Identification and Characterization of Novel Flavivirus Antivirals

黄病毒是许多重要的公共卫生疾病的病原体，包括西 尼罗河脑炎、黄热病、登革热和日本脑炎。然而，目前的治疗方案 因此真实的需要新的抗病毒剂， 这些病毒。该项目的长期目标是通过确定和描述 具有抗医学上重要的F 1病毒的广谱活性的新的小分子化合物。的 我们提出的项目有三个具体目标。 具体目标1：新型先导化合物的高通量筛选鉴定。 使用我们开发的西尼罗河病毒（WNV）病毒样颗粒（VLP）检测方法， 为了在低生物防护条件下使用，我们将对一个大的文库进行高通量筛选， 在国家筛选实验室的区域卓越中心的化合物。数据 将对筛选期间产生的抗病毒化合物进行分析，并确定最有希望的抗病毒化合物。的 所选化合物对西尼罗河病毒的活性将使用活病毒测定来证实， 在用登革病毒和黄热病毒的活病毒测定中测定的谱抗黄病毒活性。 具体目标2：药物化学修饰和迭代体外抗病毒测试。 基于它们对所有三种黄病毒的活性、毒性特征和化学修饰的潜力， 三种化合物将被确定为先导化合物，并将在 UTMB的平行合成和药物化学设施每一轮化学修饰都会 涉及少量化合物的合成，然后这些化合物将经历快速的体外毒性， 活病毒检测中的抗病毒活性测试。这个过程将导致几轮受控迭代 合成和评估，我们预计将导致更有效的抗病毒化合物的鉴定。 具体目标3：在WNV脑炎小动物模型中的活性体内评价 在这一目标中，在化学改性研究中确定的两种最有前途的化合物将是 使用WNV脑炎的小鼠模型评估体内活性。 因此，到项目期结束时，我们预计已经确定了一些新的化合物， 对具有医学重要性的黄病毒的广谱活性，并已进行初步研究， 在明确定义的病毒学和病理学动物模型中评估它们的潜在治疗效用， 与临床情况相关的终点。

Flaviviruses are the causative agents of a number of diseases of public health importance including West Nile encephalitis, yellow fever, dengue fever and Japanese encephalitis. However, current treatment options for these diseases are limited and there is a real need for new antiviral agents with specific activity against these viruses. The long-term objectives of this project are to meet this need by identifying and characterizing new small molecule compounds with broad spectrum activity against medically important f1aviviruses. The project that we propose has three specific aims. Specific Aim 1: High Throughput Screening Identification of Novel Lead Compounds. Using a West Nile virus (WNV) virus-like particle (VLP) assay that we have developed and, which is suitable for use under low bio-containment conditions we will conduct high-throughput screening of a large library of compounds at the National Screening Laboratory for the Regional Centers of Excellence. The data generated during screening will be analyzed and the most promising antiviral compounds identified. The activity of the selected compounds against WNV will be confirmed using live virus assays and, their broad spectrum anti-flavivirus activity determined in live virus assays with dengue virus and yellow fever virus. Specific Aim 2: Medicinal Chemistry Modification and Iterative in vitro Antiviral Testing. Based on their activity against all three flaviviruses, toxicity profiles and potential for chemical modification, three compounds will be identified as leads and will undergo a series of medicinal chemical modifications in the Parallel Synthesis and Medicinal Chemistry facility at UTMB. Each round of chemical modifications will involve the synthesis of a small number of compounds which will then undergo rapid in vitro toxicity and antiviral activity testing in live virus assays. This process will result in several rounds of controlled iterative synthesis and evaluation that we expect will lead to the identification of more potent antiviral compounds. Specific Aim 3: In Vivo Evaluation of Activity in a Small Animal Model of WNV Encephalitis In this aim the two most promising compounds identified during the chemical modification studies will be evaluated for activity in vivo using a mouse model of WNV encephalitis. Thus by the end of the project period we expect to have identified a number of new compounds with broad spectrum activity against medically important flaviviruses and to have undertaken preliminary studies to evaluate their potential therapeutic utility in a well defined animal model with virologic and pathologic endpoints that are relevant to those seen in the clinical situation.