Phage Display with Two Genetically Incorporated Noncanonical Amino Acids

具有两种基因掺入的非规范氨基酸的噬菌体展示

• 批准号: 8658699
• 负责人: Wenshe Ray Liu
• 金额: $ 28.85万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8658699
• 关键词: Affinity; Amino Acids; Angiogenesis Inhibitors; Bacteriophages; Binding; Chemical Structure; Chemicals; Data; Disease; Escherichia coli; Genetic; Genotype; Goals; Growth; Health; Image; Ions; Lead; Libraries; Ligands; Malignant Neoplasms; Membrane Proteins; Methods; Mission; Modification; Mole the mammal; Outcome; Peptides; Phage Display; Pharmaceutical Preparations; Phenotype; Proteins; Public Health; Research; Side; Site; Structure; Surface; Synthesis Chemistry; Techniques; Therapeutic; United States National Institutes of Health; Variant; Vascular Endothelial Growth Factors; Work; base; cancer diagnosis; cancer prevention; cancer therapy; carcinogenesis; combat; combinatorial chemistry; drug discovery; innovation; novel therapeutics; overexpression; screening; small molecule; therapeutic target; tumor growth

DESCRIPTION (provided by applicant): Phage display and combinatorial chemistry are two high throughput approaches for identification of targeted therapeutics of cancer. However, a phage display library has a limited structural diversity and small molecules generated by combinatorial chemistry generally cannot be pooled in large numbers for efficient screening. Our long term goal is to combine the rapid-screening feature of phage display and the diversity- generating power of synthetic chemistry to assemble approaches for high throughput identification of small- molecule antiangiogenic agents and apply these molecules to cancer diagnosis, profiling, imaging, and therapy. The primary objective of this particular application is to develop methods for constructing phage display libraries with expanded chemical diversities and screening these libraries to identify ligands specific for vascular endothelial growth factor (VEGF), a key target of antiangiogenic drugs. Our central hypothesis is that the chemical diversity of a phage display library can be significantly expanded by genetically incorporating two different noncanonical amino acids (NAAs) into the library, varying the identities of NAAs, and/or chemically modifying the incorporated NAAs. The rationale for the proposed research is that, once these unnatural phage display libraries are constructed, a large variety of unnatural peptides with diversified chemical structures can be rapidly screened against many cancer therapeutic targets, leading to identification of new therapeutics for cancer treatment and prevention. Guided by strong preliminary data, the objective of this application will be attained by pursuing three specific aims: 1) Optimize M13KE phage for the unbiased display of 20 natural amino acids and 2 NAAs; 2) Construct unnatural phage display libraries incorporated with different combinations of 2 NAAs and screen these libraries to identify VEGF-specific ligands; and 3) Expand the chemical diversity of unnatural phage display libraries by expanding the pool of genetically encoded NAAs and selectively modifying the incorporated NAAs. The research proposed in this application is innovative, because it will expand the phage display technique significantly by amending the displayed peptides with diversified structure moieties desirable for drug discovery. The proposed research is significant because it will accelerate the current drug discovery progresses and contribute to fulfilling the NIH mission in promoting health and combating diseases.

描述（由申请人提供）：噬菌体显示和组合化学是两种高通量方法，用于鉴定靶向癌症的靶向疗法。但是，噬菌体展示库的结构多样性有限，组合化学产生的小分子通常不能大量汇总以进行有效的筛选。我们的长期目标是结合噬菌体显示的快速筛查特征，以及合成化学的多样性产生能力，以组装方法，以高吞吐量鉴定小分子抗血管生成剂，并将这些分子应用于癌症诊断，分析，成像，成像和治疗。该特定应用的主要目的是开发用于构建具有扩展化学多样性的噬菌体显示文库的方法，并筛选这些文库，以识别针对抗高基因药物的主要目标血管内皮生长因子（VEGF）特异性的配体。我们的中心假设是，可以通过将两个不同的非规范氨基酸（NAAS）纳入文库中，可以显着扩展噬菌体展示文库的化学多样性，从而改变NaAS的身份和/或化学修改掺入的NAA。拟议的研究的理由是，一旦这些不自然的噬菌体展示库构建，可以迅速筛选出许多具有多样化化学结构的非天然肽，以与许多癌症治疗靶标相对，从而鉴定出用于癌症治疗和预防的新疗法。在强大的初步数据的指导下，将通过追求三个特定目的来实现本应用的目的：1）优化M13KE噬菌体，以无偏见的20个天然氨基酸和2个NAA； 2）构造与2个NAA的不同组合的不自然噬菌体展示库，并筛选这些库以识别VEGF特异性配体； 3）通过扩大遗传编码的NaAs的池并选择性地修改Incorporated NAAS，扩大了不自然的噬菌体展示文库的化学多样性。本应用程序中提出的研究具有创新性，因为它将通过修改具有多种结构部分的肽来显着扩展噬菌体显示技术。拟议的研究之所以重要，是因为它将加速当前的药物发现的进展，并有助于履行NIH促进健康和打击疾病的任务。