In Vitro Screening Strategies

体外筛选策略

• 批准号: 8465866
• 负责人: Patricia Helen McDonald
• 金额: $ 41.81万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8465866
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Abstinence; Agonist; Animal Model; Biological Assay; Calcium; Cell Line; Cells; Cyclic AMP; Data; Development; Discrimination; Drug Addiction; Drug Kinetics; Dyes; Enzymes; Exhibits; Fluorescence; GTP-Binding Proteins; Goals; Image; In Vitro; Inositol Phosphates; Instruction; Kinetics; Label; Lead; Life; Ligands; MAP Kinase Gene; Measurement; Measures; Mediating; Molecular Bank; Monitor; Neurons; Nicotine Dependence; Pharmaceutical Chemistry; Pharmacology; Phosphorylation; Physiological; Production; Property; Reader; Receptor Activation; Recombinants; Relapse; Rodent; Series; System; Technology; Therapeutic Intervention; Time; Triage; base; cell type; cigarette smoking; counterscreen; design; drug discovery; effective therapy; electric impedance; high throughput screening; hypocretin; in vivo; interest; new technology; novel; novel therapeutics; orexin 1 receptor; orexin B receptor; programs; receptor; response; sample collection; screening

The objectives of Project 2 are the refinement and application of G protein-dependent cell-based functional assays for monitoring Orexin 1 receptor (0X1R) activity, suitable to determine potency and mechanism of action of 0X1R antagonists that have the potential to be developed as novel therapeutics for the treatment of nicotine dependence. In addition, a series of G protein-independent cell-based functional assays will be developed and optimized to further characterize 0X1R ligands and allow for identification of compounds exhibiting 'functional selectivity'. Compounds synthesized in Project 1 and compounds emerging from the OXIR high throughput screening (HTS) campaign of the 'Molecular Libraries Probe Production Centers Network' (MLPCN) sample collection will be characterized in the aforementioned assays against recombinant 0X1R in high throughput fashion. Counterscreens to assess selectivity of high priority potent modulators will also be developed and activity of interesting compounds will be confirmed using primary neuronal cells to help bridge the gap between in vitro and in vivo pharmacology. This multiple assay approach together with pharmacokinetic data generated in Project 3 is designed to drive an iterative medicinal chemistry program (Project 1) aimed at identifying potent, selective, cell penetrant 0X1R antagonists that will advance to in vivo efficacy studies in animal models of nicotine dependence (Project 4).

项目2的目标是G蛋白依赖性细胞功能的完善和应用 用于监测食欲素 1 受体 (0X1R) 活性的测定，适用于确定食欲素 1 受体 (0X1R) 活性的效力和机制 0X1R拮抗剂的作用有可能被开发为治疗新疗法 尼古丁依赖。此外，一系列不依赖于 G 蛋白的基于细胞的功能测定将被 开发和优化以进一步表征 0X1R 配体并允许识别化合物 表现出“功能选择性”。项目 1 中合成的化合物以及从项目 1 中出现的化合物 “分子库探针生产中心”的 OXIR 高通量筛选 (HTS) 活动 Network'（MLPCN）样本收集将在上述分析中进行表征 以高通量方式重组 0X1R。反筛选以评估高优先级有效的选择性 还将开发调节剂，并使用初级技术确认感兴趣化合物的活性 神经元细胞有助于弥合体外和体内药理学之间的差距。这种多重检测 该方法与项目 3 中生成的药代动力学数据一起旨在驱动迭代 药物化学计划（项目 1）旨在识别有效、选择性、细胞渗透剂 0X1R 拮抗剂，将进入尼古丁依赖动物模型的体内功效研究（项目 4）。