Broad spectrum inhibitors of paramyxovirus envelope proteins

摘要 人副流感病毒（HPIV）引起儿童哮吼、细支气管炎和 肺炎在美国和世界各地，但没有有效的药物或疫苗。致命的尼帕病毒 (NiV)和亨德拉（HeV）病毒，面对NiV的人际传播，全球存在风险 传播以及意外或故意暴露。尽管存在用于被动免疫的抗体制剂， 尽管免疫预防是有效的，但没有可行的方法来预防或治疗人类疾病。这些病毒 作为副粘病毒的原型。有了这个应用程序，我们建立在最近的突破， 导致了一种新的广谱抗病毒策略，该策略基于在病毒进入期间抑制融合。我们将联合收割机 融合抑制脂肽的基于结构的优化，通过部分取代 α-氨基酸残基与非天然β-氨基酸残基，以提高半衰期，并添加脂质 成分以增强抗病毒功效。使用病毒学，体外和体内实验，我们将开发 可以通过吸入给药的强效新型HPIV 3/NiV/HeV融合抑制剂。验证我们的 循环呼吸道病毒和临床NiV病毒株的体外和体内策略将使我们 在临床环境中有效和实用的抗病毒剂。 目标1。靶向副粘病毒融合复合物：体外评价进入抑制剂方法。 我们将开发融合抑制脂肽，结合传统的肽设计与骨架- 修饰和膜靶向，以优化对NiV、HeV、HPIV的广泛融合抑制活性， 优化先导脂肽的效力、蛋白酶抗性和组织靶向。我们将评估抗病毒药物 在真实的离体模型中的抗病毒活性和抗病毒抗性的潜力。人类气道和大脑模型将 在离体进化实验中用于引发抗性病毒，以研究 抗病毒活性和对先导肽的抗性潜力。 目标2.药物动力学、生物分布和体内抗病毒制剂的效力。 我们将通过鼻或吸入评估生物分布和免疫原性、急性毒性和PK/PD 在小动物模型中递送先导候选物系列，并评估先导肽对NiV/HeV的功效， 金黄仓鼠和棉鼠中的HPIV。将在非洲开展NiV电极导线的有效性研究 绿色猴子。 我们认为，肽骨架修饰和膜靶向策略将产生广泛的抗- 副粘病毒剂，其可以通过吸入递送，具有前所未有的功效和药代动力学 特性.

Abstract Human parainfluenza viruses (HPIV) cause a significant portion of childhood croup, bronchiolitis and pneumonia in the U.S. and worldwide, yet no effective drugs or vaccines are available. For the lethal Nipah (NiV) and Hendra (HeV) viruses, in the face of human-to-human transmission of NiV, there is risk for global spread as well as accidental or deliberate exposure. Despite the existence of antibody preparations for passive immunoprophylaxis, there is no feasible approach to prevent or treat the human disease. These viruses serve as prototypes for emerging paramyxoviruses. With this application we build on recent breakthroughs that have led to a new broad-spectrum antiviral strategy based on inhibiting fusion during viral entry. We combine structure-based optimization of fusion inhibitory lipopeptides, backbone modification via partial replacement of α-amino acid residues with unnatural β-amino acid residues to enhance half-life, and addition of lipid components to enhance antiviral efficacy. Using virologic, ex vivo, and in vivo experiments we will develop powerful novel HPIV3/NiV/HeV fusion inhibitors that can be administered by inhalation. Validation of our strategies with circulating respiratory viruses and clinical NiV virus strains in vitro and in vivo will lead us to antiviral agents that are effective and practical in the clinical setting. Aim 1. Targeting the paramyxovirus fusion complex: Evaluate entry inhibitor approaches in vitro. We will develop fusion-inhibitory lipopeptides, combining conventional peptide design with backbone- modification and membrane targeting to optimize broad fusion inhibition activity for NiV, HeV, HPIV, and optimize potency, protease resistance, and tissue targeting of lead lipopeptides. We will evaluate antiviral activity and potential for antiviral resistance in authentic ex vivo models. Human airway and brain models will be employed to elicit resistant viruses in ex vivo evolution experiments, to study the molecular bases for antiviral activity and potential for resistance to lead peptides. Aim 2. Pharmacokinetics, biodistribution and efficacy of formulated lead antivirals in vivo. We will assess the biodistribution and immunogenicity, acute toxicity, and PK/PD via nasal or inhalation delivery of lead candidate series in small animal models, and evaluate efficacy of lead peptides for NiV/HeV in golden hamsters and for HPIV in cotton rats. An efficacy study for the NiV lead will be conducted in African green monkeys. We propose that peptide backbone modification and membrane targeting strategies will generate broad anti- paramyxovirus agents that can be delivered via inhalation with unprecedented efficacy and pharmacokinetic properties.