DRUG DISCOVERY BY DIRECTED EVOLUTION IN MAMMALIAN CELLS

通过哺乳动物细胞定向进化发现药物

基本信息

批准号：
10644749
负责人：
Daniel Dewran Kocak
金额：
$ 9.82万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10644749
关键词：
Address Adopted Animals Archaea Arrestins Bacteria Bacteriophages Biological Assay Biology Cell Line Cells Chemicals Clustered Regularly Interspaced Short Palindromic Repeats Coupled Cryoelectron Microscopy DNA Viruses DNA-Directed RNA Polymerase Darkness Development Directed Molecular Evolution Docking Drug Targeting Endowment Engineering Environment Evolution Failure Family Family member Fertility G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Generations Genes Genetic Genetic Transcription Genetic Variation Growth HTR2A gene Human In Vitro Innate Immune System Interferons Laboratories Lead Libraries Mammalian Cell Methodology Methods Modification Molecular Molecular Conformation Molecular Evolution Nature Organism Orphan Output Pharmaceutical Preparations Plants Prokaryotic Cells Property Proteins RNA RNA Viruses Replicon Ribosomes Signal Transduction Specificity Structure System Technology Testing Tetracyclines Therapeutic Therapeutic antibodies Time Togaviridae Trans-Activators Transducers Transplantation Viral Virus Work Yeasts antagonist clinical development computerized data processing cost direct application drug candidate drug development drug discovery extracellular human disease in silico insight member nanobodies novel pressure receptor serotonin receptor tissue culture tool

项目摘要

Project Summary Directed evolution, which adopts principles of natural evolution to the laboratory, is singular in its impact on molecular engineering. As one example, it is responsible for the generation of the majority of approved therapeutic antibodies and those still in clinical development. Despite its progress, a valuable therapeutic niche remains outside its scope: the mammalian cell. While in vitro systems, phage, bacteria, and yeast have lent themselves to laboratory manipulation, mammalian cells have proven less tractable. Consequently, the power of evolution remains inaccessible to drug development pipelines that seek to modulate mammalian cell signaling. Further, many directed evolution campaigns result in biomolecules that fail in critical ways when transplanted to human cells. To address these limitations and advance methods in drug discovery, I will explore and mine viral diversity to create a novel system for molecular evolution in mammalian cells. Next, focusing on the 5-HT2A serotonin receptor, I will create extracellular nanobodies to template a drug discovery campaign via structural determination and in silico docking. The insights gained by these studies will be applied toward the directed evolution of state-specific nanobodies against dark GPCRs. This work will result in a general method for directed evolution in mammalian cells, chemical matter against HTR2A, and new paths forward for the deorphanization of GPCRs.

项目概要定向进化将自然进化的原理带入实验室，它对人类的影响是独一无二的。分子工程。举个例子，它负责生成大多数已批准的治疗性抗体和仍在临床开发中的抗体。尽管取得了进展，但仍存在有价值的治疗利基仍然超出了它的范围：哺乳动物细胞。虽然在体外系统中，噬菌体、细菌和酵母已经借事实证明，对于实验室操作而言，哺乳动物细胞不太容易处理。因此，功率寻求调节哺乳动物细胞的药物开发管道仍然无法获得进化的知识发信号。此外，许多定向进化运动会导致生物分子在关键时刻失败。移植到人体细胞中。为了解决药物发现中的这些限制并推进方法，我将探索和挖掘病毒多样性，以创建哺乳动物细胞分子进化的新系统。下一个，专注于 5-HT2A 血清素受体，我将创建细胞外纳米抗体来模板药物发现通过结构确定和计算机对接进行活动。这些研究获得的见解将是应用于针对暗 GPCR 的状态特异性纳米抗体的定向进化。这项工作将导致哺乳动物细胞定向进化的通用方法、针对 HTR2A 的化学物质以及新途径推进 GPCR 的去孤儿化。