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 目前，世界正受到一种新型正链RNA病毒-严重急性呼吸道综合征-的传播的困扰。 冠状病毒2型综合征（SARS-CoV-2）。所有的正链RNA病毒都需要它们的病毒编码RNA- 依赖性RNA聚合酶（RdRp）合成RNA，作为mRNA用于产生病毒蛋白 或作为子代病毒体的基因组RNA。病毒聚合酶是抗病毒治疗的公认靶标， 广谱活动的明显潜力。丙型肝炎病毒（HCV）的RdRp，另一种正链RNA 病毒，是开发用于治愈HCV感染的抗病毒鸡尾酒的关键靶标。最有效的抑制剂 具有广谱活性的病毒聚合酶是核苷或核苷酸类似物。这类化合物 与病毒聚合酶的核苷酸底物结合口袋结合并掺入新生RNA中， 导致终止、诱变或重组，所有这些都可能对病毒是致命的。鉴于保守的 病毒RdRps的核苷酸底物结合口袋的结构和序列要求，一般来说， 开发用于治疗一种病毒的核苷酸类似物将显示抗病毒活性， 另体外和/或体内的效力可以不同。体内的问题往往与化合物的失效有关 在相关组织/器官中的激活和/或积累。冠状病毒特异性并发症的使用 核苷酸类似物的一个重要特征是存在3 '→ 5'核糖核酸外切酶活性，其不仅能够切除错配的 还有一些抗病毒的核苷酸该项目的主要动机是， 斯坦福大学AViDD中心（SyneRx）正在开发安全、有效的核苷/核苷酸候选物，用于 治疗和/或预防SARS CoV-2感染以及可能的其他呼吸道病毒病原体， 系统也是。我们项目的一项独特资产是一组具有抗SARS活性的核苷酸类似物- CoV-2 RdRp是通过筛选Riboscience LLC拥有的一个图书馆发现的。为了提高这些药物的效力 候选人，我们将通过抑制核糖核酸外切酶来拮抗切除。为了实现这些目标，我们将 具体目标如下：（1）鉴定用于开发的核苷酸类似物。(2)制定对抗策略 核糖核酸外切酶(3)合成和表征用于治疗癌症的磷酰胺酯前体和第二代核苷酸类似物 用作抗冠状病毒治疗剂。(4)聚合酶和核糖核酸外切酶抑制剂的生物学分析。 总之，这些研究有可能发现安全、有效的核苷酸类似物，用于治疗癌症。 治疗SARS-CoV-2感染和可能的其他呼吸系统病毒感染。