Development of antivirals for newly emerging pathogens Ebola and Marburg.

描述（由申请方提供）：埃博拉病毒（EBOV）和马尔堡病毒（MARV）是构成丝状病毒科的两种包膜病毒属。它们是最致命的人类病原体之一，引起严重的出血热，EBOV-Zaire的死亡率高达90%。没有疫苗或特定的抗病毒药物可供人类使用。我们建议将我们的基础研究成果应用于开发一种新的丝状病毒感染的抗病毒策略。该提议是基于我们最近的发现，即胆固醇基团与肽融合抑制剂的连接产生3个主要优点：（1）增加的效力，（2）肽与病毒在融合活化位点的定位，和（3）增强的药代动力学性质。通过将序列优化与胆固醇标记相结合，我们提出开发在体外抑制丝状病毒的高效肽融合抗病毒药物，并在丝状病毒感染的相关动物模型中测试其治疗潜力。这些融合/进入抑制肽将被设计成“跟随”病毒到与细胞膜融合的位点，从而克服了将肽抑制剂用于在细胞内区室中融合的病毒的主要障碍。我们建议开发丝状病毒融合抑制剂：目的1。设计并测试靶向质膜的丝状病毒融合抑制肽，使用（a）生物物理分析指导的序列优化以增强肽与靶序列的相互作用，和（B）膜靶向融合激活位点。目标二。评估融合抑制剂在致死性感染的鼠模型中保护免于EBOV感染的有效性。我们将确定（a）靶向肽的生物利用度，和（B）在激发实验中的功效。因此，我们将获得有效性的原理证明，从而使我们能够选择具有最大体内潜力的肽进行改进。 这一提议的总体高度影响在于将为融合在内体中的包膜病毒建立的概念验证。新的策略将克服使用肽类抗病毒药物治疗在细胞内融合的病毒的障碍，这些病毒包括orhomyxoviruses（流感）、flaviruses（西尼罗河病毒、登革热病毒）、coronaviruses（严重急性呼吸道综合征）、arenaviruses（朱宁、拉沙病毒），所有这些病毒都具有类似的融合机制。 公共卫生相关性：丝状病毒（即埃博拉病毒和马尔堡病毒）是对人类的真实的和明确的威胁，可以并且已经被武器化。我们将开发基于肽的丝状病毒融合/进入抑制剂，这些肽被设计成“跟随”病毒到其与细胞膜融合的位点。该建议旨在为开发治疗药物奠定基础，因此本身具有临床意义，因为迫切需要治疗出血热和相关疾病。该提案的总体高度影响在于将为在内体中融合的包膜病毒建立概念验证，包括orhomyxoviruses（流感病毒），黄病毒（西尼罗河病毒，登革热病毒），冠状病毒（严重急性呼吸道综合征），沙粒病毒（朱宁，拉沙病毒），所有病毒都共享类似的融合机制。

DESCRIPTION (provided by applicant): Ebola viruses (EBOV) and Marburg virus (MARV) are two genera of enveloped viruses that constitute the family of Filoviridae. They are among the most lethal human pathogens, causing a severe hemorrhagic fever with mortality rates up to 90% for EBOV-Zaire. No vaccine or specific antiviral is available for human use. We propose to apply the results of our fundamental research to the development of a new antiviral strategy for filovirus infections. The proposal is based on our recent discovery that attachment of a cholesterol group to a peptide fusion inhibitor yields 3 major advantages: (1) increased potency, (2) localization of the peptide with the virus at the site of fusion activation, and (3) enhanced pharmacokinetic properties. By combining sequence optimization with cholesterol tagging, we propose to develop highly effective peptide fusion antivirals that inhibit filoviruses in vitro, and to test their therapeutic potential in a relevant animal model of filovirus infection. These fusion/entry inhibitory peptides will be designed to "follow" the virus to the site of fusion with the cell membrane, thus overcoming the major obstacle to the use of peptide inhibitors for viruses that fuse in intracellular compartments. We propose to develop filovirus fusion inhibitors by: Aim 1. Design and test filovirus fusion inhibitory peptides that are targeted to the plasma membrane, using (a) biophysical analysis-guided sequence optimization to enhance peptide interaction with the target sequence, and (b) membrane targeting to the site of fusion activation. Aim 2. Assess the effectiveness of the fusion inhibitors at protecting against EBOV infection in a murine model of lethal infection. We will determine (a) bioavailability of the targeted peptides, and (b) efficacy in challenge experiments. We will thereby obtain proof of principle for efficacy, allowing us to select peptides for advancement that have the most in vivo potential. The overarching high impact of this proposal lies in the proof-of-concept that will be established for enveloped viruses that fuse in the endosome. The new strategy would overcome the barrier to use of peptide antivirals for viruses that fuse in intracellular compartments, including orhomyxoviruses (influenza), flaviruses (West Nile virus, Dengue virus), coronaviruses (severe acute respiratory syndrome), arenaviruses (Junin, Lassa), all sharing a similar fusion machinery. PUBLIC HEALTH RELEVANCE: Filoviruses (i.e. Ebola and Marburg virus) are a real and clear threat to humans that can be and have been weaponized. We will develop filovirus fusion/entry inhibitors based on peptides that are designed to "follow" the virus to its site of fusion with the cell membrane. This proposal aims to lay the groundwork for developing a therapeutic and is therefore of clinical relevance in itself, since therapy for hemorrhagic fevers and associated diseases is urgently needed. The overarching high impact of this proposal lies in the proof-of-concept that will be established for enveloped viruses that fuse in the endosome, including orhomyxoviruses (influenza), flaviruses (West Nile virus, Dengue virus), coronaviruses (severe acute respiratory syndrome), arenaviruses (Junin, Lassa), all sharing a similar fusion machinery.