Measles infection blockage by intranasal delivery of F targeting peptides

描述(申请人提供)：麻疹病毒(MV)感染会导致一种急性儿童疾病，在某些情况下，可能会导致严重的神经后遗症和死亡。迫切需要针对麻疹的特定抗病毒治疗，以补充疫苗接种和加强全球控制麻疹的努力。这项研究计划的长期目标是开发一种安全高效的鼻腔融合蛋白(F)抑制剂，用于预防未接种疫苗的高危人群。在初步工作中，我们已经通过二聚化来自F的C-末端七重复区的多肽并将它们与胆固醇结合来产生有效的MV融合抑制物。我们已经证明，预防性鼻腔注射我们的铅F融合抑制剂可以有效地保护乳鼠在鼻腔感染MV后发生致死性脑炎。通过将融合抑制剂的结构优化和脂类成分的修饰与病毒学和体内试验相结合，我们的目标是鉴定和表征可经鼻给药并显著提高抗MV效果的新型MV F融合抑制剂。为了实现我们的目标，我们提出了两个目标：1.利用结构导向突变和蛋白质工程来鉴定和开发优化的F肽融合抑制剂。一种系统的突变、生物物理和结构方法将识别和结合抑制剂结合界面上的特定残基取代，以增加额外的结合能。我们将进行体外和体外研究，以评估工程F肽融合抑制剂的抗病毒活性。2.评价优化的融合抑制剂对小鼠MV感染的保护作用。我们将评估这些抑制剂的生物分布和毒性特性，并使用转基因小鼠模型来评估它们在体内的抗MV效力。在迭代过程中，实验结果将指导进一步的优化，产生一组有前景的抗MV药物。

DESCRIPTION (provided by applicant): Measles virus (MV) infection causes an acute childhood disease and, in some cases, can lead to severe neurological sequelae and death. Specific antiviral therapy for measles is urgently needed to complement vaccination and strengthen global efforts to control measles. The long-term objective of this research plan is to develop a safe and highly effective intranasal fusion protein (F) inhibitor as prophylaxis for use in high- risk unvaccinated populations. In preliminary work, we have generated potent MV fusion inhibitors by dimerizing peptides derived from the C-terminal heptad-repeat region of F and conjugating them to cholesterol. We have shown that prophylactic intranasal administration of our lead F fusion inhibitor efficiently protects suckling transgenic mice from developing fatal encephalitis after intranasal MV infection. By combining structure-based optimization of fusion inhibitors and modification of lipid components with virologic and in vivo assays, we aim to identify and characterize novel MV F fusion inhibitors that can be administered intranasally and have significantly improved anti-MV efficacy. To achieve our goals we propose two aims: 1. To use structure-guided mutagenesis and protein engineering to identify and develop optimized F peptide fusion inhibitors. A systematic mutational, biophysical and structural approach will identify and incorporate specific residue substitutions at the inhibitor-binding interface to impar additional binding energy. We will conduct in vitro and ex vivo studies to assess the antiviral activity of engineered F peptide fusion inhibitors. 2. To evaluate the protection afforded by optimized fusion inhibitors against MV infection in mice. We will evaluate the biodistribution and toxicity properties of the inhibitors, and use transgenic mouse models to assess their in vivo anti-MV potency. In an iterative process, the outcome of the experiments will guide further optimization, yielding a set of promising investigational anti-MV agents.