Antivirals Targeting Flavivirus Envelope Proteins

描述(由申请人提供)：该项目的总体目标是开发治疗危及生命的黄病毒感染的治疗性抗病毒药物。恩福韦肽是一种临床上成功的多肽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，我们筛选了与重组西尼罗河病毒(WN)病毒包膜蛋白结合的蛋白质和多肽的噬菌体展示文库。从包膜蛋白结构中筛选出与包膜蛋白有亲和力的WN病毒多肽。这些研究确定了6个专有多肽，它们与重组WN病毒囊膜蛋白结合，并抑制WN病毒对培养细胞的感染。在这个项目中，我们将识别更多阻止WN和登革热病毒感染的多肽。选定的多肽将被改造，以产生针对多种黄病毒的广谱多肽。该项目还将启动抗病毒小分子化合物库的高通量筛选，这些小分子与中和肽竞争与包膜蛋白结合。将评估候选抗病毒化合物中和黄病毒，包括4种登革热病毒血清型，以及保护小鼠免受登革热和WN病毒感染的能力。这一实验方法将在第二阶段扩大，以开发一组可作为人类治疗药物开发的修饰多肽和小分子。该项目将为临床医生和公共卫生官员提供管理自然、意外和故意爆发的黄病毒的新手段。

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.