Novel assay to identify non-ATP competitive protein kinase inhibitors

鉴定非 ATP 竞争性蛋白激酶抑制剂的新方法

基本信息

批准号：
7271467
负责人：
Mark W Nowak
金额：
$ 20.13万
依托单位：
OMNEURON, INC.
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-01 至 2007-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7271467
关键词：
Adopted Antibodies Binding Binding Sites Biological Assay Biological Models Cell physiology Central Nervous System Diseases Cessation of life Chronic Myeloid Leukemia Cobalt Complementary DNA Development Disease Drug Design Environment Etiology Fluorescence Gleevec Human Human Genome Imatinib Inflammation Kinetics Label Ligands Malignant Neoplasms Mammalian Cell Measurement Measures Methodology Methods Molecular Molecular Conformation Monitor Oncogenic Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Protein Biosynthesis Protein Kinase Protein Kinase Inhibitors Proteins Range Research Personnel Role Safety Screening procedure Sepharose Signal Pathway Signal Transduction Site Structure Substrate Interaction System Technology Therapeutic Translations aqueous base concept drug discovery fluorophore high throughput screening in vitro Assay in vivo inhibitor/antagonist kinase inhibitor mutant novel protein aggregation protein kinase inhibitor small molecule

项目摘要

DESCRIPTION (provided by applicant): Protein kinases play a central role in many signaling pathways and are key effectors of cellular functions. As such, protein kinases inhibitors can potentially be developed as therapeutics for many diseases, including cancer, central nervous system disorders and inflammation. Drug discovery efforts have primarily identified small molecule ligands that compete directly with ATP binding. However, molecules that inhibit one kinase by binding at the highly conserved ATP site often inhibit other kinases, resulting in potential drug safety issues. We propose to develop a novel method for in vivo measurement of protein aggregation by monitoring the signals from small fluorophores that have been site-specifically incorporated into the protein during protein synthesis using nonsense suppression methodology. We propose to develop a novel high throughput screening assay to identify non-ATP competitive protein kinase inhibitors with potentially better selectivity and safety profiles. Our assay will identify compounds that allosterically inactivate protein kinases (i.e. stabilize the inactive or "off" state). We will employ a proven model system, Abelson ("Abl") kinase, for the "Proof of Concept" studies. We will develop and optimize the assay to detect inactive and active states of Abl.

描述（由申请人提供）：蛋白激酶在许多信号传导途径中发挥核心作用，并且是细胞功能的关键效应器。因此，蛋白激酶抑制剂有可能被开发为许多疾病的治疗方法，包括癌症、中枢神经系统疾病和炎症。药物发现工作主要确定了与 ATP 结合直接竞争的小分子配体。然而，通过在高度保守的 ATP 位点结合来抑制一种激酶的分子通常会抑制其他激酶，从而导致潜在的药物安全问题。我们建议开发一种体内测量蛋白质聚集的新方法，通过使用无义抑制方法监测在蛋白质合成过程中特定位点掺入蛋白质中的小荧光团的信号。我们建议开发一种新型高通量筛选方法来鉴定具有潜在更好选择性和安全性的非 ATP 竞争性蛋白激酶抑制剂。我们的测定将识别变构失活蛋白激酶的化合物（即稳定非活性或“关闭”状态）。我们将采用经过验证的模型系统 Abelson（“Abl”）激酶来进行“概念验证”研究。我们将开发和优化检测 Abl 的非活性和活性状态的检测方法。