Discovery of new antiviral methylase, protease, and helicase inhibitors of corona-, flavi-, and alphaviruses

发现冠状病毒、黄病毒和甲病毒的新型抗病毒甲基化酶、蛋白酶和解旋酶抑制剂

基本信息

批准号：
10513923
负责人：
THOMAS TUSCHL
金额：
$ 570.11万
依托单位：
HACKENSACK UNIVERSITY MEDICAL CENTER
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/10513923
关键词：
2019-nCoV Aerosols Alphavirus Animal Model Animals Antiviral Agents Biochemical Biological Assay Biological Availability COVID-19 COVID-19 outbreak Caspase Cell Membrane Permeability Cells Cessation of life Characteristics Chemicals Chikungunya virus China Collection Combined Modality Therapy Complex Consultations Contracts Coronavirus Development Disease Disease Outbreaks Distant Drug Kinetics Enzymes Family Flaviviridae Future Goals Human In Vitro Individual Industry Infection Innate Immune Response Institutes Laboratories Lead Libraries Medicine Metabolic Methyltransferase Middle East Respiratory Syndrome Coronavirus Monitor Mutation Names Oral Patients Peptide Hydrolases Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Polyproteins Preparation Program Development Property Protein Biosynthesis RNA Helicase RNA Viruses Recombinant Proteins Recording of previous events Reporting Resources Rice SARS-CoV-2 protease SH2D3A gene SH2D3C gene Series Solubility Specificity Structure Surface Therapeutic Togaviridae Toxic effect Universities Viral Viral Pneumonia Viral Proteins Virus Virus Replication ZIKA Zoonoses assay development base clinical candidate clinical development combat design drug development drug resistant virus falls helicase high throughput screening improved in vivo inhibitor insight lead optimization lead series luminescence member metropolitan mosquito-borne novel operation pandemic disease programs resistance mutation small molecule structural biology virology

项目摘要

Coronavirus Disease 2019 (COVID-19) represents an ongoing pandemic caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) and a current death toll of about 5 million. There are only limited antiviral treatment options for patients who contracted the disease. In March 2020, we initiated collaborative small molecule drug development including the laboratories of Dr. Tuschl (high-throughput screening (HTS) assay development), Dr. Glickman (HTS operations), Dr. Patel (structural biology), Dr. Rice (virology), and the Tri- Institutional Therapeutics Discovery Institute (medicinal chemistry) focusing on the two SARS-CoV-2 proteases, NSP3/PLpro and NSP5/3CLpro, required for viral polyprotein processing, the cap N7-G methyltransferase (MTase) NSP14, required for efficient viral protein synthesis and escape from host cell innate immune response, and the RNA helicase NSP13 important for viral replication. We developed robust, HTS fluorescent-substrate-based assays for monitoring PLpro, 3CLpro, and NSP13 activities as well as a luminescence-based assay for monitoring NSP14 N7-G-MTase activity. By HTS of the diverse library of 430,000 compounds available at Rockefeller University (RU), we identified hits in all assays, validated hits, and pursued the most promising non-covalent hits in medicinal chemistry programs. Our lead compounds for NSP14 and PLpro show IC50 values of 2 nM and 100 nM, respectively, and cell-based inhibition of SARS-CoV-2 replication with EC50 values of 80 nM and 7 µM, without cellular toxicity. Here, we propose further lead optimization to improve inhibitor potency and drug-like properties, as well as apply our HTS expertise to target proteases, MTases, and helicases of other RNA viruses with pandemic potential, including MERS, flavi- and alphaviruses. Our specific aims are: (1) Development of SARS-CoV-2 NSP14 and PLpro clinical candidates. Guided by structural insights from inhibitor-PLpro and NSP14 complexes, we will further optimize inhibitor potency and drug-like characteristics with industry-quality medicinal chemistry support from TDI and this program, and the goal to develop 3-5 advanced leads that are subjected to in vitro and in vivo DMPK/ADME-Tox studies utilizing all scientific cores designated for hit-to-lead and lead optimization (2) Develop antiviral lead series against MTases, proteases and helicases of select members of the Coronaviridae, Flaviviridae and Togaviridae families. We will adapt and develop HTS assays using recombinant proteins and identify covalent and non-covalent inhibitors in the RU drug collection and accompany the medicinal chemistry lead development to provide 3-5 leads per target for comprehensive DMPK/ADME-Tox analysis. (3) Monitor the selectivity of novel small molecule antivirals in biochemical assays (using increasingly distant viral and human recombinant proteins) and cell-based viral propagation, as well as resistance mutation identification. We will express recombinant proteins or exploit assays from other projects and scientific cores to define specificity of viral enzyme and replication inhibition and viral mutational escape.

冠状病毒疾病2019（COVID-19）代表了严重急性呼吸系统引起的持续大流行综合征冠状病毒2（SARS-COV-2），目前的死亡人数约为500万。只有有限的抗病毒患有疾病的患者的治疗选择。 2020年3月，我们启动了合作小型分子药物开发包括Tuschl博士的实验室（高通量筛查（HTS）测定法开发），Glickman博士（HTS操作），Patel博士（结构生物学），赖斯博士（病毒学）和三机构治疗学发现研究所（药物化学），重点是两个SARS-COV-2蛋白酶， NSP3/PLPRO和NSP5/3CLPRO，病毒多蛋白加工所需（MTase）NSP14，需要有效的病毒蛋白合成并摆脱宿主细胞先天免疫反应所必需的， RNA解旋酶NSP13对于病毒复制很重要。我们开发了强大的HTS荧光基底物测定法，以监测PLPRO，3ClPRO和 NSP13活性以及用于监测NSP14 N7-G-rtase活性的基于发光的测定。由HTS 洛克菲勒大学（RU）提供了430,000种化合物的潜水图书馆，我们确定了所有测定的命中经过验证的热门歌曲，并在药物化学计划中追求了最有希望的非共价打击。我们的领导 NSP14和PLPRO的化合物分别显示为2 nm和100 nm的IC50值，以及基于细胞的抑制作用具有80 nm和7 µm的EC50值的SARS-COV-2复制的，没有细胞毒性。在这里，我们提出了进一步的铅优化，以提高抑制剂效力和类似药物的特性因为将我们的HTS专业知识应用于其他大流行的RNA病毒的蛋白酶，MTase和Helicases 潜在的，包括MERS，黄霉菌和α病毒。我们的具体目的是：（1）开发SARS-COV-2 NSP14和PLPRO临床候选物。以抑制剂-PLPRO和NSP14的结构见解为指导复合物，我们将通过行业质量医学进一步优化抑制剂效力和类似药物的特征 TDI和该计划的化学支持，以及开发3-5个高级线索的目标体外和体内DMPK/ADME-TOX研究利用所有指定的科学核心优化（2）针对精选成员的MTase，蛋白酶和解旋酶开发抗病毒铅系列 Coronaviridae，Flaviviridae和Togaviridae家族的家族。我们将使用重组蛋白质并鉴定RU药物收集中的共价和非共价抑制剂，并涉及医学化学铅开发为综合DMPK/ADME-TOX提供3-5条线索提供3-5条线索分析。（3）监测新型小分子抗病人在生化测定中的选择性（使用越来越远的病毒和人类重组蛋白）和基于细胞的病毒传播以及抗性突变鉴定。我们将从其他项目中表达重组蛋白或利用测定法和科学核心，以定义病毒酶的特异性和复制抑制和病毒突变逃生。