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.

项目摘要指示进化是对实验室采用自然进化原则的，对其对分子工程。作为一个例子，它负责大多数批准的生成治疗性抗体以及仍在临床发育中的抗体。尽管取得了进步，但有价值的治疗层保持其范围之外：哺乳动物细胞。在体外系统中，噬菌体，细菌和酵母有借出哺乳动物细胞本身进行了实验室操纵，事实证明，疗法较少。因此，力量试图调节哺乳动物细胞的药物开发管道的进化仍无法访问信号。此外，许多定向的进化运动导致生物分子以关键的方式失败移植到人类细胞。为了解决这些限制和药物发现中的进步方法，我将探索和地雷病毒多样性，以创建一个新型的哺乳动物细胞分子进化系统。下一个，重点关注5-HT2A 5-羟色胺受体，我将创建细胞外纳米构造来模板发现药物通过结构性决心和硅对接进行运动。这些研究获得的见解将是应用于针对黑暗GPCR的状态特异性纳米型的定向演变。这项工作将导致哺乳动物细胞，针对HTR2A的化学物质和新路径的通用方法向GPCR进行去畸形的前进。