In Vitro Screening Strategies

项目2的目标是完善和应用G蛋白依赖的细胞为基础的功能 用于监测食欲素1受体（0X1R）活性的测定，其适用于确定 具有开发为治疗以下疾病的新治疗剂的潜力的0X1R拮抗剂的作用 尼古丁依赖此外，一系列G蛋白非依赖性细胞功能测定将在 开发和优化，以进一步表征0X1R配体并识别化合物 表现出“功能选择性”。在项目1中合成的化合物和在项目2中产生的化合物 “分子库探针生产中心”的OXIR高通量筛选（HTS）活动 将在上述试验中对“MLPCN”（MLPCN）样本采集进行表征， 重组0X1R。评估高优先级有效药物的选择性的反筛选 还将开发调节剂，并且将使用初级药物来确认感兴趣的化合物的活性。 神经元细胞，以帮助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设计用于驱动迭代 药物化学计划（项目1），旨在确定有效的，选择性的，细胞渗透剂0X1R 拮抗剂将推进到尼古丁依赖动物模型的体内疗效研究（项目4）。

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).