SYNTHESIS OF INHIBITORS OF BOTULINUM METALLOPROTEASES

描述：(改编自申请人的摘要)本提案 描述了合成有效的非竞争性抑制剂的策略。 肉毒神经毒素金属蛋白水解酶。这些 金属蛋白水解酶是麻风的分子效应因子 这些毒素的作用是已报道的最具选择性的蛋白酶，因为 底物的一部分对酶的变构激活。 已知底物“激活”的构象受限类似物 序列将被合成以开发多肽衍生的抑制剂 在酶上的非竞争性位置进行抑制。酵母双杂交 实验将被用来鉴定在 激活站点。当生物活性构象被阐明后，它 将有可能开发用于创作小说的非肽模板 这些酶的抑制物。在第一阶段，申请者将 合成一组构象受限的化合物并确定 用所建立的方法测定它们的抑制活性。已知底物 被毒素金属蛋白酶切割，产生带有 电泳迁移率不同于未切割的 底物。这些化合物有望发挥药理作用。 拮抗剂，并作为神经生物学的重要探针 BONT逆转失弛缓症疗效的辅助性研究 和人类的相关疾病，以及作为人类体内重要的先导化合物 抗毒素药物开发。

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.