Project 4 - Inhibitors of Flavivirus Replication

黄病毒属的新成员和地方性成员构成了重大的全球公共卫生威胁。的 登革热病毒、日本脑炎病毒、西尼罗河病毒（WNV）、蜱传脑炎的全球范围 病毒和ZIKV最近已经扩大。据估计，登革热病毒感染约4亿人， 100万，每年杀死21,000人。黄热病病毒仍然是34个非洲国家的公共卫生问题 13个南美国家新出现的黄病毒包括Powassan病毒（POWV）（美国东北部 和加拿大）和Alpentu病毒（欧洲）。人类只有一些黄病毒的疫苗，但它们 并不总是使用。少数个体在接种活的17D黄色疫苗后发生多器官疾病 典型的致命性发热疫苗。目前还没有有效的黄病毒特异性小分子 用于治疗人黄病毒感染的治疗剂。项目4提出的研究目标是 AC/DC将开发可口服、具有广泛抗黄病毒作用的临床候选化合物 活性并直接作用于病毒成分。在目标1下，我们将开发四种核糖核苷类似物 由Core合成 B，已经获得了表明抗黄病毒活性的初步数据， 采用Cores B和C的重复模拟合成-生物学测试策略。将对化合物进行检测， 对一组黄病毒的抗病毒活性和在原代小鼠和人巨噬细胞中的细胞毒性， 神经细胞还将分析作用模式和耐药性的发展。在目标2下，报告人 将生成POWV和WNV病毒，并用于高通量筛选约300，000个独特的 在BSL 3控制下的化合物。将进一步开发经反筛选验证的命中率 在目标1下。将测试满足进展标准的化合物在黄病毒小鼠中的体内功效。 和仓鼠感染模型以及人类皮质类器官中。肠道微生物组对活动的影响 将与项目7合作评估黄病毒抗病毒化合物。抗病毒药物的影响 治疗POWV和WNV感染动物的急性和慢性病毒诱导的神经特征， 将与项目2合作评估类器官。核心D将提供病毒的全基因组测序 在先导化合物的病毒耐药性分析期间产生的突变体。

Emerging and endemic members of the genus Flavivirus pose significant worldwide public health threats. The global ranges of dengue viruses, Japanese encephalitis virus, West Nile virus (WNV), tick-borne encephalitis virus and ZIKV have recently expanded. Dengue virus is estimated to infect ~400 million, cause illness in ~100 million, and kill ~21,000 people a year. Yellow fever virus remains a public health concern in 34 African countries and 13 South American countries. Newly emerging flaviviruses include Powassan virus (POWV) (northeast US and Canada) and Usutu virus (Europe). Vaccines are available for humans for only some flaviviruses, but they are not always used. A few individuals develop multiorgan disease after vaccination with the live 17D yellow fever vaccine which is typically fatal. There are currently no effective flavivirus-specific small molecule therapeutics for treatment of humans with flavivirus infections. The goal of the research proposed in Project 4 of the AC/DC is to develop clinical candidate compounds that can be delivered orally, have broad anti-flavivirus activity and directly act on a viral component. Under Aim 1, we will develop four ribonucleoside analogs synthesized by Core B, for which preliminary data indicating anti-flavivirus activity has already been obtained, by a reiterative analog synthesis-biological testing strategy with Cores B and C. Compounds will be tested for antiviral activity against a panel of flaviviruses and cytotoxicity in primary mouse and human macrophages and neural cells. Mode of action and development of drug resistance will also be analyzed. Under Aim 2, reporter POWV and WNV viruses will be generated and used in high throughput screening of ~300,000 unique compounds under BSL3 containment by Core F. Hits validated by counter-screening will be further developed under Aim 1. Compounds meeting the advancement criteria will be tested for in vivo efficacy in flavivirus mouse and hamster infection models and in human cortical organoids. The effect of the gut microbiome on the activity of lead flavivirus antiviral compounds will be assessed in collaboration with Project 7. The impact of antiviral treatment on acute and chronic virus-induced neurological signatures in POWV and WNV infected animals and organoids will be evaluated in collaboration with Project 2. Core D will provide whole genome sequencing of viral mutants generated during viral resistance profiling of lead compounds.