Novel Small Molecule Inhibitors of Botulinum Neurotoxin A

新型肉毒杆菌神经毒素 A 小分子抑制剂

• 批准号: 7494157
• 负责人: Terry L. Bowlin
• 金额: $ 161.15万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-15 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7494157
• 关键词: Active Sites; Aerosols; Antibodies; Binding; Bioavailable; Biological Assay; Biological Availability; Blood; Bontoxilysin; Botulinum Toxin Type A; Botulinum Toxins; Breathing; Cause of Death; Cells; Cessation of life; Chemicals; Cholinergic Agents; Class; Clinical; Clinical Trials; Clostridium; Data; Databases; Development; Development Plans; Dose; Drug Design; Drug Kinetics; Endopeptidases; Ethylmaleimide; Evaluation; Exhibits; Exocytosis; Food; Goals; Human; Hydroxamic Acids; Immune; In Vitro; Inhibitory Concentration 50; Intoxication; Investigational Drugs; Investigational New Drug Application; Lead; Libraries; Light; Liver Microsomes; Lung; Measures; Mechanical ventilation; Metabolism; Military Personnel; Mining; Modeling; Multienzyme Complexes; Mus; Nerve Endings; Neurons; New Drug Approvals; Oral; Peripheral; Permeability; Pharmaceutical Preparations; Pharmacology; Phase; Poison; Population; Process; Property; Protein Isoforms; Proteins; Proteolysis; Qualifying; Range; Rattus; Reproduction spores; Roentgen Rays; Safety; Screening procedure; Security; Series; Serotyping; Specificity; Stream; Structure; Structure-Activity Relationship; Supportive care; Techniques; Testing; Therapeutic; Toxic effect; Toxicology; Toxin; United States; Vertebral column; X-Ray Crystallography; Zinc; absorption; autonomic nerve; base; biodefense; botulinum; cholinergic; combinatorial chemistry; improved; in vivo; indexing; inhibitor/antagonist; molecular modeling; mouse model; neurotransmission; novel; novel therapeutics; pharmacophore; pre-clinical research; prevent; programs; receptor; scaffold; scale up; small molecule; small molecule libraries

DESCRIPTION (provided by applicant): The overall project goal is to develop an orally active small molecule BoNT/A inhibitor. The neuronal substrate for BoNT/A LC is synaptosomal-associated protein of 25.000MW (SNAP-25). Our hypothesis is that small molecule BoNT/A inhibitors which can penetrate the neuron, preventing SNAP-25 cleavage by the BoNT LC, will provide the potential for treatment of both pre- and post-toxin exposure. Our strategy has been to identify "drug-like" small molecule BoNT/A LC inhibitor scaffolds, through a combination of chemical library screening and 3D sub-structure database mining, suitable for further chemical refinement in a rational drug design program. Through this process, we have identified several BoNT/A inhibitors with low uM potencies in enzymatic and cell based assays, which prevent BoNT/A intracellular proteolysis of its target substrate SNAP-25. The identified inhibitor classes are novel and devoid of non-drug like features such as hydroxamic acid and peptidic backbones. These validated chemical "hit" series will form the basis for development of efficacious, safe and orally bioavailable drugs against BoNT/A. We will apply proven techniques of medicinal and combinatorial chemistry; inhibitor-enzyme complex X-ray crystallography and structure-based drug design (SBDD) to rapidly synthesize and evaluate derivatives of these new validated BoNT/A inhibitor scaffolds. In an iterative process, we will probe focused compound libraries for features contributing to tighter binding and more potent inhibition of BoNT/A by measuring the enzymatic and cellular activity and specificity of derivatives. We will determine the X-ray structures of improved inhibitors bound to the BoNT active site, and use the data to develop refined pharmacophore models to guide further probing of the structure activity relationship (SAR). We will assess compounds for optimal ADME (Absorption, Distribution, Metabolism, Elimination), pharmacokinetic and bioavailability properties. Compounds with sufficient enzymatic, cellular potency and ADME properties will be scaled-up and tested for efficacy in a BoNT induced rat death model. Successful rescue of rats from BoNT-induced death will qualify compounds as in vivo-validated leads. We will evaluate these leads for investigational new drug (IND) enabling toxicity and safety pharmacology in order to develop them into pre-IND clinical candidates, suitable for human clinical trials. By the end of this proposed research and pre-clinical development plan, our overall project milestone is to file an IND for the clinical human safety evaluation of an orally active small molecule BoNT/A inhibitor.

描述（由申请人提供）：项目总体目标是开发一种口服活性小分子 BoNT/A 抑制剂。 BoNT/A LC 的神经元底物是 25.000MW 的突触体相关蛋白 (SNAP-25)。我们的假设是，小分子 BoNT/A 抑制剂可以穿透神经元，防止 SNAP-25 被 BoNT LC 裂解，将为治疗毒素暴露前和毒素暴露后提供潜力。我们的策略是通过化学库筛选和3D子结构数据库挖掘相结合，识别“类药物”小分子BoNT/A LC抑制剂支架，适合在合理的药物设计方案中进一步化学精制。通过这一过程，我们在酶促和细胞检测中鉴定了几种具有低 uM 效力的 BoNT/A 抑制剂，这些抑制剂可防止 BoNT/A 其靶底物 SNAP-25 的细胞内蛋白水解。所确定的抑制剂类别是新颖的并且缺乏非药物特征，例如异羟肟酸和肽主链。这些经过验证的化学“热门”系列将构成开发有效、安全和口服生物可利用的 BoNT/A 药物的基础。我们将应用经过验证的药物和组合化学技术；抑制剂-酶复合物 X 射线晶体学和基于结构的药物设计 (SBDD)，可快速合成和评估这些新的经过验证的 BoNT/A 抑制剂支架的衍生物。在迭代过程中，我们将通过测量衍生物的酶和细胞活性以及特异性，探索重点化合物库，以寻找有助于更紧密结合和更有效抑制 BoNT/A 的特征。我们将确定与 BoNT 活性位点结合的改进抑制剂的 X 射线结构，并利用这些数据开发精细的药效团模型，以指导进一步探讨结构活性关系 (SAR)。我们将评估化合物的最佳 ADME（吸收、分布、代谢、消除）、药代动力学和生物利用度特性。具有足够的酶、细胞效力和 ADME 特性的化合物将被放大并在 BoNT 诱导的大鼠死亡模型中测试其功效。成功地将大鼠从 BoNT 诱导的死亡中拯救出来将使化合物成为体内验证的先导化合物。我们将评估这些研究性新药 (IND) 的先导化合物，以实现毒性和安全性药理学，以便将它们开发为适合人体临床试验的 IND 前临床候选药物。在这项拟议的研究和临床前开发计划结束时，我们的总体项目里程碑是提交一份 IND，用于口服活性小分子 BoNT/A 抑制剂的临床人体安全性评估。