Optimizing nucleoside analog efficacy with novel exonuclease inhibitors

使用新型核酸外切酶抑制剂优化核苷类似物的功效

• 批准号: 10514274
• 负责人: CRAIG E. CAMERON
• 金额: $ 627.12万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 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/10514274
• 关键词: 2019-nCoV; 5' -exoribonuclease; Amino Acid Substitution; Antiviral Therapy; Binding; Biochemical; Biological; Biological Assay; COVID-19 treatment; Cell Culture Techniques; Cells; Complication; Connecticut; Coronavirus; DNA-Directed RNA Polymerase; Development; Drug Exposure; Drug resistance; Excision; Exhibits; Exonuclease; Exoribonucleases; Failure; Generations; Genetic Recombination; Goals; Health Sciences; Hepatitis C; Hepatitis C virus; Hydrolysis; In Vitro; Ions; Laboratories; Lead; Libraries; Measures; Messenger RNA; Metabolism; Metals; Mitochondrial RNA; Motivation; Mutagenesis; Mutation; North Carolina; Nucleosides; Nucleotides; Organ; Outpatients; Permeability; Phenotype; Polymerase; Prevention; Production; RNA; RNA Viruses; RNA-Directed RNA Polymerase; Resistance; Respiratory System; SARS-CoV-2 infection; SARS-CoV-2 inhibitor; Specificity; Structure; Structure-Activity Relationship; Testing; Tissues; Universities; Viral; Viral Genome; Viral Proteins; Viral Respiratory Tract Infection; Virion; Virus; Virus Inhibitors; analog; base; coronavirus therapeutics; deep sequencing; drug candidate; drug isolation; genomic RNA; in vivo; inhibitor; mutant; novel; nucleoside analog; nucleoside triphosphate; nucleotide analog; pandemic disease; pathogenic virus; programs; reverse genetics; screening; single molecule; viral RNA

Project Abstract The world is currently under siege by the spread of a novel positive-strand RNA virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). All positive-strand RNA viruses require their virus-encoded RNA- dependent RNA polymerase (RdRp) to synthesize RNA to serve as either mRNA for production of viral proteins or as genomic RNA for progeny virions. Viral polymerases are well-established targets for antiviral therapy with clear potential for broad-spectrum activity. The RdRp from hepatitis C virus (HCV), another positive-strand RNA virus, is a key target of the antiviral cocktails developed to cure HCV infection. The most efficacious inhibitors of viral polymerases with broad-spectrum activity are nucleoside or nucleotide analogs. This class of compounds binds to the nucleotide substrate-binding pocket of the viral polymerase and is incorporated into nascent RNA, leading to termination, mutagenesis, or recombination, all of which can be lethal to the virus. Given the conserved structural and sequence requirements of the nucleotide substrate-binding pocket of viral RdRps, in general, it is not at all surprising that a nucleotide analog developed to treat one virus would exhibit antiviral activity against another. Potency in vitro and/or in vivo can differ. The problem in vivo often relates to failure of compound activation and/or accumulation in the relevant tissue/organ. A coronavirus-specific complication for use of nucleotide analogs is the presence of a 3’→5’ exoribonuclease activity capable of excising not only mismatched basepairs but also some antiviral nucleotides. The primary motivation of this project, in alignment with the Stanford AViDD Center (SyneRx), is development of safe, effective nucleoside/nucleotide candidates for use in the treatment and/or prevention of SARS CoV-2 infection and perhaps other viral pathogens of the respiratory system as well. A unique asset of our program is a panel of nucleotide analogs with activity against the SARS- CoV-2 RdRp discovered by screening a library owned by Riboscience LLC. To increase the potency of these candidates, we will antagonize excision by inhibiting the exoribonuclease. To achieve these goals we will pursue the following specific aims: (1) Identify nucleotide analogs for development. (2) Develop strategies to antagonize the exoribonuclease. (3) Synthesize and characterize ProTides and second-generation nucleotide analogs for use as anti-coronavirus therapeutics. (4) Biological analysis of polymerase and exoribonuclease inhibitors. Together, these studies have the potential to discover safe, efficacious nucleotide analogs for use in the treatment of SARS-CoV-2 infection and perhaps other viral infections of the respiratory system.

项目摘要 当前，通过新型阳性RNA病毒，严重的急性呼吸道的传播，世界正在围困。 综合征冠状病毒2（SARS-COV-2）。所有阳性链RNA病毒都需要病毒编码的RNA- 依赖性RNA聚合酶（RDRP）合成RNA，作为生产病毒蛋白的mRNA 或作为后代病毒的基因组RNA。病毒聚合酶是抗病毒治疗的靶标 明确的广谱活动潜力。来自丙型肝炎病毒（HCV）的RDRP，另一种阳性RNA 病毒是为治疗HCV感染而开发的抗病毒药鸡尾酒的关键靶标。最有效的抑制剂 具有广谱活性的病毒聚合酶是核外侧或核侧类似物。这类化合物 结合病毒聚合酶的核前底底物结合袋，并掺入新生的RNA， 导致终止，诱变或重组，所有这些都可能对病毒致命。给定保守 通常 毫不奇怪的是，开发用于治疗一种病毒的核丁基类似物会表现出针对的抗病毒活性 其他。体外和/或体内的效力可能会有所不同。体内问题通常与化合物的失败有关 相关组织/器官中的激活和/或积累。冠状病毒特异性并发症 核苷酸类似物是3'→5'驱核核酸酶活性的存在，不仅可以切除不匹配 底培和一些抗病毒核苷酸。该项目的主要动机，与 Stanford Avidd Center（Synerx）是开发安全，有效的核苷/核苷酸候选者 SARS COV-2感染以及呼吸道的其他病毒病原体的治疗和/或预防 系统。我们计划的独特资产是一组核苷酸类似物，具有对SARS-的活性 COV-2 RDRP通过筛选Riboscience LLC拥有的库发现。增加这些效力 候选人，我们将通过抑制驱虫夹来对抗惊喜。为了实现这些目标，我们将追求 以下具体目的：（1）确定开发的核苷酸类似物。 （2）制定对抗的策略 驱虫核酸酶。 （3）合成并表征蛋白酶和第二代核苷酸类似物的类似物 用作抗癌病毒疗法。 （4）聚合酶和驱虫酶抑制剂的生物学分析。 这些研究共同发现安全，有效的核苷酸类似物用于 治疗SARS-COV-2感染以及呼吸系统的其他病毒感染。