Marine Microbial Products as Novel Agents Against Neurotropic Arboviruses

描述（由申请人提供）：蚊媒病毒或虫媒病毒是全球最重要的新兴病原体之一，对人类健康具有重大潜在影响，也是NIAID A、B和C类优先病原体列表的主要组成部分。将许多虫媒病毒列为高优先级病原体的部分原因是它们的毒力、媒介介导传播的可能性以及公众对虫媒病毒感染的关注。此外，尽管虫媒病毒在世界范围内产生影响，但目前几乎没有有效的治疗方法。本提案的目的是利用西方马脑炎病毒（WEEV），一种B类虫媒病毒，鉴定和开发新的抗病毒化合物，从海洋沉积物衍生的微生物产生的天然产物提取物分离。来源于陆地和海洋微生物的化合物在历史上为针对一系列微生物的新型治疗剂提供了丰富的来源，但迄今为止在抗病毒剂的开发中未得到充分利用。我们已经开发、验证并采用了基于WEEV复制子的测定来完成针对来自多种海洋微生物物种的> 16，000种预分级提取物的文库的高通量筛选。此外，我们已经完成了两个不同的提取物与验证的抗病毒活性的分析和制备的初步色谱分离程序。本提案的具体目的是完成从这两种候选提取物中分离和功能表征活性化合物，包括结构检查和初始病毒靶标鉴定，并扩大具有抗虫媒病毒活性的微生物源天然产物的鉴定。本研究项目的长期目标是开发多种海洋微生物抗病毒化合物，表征其结构和抗病毒活性，并完成其在虫媒病毒介导疾病动物模型中的临床前试验，最终目标是这些候选新药的临床实施。本探索性项目提案中概述的实验将用于为实现这些目标奠定基础。

DESCRIPTION (provided by applicant): Mosquito-borne viruses, or arboviruses, are among the most important emerging pathogens worldwide with a significant potential impact on human health, and are also prominent components of the NIAID Category A, B, and C priority pathogens lists. The inclusion of many arboviruses as high priority pathogens is due in part to their virulence, the potential for vector-mediated dissemination, and the public concern regarding insect-borne viral infections. In addition, despite the worldwide impact of arboviruses, few effective treatments are currently available. The objective of this proposal is to use western equine encephalitis virus (WEEV), a category B arbovirus, to identify and develop novel antiviral compounds isolated from natural product extracts produced by marine sediment-derived microbes. Compounds derived from terrestrial and marine microorganisms have historically provided a rich source for novel therapeutics against a range of microorganisms, but have thus far been underutilized in the development of antiviral agents. We have already developed, validated, and employed a WEEV replicon-based assay to complete high-throughput screens against a library of >16,000 pre- fractionated extracts derived from a diverse array of marine microbial species. Furthermore, we have completed both analytical and preparative preliminary chromatographic fractionation procedures for two distinct extracts with validated antiviral activity. The specific aims of this proposal are designed to complete the isolation and functional characterization of active compounds from these two candidate extracts, including structural examination and initial viral target identification, and to expand the identification of microbe-derived natural products with anti-arboviral activity. The long term goals of this research project are to develop a wide range of antiviral compounds derived from marine microbes, characterize their structures and antiviral activities, and complete their preclinical testing in animal models of arbovirus-mediated disease, with the ultimate objective being clinical implementation of these candidate novel drugs. The experiments outlined in this exploratory project proposal will be used to establish the groundwork to accomplish these goals.