Rapid Kinase Profiling with Luminescent Reporters

描述（由申请人提供）：已在人体中鉴定出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ATP的3-磷酰基转移到特定蛋白质底物上的羟基而发挥作用。激酶在疾病中的中心作用和它们在人类生物学中的基本作用已经促使开发有效的和选择性的抑制剂作为治疗剂和作为用于在信号转导途径的背景下剖析激酶功能的新工具。由于许多人类激酶在遗传和结构上都是如此密切相关，因此许多抑制剂对单一靶标缺乏特异性，并且通常通过竞争活性位点中的ATP而作用于多个激酶。因此，需要全面的激酶面板和工具来评估药物的选择性。2008年，激酶筛选和分析市场估计为1.06亿美元，需求每年都在稳步增长。本申请的目的是开发一种基于发光的、均质的、非放射性的、灵敏的、无细胞的测定法，其可以快速适用于激酶的小分子拮抗剂的高通量（HTP）筛选和分析。将证明该测定针对市售药物/dug候选物的实用性以及这些分子针对激酶组的选择性。 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。我们的应用程序的目的是开发一种方法，以确定选择性药物的分析对激酶面板。

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.