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

摘要 神经降压素系统作为治疗药物成瘾的潜在靶点已经引起了人们的广泛关注。 其调节多巴胺信号和传输的能力。然而，尽管美国政府做出了广泛努力 医药行业和学术界过去30年来，仍然没有强有力的、选择性的NTSR1非 多肽激动剂或增强剂和文献中仅描述的几种拮抗剂。从高吞吐量 筛选(HTS)视角NTSR1受体之间的相互作用一直是人们关注的焦点 并通过Gq蛋白偶联信号转导途径进行信号转导。然而，现在很明显， NTSR1可以与多个G蛋白偶联，甚至可能以不依赖G蛋白的方式发出信号。因此， 该提案的总体目标是使用多个化验方法来推动迭代的药物化学 旨在识别有效的、选择性的、细胞穿透性的NTSR1正向调节剂的计划。为避免遗漏 潜在的有价值的化合物我们的方法是在多个细胞分析中筛选化合物 不同的功能读数。这可以被视为撒下了一张大网，以捕获调节 受体通过不同的机制，然后让功效在功能分析中使我们能够创造 每种化合物的细胞反应图谱或“功能指纹”。这一体外功能指纹将 对于未来体内化合物的评估是非常宝贵的，挑战是识别生理学的 NTSR1的功能选择性的后果。因此，我们的多重化验方法是一个巨大的改进。 通过制药业通常采用的单一[Ca~(2+)]/FLIPR分析方法。

Summary 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.