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

总结 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， 调节多巴胺信号传导的能力。然而，尽管作出了广泛的努力， 在过去的30年里，制药业和学术界仍然没有有效的，选择性的NTSR 1非- 肽激动剂或增效剂以及文献中描述的仅少数拮抗剂。从高吞吐量 从高通量筛选（HTS）的角度来看，大多数注意力都集中在NTSR 1受体之间的相互作用 以及其通过Gq蛋白偶联信号转导途径的信号传导。然而，现在很明显， NTSR 1可以与多种G蛋白偶联，甚至可以以不依赖于G蛋白的方式发出信号。所以 本提案的总体目标是采用多种测定方法来推动迭代药物化学 该项目旨在鉴定NTSR 1的强效、选择性、细胞渗透性阳性调节剂。避免错过 潜在的有价值的化合物我们的方法是在多个基于细胞的测定中筛选化合物， 不同的功能读数。这可以被看作是撒下一张大网来捕获调节的化合物。 受体通过不同的机制，然后让功效在功能测定，使我们能够创造 每种化合物的细胞反应谱或“功能指纹”。这种体外功能指纹将 对于将来在体内评价化合物是非常宝贵的，挑战是识别生理活性。 NTSR 1的功能选择性的结果。因此，我们的多重测定方法是一个巨大的进步， 相对于制药行业通常采用的单一[Ca 2 +]/FLIPR测定方法。

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.