SYNTHESIS OF INHIBITORS OF BOTULINUM METALLOPROTEASES

肉毒杆菌金属蛋白酶抑制剂的合成

• 批准号: 2024568
• 负责人: Alexey G Zdanovsky
• 金额: $ 9.97万
• 依托单位: PROMEGA CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-06-01 至 1998-05-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2024568
• 关键词: Clostridium; active sites; botulinum toxins; chemical synthesis; enzyme inhibitors; metalloendopeptidases; peptides; yeast two hybrid system

DESCRIPTION: (Adapted from the applicant's abstract) This proposal describes strategies to synthesize potent non-competitive inhibitors of botulinum neurotoxin metalloproteases {BoNT protease}. These metalloproteases which are the molecular effectors of the paralytic effects of these toxins are the most selective proteases reported due to allosteric activation of the enzyme by a portion of the substrate. Conformationally-restricted analogs of known substrate "activation" sequences will be synthesized to develop peptide- derived inhibitors that inhibit at a non-competitive site on the enzyme. Yeast two-hybrid experiments will be used to identify the enzyme residues at the activation site. When the bioactive conformation has been elucidated, it will become possible to develop non-peptide templates for creating novel inhibitors of these proteases. During Phase I, the applicants will synthesize a set of conformationally restricted compounds and determine their inhibitory activity using the established assay. Known substrates are cleaved by the toxin metalloprotease to generate fragments with an electrophoretic mobility different from those of the uncleaved substrates. These compounds are expected to function as pharmacological antagonists and to serve as important probes useful for neurobiological research as adjuncts for reversing effects of BoNT in treating achalasia and related disorders in humans and as important lead compounds in antitoxin drug development.

描述：（改编自申请人的摘要）该提案 描述了合成有效的非竞争性抑制剂的策略 肉毒杆菌神经毒素金属蛋白酶{BoNT蛋白酶}。 这些 金属蛋白酶是麻痹症的分子效应器 据报道，这些毒素的影响是最具选择性的蛋白酶，因为 一部分底物对酶的变构激活。 已知底物“激活”的构象限制类似物 将合成序列以开发肽衍生的抑制剂 抑制酶的非竞争性位点。 酵母二杂交体 实验将用于鉴定酶残基 激活站点。 当生物活性构象被阐明后， 将有可能开发非肽模板来创造新的 这些蛋白酶的抑制剂。 在第一阶段，申请人将 合成一组构象限制化合物并确定 使用已建立的测定法检测它们的抑制活性。 已知基材 被毒素金属蛋白酶裂解产生带有 电泳迁移率与未切割的不同 基材。 这些化合物有望发挥药理学作用 拮抗剂并作为神经生物学的重要探针 作为逆转 BoNT 治疗贲门失弛缓症的辅助作用的研究 和人类相关疾病以及作为重要的先导化合物 抗毒素药物开发。

项目成果

Synthesis of substrates and inhibitors of botulinum neurotoxin type A metalloprotease.

A 型肉毒杆菌神经毒素金属蛋白酶的底物和抑制剂的合成。

• DOI: 10.1111/j.1399-3011.2004.00124.x
• 发表时间: 2004
• 期刊: The journal of peptide research : official journal of the American Peptide Society
• 作者: Sukonpan,C;Oost,T;Goodnough,M;Tepp,W;Johnson,EA;Rich,DH
• 通讯作者: Rich,DH