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

基本信息

批准号：
8337283
负责人：
CHARLES J. BIEBERICH
金额：
$ 15.19万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE COUNTY
依托单位国家：
美国
项目类别：
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/8337283
关键词：
Alzheimer&apos 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

项目摘要

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.

描述（由申请人提供）：激酶抑制是一种可行的治疗方法，用于多种疾病，包括癌症，糖尿病和阿尔茨海默氏症。尽管小分子抑制剂在激酶抑制领域占据了中心阶段，但毒性和抗性出现的挑战的有效性和影响有限。人们对将代理相结合以靶向多种信号通路以治疗癌症的需求的认识日益认识到这些已经重大的挑战。必须探索开发有效的新激酶抑制剂的每种可能的途径。干扰激酶 - 基底相互作用的肽具有低毒性，高特异性剂的潜力，可以满足该区域的关键未满足需求。我们建议将多种创新技术结合起来，以开发和测试，在体内，新型致癌激酶的肽抑制剂。将使用反向内凝胶激酶测定法与新型高分辨率染色体蛋白分离结合使用，将鉴定出具有最高观察到的催化效率的天然存在的蛋白激酶CK2底物。磷酸acceptor位点将通过电子转移解离辅助质谱法确定，并且蛋白质 - 蛋白质相互作用的位点将通过化学移动扰动介导的核磁共振光谱探测。将在体外和体内设计和分析底物竞争性假基质级肽抑制剂和对接现场肽抑制剂的对接现场肽抑制剂。对于体内分析，将使用一种新的方法来监测激酶抑制活性，以确定小鼠肠胃外治疗后CK2和外周血单核细胞中底物的噬菌体状态。这些目标的成功完成将提供原则上的证明，即现有的IMAT资助技术的创新应用可以促进新的肽激酶抑制剂的发展，以治疗异常激酶活性构成疾病引发或进展的癌症和其他疾病。