杆菌产生的所有七种血清型中，我们选择了肉毒杆菌神经毒素B（BONT B）作为抑制剂发育的初始靶标，因为它是最常见的形式之一，并且由于结构上已经良好。原位点击化学是一种探索性新技术，其中目标蛋白将从简单的前体中组装出自己的抑制剂，以进行铅发现和优化。该项目的主要目标是（a）调查使用原位点击化学来发现和优化BONT B抑制剂； （b）通过结构活性分析，分子建模和X射线晶体学对BONT B抑制的结构要求详细了解； （c）通过原位点击化学形式开发化合物，该化学抑制具有高亲和力的BONT B（<1 mu/m）。具体目的如下：（1）搜索锚定分子：生产和筛选几个拟肽化合物的库，这些化合物的靶向对抗BONT B金属蛋白酶； （2）通过原位点击化学对BONT B抑制剂进行目标引导的合成； （3）通过原位点击化学并进行基于结构的研究来研究结构活性关系。