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: 8153081
• 负责人: CHARLES J. BIEBERICH
• 金额: $ 12.65万
• 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 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8153081
• 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. PUBLIC HEALTH RELEVANCE: The successful completion of this project will fill a need for new technologies to support the development of peptide kinase inhibitors. It will provide proof-of- principle that the reverse in-gel kinase assay can identify candidate substrates from which inhibitory peptides with therapeutic potential can be derived.

描述（由申请人提供）：激酶抑制是多种疾病（包括癌症、糖尿病和阿尔茨海默病）的可行治疗方法。尽管小分子抑制剂已在激酶抑制竞技场占据中心舞台，但毒性挑战和耐药性的出现具有有限的有效性和影响。越来越多的人认识到需要联合收割机药物靶向多种信号通路来治疗癌症，这使这些已经很大的挑战更加复杂。迫切需要迅速探索开发有效的新激酶抑制剂的每一条可能途径。干扰激酶-底物相互作用的肽具有作为低毒性、高特异性试剂的潜力，以满足该领域中关键的未满足的需求。我们建议联合收割机多个创新技术开发和测试，在体内，新的肽抑制剂的致癌激酶。使用反向凝胶内激酶测定结合新型高分辨率基于色谱聚焦的蛋白质分离，将鉴定具有最高观察到的催化效率的天然存在的蛋白激酶CK 2底物。将通过电子转移解离辅助质谱法测定磷酸受体位点，并通过化学位移扰动介导的核磁共振光谱法探测蛋白质-蛋白质相互作用的位点。将设计底物竞争性假底物类肽抑制剂和对接位点类肽抑制剂，并分析其体外和体内激酶抑制活性。对于体内分析，将使用一种新方法监测激酶抑制活性，以确定小鼠胃肠外给药后外周血单核细胞中CK 2和底物的磷酸化状态。这些目标的成功完成将提供原则证明，即现有IMAT资助技术的创新应用可以促进新肽激酶抑制剂的开发，以治疗癌症和其他疾病，其中异常激酶活性是疾病开始或进展的基础。 公共卫生相关性：该项目的成功完成将满足对新技术的需求，以支持肽激酶抑制剂的开发。它将提供反向凝胶内激酶测定可以鉴定候选底物的原理证明，从该候选底物可以衍生具有治疗潜力的抑制肽。