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Genetically encoded bicyclic peptide libraries for the discoveryof novel antiviral agents

用于发现新型抗病毒药物的基因编码双环肽库

基本信息

批准号：
10189880
负责人：
Jeffery Micheal Tharp
金额：
$ 9.52万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-02 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10189880
关键词：
2019-nCoV Address Adoption Affinity Amines Amino Acids Amino Acyl-tRNA Synthetases Antibodies Antiviral Agents Bacteriophage M13 Bacteriophages Binding Cells Chemicals Communicable Diseases Coronavirus Coronavirus Infections Coupled Cyclic Peptides Cyclization Cysteine Development Development Plans Diagnosis Diagnostic Directed Molecular Evolution Disease Drug Targeting Engineering Evolution Future Genetic Code Goals Human In Vitro Laboratories Libraries Ligands Lung diseases Mediating Membrane Fusion Mentors Methods Middle East Respiratory Syndrome Coronavirus Modeling Molecular Conformation Peptide Conformation Peptide Library Peptides Phage Display Phase Prevention Property Proteins Reaction Research Ribosomes Route SARS coronavirus Side Supervision Surface System Techniques Technology Therapeutic Training Transfer RNA Translations Tyrosine-tRNA Ligase Virus Work base career career development combinatorial functional group human coronavirus in vivo inhibitor/antagonist novel novel coronavirus novel therapeutics pandemic disease peptide drug prevent programs protein protein interaction scaffold screening skills small molecule technology development thioether tool

项目摘要

PROJECT SUMMARY Bicyclic peptides are conformationally constrained peptides comprised of two macrocyclic rings. Owing to their increased rigidity, bicyclic peptides can bind to protein targets with greater affinity and selectivity than their linear and monocyclic counterparts. As a result, these molecules are highly desirable scaffolds for the development of peptide-based therapeutics. Phage display is a laboratory evolution technique that enables the discovery of high- affinity peptide ligands from large, combinatorial peptide libraries. Although initially limited to linear peptides, this technique has been adapted for the discovery of bicyclic peptide ligands. Most often, phage-displayed bicyclic peptides are generated by chemically modifying linear peptides using cysteine-reactive small molecules; how- ever, this method is technically challenging. As a result, progress in this field has been limited. Recently, several studies have used genetic code expansion to co-translationally install cysteine-reactive noncanonical amino acids (ncAAs) into phage-displayed peptides to produce libraries of cyclic peptides. This strategy has significant advantages over the chemical cyclization approach, but is currently limited to monocyclic peptides. The over- arching objective of this proposal is to develop technology that enables phage display of bicyclic pep- tides using genetic code expansion. Our central hypothesis is that bifunctional ncAAs, i.e. ncAAs containing two cysteine-reactive functional groups, can be used to generate ribosomally synthesized bicyclic peptides by intramolecular reaction with cysteine residues. To realize our objective, we will pursue three Specific Aims. In Aim 1 (K99 Phase) we will engineer an aminoacyl-tRNA synthetase that recognizes bifunctional ncAAs contain- ing two cysteine-reactive moieties. This will be accomplished using traditional and state-of-the-art methods of directed evolution. In Aim 2 (K99/R00 Phase) we will develop a phage display system that is compatible with co- translational installation of bifunctional ncAAs and we will optimize this system for bicyclic peptide formation. We will then validate this system by selecting and characterizing ligands for model targets. In Aim 3 (R00 Phase) we will use phage display to identify bicyclic peptides that bind to the spike protein of human coronaviruses and inhibit virus-host membrane fusion. By targeting proteins from various coronaviruses, we will strive to identify inhibitors with broad-spectrum antiviral activity. The proposed work will provide a facile route for generating bi- cyclic peptide libraries thereby greatly accelerating the discovery of therapeutic peptide leads. The Candidate, Dr. Jeffery Tharp’s long-term career goal is to establish an independent research program that uses genetic code expansion and phage display to develop antiviral peptides for the diagnosis, treatment, and prevention of infectious diseases. Herein we propose a detailed five-year Career Development Plan supervised by mentors Drs. Dieter Söll and Craig Wilen, and a team of subject-matter experts. This plan will augment previous training and address key training gaps to prepare Dr. Tharp for accomplishing his long-term career goal.

项目总结