Rapid Kinase Profiling with Luminescent Reporters

描述（由申请人提供）：大约518种激酶由人类基因组编码，并作为磷酸化事件引起的信号转导的关键介质。已知许多激酶参与糖尿病、炎症、心血管疾病、肿瘤细胞增殖和转移，因此是经验证的治疗干预靶标。目前有10种已上市的激酶药物，另外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。这些激酶在活性位点（ATP结合结构域）具有相对相似的结构，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，预计会引起不必要的副作用。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谱，还有助于确定旧化合物的新靶点。在这个第二阶段的应用中，我们将开发我们的分裂荧光素酶为基础的发光分析的激酶组范围内的配置文件。目前，由于针对激酶的分析成本高，通常在药物开发后期获得选择性分析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。我们的目标是使这些分析测定能够负担得起，以便可以更早地进行化合物分析，从而早期识别失败，并带来更多的成功机会。 公共卫生相关性：激酶是信号转导途径的重要介质，其在细胞内的活性受到严格调节。激酶的调节异常与许多疾病有关，从而证实它们是治疗靶点。设计针对激酶的药物的挑战来自于它们的交叉反应性，这是由于许多激酶在ATP结合位点的相似结构而产生的。针对大量激酶筛选化合物可以帮助开发选择性指纹，这可以用于做出将化合物推向临床的重要决定。本申请的目的是开发用于药物发现的低成本、灵敏、基于发光的激酶测定。

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.