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（由申请人提供）：由于已纯化的激酶数量有限且缺乏可用的高通量测定法来筛选潜在的候选药物，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生化筛选平台，以分析伴随平行合成库中新型激酶抑制剂的选择性。该应用的总体目标是开发恶性疟原虫激酶展示 T7 噬菌体平台模型和适用于高通量筛选的生物层干涉测量 (BLI) 测定。本申请的中心假设是，可以开发恶性疟原虫激酶展示噬菌体来筛选假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检验我们的中心假设并实现本申请的目标：（1）开发恶性疟原虫激酶的噬菌体展示平台； (2) 开发一种适合高通量的生化测定，能够直接测定激酶抑制剂的结合亲和力。开发适用于高通量的 BLI 筛选测定的基本原理是，与寄生增殖测定相结合，它将能够识别必需的疟疾激酶，并为平行合成药物库提供筛选平台。拟议工作的预期成果是，将开发必要的支持技术，以使用噬菌体和 BLI 建立疟疾激酶筛选平台。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标。此外，随后可以筛选推定的抗疟疾激酶抑制剂的伴随平行合成文库，以确定其对特定恶性疟原虫激酶的特异性。 公共健康相关性：本申请的总体目标是开发一种展示恶性疟原虫激酶的 T7 噬菌体平台模型和一种可修正用于高通量筛选的生物层干涉测量 (BLI) 测定法。拟议工作的预期成果是，将开发必要的支持技术，以使用噬菌体和 BLI 建立疟疾激酶筛选平台。我们预计这项技术将允许对来自并行合成激酶抑制剂库的抗疟候选药物进行分析，以了解相关疟疾激酶靶点的亲和力和特异性。