Novel Drug Screening Assays for Orphan GPCRs

描述（由申请人提供）：gpcr是一个细胞表面蛋白家族，参与介导大脑中神经递质的许多生物行为。它们在中枢神经系统生理学中起着至关重要的作用，也是制药行业药物发现的主要目标。大脑中的大多数gpcr都是孤儿，没有已知的配体或功能。孤儿gpcr （oGPCR）可能为开发新药提供重大机会，以更好地治疗常见和普遍的精神障碍。发现靶向这些受体的药物的一个主要问题是缺乏适当的化合物文库筛选技术。我们建议在这笔拨款中开发一种技术，允许发现针对ogpcr的药物。这笔拨款将由三家专注于GPCR药物发现的生物技术公司共同努力。Patobios的科学家们发现，在gpcr的近端c端区域插入NLS会导致受体从细胞膜表面转移到细胞质中。当配体通过NLS与GPCR结合时，它使受体保留在细胞表面防止易位，从而提供了与受体功能无关的高亲和力配体与GPCR结合的读数。DiscoveRx开发了一种酶片段互补（EFC）技术，可用于检测HTS格式的gpcr表面表达。在这笔拨款中，我们将合并这些技术来开发一种HTS筛选试验，以确定配体与ogpcr的结合。第三家公司Nura利用基因ko技术鉴定了三种ogpcr，其表型表明它们可能在精神障碍中起作用。我们将把Patobios/DiscoveRx HTS技术应用于这些孤儿，以便在未来的研究中，我们将利用Nura的药物发现能力来识别针对这些受体的高亲和力配体。这将达到两个目的；1)它将验证配体筛选试验在GPCR药物发现中的实用性；2)可能会导致针对这些受体的新型药物的开发，以治疗精神障碍。该项目将开发一种新技术，该技术将允许发现针对孤儿G蛋白连接受体的独特药物。目前，还没有类似的技术可以针对这一大家族受体进行药物筛选。针对这些受体发现和开发的药物有可能治疗包括焦虑和肥胖在内的许多精神障碍。

DESCRIPTION (provided by applicant): GPCRs are a family of cell surface proteins involved in mediating many of the biological actions of neurotransmitters in the brain. They play a critical role in CNS physiology and are a major target for drug discovery in the pharmaceutical industry. The majority of GPCRs in the brain are orphans, with no known ligand or function. Orphan GPCRs (oGPCR) may provide major opportunities in the development of new drugs to better treat common and pervasive mental disorders. A major problem in discovering drugs targeting these receptors is the lack of appropriate technologies for compound library screening. We propose in this grant to develop a technology that will allow for discovery of drugs against oGPCRs. This grant will be a joint effort between three biotechnology companies focused on GPCR drug discovery. Scientists at Patobios have discovered that inserting an NLS in the proximal C-terminal region of GPCRs causes the receptor to translocate from the cell surface membrane to the cytosol. When a ligand binds to the GPCR with the NLS, it causes the receptor to be retained at the cell surface preventing translocation providing a readout of binding of high affinity ligands to the GPCR independent of the receptors function. DiscoveRx has developed an enzyme fragment complementation (EFC) technology that can be used to detect surface expression of GPCRs in a HTS format. In this grant we will merge these technologies to develop a HTS screening assay to identify the binding of ligands to oGPCRs. The third company, Nura has employed gene ko technologies to identify three oGPCRs with phenotypes that suggest they may have roles in mental disorders. We will adapt the Patobios/DiscoveRx HTS technologies to these orphans so that in future studies we will employ the drug discovery capabilities of Nura to identify high affinity ligands targeting these receptors. This will serve two purposes; 1) it will validate the utility of the ligand screening assay for GPCR drug discovery and; 2) it may lead to the development of novel drugs targeting these receptors to treat mental disorders. This project will develop a novel technology that will allow for the discovery of unique drugs targeting orphan G protein linked receptors. At present, there is no comparable technology that will allow for drug screening against this large family of receptors. Drug discovered and developed against these receptors could have the potential for treating a number of mental disorders including anxiety and obesity.