Rapid Kinase Profiling with Luminescent Reporters

描述(申请人提供)：大约518个激酶由人类基因组编码，是由磷酸化事件带来的信号转导的关键媒介。已知许多激酶与糖尿病、炎症、心血管疾病、肿瘤细胞增殖和转移有关，因此是治疗干预的有效靶点。目前有10种激酶药物上市，另有80种抑制剂正在临床试验中，还有更多的药物正在临床前阶段进行评估。这些激酶在活性部位(ATP结合域)具有相对相似的结构，这使得选择性成为药物发现和开发中的一个问题。一种混杂的或“肮脏的”药物，它与许多激酶结合，预计会产生不想要的不良反应。与此同时，在某些情况下，在信号转导途径中选择性地抑制多个靶点，如激酶，可能是治疗疾病的理想策略。因此，针对一大组激酶分析候选药物不仅有助于预测毒性分布，还有助于为旧化合物确定新的靶点。在这个第二阶段的应用中，我们将开发我们的基于分裂荧光素酶的发光分析方法，用于全基因组分析。目前，由于针对激酶的图谱分析成本较高，通常在药物开发的后期获得选择性图谱，以验证先导化合物的特异性。此外，许多研究人员经常被拒之门外。我们的目标是让这些分析方法变得负担得起，这样就可以更早地进行复合分析，从而及早识别故障，并带来更多成功的机会。 与公共健康相关：激酶是信号转导途径的重要中介，它们在细胞内的活动受到严格调控。激酶调节失调与许多疾病有关，证实了它们是治疗的靶点。设计针对激酶的药物的挑战来自于它们的交叉反应，这是由于许多激酶在ATP结合部位的结构相似而引起的。筛选针对大量激酶的化合物可以帮助开发选择性指纹，这些指纹可用于为将化合物推向临床做出重要决定。我们应用的目的是开发低成本、灵敏、基于发光的激酶分析方法用于药物发现。

DESCRIPTION (provided by applicant): Approximately 518 kinases are encoded by the human genome and serve as critical mediators of signal transduction brought about by a phosphorylation event. Many kinases are known to be involved in diabetes, inflammation, cardiovascular diseases, tumor cell proliferation and metastasis and are therefore validated targets for therapeutic intervention. Currently there are 10 marketed kinase drugs, another 80 inhibitors are in clinical trials and many more are being evaluated in the preclinical stage. The kinases share a relatively similar architecture at the active-site (ATP-binding domain), making selectivity an issue in drug discovery and development. A promiscuous or 'dirty' drug, which binds to many kinases, is expected to give rise to unwanted adverse-effects. At the same time, in some cases, inhibiting multiple targets, like kinases, selectively in a signal transduction pathway might be a desired strategy for treating a disease. Profiling drug candidates against a large panel of kinases can therefore not only aid in anticipating toxicity profiles, but also help in identifying new targets for old compounds. In this Phase II application, we will develop our split-luciferase based luminescent assays for kinome-wide profiling. Currently due to the high costs of profiling against kinases, selectivity profiles are typically obtained later in drug development to verify the lead compound's specificity. In addition, many researchers are shut out on a routine basis. Our goal is to make these profiling assays affordable, so that compound profiling can be done earlier leading to early identification of failures and resulting in many more opportunities for success. PUBLIC HEALTH RELEVANCE: Kinases are important mediators of signal transduction pathways and their activity inside cells is tightly regulated. Dysregulation of kinases has been implicated in many diseases, validating them as therapeutic targets. The challenge in designing drugs against kinases comes from their cross-reactivity, which arises due to similar architecture of many kinases at the ATP-binding site. Screening compounds against a large number of kinases can help develop a selectivity fingerprint, which can be used to make important decisions for advancing a compound into the clinic. The purpose of our application is to develop low-cost, sensitive, luminescence based kinase assays for drug discovery.