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) 是南美洲和中美洲爆发性感染大流行的原因， 加勒比地区和美国东南部地区的 ZIKV 是一种黄病毒，属于小阳性病毒家族。 链RNA病毒，包括登革热病毒（DENV）、西尼罗河病毒（WNV）、黄热病病毒（YFV）和 丙型肝炎病毒 (HCV) 大部分通过伊蚊属传播给人类。 ZIKV 会引起类似登革热的轻微症状；然而，ZIKV 在美洲的感染率很高。 与先天性小头畸形和神经性格林-巴利综合征有关。 世界卫生组织宣布寨卡病毒为“国际关注的突发公共卫生事件”。 预防性或治疗性治疗的重大医疗需求，没有疫苗或药物可以 因此，用于预防或治疗急性 ZIKV 的新治疗药物已被批准。 为了解决这一未满足的重大医疗需求，我们将利用稳定的 ZIKV 复制子。 细胞系进行高通量筛选，以确定 ZIKV 复制的新型抑制剂，这将是 我们的方法基于强有力的科学前提。 最近，黄病毒复制子在药物发现和开发方面取得了长期成功。 我们的合作者Pei-Yong Shi（德克萨斯大学医学分部）实验室报告了建设和 ZIKV 复制子细胞系 (ZIKV Rep-neo) 的验证 在初步研究中，我们使用了 ZIKV Rep-neo。 细胞系成功地对 FDA 批准的化合物进行了小规模筛选，证明了 在第一阶段，我们将应用 ZIKV 复制子检测来筛选 ≥350,000 个文库。 用于抑制复制的小分子化合物，并将使用计数器屏幕和 PAINS 过滤器来 我们将根据结果优先考虑非特异性抑制剂和细胞毒性化合物。 评估针对传染性 ZIKV 的效力、针对 ZIKV 的活性谱的二次测定组 黄病毒、细胞毒性和这些化合物的类药特性的作用机制。 将研究优先化合物，以验证优先命中目标病毒蛋白，并初步确定 将评估符合严格列出标准的抑制剂。 每个具体目标的里程碑将在后续的第二阶段项目中进行优化。 在该项目的第一阶段，我们将实现以下具体目标 1. 识别有效的抑制剂。 高通量筛选中的 ZIKV 复制子 目标 2. 根据效力、特异性、优先级确定已确认的命中。 目标 3. 确定优先抑制剂的作用机制。 目标 4. 建立。 优先抑制剂的初步结构活性关系。