In Vitro Evaluation and Characterization of Novel Lead Therapeutic Candidates for Encephalitic Alphavirus Treatment

计划摘要-项目3 脑炎甲型病毒(即委内瑞拉、西部和东部马脑炎病毒)可引起 人类和马类的破坏性脑炎。已经进行了几项药物发现努力，以实现 对这些病毒的有效抗病毒药物的需求尚未得到满足；然而，它们仍然无法治疗。在这里我们 建议基于我们的两个新的先导开发治疗脑炎甲型病毒的候选药物 系列片。每个系列中的原型在体外和体内都显示出强大的抗病毒活性，没有不良反应。 在目标1中，我们将在体外评估新设计的化合物作为抗VEEV，EEEV的潜力 治疗公司支持研究项目1的领导优化，并为选择将成为 在研究项目2中进行了活体测试。在目标2中，我们将为以下方面的实验研究和文献工作做出贡献 为准备非正式和预科考试而选择的主要候选人的非临床病毒学研究报告 与Leidos合作召开IND会议。我们将通过实验证明，甲型病毒RNA复制酶是 主要候选化合物的靶标，采用体外病毒RNA合成试验。具体来说，我们将 评估抗V/W/EEEV在生物相关系统中的活性，重要的是测量抑制 商数来证明人类血液中的抗病毒活性。我们还将确定体外安全性和 先导候选化合物的治疗指数，并定义抗药性病毒的抗药性阈值。

Program Abstract- Project 3 Encephalitic alphaviruses (i.e., Venezuelan, Western, and Eastern Equine Encephalitis viruses) can cause devastating encephalitis in humans and equids. Several drug discovery efforts have been pursued to fulfill the unmet needs for effective antivirals for these viruses; however, they are still not available for treatment. Here we propose the development of a therapeutic candidate for encephalitic alphaviruses based on our two novel lead series. Prototypes within each series showed potent antiviral activity in vitro and in vivo without adverse effects. In Aim 1, we will evaluate newly designed compounds in vitro for their potentials as anti-VEEV, EEEV therapeutics to support the lead optimization of Research Project 1 and contribute to the selection of leads to be tested in vivo in Research Project 2. In Aim 2, we will contribute to experimental studies and documentation for the Non-Clinical Virology Study Report for the selected lead candidate in preparation for the informal and Pre- IND meetings in collaboration with Leidos. We will experimentally establish that the alphaviral RNA replicase is the target of the lead candidate compounds, using in vitro viral RNA synthesis assays. Specifically, we will evaluate the anti-V/W/EEEV activity in biologically relevant systems, and importantly, measure the Inhibitory Quotient to demonstrate antiviral activity in human blood. We will also determine the in vitro safety and therapeutic index of the lead candidate compounds and define the resistance threshold of resistant viruses.