Parallel Multimodal High-throughput screening to identify activators of the orexin receptors

并行多模式高通量筛选鉴定食欲素受体激活剂

• 批准号: 10705475
• 负责人: Theodore M Kamenecka
• 金额: $ 8.25万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-05 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10705475
• 关键词: Parallel; Multimodal; throughput; screening; identify

PROJECT SUMMARY The GPCR superfamily of ligand regulated receptors has proven to be a rich source of targets for development of therapeutics for a myriad of human diseases. The orexin 1 and orexin 2 receptors are class A GPCR’s differentially expressed in the central nervous system. The orexin 1 receptor is most abundantly expressed in the locus coeruleus and is thought to control aspects of emotion, reward, and the autonomic nervous system, whereas the orexin 2 receptor is expressed in regions controlling arousal such as the tubermammillary nucleus, an important site for the regulation of sleep/wakefulness. Almost 20 years after their initial discovery, the first potent dual orexin 1 / orexin 2 receptor antagonist therapeutic has been brought to market for insomnia (Belsomera ®, suvorexant, Merck). Despite a massive effort to identify antagonists of the the orexin receptor, almost no reports of small molecule agonists appear in the primary or patent literature. Elegant genetic ablation experiments using orexin knock-out animals and experiments with intracerebroventricular dosing of orexin A or orexin B peptides suggests a role for orexin receptor agonists or potentiators for a number of potential indications including: 1) depression; 2) learning and memory (cognition); 3) attention deficit hyperactivity disorder (ADHD); 4) treatment for colon cancer; and 5) sleep disorders including narcolepsy. While the orexin peptides are potent agonists of both orexin receptors, they are not selective nor do they cross the blood brain barrier well, making them poor probe substrates for in vivo pharmacology studies. This provides the impetus to identify small molecule, brain penetrant activators of the orexin receptors to interrogate the receptors function in the context of disease states. We have optimized a cell-based high-throughput screening compatible primary assay that specifically measures the functional activity of orexin 1 and orexin 2. A preliminary 10k pilot screen was performed leading to the identification of a number of small molecule agonists of these receptors demonstrating this assay to be robust and reproducible. A full HTS screening campaign of the Scripps Institutional Drug Discovery Library (~640k compounds) will lead to the identification of multiple classes of orexin receptor agonists and potentiators for further development. In an iterative process, these validated hits will be characterized through a cascade of in vitro cell-based assays to determine potency, selectivity and mechanism of action. Preliminary medicinal chemistry lead optimization will identify early structure activity relationships and in vitro drug metabolism will be assessed to confirm tractibility of early leads. These efforts will provide first in-class chemical tools to be used to further probe orexin receptor function in animal models of disease.

项目摘要 事实证明，配体调节受体的GPCR超家族是丰富的目标来源 多种人类疾病的理论发展。 Orexin 1和Orexin 2受体是A类 GPCR在中枢神经系统中差异表达。 Orexin 1接收器最丰富 在核座基因座中表达，被认为可以控制情感，奖励和自主神经的各个方面 神经系统，而Orexin 2受体在控制唤醒的地区表达 肺结核核，这是调节睡眠/清醒的重要部位。他们近20年后 最初发现，第一个潜在的双Orexin 1 / Orexin 2受体拮抗剂疗法已被带到 失眠市场（Belsomera®，Suvorexant，Merck）。尽管竭尽全力识别 Orexin受体，几乎没有关于小分子激动剂的报道出现在主要或专利文献中。优雅的 使用Orexin敲除动物和脑室内剂量的实验的遗传消融实验 Orexin A或Orexin B Petides的作用是Orexin接收器激动剂或潜在主义者的作用 指示包括：1）抑郁； 2）学习和记忆（认知）； 3）注意力缺陷多动障碍 （多动症）; 4）结肠癌治疗； 5）睡眠障碍，包括睡病。而Orexin Petides是 两种Orexin受体的有效激动剂，它们都不是选择性的，也不会越过血脑屏障， 使它们对体内药理学研究的探针底物很差。这提供了识别小的动力 分子，Orexin受体的大脑渗透激活剂，以询问受体的功能 疾病状态。我们已经优化了基于细胞的高通量筛选兼容的主要测定法 专门测量Orexin 1和Orexin 2的功能活性。初步的10K试验屏幕是 进行了这些受体的许多小分子激动剂的鉴定，证明了 该测定法是可重复的。 Scripps机构药物的完整HTS筛查运动 发现库（〜640K化合物）将导致多种类Orexin受体激动剂的识别 和进一步发展的潜在者。在迭代过程中，这些经过验证的命中将被表征 通过一系列基于体外细胞的测定，以确定效力，选择性和作用机理。 初步的医学化学铅优化将确定早期的结构活动关系和体外 将评估药物代谢以确认早期铅的疗程。这些努力将提供第一类 化学工具用于进一步探测疾病动物模型中的Orexin受体功能。