Mechanism of RVFV nucleocapsid targeting drugs

描述(申请人提供)：裂谷热病毒(RVFV)是一种由蚊子传播的新出现的传染病，可能对人类和农业动物的健康造成重大影响。它是美国疾病控制与预防中心/美国农业部A类选择剂，已被世界动物卫生组织认定为具有全球传播潜力的高后果病原体。目前还没有经过验证的针对RVFV感染的治疗方法，也没有已被证明安全有效的许可疫苗。我们最近在一个新的分子靶标高通量筛选中发现了78个化合物，它们干扰RVFV核衣壳蛋白N和靶RNA的相互作用。这些化合物来自现有的药物和类药物分子库，其中几个化合物在细胞培养中显示出明显的抑制RVFV复制的作用。我们建议确定几个最有希望的化合物的分子作用机制。我们将使用生化、生物物理和遗传学方法来确定这些化合物在N上的结合位点(S)，并确定它们是抑制特定的核糖核酸结合活性，还是抑制通用的核糖核酸结合活性 具体地说，我们将使用RNA和药物与野生型和突变型核衣壳蛋白的结合分析来确定抑制化合物是否直接干扰RNA结合，干扰核壳蛋白齐聚，或者具有不同于RNA-蛋白质或蛋白质-蛋白质界面的靶点。这项研究扩大了我们对病毒生长和抑制的理解，并在对抗RVFV感染的新药疗法方面取得了进展，有望为生物医学和农业科学领域带来好处。

DESCRIPTION (provided by applicant): Rift Valley fever virus (RVFV) is a mosquito-borne emerging infectious agent with potential for major impacts on human and agricultural animal health. It is a CDC/USDA Category A Select Agent, and has been recognized by the World Organization for Animal Health as a high consequence pathogen with potential for global spread. There are no proven treatments for RVFV infections and there is no licensed vaccine that has been proven safe and effective. We recently identified 78 compounds in a novel molecular target high throughput screen that interfere with the interaction of RVFV nucleocapsid protein, N, and a target RNA. These compounds come from existing drug and drug-like molecule libraries and several of the compounds show measurable inhibition of RVFV replication in cell culture. We propose to ascertain the molecular mechanism of action of several of the most promising compounds. We will use biochemical, biophysical and genetic approaches to determine the binding site(s) for the compounds on N and we will determine whether they inhibit the specific RNA binding activity or the generic RNA binding activity that are both intrinsic to N. Specifically, we will determine whether the inhibitory compounds interfere directly with RNA binding, interfere with nucleocapsid protein oligomerization, or have a target that is distinct from either the RNA-protein or protein-protein interfaces using RNA and drug binding assays with wild-type and mutant nucleocapsid protein. The research promises benefits to the fields of biomedicine and agricultural science by expanding our understanding of virus growth and inhibition, and by making advances toward new drug therapies to combat RVFV infections.