Project 2 - Novel Therapeutics for Emerging Alphavirus

项目摘要 蚊子传播的甲病毒引起急性和慢性关节炎的爆发， 威胁脑炎致关节炎甲病毒包括重新出现的基孔肯雅病毒（CHIKV）， Mayaro和Ross River病毒。自2004年以来，CHIKV已经感染了数百万人，并扩大到 欧洲、亚洲和美洲。由于关节炎可以持续数月至数年后，感染关节炎原 甲型病毒，大规模流行具有严重的经济后果。脑炎甲病毒包括 美洲特有的东部、委内瑞拉（VEEV）和西部马脑炎病毒， 感染可导致死亡或长期神经后遗症。目前还没有批准的甲病毒- 特定疫苗或抗病毒剂。与南方研究合作，我们使用HTS来识别非 抑制CHIKV和VEEV复制的毒性小分子。此外，我们还鉴定了 显示出对甲病毒以及其他病原性病毒的广泛抑制活性的化合物类别， 人病毒（例如，黄病毒）。我们还绘制了针对两种高活性化合物的抗性突变体 并鉴定了甲病毒nsP 2解旋酶结构域和nsP 3宏结构域作为潜在的抗病毒靶标。在 除了这项工作外，我们还与埃默里药物开发研究所合作， 靶向RNA依赖性RNA聚合酶的核苷，对甲病毒具有有效的抗病毒活性 感染因此，我们已经确定了针对三种不同分子靶标的有效抗病毒化合物， for alphavirus甲病毒replication复制.这个高度互动的抗病毒药物发现项目2的目标， 开发中心（AD 3C）是开发新的治疗策略，抑制甲病毒复制，预防 选择耐药性，并降低甲病毒疾病的严重程度。在具体目标1中，先导化合物将 与核心A和核心B合作，通过迭代药物化学、基于细胞培养 抗病毒和细胞毒性测定，以及体内药代动力学研究，以提高它们的功效，选择性， 溶解度和生物利用度。在具体目标2中，我们将与其他AD 3C项目合作， 抗病毒活性和细胞类型特异性，通过耐药图谱的分子靶标， 结构/计算分析和鉴定具有有效活性的化合物的协同作用概况。在特定 目的3，将使用完善的临床前小鼠模型评估优化的化合物的体内功效。 甲病毒感染的模型。由于针对病毒生命周期不同方面的治疗可能会 显示出改善的疗效并限制耐药性的发展，联合疗法也将进行测试。 最后，由于甲病毒诱导的疾病包括炎性免疫病理学，我们将评估甲病毒诱导的疾病的免疫病理学。 联合抗病毒治疗与抗炎治疗方案的治疗潜力。这项工作将促进 开发具有改善人类健康潜力的新的抗病毒治疗策略。

PROJECT SUMMARY Mosquito-transmitted alphaviruses cause outbreaks of incapacitating acute and chronic arthritis and life- threatening encephalitis. The arthritogenic alphaviruses include the re-emerging chikungunya (CHIKV), Mayaro, and Ross River viruses. Since 2004, CHIKV has infected millions of people and expanded into Europe, Asia, and Americas. As arthritis can endure for months to years after infection with an arthritogenic alphavirus, large epidemics have severe economic consequences. The encephalitic alphaviruses include Eastern, Venezuelan (VEEV), and Western equine encephalitis viruses that are endemic to the Americas and infection can lead to mortality or long-term neurological sequelae. There are currently no approved alphavirus- specific vaccines or antiviral agents. In collaboration with Southern Research, we used HTS to identify non- toxic small molecules that inhibit CHIKV and VEEV replication. Additionally, we identified chemically distinct compound classes that display broad inhibitory activity against alphaviruses, as well as other pathogenic human viruses (e.g., flaviviruses). We also mapped resistance mutants against two highly active compounds and identified the alphavirus nsP2 helicase domain and the nsP3 macrodomain as potential antiviral targets. In addition to this work, in collaboration with the Emory Institute for Drug Development, we identified novel nucleosides, which target RNA-dependent RNA polymerases, with potent antiviral activity against alphavirus infection. Thus, we have identified potent antiviral compounds against three distinct molecular targets essential for alphavirus replication. The goal of this highly interactive Project 2 of the Antiviral Drug Discovery and Development Center (AD3C) is to develop new therapeutic strategies that inhibit alphavirus replication, prevent selection for drug resistance, and reduce alphavirus disease severity. In Specific Aim 1, lead compounds will be optimized in collaboration with Core A and Core B by iterative medicinal chemistry, cell culture-based antiviral and cytotoxicity assays, and in vivo pharmacokinetic studies to improve their efficacy, selectivity, solubility, and bioavailability. In Specific Aim 2, we will work with the other AD3C Projects to define the breadth of antiviral activity and cell type specificity, molecular targets through resistance mapping and structural/computational analyses and identify synergy profiles for compounds with potent activity. In Specific Aim 3, optimized compounds will be evaluated for in vivo efficacy using well-established pre-clinical mouse models of alphavirus infection. Since therapies that target different aspects of the viral life cycle are likely to show improved efficacy and limit the development of drug resistance, combination therapies also will be tested. Last of all, since alphavirus-induced disease includes inflammatory immune pathology, we will evaluate the therapeutic potential of combining antiviral therapy with anti-inflammatory regimens. This work will promote the development of new antiviral therapeutic strategies that have the potential to improve human health.