Complementation Assay for G protein linked Receptors

描述（由申请人提供）：G蛋白偶联受体（GPCR）是介导神经递质和激素生物学作用的膜蛋白家族，是药物开发的主要靶点。在前200种最畅销的药物中，有40多种针对GPCR，这些药物产生了超过200亿美元的收入。至少5%的人类基因组表达受体，其中绝大多数是孤儿受体，其功能和选择性配体尚未确定。这些孤儿受体可以为药物开发提供重要的新靶点家族。然而，有几个，如果有的话，可以成功地用于识别针对这些受体的药物的测定。在我们的第一阶段SBIR资助中，我们开发了一种基于细胞的检测方法，可以使用我们专有的CEDIA技术检测GPCR激活。具体而言，我们开发了一种测定NFkB/IkB信号传导的受体活化的测定法。与NFkB/IkB信号传导相关的细胞内信号传导途径的混杂性为测定孤儿受体的测定提供了理想的基础。SBIR第二阶段资助的目的是将这种受体检测方法发展到商业化的程度。我们将确定我们的GPCR测定法的适用性，以测量通过不同的第二信使信号传导途径作用的受体的刺激以及测量对孤儿受体激活的反应。我们将优化我们的检测试剂盒，以HTS格式进行测量。为了扩大我们的GPCR测定的实用性，我们将调整该测定以直接测量NFkB向细胞核的易位作为对GPCR刺激的响应，使得整个测定可以以非常简单且易于使用的形式在完整细胞中进行。除了测量对GPCR活化的响应之外，我们的NFkB/IkB信号传导测定测量了生长因子受体活化的刺激。在这项资助中，我们将开发和优化我们的检测方法，以测量对生长因子的反应，使第一个生长因子受体拮抗剂的HTS检测方法商业化。DiscoveRx的目标是开发一套技术，帮助发现新一代治疗上重要的药物，这些药物作用于孤儿GPCR和生长因子受体的大家族。 拟议的商业应用：我们将开发一种HTS检测方法，以发现针对孤儿GPCR大家族的药物。 这个市场是在数十亿美元。 我们也将能够商业化的一个大家族的生长因子受体，这是一个规模相当的市场。 最后，我们的核易位试验将作为GPCR筛选商业化，但也将在发现针对转录因子的药物和修饰基因表达的药物中应用。

DESCRIPTION (provided by applicant): G protein coupled receptors (GPCRs) are a family of membrane proteins that mediate the biological actions of neurotransmitters and hormones and are a major target for pharmaceutical drug development. Of the top 200 best selling drugs, over 40 of them target GPCRs and those drugs generate over $20 billion. At least 5 percent of the Human Genome expresses receptors, the vast majority of which are orphan receptors for which functions and selective ligands have not been identified. These orphan receptors could provide an important new family of targets for drug development. However, there are few, if any, assays that can be successfully employed to identify drugs against these receptors. In our Phase I SBIR grant, we developed a cell-based assay that can detect GPCR activation using our proprietary CEDIA technology. Specifically, we developed an assay to measure receptor activation of NFkB/IkB signaling. The promiscuity of intracellular signaling pathways linked to NFkB/IkB signaling provides an ideal basis for an assay to measure orphan receptors. The objective of this Phase II SBIR grant is to develop this receptor assay to the point of commercialization. We will determine the breath of applicability of our GPCR assay to measure stimulation of receptors acting via different second messenger signaling pathways as well as measuring responses to orphan receptor activation. We will optimize our assay to be measured in HTS format. To expand the utility of our GPCR assay, we will adapt the assay to directly measure the translocation of NFkB to the nucleus as a response to GPCR stimulation so that the entire assay can be done in intact cells in a very simple and easy-to-use format. In addition to measuring responses to GPCR activation, our NFkB/IkB signaling assay measured the stimulation of growth factor receptor activation. In this grant we will develop and optimize our assay to measure responses to growth factors to commercialize the first HTS assay for growth factor receptor antagonists. The goal of DiscoveRx is to develop a suite of technologies that can aid in the discovery of a new generation of therapeutically important drugs that act upon the large family of orphan GPCRs and growth factor receptors. PROPOSED COMMERCIAL APPLICATION: We will develop a HTS assay to discover drugs against the large family of orphan GPCRs. This market is in the billions of dollars. We will also be able to commercialize an assay for the large family of growth factor receptors which is a comparable size market. Finally, our nuclear translocation assay will be commercialized as a GPCR screen, but will also have applications in the discovery of drugs against transcription factors and drugs to modify gene expression.