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

项目摘要-项目3 脑炎甲病毒（即，委内瑞拉、西部和东部马脑炎病毒）可引起 人类和马科动物的毁灭性脑炎。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未得到满足;然而，这些药物仍然无法用于治疗。这里我们 基于我们的两种新型先导化合物，提出开发脑炎甲病毒的治疗候选药物 系列.每个系列中的原型在体外和体内均显示出有效的抗病毒活性，且无不良反应。 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 治疗，以支持研究项目1的电极导线优化，并有助于选择电极导线 在研究项目2中进行了体内测试。在目标2中，我们将致力于以下方面的实验研究和文献记录： 选定的主要候选人的非临床病毒学研究报告，为非正式和预 与Leidos合作举行IND会议。我们将通过实验确定甲病毒RNA复制酶是 先导候选化合物的靶标，使用体外病毒RNA合成测定。具体来说，我们将 评价生物相关系统中的抗V/W/EEEV活性，重要的是，测量抑制性 在人血液中证明抗病毒活性的商。我们还将确定体外安全性， 确定先导候选化合物的治疗指数，并确定耐药病毒的耐药阈值。

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.