Development of Chemical Probes to Investigate the Role of NTSR1 in CNS Disorders

描述（由申请人提供）：神经紧张素系统由于其调节多巴胺信号和传递的能力，作为治疗药物成瘾的潜在靶点引起了很多关注。然而，尽管在过去的30年里，制药行业和学术界进行了广泛的努力，仍然没有有效的、选择性的NTSR1非肽激动剂或增强剂，只有文献中描述的几种拮抗剂。从高通量筛选（high - throughput screening， HTS）的角度来看，人们大多关注的是NTSR1受体及其信号通过Gq蛋白偶联信号转导途径之间的相互作用。然而，现在很清楚，NTSR1可以与多种G蛋白偶联，甚至可能以独立于G蛋白的方式发出信号。因此，本提案的总体目标是采用多种分析方法来驱动迭代的药物化学计划，旨在鉴定NTSR1的有效的、选择性的、细胞渗透性的阳性调节剂。为了避免遗漏潜在的有价值的化合物，我们的方法是在多种基于细胞的分析中筛选具有不同功能读数的化合物。这可以看作是撒下一张大网，捕捉通过不同机制调节受体的化合物，然后让功能分析中的功效使我们能够为每种化合物创建细胞反应谱或“功能指纹”。这种体外功能指纹对于未来体内化合物的评估将是无价的，挑战在于确定NTSR1功能选择性的生理后果。因此，我们的多重分析方法是一个巨大的改进，比单一的[Ca2+]/FLIPR分析方法通常追求制药行业。

DESCRIPTION (provided by applicant): The neurotensin system has attracted a lot of attention as a potential target to treat drug addiction due to its ability to modulate dopamine signaling and transmission. However, despite extensive effort by the pharmaceutical industry and academia over the past 30 years, there are still no potent, selective NTSR1 non- peptide agonists or potentiators and only a few antagonists described in the literature. From a high throughput screening (HTS) perspective most of the attention has been paid to interactions between the NTSR1 receptor and its signaling through the Gq protein-coupled signal transduction pathway. However, it is now clear that NTSR1 can couple to multiple G proteins and may even signal in a G-protein independent fashion. Hence, the overall goal of this proposal is to employ a multiple assay approach to drive an iterative medicinal chemistry program aimed at identifying potent, selective, cell penetrant positive modulators of NTSR1. To avoid missing potential valuable compounds our approach is to screen compounds in multiple cell-based assays with different functional readouts. This can be viewed as casting a wide net to capture compounds that modulate the receptor via different mechanisms and then letting the efficacy in the functional assays enable us to create a cellular response profile or 'functional fingerprint' for each compound. This in vitro functional fingerprint will be invaluable for future evaluation of compounds in vivo, the challenge being to identify physiological consequences of functional selectivity at NTSR1. Thus, our multiple assay approach is a vast improvement over the single [Ca2+]/FLIPR assay approach typically pursued by the pharmaceutical industry. PUBLIC HEALTH RELEVANCE: The neurotensin 1 receptor (NTSR1) is expressed peripherally and in the CNS and has attracted a lot of attention as a potential target to treat drug addiction due to its ability to modulate dopamine signaling and transmission. This proposal seeks to develop a series of cell-based functional assays to drive an iterative medicinal chemistry program aimed at identifying potent, selective, cell penetrant positive modulators of NTSR1.