Application of an Innovative Technology to Develop Low Toxicity Kinase Inhibitors

应用创新技术开发低毒性激酶抑制剂

• 批准号: 8494129
• 负责人: CHARLES J. BIEBERICH
• 金额: $ 5.41万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE COUNTY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-22 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8494129
• 关键词: Alzheimer' s Disease; Area; Biological; Biological Assay; Cells; Chemicals; Complex; Data; Development; Diabetes Mellitus; Disease; Dissociation; Docking; Effectiveness; Electron Transport; Funding; Gel; Goals; Human; Kinetics; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Modality; Monitor; Mus; NMR Spectroscopy; Oncogenic; Outcome; Peptides; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phosphorylation; Phosphotransferases; Proteins; Proteome; Research; Resistance; Resolution; Route; Signal Pathway; Site; Source; Specificity; Staging; Substrate Interaction; Surveys; Technology; Testing; Therapeutic; Toxic effect; base; casein kinase II; design; in vitro activity; in vivo; inhibitor/antagonist; innovation; innovative technologies; kinase inhibitor; new technology; novel; novel strategies; novel therapeutics; protein protein interaction; small molecule; success

DESCRIPTION (provided by applicant): Kinase inhibition is a viable treatment approach for multiple diseases including cancer, diabetes, and Alzheimer's. Although small molecule inhibitors have occupied center stage in the kinase inhibition arena, challenges with toxicity and emergence of resistance have limited effectiveness and impact. The growing recognition of the need to combine agents to target multiple signaling pathways to treat cancers compounds these already significant challenges. It is imperative that every possible route to developing effective new kinase inhibitors be explored with alacrity. Peptides that interfere with kinase-substrate interactions have the potential to serve as low toxicity, high specificity agents to fill a critical unmet need in this area. We propose to combine multiple innovative technologies to develop and test, in vivo, novel peptide inhibitors of an oncogenic kinase. Using the reverse in-gel kinase assay in combination with novel high-resolution chromatofocusing-based protein separation, naturally occurring Protein Kinase CK2 substrates with the highest observed catalytic efficiency will be identified. Phosphoacceptor sites will be determined by Electron Transfer Dissociation-assisted mass spectrometry, and sites of protein-protein interaction will be probed by Chemical Shift Perturbation-mediated Nuclear Magnetic Resonance spectroscopy. Substrate- competitive pseudosubstrate-class peptide inhibitors, and docking site-class peptide inhibitors will be designed and analyzed for kinase inhibitory activity in vitro and in vivo. For in vivo analyses, kinase inhibitory activity will be monitored using a novel approach to determine the phophorylation state of CK2 and substrates in peripheral blood mononuclear cells after parenteral administration in mice. The successful completion of these aims will provide proof-of-principle that the innovative application of an existing IMAT-funded technology can facilitate the development of new peptide kinase inhibitors to treat cancers and other diseases where aberrant kinase activity underlies disease initiation or progression.

说明(申请人提供)：激酶抑制是一种可行的治疗多种疾病的方法，包括癌症、糖尿病和阿尔茨海默氏症。尽管小分子抑制剂已经占据了激酶抑制领域的中心舞台，但具有毒性的挑战和出现耐药的有效性和影响有限。越来越多的人认识到需要联合药物来靶向多个信号通路来治疗癌症，这加剧了这些本已重大的挑战。当务之急是迅速探索开发有效的新激酶抑制剂的每一种可能途径。干扰酶-底物相互作用的多肽有可能作为低毒、高特异性的试剂来填补这一领域尚未满足的关键需求。我们建议结合多种创新技术，在体内开发和测试致癌激酶的新型多肽抑制剂。将反向凝胶内蛋白激酶分析与基于新型高分辨率色谱聚焦的蛋白质分离相结合，将识别出具有最高观察催化效率的自然产生的蛋白激酶CK2底物。用电子转移解离辅助质谱仪确定磷受体的位置，用化学位移微扰介导的核磁共振波谱探测蛋白质-蛋白质相互作用的位置。将设计底物竞争性拟底物类多肽抑制剂和对接部位类多肽抑制剂，并对其进行体外和体内的激酶抑制活性分析。对于体内分析，将使用一种新的方法来监测KK2的活性，以确定小鼠肠外给药后外周血单核细胞中CK2和底物的磷酸化状态。这些目标的成功实现将提供原则证明，现有由IMAT资助的技术的创新应用可以促进新的肽激酶抑制剂的开发，以治疗癌症和其他疾病，这些疾病的异常激酶活性是疾病发生或发展的基础。