Rapid Kinase Profiling with Luminescent Reporters

描述(申请人提供)：已在人类中鉴定出500多种不同的蛋白激酶，占人类基因组的1.7%。蛋白激酶通过催化三磷酸腺苷的3-磷酸基转移到特定蛋白质底物上的羟基来发挥作用。激酶在疾病中的核心作用及其在人类生物学中的基础作用促使了有效和选择性的抑制剂的发展，作为治疗药物和在信号转导途径背景下剖析激酶功能的新工具。由于许多人的激酶在基因和结构上都是如此密切相关，许多抑制剂缺乏对单个靶点的特异性，并且作用于不止一个激酶，通常是通过在活性部位竞争超过ATP来实现的。因此，需要用于药物选择性评估的全面的激酶小组和工具。2008年，激酶筛查和特征分析市场估计为1.06亿美元，需求每年都在稳步上升。我们应用的目的是开发一种基于发光的、均匀的、非放射性的、灵敏的、无细胞的检测方法，该方法可以快速适用于高通量(HTP)筛选和分析小分子激动酶拮抗剂。我们将展示这种检测方法对商业上可用的药物/DUG候选药物的实用性，以及这些分子对一组激活酶的选择性。 与公共卫生相关：激酶占基因组的1.7%，与从癌症到糖尿病的各种疾病有关。评价药物对激酶的选择性是评价其生理安全性的关键。我们应用程序的目的是开发一种方法来识别选择性药物，方法是将它们与激酶面板进行比对。

DESCRIPTION (provided by applicant): More than 500 different protein kinases have been identified in humans and represent 1.7% of the human genome. Protein kinases function by catalyzing the transfer of the 3-phosphoryl group of ATP to the hydroxyl group on a specific protein substrate. The central role of kinases in disease and their fundamental role in human biology have prompted the development of potent and selective inhibitors as therapeutic agents and as novel tools for dissecting kinase function in the context of signal transduction pathways. Because many human kinases are so closely related both genetically and structurally, many inhibitors lack specificity for a single target and act on more than one kinase, typically by out-competing ATP in the active site. Hence there is a need for comprehensive kinase panels and tools for selectivity assessment of drugs. The kinase screening and profiling market was estimated to be $106 MM in 2008 and the demand is steadily rising every year. The purpose of our application is to develop a luminescence-based, homogeneous, non- radioactive, sensitive, cell-free assay that can be rapidly adapted for high throughput (HTP) screening and profiling of small molecule antagonists of kinases. The utility of this assay against commercially available drugs/dug candidates and the selectivity of these molecules against a kinase panel will be demonstrated. PUBLIC HEALTH RELEVANCE: Kinases constitute 1.7% of the genome and are implicated in various diseases, from cancer to diabetes. Assessment of selectivity of drugs against kinases is critical to evaluate their physiological safety. The purpose of our application is to develop a method to identify selective drugs by profiling them against a kinase panel.