Self-assembling nanoparticles for intranasal delivery of influenza fusion inhibitors

用于鼻内递送流感融合抑制剂的自组装纳米颗粒

• 批准号: 9441694
• 负责人: Matteo Porotto
• 金额: $ 64.49万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-10 至 2020-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9441694
• 关键词: Acute; Address; Affinity; Anti-influenza Agent; Antiviral Agents; Antiviral Therapy; Awareness; Binding; Biodistribution; Biological Assay; Biological Availability; Biomedical Engineering; C-terminal; Cell Culture Techniques; Cell membrane; Cells; Chronic; Clinical; Complement; Cotton Rats; Couples; Disease; Dominant-Negative Mutation; Endosomes; Engineering; Epidemic; Evaluation; Glutamine; Glycoproteins; Goals; Hemagglutinin; Human; In Vitro; Influenza; Influenza Hemagglutinin; Influenza prevention; Intranasal Administration; Investigation; Lead; Leucine; Life Cycle Stages; Lipids; Mediating; Membrane; Membrane Fusion; Modeling; Molecular Conformation; Mutagenesis; Neuraminidase inhibitor; Outcome; Peptides; Pharmaceutical Preparations; Population; Process; Property; Prophylactic treatment; Protein Engineering; Proteins; Public Health; Regimen; Research; Resistance; Scanning; Solubility; Structure; Surface; Testing; Toxic effect; Vaccination; Viral; Viral Drug Resistance; Virus Diseases; Work; airway epithelium; analog; anti-influenza; aqueous; biophysical analysis; biophysical properties; cytotoxicity; design; experimental study; falls; high risk; immunogenicity; improved; in vivo; influenzavirus; inhibitor/antagonist; nanomolar; nanoparticle; novel; pandemic disease; particle; peptide analog; prevent; protein aminoacid sequence; public health relevance; resistant strain; self assembly; trimer core; uptake; zanamivir

 DESCRIPTION (provided by applicant): New antiviral therapy for influenza is urgently needed to complement vaccination and existing drugs, and to strengthen global efforts to control epidemics and potential new pandemics. New anti-influenza treatments are critical in the face of emerging antiviral resistance to the existing drugs. The long-term objective of this research plan is to develop a safe and highly effective intranasal influenza hemagglutinin (HA)-derived inhibitor as prophylaxis for use in high-risk unvaccinated populations. In preliminary work, we have generated influenza fusion inhibitors by conjugating lipid to specific peptides derived from the C-terminal region of HA and adding cell penetrating peptide sequences for intracellular targeting. We showed that intranasal administration of our lead fusion inhibitor provides antiviral prophylaxis as efficient as the approved drug Relenza(c) in vivo. The fusion inhibitory peptides self-assemble into ~30- 50 nm nanoparticles and are internalized by the target cells. We plan to optimize the candidate antiviral peptides and assess them in vivo to lay the groundwork for human use. To do so, we propose to enhance: 1) antiviral potency; 2) peptide self-assembly; 3) target cell membrane insertion; 4) in vivo biodistribution. We aim to generate candidate peptides that, when administered intranasally, protect the human airway epithelium and prevent viral infection. These goals will be accomplished with two specific aims: 1. Use structure-guided mutagenesis and protein engineering to optimize the antiviral potency and bioavailability of influenza peptide fusion inhibitors. A systematic structural approach will be used to incorporate specific residue substitutions at the inhibitor binding interface that increase the binding energy f the inhibitor to its target. Using a combination of biophysical analysis and bioengineering we will optimize the inhibitors' features -- including particle stability and endosomal localization -- in order to lower the IC50 to nanomolar values. We will conduct in vitro and ex vivo studies to assess the antiviral activity of the engineered inhibitory peptides. 2. Evaluate the protection against influenza infection afforded by optimized self- assembling peptides in cotton rats. We will evaluate the bio-distribution and toxicity properties of the optimized nanoparticles, and assess their in vivo anti-influenza potency. In an iterative process, the outcome of the experiments will guide further optimization, yielding a set of promising investigational anti-influenza agents.

 描述（由申请人提供）：迫切需要新的流感抗病毒疗法来补充疫苗接种和现有药物，并加强全球控制流行病和潜在新大流行病的努力。面对现有药物出现的抗病毒耐药性，新的抗流感治疗至关重要。该研究计划的长期目标是开发一种安全高效的鼻内流感血凝素（HA）衍生抑制剂，作为高危未接种人群的预防药物。在前期工作中，我们通过将脂质与HA C末端区域衍生的特定肽缀合并添加用于细胞内靶向的细胞穿透肽序列来产生流感融合抑制剂。我们表明，鼻内施用我们的先导融合抑制剂可提供抗病毒作用 体内预防效果与已批准的药物 Relenza(c) 一样有效。融合抑制肽自组装成约 30-50 nm 纳米粒子，并被靶细胞内化。我们计划优化候选抗病毒肽并对其进行体内评估，为人类使用奠定基础。为此，我们建议增强：1）抗病毒效力； 2）肽自组装； 3) 靶细胞膜插入； 4)体内生物分布。我们的目标是产生候选肽，当鼻内给药时，可以保护人类呼吸道上皮并预防​​病毒感染。这些目标将通过两个具体目标来实现： 1. 使用结构引导诱变和蛋白质工程来优化流感肽融合抑制剂的抗病毒效力和生物利用度。将使用系统的结构方法在抑制剂结合界面处纳入特定的残基取代，以增加抑制剂与其靶标的结合能。结合生物物理分析和生物工程，我们将 优化抑制剂的特性——包括颗粒稳定性和内体定位——以将IC50降低至纳摩尔值。我们将进行体外和离体研究，以评估工程抑制肽的抗病毒活性。 2. 评估优化的自组装肽对棉鼠的流感感染保护作用。我们将评估优化纳米粒子的生物分布和毒性特性，并评估其体内抗流感效力。在迭代过程中，实验结果将指导进一步优化，产生一组有前途的研究性抗流感药物。