Development of a Malarial Kinase-on-Phage Screening Platform

疟疾激酶噬菌体筛选平台的开发

• 批准号: 8240362
• 负责人: Clifford Berkman
• 金额: $ 21.89万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8240362
• 关键词: Achievement; Affinity; Antimalarials; Bacteriophage T7; Bacteriophages; Binding; Biochemical; Biological Assay; Capsid; Chimeric Proteins; Companions; Competitive Binding; Cyclin-Dependent Kinases; DNA biosynthesis; Data; Development; Drug Delivery Systems; Enzymes; Genomics; Goals; Human; Interferometry; Libraries; Malaria; Measures; Methods; Modeling; Outcome; Parasites; Phage Display; Pharmaceutical Preparations; Phosphotransferases; Plasmodium falciparum; Protein Kinase; Proteome; Recombinants; Regulation; Screening procedure; Specificity; Surface Plasmon Resonance; System; Technology; Testing; Work; asexual; base; drug candidate; high throughput screening; inhibitor/antagonist; kinase inhibitor; novel; novel therapeutics; sensor; small molecule; tool; treatment strategy; vector

DESCRIPTION (provided by applicant): The identification of new Plasmodium falciparum kinase targets have been hampered due to the limited numbers of kinases that have been purified and the lack of an available high-throughput assay to screen potential drug candidates. Our long-term goal is to develop a high-throughput biochemical screening platform to profile the selectivity of novel kinase inhibitors from companion parallel-synthetic libraries. The overall objective of this application is to develop both a model P. falciparum kinase-displaying T7 phage platform and a Bio-Layer Interferometry (BLI) assay amendable for high-throughput screening. The central hypothesis of this application is that P. falciparum kinase-displaying phage can be developed to screen putative anti-malarial P. falciparum kinase inhibitors. We plan to test our central hypothesis and accomplish the objective of this application by pursuing the following two specific aims: (1) Develop a phage display platform for P. falciparum kinases; and (2) Develop a biochemical assay amendable to high-throughput capable of direct determination of binding affinity of kinase inhibitors. The rationale for developing a BLI screening assay amendable for high- throughput is that in combination with parasitic proliferation assays it will allow for both the identification of essential malarial kinases and provide a screening platform for parallel-synthetic drug libraries. The expected outcome of the proposed work is that the necessary enabling technology will be developed for establishing a malarial kinase screening platform using phage and BLI. In combination with antimalarial assays it will then become possible to identify key malarial kinases as new drug targets. In addition, companion parallel-synthetic libraries of putative antimalarial kinase inhibitors can be later screened for specificity to particular P. falciparum kinases. PUBLIC HEALTH RELEVANCE: The overall objective of this application is to develop both a model Plasmodium falciparum kinase-displaying T7 phage platform and a Bio-Layer Interferometry (BLI) assay amendable for high-throughput screening. The expected outcome of the proposed work is that the necessary enabling technology will be developed for establishing a malarial kinase screening platform using phage and BLI. We anticipate this technology would allow antimalarial drug candidates from companion parallel synthetic kinase inhibitor libraries to be profiled for both affinity and specificity for relevant malarial kinase targets.

描述（由申请方提供）：由于纯化的激酶数量有限，且缺乏筛选潜在候选药物的高通量检测方法，因此新的恶性疟原虫激酶靶标的鉴定受到阻碍。我们的长期目标是开发一个高通量的生化筛选平台，从伴侣平行合成库中分析新型激酶抑制剂的选择性。本申请的总体目标是开发模型恶性疟原虫激酶展示T7噬菌体平台和可用于高通量筛选的生物层干涉（BLI）测定。本申请的中心假设是恶性疟原虫激酶展示噬菌体可以被开发以筛选推定的抗疟疾恶性疟原虫激酶抑制剂。我们计划通过追求以下两个具体目标来测试我们的中心假设并实现本申请的目标：（1）开发用于恶性疟原虫激酶的噬菌体展示平台;以及（2）开发能够高通量地直接测定激酶抑制剂的结合亲和力的生物化学测定。开发可用于高通量的BLI筛选测定的基本原理是，与寄生虫增殖测定组合，其将允许鉴定必需的疟疾激酶并为平行合成药物文库提供筛选平台。拟议工作的预期成果是，将开发必要的使能技术，以建立一个使用噬菌体和BLI的疟疾激酶筛选平台。与抗疟疾检测相结合，将有可能确定关键的疟疾激酶作为新的药物靶点。此外，可以随后筛选推定的抗疟激酶抑制剂的伴随平行合成文库对特定恶性疟原虫激酶的特异性。 公共卫生相关性：本申请的总体目标是开发一个模型恶性疟原虫激酶展示T7噬菌体平台和生物层干涉（BLI）测定可用于高通量筛选。拟议工作的预期成果是，将开发必要的使能技术，以建立一个使用噬菌体和BLI的疟疾激酶筛选平台。我们预计，这项技术将允许从伴侣平行合成激酶抑制剂库的抗疟药物候选人，以分析相关的疟疾激酶靶点的亲和力和特异性。