Biochemical Strategy to Avert Microbial Drug Resistance

避免微生物耐药性的生化策略

• 批准号: 10645218
• 负责人: Ronald T Raines
• 金额: $ 15.51万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-14 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10645218
• 关键词: 2019-nCoV; Active Sites; Amino Acid Sequence; Antiviral Agents; Biochemical; Cause of Death; Cells; Clinical; Communicable Diseases; Cytotoxin; Enzyme Precursors; Enzymes; Equus caballus; Esterification; Genome; Goals; Human; Life Cycle Stages; Link; Lung; Mediating; Microbial Drug Resistance; Modeling; Pathogenicity; Patients; Peptide Hydrolases; Periodicity; Prodrugs; Proteins; RNA; RNA Splicing; RNA Viruses; Research; Resistance; Ribonucleases; SARS-CoV-2 protease; Specificity; System; Variant; Viral reservoir; Virus; antimicrobial; catalyst; cytotoxic; frontier; infectious disease treatment; intein; microbial; microbial disease; pathogen; pathogenic microbe; public health relevance; resistance mechanism

PROJECT SUMMARY/ABSTRACT The goal of the proposed research is to develop a generalizable antimicrobial strategy that averts the onset of resistance. The proposed antiviral agent has a mechanism of action that relies on the function of a pathogenic protein rather than its inhibition. Specifically, the agent is a cytotoxic variant of a human enzyme, ribonuclease 1 (RNase 1), that is cloaked and thus inactive as a catalyst. The cloak is removable only by a pathogenic protease that is essential for the lifecycle of the pathogen. SARS-CoV-2 will serve as a model pathogen in the study. The goal will be attained by (1) identifying an optimal amino acid sequence that is cleaved by the 3CLpro protease of SARS-CoV-2 but not by endogenous human proteases, (2) using that sequence and intein-mediated cis- splicing to create a cyclic RNase 1 zymogen, and (3) demonstrating the ability of the zymogen to kill human lung cells that express 3CLpro and that are infected by SARS-CoV-2. Because the activated zymogen can also destroy the genome of an RNA virus like SARS-CoV-2, it could eradicate the viral reservoir in patients. A notable feature of the strategy is its modularity: the protease-cleavage sequence is a cassette that can be altered to match the specificity of pathogenic proteases for the treatment of other infectious diseases.

项目概要/摘要 拟议研究的目标是开发一种通用的抗菌策略 从而避免抵抗的发生。所提出的抗病毒剂的机制是 依赖于致病蛋白的功能而不是其抑制的作用。 具体来说，该试剂是人类酶核糖核酸酶 1 的细胞毒性变体 （RNase 1），它被隐藏起来，因此没有催化剂活性。斗篷只能通过 一种对病原体生命周期至关重要的致病性蛋白酶。 SARS-CoV-2 将 作为研究中的模型病原体。该目标将通过以下方式实现：(1) 确定 SARS-CoV-2 的 3CLpro 蛋白酶切割的最佳氨基酸序列，但 不是通过内源性人类蛋白酶，（2）使用该序列和内含肽介导的顺式- 剪接以产生环状 RNase 1 酶原，以及 (3) 证明 酶原杀死表达 3CLpro 并被感染的人肺细胞 SARS-CoV-2。因为激活的酶原也会破坏RNA的基因组 像SARS-CoV-2这样的病毒，它可以根除患者体内的病毒库。一个显着的特点 该策略的关键在于其模块化：蛋白酶切割序列是一个盒式磁带，可以 改变以匹配致病性蛋白酶的特异性，用于治疗其他疾病 传染病。