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，并作为磷酸化事件带来的信号转导的关键介质。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。一种混杂或“肮脏”的药物，与许多激酶结合，预计会产生意想不到的副作用。同时，在某些情况下，选择性地抑制信号转导途径中的多个靶点，如激酶，可能是治疗疾病的理想策略。因此，分析针对大量激酶的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II期申请中，我们将开发基于分裂荧光素酶的发光分析，用于全基因组分析。目前，由于针对激酶的谱分析成本高昂，通常在药物开发后期获得选择性谱，以验证先导化合物的特异性。此外，许多研究人员在日常工作中被拒之门外。我们的目标是使这些分析分析能够负担得起，这样就可以更早地进行复合分析，从而早期识别故障，并获得更多成功的机会。

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.