Rapid Kinase Profiling with Luminescent Reporters

使用发光报告基因快速分析激酶

• 批准号: 8124548
• 负责人: REENA ZUTSHI
• 金额: $ 91.79万
• 依托单位: LUCEOME BIOTECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8124548
• 关键词: ATP phosphohydrolase; Active Sites; Adverse effects; Architecture; Area; Binding; Binding Sites; Biological Assay; Cardiovascular Diseases; Cell Proliferation; Cells; Clinic; Clinical Trials; Collection; Communities; Complex; Dasatinib; Diabetes Mellitus; Disease; Drug Design; Early identification; Enzymes; Event; Failure; Family; Fingerprint; Goals; Gray unit of radiation dose; Human Genome; Individual; Inflammation; Lead; Link; Lipids; Luciferases; Luminescent Measurements; Malignant Neoplasms; Market Research; Marketing; Mediating; Mediator of activation protein; Metabolic Diseases; Methods; Neoplasm Metastasis; Pharmaceutical Preparations; Pharmacologic Substance; Phase; Phosphorylation; Phosphotransferases; Protein Kinase; Protein Tyrosine Kinase; Provider; Reporter; Research Personnel; Resources; Scaffolding Protein; Screening procedure; Services; Signal Transduction; Signal Transduction Pathway; Specificity; Staging; Staurosporine; Technology; Therapeutic; Therapeutic Intervention; Time; Toxic effect; Validation; Work; base; cost; cross reactivity; drug candidate; drug development; drug discovery; inhibitor/antagonist; kinase inhibitor; luminescence; meetings; mutant; neoplastic cell; phase 1 study; pre-clinical; small molecule; success; therapeutic target

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.

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标。目前已上市的激酶药物有10种，另有80种抑制剂正在进行临床试验，还有更多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，使得选择性成为药物发现和开发中的一个问题。与许多激酶结合的混杂或“肮脏”药物预计会引起不良副作用。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标（例如激酶）可能是治疗疾病的理想策略。因此，针对大量激酶分析候选药物不仅可以帮助预测毒性特征，还可以帮助确定旧化合物的新靶点。在此第二阶段应用中，我们将开发基于分裂荧光素酶的发光测定法，用于全激酶组分析。目前，由于激酶分析的成本很高，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，许多研究人员通常被拒之门外。我们的目标是使这些分析测定变得经济实惠，以便可以更早地进行化合物分析，从而及早识别失败并带来更多成功的机会。 公共健康相关性：激酶是信号转导途径的重要介质，它们在细胞内的活性受到严格调控。激酶的失调与许多疾病有关，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 结合位点具有相似的结构而产生的。针对大量激酶筛选化合物有助于开发选择性指纹，可用于做出将化合物推进临床的重要决策。我们应用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。