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 世界目前正被一种新的正链RNA病毒--严重急性呼吸道病毒--的传播所包围 SARS冠状病毒2型(SARS-CoV-2)。所有正链RNA病毒都需要病毒编码的RNA- 依赖RNA聚合酶(RdRp)合成RNA，作为生产病毒蛋白的任一种信使核糖核酸 或作为后代病毒粒子的基因组RNA。病毒聚合酶是公认的抗病毒治疗的靶点 显然存在广谱活动的潜力。丙型肝炎病毒的RdRp，另一种正链RNA 病毒，是为治疗丙型肝炎病毒感染而开发的抗病毒鸡尾酒的关键靶点。最有效的抗肿瘤药物 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 结合到病毒聚合酶的核苷酸底物结合口袋，并被结合到新生RNA中， 导致终止、突变或重组，所有这些都可能对病毒致命。考虑到保守派 病毒RdRps的核苷酸底物结合口袋的结构和序列要求，通常是 毫不奇怪，一种用于治疗一种病毒的核苷酸类似物会对 又一个。体外和/或体内的效力可以不同。体内的问题通常与化合物的失效有关。 激活和/或在相关组织/器官中积聚。使用冠状病毒特异性并发症 核苷酸类似物是存在的3‘→5’外切核糖核酸酶活性，不仅能够切除错配 但也有一些抗病毒的核苷酸。这个项目的主要动机，与 斯坦福AViDD中心(SyneRx)正在开发安全、有效的核苷酸/核苷酸候选药物，用于 治疗和/或预防SARS CoV-2感染，可能还有呼吸道的其他病毒病原体 系统也是如此。我们计划的一个独特资产是一组具有抗SARS活性的核苷酸类似物- 通过对RiBoscience LLC拥有的文库进行筛选，发现了CoV-2 RdRp。为了提高这些产品的效力 候选人，我们将通过抑制外切核糖核酸酶来对抗切除。为了实现这些目标，我们将继续努力 具体目标如下：(1)确定用于开发的核苷酸类似物。(2)制定对抗战略 外切核酸酶。(3)蛋白质及其第二代核苷酸类似物的合成和表征 用作抗冠状病毒的治疗药物。(4)聚合酶和外切核糖核酸酶抑制剂的生物分析。 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SARS-CoV-2感染，可能还有呼吸系统的其他病毒感染。