Project 4: Nuclease Inhibitors for Viruses of Pandemic Concern

项目 4：针对流行病病毒的核酸酶抑制剂

基本信息

批准号：
10522813
负责人：
Hideki Aihara
金额：
$ 304.02万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-16 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10522813
关键词：
2019-nCoV 5&apos -exoribonuclease Address Animal Model Antiviral Agents Antiviral Response Arenavirus Binding Biochemical Biological Assay Bolivian Hemorrhagic Fever Virus COVID-19 screening Cell Culture Techniques Cell model Cells Chemicals Chemistry Collaborations Complement Complex Computer Models Cryoelectron Microscopy DNA Dose Effectiveness Ensure Enzymes Exoribonucleases FDA approved Fluorescence Generations Genome Goals In Vitro Infection Interferons Ions Junin virus Kinetics Knockout Mice Lassa virus Metals Midwestern United States Molecular Mutagenesis Nonstructural Protein Nucleoproteins Pathogenicity Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacodynamics Play Property RNA RNA chemical synthesis RNA replication RNA-Directed RNA Polymerase Replicon Resistance Resolution Ribavirin Role SARS-CoV-2 infection STAT1 gene Structure Structure-Activity Relationship Technology Testing Transgenic Mice Vaccines Validation Viral Viral Hemorrhagic Fevers Viral Pathogenesis Virus Virus Diseases Virus Inhibitors Virus Replication Work X-Ray Crystallography animal facility aptamer base design drug candidate effective therapy efficacy evaluation efficacy testing gain of function high throughput screening improved inhibitor laboratory facility molecular dynamics mouse model new therapeutic target novel nuclease nucleoside analog pandemic disease pharmacokinetics and pharmacodynamics preference remdesivir resistance mutation response structural biology synergism viral RNA virtual screening

项目摘要

Project 4 – Nuclease Inhibitors for Viruses of Pandemic Concern Abstract SARS-CoV-2 (SARS2) and highly pathogenic arenaviruses including Lassa virus (LASV), Junin virus, and Machupo virus share a structurally and functionally related 3'-to-5' exoribonuclease (ExoN) domain, which plays essential roles in proofreading during RNA syntheses by the error-prone viral RNA-dependent RNA polymerase (RdRp) and suppressing host antiviral responses. Project 4 is pursuing chemical inhibition of ExoN from these viruses of pandemic concern, with the goal of blocking viral replication by lethal mutagenesis as well as mitigating viral pathogenesis by reactivating host’s interferon responses. Our team has contributed extensively to the structural and functional understanding of these viral ExoN enzymes, including the elucidation of their first atomic structures and characterization of catalytic mechanisms. We have also developed robust fluorescence-based assays to quantitatively analyze the ExoN catalytic activities, including a novel assay featuring fluorogenic RNA aptamer substrates that enables a gain-of-function readout in ultra-high-throughput screening (uHTS). Furthermore, we have used DNA-encoded chemistry technology (DEC-Tec) to obtain selective binders to SARS2 ExoN and LASV nucleoprotein (NP) containing the ExoN domain. Building on these prior and preliminary studies, we will continue to work closely with Core B and use 3 complementary approaches (uHTS, DEC-Tec, and Virtual screening) to identify first-in-class viral ExoN inhibitors. We will then leverage the deep expertise of Core C and Core D in medicinal chemistry and structural biology, respectively, to enhance the potency, selectivity, and pharmacodynamic/kinetic properties of hit compounds for detailed antiviral studies in cell and animal models by Core E. These studies will deliver antiviral drug candidates with a distinct mechanism of action to complement those developed against established antiviral targets including RdRp, helping to address the critical need for novel antiviral drugs against both SARS2 and the highly pathogenic arenaviruses that cause fatal hemorrhagic fever infections.

项目4 - 有关大流行关注病毒的核酸酶抑制剂抽象的 SARS-COV-2（SARS2）和高度致病的体育症病毒，包括Lassa病毒（LASV），JUNIN病毒和 Machupo病毒在结构和功能上相关的3'to-5'Exoriboneclease（Exon）域，播放通过易易易用的病毒RNA依赖性RNA聚合酶在RNA合成过程中校对中的基本作用（RDRP）并抑制宿主抗病毒反应。项目4正在从这些中追求化学抑制外显子大流行关注的病毒，目的是通过致命的诱变和缓解来阻止病毒复制病毒发病机理通过重新激活宿主的干扰素反应。我们的团队为对这些病毒外显子酶的结构和功能理解，包括阐明其第一个原子催化机制的结构和表征。我们还开发了强大的基于荧光的强大定量分析外显子催化活性的测定法，包括具有荧光RNA的新型测定 APATMER基板可以在超高通量筛选（UHTS）中获得功能读数。此外，我们已经使用了DNA编码的化学技术（DEC-TEC）来获得选择性的粘合剂 SARS2外显子和LASV核蛋白（NP）含有外显子结构域。以这些先验和初步为基础研究，我们将继续与核心B紧密合作，并使用3种完整的方法（UHTS，DEC-TEC，，和虚拟筛查）以识别一流的病毒外显子抑制剂。然后，我们将利用核心C和核心D分别在医学化学和结构生物学中，以提高效力， HIT化合物的选择性和药效/动力学特性，用于细胞和细胞中详细的抗病毒研究核心E的动物模型。这些研究将提供具有不同作用机理的抗病毒药候选者补充那些针对包括RDRP在内的既定抗病毒目标的人，有助于解决针对SARS2和高度致病性舞台病毒的新型抗病毒药致命的出血热感染。