Discovery of Zika virus therapeutics using a replicon assay

使用复制子测定发现寨卡病毒疗法

• 批准号: 9761977
• 负责人: Terry L. Bowlin
• 金额: $ 28.5万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-13 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9761977
• 关键词: Acute; Address; Aedes; African; Americas; Antiviral Agents; Arboviruses; Asians; Biological Assay; Caribbean region; Cell Line; Central America; Combined Modality Therapy; Congenital Abnormality; Culicidae; Data; Dengue; Dengue Virus; Development; Disease Outbreaks; Drug Combinations; Engineering; Exhibits; FDA approved; Family; Flavivirus; Genome; Guillain-Barré Syndrome; Hepatitis C virus; Human; Infection; International; Laboratories; Libraries; Mammalian Cell; Mass Vaccinations; Medical; Microcephaly; Nature; Nonstructural Protein; Nucleotides; Peptide Hydrolases; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Play; Polyproteins; Prevalence; Property; Prophylactic treatment; RNA Viruses; Replicon; Reporting; Resistance development; Role; Series; Serotyping; Small RNA; South America; Specificity; Structural Protein; Structure; Structure-Activity Relationship; Symptoms; Texas; Therapeutic; Therapeutic Agents; Therapeutic Uses; Ticks; United States; Universities; Vaccines; Validation; Viral; Viral Proteins; Virus; Virus Replication; West Nile virus; World Health Organization; Yellow fever virus; ZIKV infection; Zika Virus; base; climate change; cytotoxic; cytotoxicity; drug development; drug discovery; high throughput screening; inhibitor/antagonist; lead optimization; neurotropic; novel; novel therapeutics; pandemic disease; particle; pre-clinical; prevent; programs; prophylactic; protein structure; public health emergency; small molecule; success; transmission process

ABSTRACT The Zika virus (ZIKV) is the cause of an explosive pandemic of infections across South and Central America, the Caribbean, and the southeastern region of the United States. ZIKV is a flavivirus, a family of small positive strand RNA viruses that includes Dengue virus (DENV), West Nile virus (WNV), Yellow fever virus (YFV), and hepatitis C virus (HCV). ZIKV is transmitted to humans by mosquitoes of the Aedes genus. For the most part, ZIKV causes mild symptoms that mimic dengue fever; however, ZIKV infections in the Americas have been associated with congenital microcephaly and to the neurotropic Guillain-Barré syndrome. This prompted the World Health Organization to declare ZIKV to be a “public health emergency of international concern”. Despite the significant medical need for prophylactic or therapeutic treatments, there are no vaccines or drugs that have been approved for ZIKV. Therefore, novel therapeutic agents for prophylaxis or treatment of acute ZIKV infections are needed. To address this significant unmet medical need, we will utilize a stable ZIKV replicon cell line to carry out a high-throughput screen to identify novel inhibitors of ZIKV replication, which will be developed into drugs for ZIKV infections. Our approach is based on the strong scientific premise that is built on the longstanding success of flavivirus replicons in drug discovery and development. Recently, the laboratory of our collaborator Pei-Yong Shi (University of Texas Medical Branch) reported the construction and validation of a ZIKV replicon cell line (ZIKV Rep-neo). In preliminary studies, we have used the ZIKV Rep-neo cell line to successfully carry out a small-scale screen of FDA-approve compounds, demonstrating the feasibility of our approach. In Phase I, we will apply the ZIKV replicon assay to screen a library of ≥350,000 small molecules for compounds that inhibit replication, and will use counter screens and PAINS filters to eliminate non-specific inhibitors and cytotoxic compounds. We will prioritize the hits based on the results of a panel of secondary assays to assess the potency against infectious ZIKV, spectrum of activity against flaviviruses, cytotoxicity, and the drug-like properties of these compounds. The mechanism of action of prioritized compounds will be investigated to verify that prioritized hits target viral proteins, and preliminary structure activity relationships will be assessed. Inhibitors that meet the stringent criteria listed in the milestones for each specific aim will undergo hit to lead optimization in a subsequent Phase II project. In Phase I of this project we will accomplish the following Specific Aims. Aim 1. Identify potent inhibitors of the ZIKV replicon in a high-throughput screen. Aim 2. Prioritize confirmed hits based on potency, specificity, and drug-like properties. Aim 3. Determine the mechanism of action of prioritized inhibitors. Aim 4. Establish preliminary structure activity relationships for prioritized inhibitors.

抽象的 寨卡病毒（ZIKV）是南美各地感染的爆炸性大流行的原因， 加勒比海和美国东南部地区。 Zikv是一位黄病毒，一个小的积极家庭 包括登革热病毒（DENV），西尼罗河病毒（WNV），黄热病病毒（YFV）和包括的链RNA病毒和 丙型肝炎病毒（HCV）。 ZIKV通过伊蚊属的蚊子传播给人类。在大多数情况下， ZIKV会导致模仿风扇的轻度症状；但是，美洲的ZIKV感染一直是 与先天性的小头畸形和神经型Guillain-Barré综合征有关。这提示了 世界卫生组织宣布ZIKV为“国际关注的公共卫生紧急情况”。尽管 对预防性或治疗性治疗的重要医疗需求，没有疫苗或药物 已被批准为Zikv。因此，用于预防或治疗急性ZIKV的新型治疗剂 需要感染。为了满足这种巨大的未满足的医疗需求，我们将使用稳定的ZIKV复制品 细胞系执行高通量屏幕以识别新型ZIKV复制抑制剂，这将是 发展为ZIKV感染的药物。我们的方法基于建立的强大科学前提 关于Flavivirus复制品在药物发现和发育中的长期成功。最近， 我们合作者Pei-Yong Shi（德克萨斯大学医学分支）的实验室报告了建设和 ZIKV复制单元线（ZIKV REP-NEO）的验证。在初步研究中，我们使用了zikv rep-neo 细胞系成功地执行FDA-Approve化合物的小规模屏幕，证明了 我们方法的可行性。在第一阶段，我们将应用ZIKV副本测定法以筛选≥350,000的库 用于抑制复制的化合物的小分子，并将使用计数器屏幕和痛苦过滤器 消除非特异性抑制剂和细胞毒性化合物。我们将根据A的结果优先考虑命中 次级评估小组以评估针对感染性ZIKV的效力，活动范围 黄病毒，细胞毒性和这些化合物的药物样特性。作用机理 将研究优先的化合物，以验证优先级的攻击目标病毒蛋白和初步 结构活动关系将被评估。符合符合严格标准的抑制剂 在随后的II阶段项目中，每个特定目标的里程碑将受到铅优化的打击。 在该项目的第一阶段，我们将完成以下特定目标。目标1。确定潜在的抑制剂 高通量屏幕中的ZIKV复制品。目标2。根据效力，特异性，确认的命中优先级 和类似药物的特性。目标3。确定优先抑制剂的作用机理。目标4。建立 优先抑制剂的初步结构活动关系。