The development of inhibitors for the infection using phage library

利用噬菌体库开发感染抑制剂

• 批准号: 14380411
• 负责人: SATO Toshinori
• 金额: $ 5.18万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14380411/
• 关键词: Phage Library; Peptide; Carbohydrate recognition; Influenza virus; Inhibitor; Hemagglutinin; Molecular evolution; Liposome; インフルエンザウィルス; 分子進化法; ファージライブラリー法; 糖脂質; 感染阻害剤

Influenza HA is known to bind sialylgalactoside having α2-3- or α2-6- linkages on host cells at the first step of infection. We attempted to design peptides as a universal inhibitor that bind to the receptor-binding pocket of HA.In order to obtain peptides which are a mimic of sialyl oligosaccharide, a phage-displayed random pentadecapeptide library have been employed. phage-displayed method is a selection technology using a pool of phage. It was expected that HA-binding peptides selected from the phage library would serve as inhibitors of influenza A virus. In this study, the HA-binding peptides and glycolipid-binding peptides were selected from the phage-displayed peptide libraries, and they inhibited the infection of both HA1 and HA3 subtypes of influenza virus to MDCK cells. Although the theoretical molecular diversity of pentadecapeptide is calculated to be 3.3x10^<19>, the phage library employed has only 2.5x10^8 kinds of diversity. Therefore, in order to obtain improved the HA-binding peptides, sublibrary generated from a selected sequence was prepared. The directed evolution approach was employed to prepare the sublibraries.The two kinds of sublibraries were constructed to obtain HA1- and HA3-binding peptides. Through the affinity selections for HA1 and HA3 using the sublibraries, several peptide sequences having mutations were obtained. Many of the mutant phage clones showed higher binding affinity for both HA1 and HA3 subtypes than the original A-1 phage. The evoluted peptides inhibited the infection of influenza virus to MDCK cells more efficiently than the original A-1 peptide.

众所周知，流感HA在感染的第一步结合宿主细胞上具有α2-3-或α2-6-链接的硫乳糖苷。我们试图将肽设计为一种通用抑制剂，该肽与HA的接收器结合袋结合，以获取肽的肽，这些肽是siAllyl寡糖的模拟物，已聘请了噬菌体斑点的随机五二肽文库。噬菌体播放方法是使用噬菌体池的选择技术。可以预期，从噬菌体文库中选择的HA结合宠物将用作影响力病毒的抑制剂。在这项研究中，从噬菌体播种的肽库中选择了HA结合的宠物和糖脂结合的宠物，它们抑制了影响MDCK细胞的HA1和HA3亚型的感染HA1和HA3亚型。尽管五肽的理论分子多样性计算为3.3x10^<19>，但噬菌体文库仅为2.5x10^8种多样性。因此，为了获得改善的HA结合肽，制备了从选定序列产生的sublibrary。进行了定向的进化方法以制备sublibraries。构建了两种sublibraries以获得HA1-和HA3结合肽。通过使用sublibraries对HA1和HA3的亲和力选择，获得了一些具有突变的肽序列。许多突变噬菌体克隆对HA1和HA3亚型的结合亲和力都比原始A-1噬菌体更高。与原始的A-1辣椒相比，进化的肽抑制了影响ZA病毒对MDCK细胞的影响。

项目成果

インフルエンザウイルス感染抑制剤

流感病毒感染抑制剂

• 发表时间: 2004

Inhibition of Influenza Virus Infection by Hemagglutinin-Binding Peptides

血凝素结合肽抑制流感病毒感染

• 发表时间: 2002
• 期刊: Peptide Science 2001
• 作者: T.Sato;M.Sumi;K.Ogino;T.Taki
• 通讯作者: T.Taki

ヘマグルチニン結合ペプチド

血凝素结合肽

• 发表时间: 2004

T.Sato et al.: "Inhibition of Influenza Virus Infection by Hemagglutinin-Binding Peputides"Peptide Science 2001. 329-330 (2002)

T.Sato 等人：“血凝素结合肽对流感病毒感染的抑制”肽科学 2001. 329-330 (2002)

佐藤 智典: "グライコーム"化学フロンティア. 171-179 (2002)

Tomonori Sato：“Glycome”化学前沿 171-179 (2002)