Antivirals Targeting Flavivirus Envelope Proteins

描述（由申请人提供）：本项目的总体目标是开发治疗性抗病毒药，以治疗危及生命的黄病毒感染。恩夫韦肽是一种临床上成功的肽HIV融合抑制剂，是一类新型抗病毒药物的原型，可抑制病毒进入宿主细胞所必需的病毒包膜蛋白结构重排。在初步实验中，我们筛选了噬菌体展示库的蛋白质和肽结合重组西尼罗（WN）病毒包膜蛋白。从病毒囊膜蛋白的结构中合理地筛选出与囊膜蛋白具有亲和性的WN病毒肽段。这些研究鉴定了6种结合重组WN病毒包膜蛋白并抑制WN病毒感染培养细胞的专有肽。在这个项目中，我们将确定其他肽阻断WN和登革病毒感染。选择的肽将被修饰以产生靶向多种黄病毒的广谱肽。该项目还将启动化合物库的高通量筛选，以寻找与中和肽竞争结合包膜蛋白的抗病毒小分子。将评价候选抗病毒化合物对黄病毒（包括登革热病毒的4种血清型）的中和作用以及保护小鼠免受登革热和WN病毒感染的能力。这种实验方法将在第二阶段扩展，以开发一组修饰的肽和小分子，这些肽和小分子是开发人类治疗药物的候选者。该项目将为临床医生和公共卫生官员提供新的手段来管理自然，意外和故意的黄病毒爆发。

DESCRIPTION (provided by applicant): The overall goal of this project is to develop therapeutic antivirals to treat life-threatening flavivirus infections. Enfuvirtide, a clinically successful peptide HIV fusion inhibitor, is the prototype for a new class of antivirals that inhibit viral envelope protein structural rearrangements essential for viral entry into host cells. In preliminary experiments, we screened a phage display library for proteins and peptides that bind recombinant West Nile (WN) virus envelope protein. WN virus peptides with affinity for envelope protein were also rationally selected from the envelope protein structure. These studies identified 6 proprietary peptides that bind recombinant WN virus envelope protein and inhibit WN virus infection of cultured cells. In this project, we will identify additional peptides that block WN and dengue virus infections. Selected peptides will be modified to produce broad-spectrum peptides that target multiple flaviviruses. The project will also initiate high throughput screening of chemical compound libraries for antiviral small molecules that compete with the neutralizing peptides for binding to envelope protein. Candidate antiviral compounds will be evaluated for neutralization of flaviviruses, including the 4 serotypes of dengue virus, and for ability to protect mice against dengue and WN virus infections. This experimental approach will be expanded in Phase II to develop a panel of modified peptides and small molecules that are candidates for development as human therapeutics. This project will provide clinicians and public health officials with new means to manage natural, accidental and intentional flavivirus outbreaks.