Quinoline-based Inhibitors of BoNT/A LC

基于喹啉的 BoNT/A LC 抑制剂

• 批准号: 8057390
• 负责人: John D Williams
• 金额: $ 29.97万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8057390
• 关键词: Aerosols; Bacteria; Binding; Biological; Biological Assay; Bioterrorism; Blood; Bontoxilysin; Botulinum Toxin Type A; Breathing; Cause of Death; Cell model; Cells; Chemicals; Clinical; Clostridial Neurotoxin; Clostridium; Clostridium botulinum; Clostridium butyricum; Clostridium tetani; Complex; Development; Dose; Drug Kinetics; Endopeptidases; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Exhibits; Exocytosis; Exposure to; Fluorescence Resonance Energy Transfer; Goals; High Pressure Liquid Chromatography; Human; In Vitro; Inhibitory Concentration 50; Interstitial Collagenase; Intoxication; Intravenous; Lead; Libraries; Life; Light; Liver Microsomes; Lung; Measures; Metalloproteases; Military Personnel; Molecular Models; Nerve Endings; Neurons; Neurotoxins; Paralysed; Peripheral; Permeability; Pharmaceutical Chemistry; Pharmaceutical Preparations; Pharmacology; Phase; Phase III Clinical Trials; Poisoning; Polyamines; Population; Preparation; Process; Property; Protein Isoforms; Proteolysis; Reproduction spores; Rodent Model; SNAP receptor; Safety; Screening procedure; Series; Serotyping; Specificity; Stream; Structure; Synthesis Chemistry; Techniques; Therapeutic; TimeLine; Toxic effect; X-Ray Crystallography; Zinc; autonomic nerve; base; botulinum; botulinum toxin type G; cholinergic; design; drug discovery; improved; in vitro activity; in vivo; inhibitor/antagonist; lead series; member; molecular modeling; mouse model; neurotransmission; novel; pre-clinical; programs; quinoline; small molecule; tetanospasmin

DESCRIPTION (provided by applicant): The botulinum neurotoxins (BoNTs) are the most poisonous biological substances known. The lethal intravenous dose of BoNT serotype A (BoNT/A) in humans is 1-5 ng/kg. If accidental exposure to BoNT occurs (e.g., from contaminated foodstuffs), loss of life or life-threatening paralysis can occur. Most importantly, the BoNTs have already been "weaponized" in a highly toxic aerosol form, and they consequently pose a significant threat to both civilian and military populations. Once ingested, BoNTs target the peripheral cholinergic nerve endings and cause death by interrupting autonomic nerve function. The zinc-dependent endopeptidase light chain (LC) portion of BoNTs impairs neuronal exocytosis through proteolysis of essential SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) components of neurotransmission. The overall goal or this project is to develop small molecule inhibitors of the BoNT/A light chain (LC) metalloprotease activity to treat botulinum poisoning. Our strategy is to optimize a novel chemotype of BoNT/A LC inhibitors to improve potency, selectivity, and drug-like properties. In preliminary studies, we refined the structures to provide a new starting point for medicinal chemistry based on a chemical core with more drug-like properties than the original compound series and with ten-fold better potency. Significant activity was also observed in a chick neuronal cell model of BoNT/A intoxication. These validated early lead compounds are more suitable lead compounds for BoNT/A inhibitor drug discovery than were the original series, and they form the basis for this optimization program to generate an advanced lead compound. Our approach in Phase I is to use proven techniques of medicinal and parallel synthetic chemistry to optimize the potency and selectivity of the lead series of compounds and produce "drug-like" molecules with the potential to advance to pre-clinical development. In an iterative process, we will probe these focused compound libraries for structural features that contribute to tighter binding and more potent inhibition of the BoNT/A metalloprotease by measuring the enzymatic and cellular activities, as well as the specificity of the enzyme inhibitors. We will correlate the in vitro activity of the compounds against BoNT/A LC with activity in a cellular model of BoNT/A action, and use the most promising members of the series to validate the inhibitors in an in vivo mouse model of BoNT/A intoxication. At the end of Phase I we will define an advanced lead compound, which will be selected from the group of in vivo-validated inhibitors that displays suitable in vitro pharmacokinetic properties for further preclinical development. In Phase II, we will further optimize and evaluate the lead for in vivo efficacy, pharmacokinetic properties, toxicity and safety pharmacology in two species, and develop it into a pre-IND clinical candidate, suitable for human clinical trials (Phase III). PUBLIC HEALTH RELEVANCE: The botulinum neurotoxins are some of the most poisonous biologic substances known. Loss of life or life-threatening paralysis can occur following exposure to these neurotoxins from contaminated foodstuffs or acts of bioterrorism. This proposal describes the preparation and development of novel drugs to treat botulinum poisoning.

描述（由申请人提供）：肉毒杆菌神经毒素（BoNT）是已知毒性最强的生物物质。A型肉毒杆菌毒素（BoNT/A）在人体内的静脉注射致死剂量为1-5 ng/kg。如果发生意外暴露于BoNT（例如，受污染的食物），可能会造成生命损失或危及生命的瘫痪。最重要的是，BONT已经以剧毒气溶胶的形式“武器化”，因此对平民和军人都构成了重大威胁。一旦摄入，BoNT靶向外周胆碱能神经末梢，并通过中断自主神经功能导致死亡。BoNT的锌依赖性内肽酶轻链（LC）部分通过神经传递的必需SNARE（可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体）组分的蛋白水解损害神经元胞吐作用。 该项目的总体目标是开发BoNT/A轻链（LC）金属蛋白酶活性的小分子抑制剂，以治疗肉毒杆菌中毒。我们的策略是优化BoNT/A LC抑制剂的新化学型以提高效力、选择性和药物样性质。在初步研究中，我们改进了结构，为药物化学提供了一个新的起点，其化学核心比原始化合物系列具有更多的药物样性质，并且效力提高了十倍。在BoNT/A中毒的鸡神经元细胞模型中也观察到显著的活性。这些经过验证的早期先导化合物比原始系列更适合用于BoNT/A抑制剂药物发现的先导化合物，并且它们形成了此优化程序的基础，以生成高级先导化合物。 我们在第一阶段的方法是使用经过验证的药物和平行合成化学技术来优化先导系列化合物的效力和选择性，并产生具有推进临床前开发潜力的“药物样”分子。在一个迭代的过程中，我们将探测这些集中的化合物库的结构特征，有助于更紧密的结合和更有效的抑制BoNT/A金属蛋白酶通过测量酶和细胞活性，以及酶抑制剂的特异性。我们将在体外活性的化合物对BoNT/A LC与活性的BoNT/A行动的细胞模型，并使用该系列的最有前途的成员，以验证在体内小鼠模型的BoNT/A中毒的抑制剂。在I期结束时，我们将确定一种先进的先导化合物，该化合物将选自体内验证的抑制剂组，这些抑制剂显示出合适的体外药代动力学特性，用于进一步的临床前开发。在II期，我们将进一步优化和评估该先导化合物在两个种属中的体内疗效、药代动力学特性、毒性和安全药理学，并将其开发为pre-IND临床候选药物，适合进行人体临床试验（III期）。 公共卫生相关性：肉毒杆菌神经毒素是已知的毒性最大的生物物质之一。在接触到这些来自受污染食品或生物恐怖主义行为的神经毒素后，可能会发生生命损失或危及生命的瘫痪。该提案描述了治疗肉毒杆菌中毒的新型药物的制备和开发。