Design of peptide entry inhibitors and delivery systems to target emerging henipa

针对新兴亨尼帕病的肽进入抑制剂和递送系统的设计

• 批准号: 7687086
• 负责人: Anne Moscona
• 金额: $ 77.81万
• 依托单位: WEILL MEDICAL COLL OF CORNELL UNIV
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-25 至 2009-09-27
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7687086
• 关键词: Acute; Address; Animal Model; Animals; Antiviral Agents; Basic Science; Binding; Biomedical Engineering; Biophysics; Bioterrorism; Cell Surface Receptors; Cell membrane; Central Nervous System Diseases; Chimeric Proteins; Clinical; Collaborations; Cultured Cells; DNA Sequence Rearrangement; Data; Development; Disease; Disease Outbreaks; Drug Formulations; Effectiveness; Engineering; Evolution; F-peptide; Family Felidae; Feedback; Felis catus; Ferrets; Health; Helix (Snails); Henipavirus; Human; Human Parainfluenza Virus 3; In Vitro; Infant; Infection; Infection prevention; Laboratories; Lead; Life; Light; Lung diseases; Medical; Membrane Fusion; Modeling; Molecular; Molecular Analysis; Molecular Conformation; Mutate; Nature; Parainfluenza; Paramyxovirus; Peptides; Process; Prophylactic treatment; Proteins; Publishing; Range; Resistance; Resources; Roentgen Rays; Structure; System; Testing; Transmembrane Domain; Validation; Variant; Viral; Viral Fusion Proteins; Virus; Virus Diseases; Work; base; biodefense; clinically relevant; design; ear helix; improved; in vivo; inhibitor/antagonist; member; mortality; multidisciplinary; nano; nanoparticle; particle; pathogen; peptide F; pressure; prevent; receptor binding; research study; virology

Paramyxoviruses cause important human illnesses that contribute significantly to global disease and mortality. Two zoonotic paramyxoviruses, Hendra (HeV) and Nipah (NiV), are of urgent concern due to their lethal and transmissible nature. HeV and NiV initiate infection by binding to cell surface receptors, and fuse directly with the cell membrane to enter. The receptor-binding molecule (G) triggers the viral fusion protein (F) to its active state, and conformational changes in F protein drive fusion. Molecular mimics of the heptad repeat (HR) regions of HeV F can prevent F from reaching fusion-ready conformation, and prevent infection. We found that a heterologous (parainfluenza 3) peptide is more effective than the homologous peptide as an anti-HeV/NiV. We propose a distinctive combination of: (1) experimental and structural analysis of the molecular mechanisms of fusion and entry inhibition, to design optimal inhibitors; (2) animal model studies to test the proposed antivirals for protection from infection; (3) bioengineering approaches to improve delivery systems for promising antivirals. A multidisciplinary collaborative team, bringing unique expertise, synergizes to study: 1. Conformational changes in HeV and NiV F-protein: Basic research to design effective peptides. 1.1 Structural analysis of the mechanism of fusion inhibition by homotypic and heterotypic HRC peptides. By combining experimental information with analysis of our X-ray crystal structures of the 6HBs of HPIV3 F, HeV F and HPIV3/HeV chimeric 6HBs, the mechanism of action of inhibitory peptides will be explored structurally and biophysically. 1.2 Design and testing of improved peptides based on crystal structure data. 1.3 Analysis of HPIV3 HRC fusion inhibition-resistant variants. 2. Effectiveness of the peptides to protect from live viral infection in vivo. Effective peptides (aim 1) will be tested for their ability to protect against infection with HeV and NiV infection in the ferret and cat models of acute HeV/NiV infection. Treatment as well as pre- and post-exposure prophylaxis will be addressed. Micro/nanoparticles will be engineered to provide sustained delivery of the most promising inhibitors, to explore the hypothesis that sustained release can improve antiviral efficacy in vivo and provide a clinical strategy that would be feasible for crisis situations. Feedback from aim 2 will lead directly to new experiments in aim 1. The results will lead to: (1) New understanding about the molecular mechanisms of virus fusion, entry, and mechanisms of action of peptide inhibitors; (2) sustained delivery systems for antivirals that may be broadly applicable and clinically relevant; (3) validation of an antiviral strategy in vivo, and identification of realistic candidate antiviral peptides. The results will be significant in light of the importance of paramyxoviruses to human health and the potential broad applicability of the new platforms, in addition to the specific clinical/biodefense relevance of these emerging zoonotic pathogens.

帕托病毒会导致重要的人类疾病，这对全球疾病和死亡率产生了重大贡献。 两个人畜共患性帕托病毒，亨德拉（HEV）和Nipah（NIV），由于其致命而引起了关注 和可传播的本性。 HEV和NIV通过与细胞表面受体结合而引发感染，并直接融合 随着细胞膜进入。受体结合分子（G）触发病毒融合蛋白（F） 活性状态和F蛋白驱动融合的构象变化。分子模拟物的重复（HR） HEV F区域可以防止F达到融合准备构象并防止感染。我们发现 异源（Parainfluenza 3）肽比同源肽作为抗HEV/NIV更有效。 我们提出了一个独特的组合：（1）分子的实验和结构分析 融合和进入抑制的机制，设计最佳抑制剂； （2）测试动物模型研究 拟议的抗病毒药以防止感染； （3）生物工程的方法，以改善 有希望的抗病毒药。一个多学科的合作团队，带来了独特的专业知识，协同研究： 1。HEV和NIV F蛋白的构象变化：设计有效肽的基础研究。 1.1同型和异型HRC融合抑制机制的结构分析 肽。通过将实验信息与对6HB的X射线晶体结构的分析相结合 HPIV3 F，HEV F和HPIV3/HEV嵌合6HB，抑制性肽的作用机理将是 在结构和生物物理上探索。 1.2基于晶体结构数据的设计和测试改进了肽。 1.3 HPIV3 HRC融合抑制抗性变体的分析。 2。肽在体内免受活病毒感染的有效性。有效的肽（AIM 1）将 在雪貂和CAT模型中预防HEV和NIV感染的感染能力测试 急性HEV/NIV感染。将解决治疗以及暴露前后的预防。 微/纳米颗粒将经过设计，以提供最有前途的抑制剂的持续交付 持续释放可以提高体内抗病毒功效的假设，并提供了一种临床策略， 对于危机情况是可行的。 AIM 2的反馈将直接导致AIM 1中的新实验。 结果将导致：（1）对病毒融合，进入和的分子机制的新了解 肽抑制剂的作用机理； （2）抗病毒药的持续输送系统可能广泛 适用和临床相关； （3）验证体内抗病毒策略，并识别现实 候选抗病毒肽。鉴于帕托伏病毒的重要性，结果将是重要的 除了具体的 这些新兴的人畜共患病原体的临床/生物浓度相关性。