Development and optimization of novel anti -flavivirus compounds

新型抗黄病毒化合物的开发和优化

• 批准号: 8465809
• 负责人: Brian Geiss
• 金额: $ 27.78万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8465809
• 关键词: Affinity; Amino Acids; Antiviral Agents; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Biological Availability; Categories; Cell Culture Techniques; Characteristics; Collaborations; Computational Biology; Computer Simulation; Culicidae; Dengue; Dengue Virus; Development; Disease; Docking; Drug Design; Drug Targeting; Enzymes; Evaluation; Exhibits; Family; Flavivirus; Flavivirus Infections; Foundations; Goals; Growth; Guanosine; Guanosine Triphosphate; Human; In Vitro; Infection; Knowledge; Laboratories; Lead; Libraries; Life; Medical; Methodology; Methyltransferase; Modeling; Molecular Models; Morbidity - disease rate; Mus; Mutation; National Institute of Allergy and Infectious Disease; Pathogenesis; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Probability; Process; Property; Protein Binding; Protein Methyltransferases; RNA; RNA Caps; Replicon; Research; Research Personnel; Research Project Grants; Resistance; Resolution; Resources; Silicon Dioxide; Structure; Structure-Activity Relationship; Techniques; Testing; Therapeutic; Therapeutic Agents; Translations; Validation; Viral; Viral Genome; Viral Hemorrhagic Fevers; Virus; West Nile virus; Yellow Fever; Yellow fever virus; analog; base; biodefense; chemotherapeutic agent; cost effective; cytotoxicity; design; drug development; fluoromethyl 2,2-difluoro-1-(trifluoromethyl)vinyl ether; high throughput screening; human morbidity; human mortality; improved; inhibitor/antagonist; insight; molecular modeling; mortality; mouse model; novel; pathogen; pre-clinical; screening; small molecule; socioeconomics; structural biology; therapy development; viral RNA; virology

Infection by flaviviruses such as dengue, yellow fever, and West Nile is a major medical and socioeconomic problem worldwide, yet effective antiviral therapeutics to treat flavivirus infection are not currently available. As such, it is imperative that potent, selective, and cost-effective antiviral compounds be identified. The overall goal of this project is the discovery of novel inhibitors of the flavivirus RNA methyltransferase (MTase) enzyme that can serve as effective broad-spectrum chemotherapeutic agents for the treatment of flavivirus infection. The MTase enzyme generates the cap structure at the 5' end of viral RNAs that is required for efficient translation of the viral genome and is essential for viral growth. We have designed and successfully implemented a simple and rapid in vitro high-throughput assay to identify compounds that interfere with RNA cap binding by the MTase. Our initial validation screen of molecule libraries at the National Screening Laboratory (NSRB) has identified a number of compounds as MTase inhibitors. In this project, we propose to expand on these results with the goal of identifying and optimizing a chemically diverse set of MTase cap-binding inhibitors in order to identify lead compounds for drug development. Specific Aim 1: We will perform additional HTS and will biochemically determine the inhibition constants and antiviral activity for additional hit compounds. Specific Aim 2: Using the resulting information, we will employ an integrated array of in silico molecular modeling techniques to identify structurally related small molecule compounds which based on our structural knowledge of the RNA cap binding site and in silico analysis will have improved affinity and cross selectivity for flavivirus MTase proteins and acceptable drug-like characteristics. We will also employ medicinal chemistry to design and synthesize derivatives when necessary to improve physiochemical properties. Specific Aim 3: The best inhibitors (in terms of breadth, potency and drug-like characteristics) will be tested for antiviral activity in cell culture and for the potential for emergence of resistance Inhibitors. Inhibitory effects for lead compounds will be determined against West Nile virus in an existing mouse model. This project takes advantage of an ongoing collaboration that brings together the expertise (virology and computational biology) of the lead investigators and will result in the rapid and efficient identification of inhibitors of flavivirus replication with the ultimate goal of describing lead compounds with drug-like properties suitable for preclinical development for the treatment flavivirus infection. This research Project fits within the RMRCE Integrated Research Focus on Viral Therapeutics, and will interact directly with RPs 3.1 and 3.8 and utilize the resources of Core C.

黄病毒感染，如登革热，黄热病和西尼罗河是一个重大的医疗和社会经济问题。 然而，治疗黄病毒感染的有效抗病毒疗法目前还没有 available.因此，迫切需要有效的、选择性的和成本有效的抗病毒化合物， 鉴定该项目的总体目标是发现新的黄病毒RNA抑制剂 甲基转移酶（MTase）酶，可作为有效的广谱化疗药物 用于治疗黄病毒感染的药剂。MTase酶在5'端产生帽结构， 病毒RNA的末端，是病毒基因组有效翻译所必需的，对病毒生长至关重要。 我们设计并成功实施了一种简单快速的体外高通量测定方法， 鉴定干扰MTase与RNA帽结合的化合物。我们的初始验证屏幕 国家筛选实验室（NSRB）的分子库已经确定了一些化合物， MTase抑制剂。在这个项目中，我们建议扩大这些结果的目标是确定和 优化MTase帽结合抑制剂的化学多样性组，以鉴定用于 药物开发具体目标1：我们将进行额外的HTS，并将通过生物化学方法确定 另外的命中化合物的抑制常数和抗病毒活性。具体目标2：使用结果 信息，我们将采用集成阵列的计算机分子建模技术，以确定 结构相关的小分子化合物，其基于我们对RNA帽的结构知识， 结合位点和计算机分析将提高黄病毒MTase蛋白的亲和力和交叉选择性 和可接受的药物样特征。我们还将利用药物化学来设计和合成 衍生物，以改善物理化学性质。具体目标3：最佳抑制剂（在 就宽度、效力和药物样特征而言）将在细胞培养物中测试抗病毒活性， 可能出现的耐药性抑制剂。铅化合物的抑制作用将是 在现有的小鼠模型中确定了针对西尼罗河病毒的作用。该项目利用了一个正在进行的 将主要研究人员的专业知识（病毒学和计算生物学）汇集在一起的合作 并将导致快速和有效地鉴定黄病毒复制的抑制剂， 描述了具有药物样性质的先导化合物，适用于临床前开发， 黄病毒感染本研究项目符合RMRCE病毒综合研究重点 治疗，并将直接与RP 3.1和3.8互动，并利用核心C的资源。