DEVELOPMENT OF RNA ANALOG INHIBITORS AGAINST VERO TOXIN PRODUCED BY E. COLI

针对大肠杆菌产生的 VERO 毒素的 RNA 类似物抑制剂的开发

• 批准号: 11680594
• 负责人: FUJII Masayuki
• 金额: $ 2.18万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11680594/
• 关键词: Vero toxin; 28SrRNA; RNA analogs; therapeutic agents; 28S rRNAアナログ; 阻害剤; RNAデコイ

Synthesis and characterization of specific inhibitor and detection probe of vero toxin (VT) produced by enterobacterium O-157 were studied. Molecular design based on detail analysis of reaction mechanisms of N-glycosidase activity of vero toxin was performed. Analogous molecules of 28S rRNA which is a substrate for vero toxin A chain and globotriose (Gb3) ceramide which is a glycoreceptor on cell surface for vero toxin B chain. Inhibitory effects of some analogous RNA molecules which have GAGA tetraloop structure in common were evaluated by conventional experiments and showed that 2'-OMe anaolgs and phosphorothioate analogs were promising structure for drug application because those structures are known to be stable in physiological conditions in cells and even in vivo. Gb3 analogs bearing biotin moiety were prepared and immobilized on SPR sensor tip. Obsevations of binding affnities of the Gb3 analogs and abrin C which has the same specificity of sugar binding as vero toxin suggested that synthesized analogs were good detection probes for vero toxin. Fluorecein labeled Gb3 analogs and some other sugar derivatives were revealed that they are also good probes for the detection of VT using a fluorospectrometer.

对肠杆菌O-157产生的Vero毒素（VT）特异性抑制剂和检测探针的合成、表征进行了研究。在详细分析vero毒素N-糖苷酶活性反应机理的基础上进行了分子设计。28 S rRNA（vero毒素A链底物）和globotriose（Gb 3）神经酰胺（vero毒素B链细胞表面糖受体）的类似分子。通过常规实验评估了一些共同具有GAGA四环结构的类似RNA分子的抑制作用，并显示2 '-OMe anaolgs和硫代磷酸酯类似物是用于药物应用的有前景的结构，因为已知这些结构在细胞中甚至在体内的生理条件下是稳定的。制备带有生物素部分的Gb 3类似物并将其固定在SPR传感器尖端上。Gb 3类似物与相思豆毒素C的结合亲和力表明，Gb 3类似物是一种很好的检测vero毒素的探针。荧光素标记的Gb 3类似物和一些其他糖衍生物也显示它们是使用荧光光谱仪检测VT的良好探针。

项目成果

藤井政幸ら11名: "Design, Synthesis and Characterization of DNA Binding Peptide"Peptide Science. 2000. 109-112 (2001)

Masayuki Fujii 等 11 人：“DNA 结合肽的设计、合成和表征”Peptide Science 2000. 109-112 (2001)。

藤井政幸ら2名: "Synthesis and Properties of Novel Acyebi Nucleosides"Nucleic Acids Res. Symp. Ser.. 44. 47-48 (2000)

Masayuki Fujii 等：“新型 Acyebi 核苷的合成和特性”Nucleic Acids Ser.. 44. 47-48 (2000)

Masaguki Fujii 他1名: "A novel solid phase synthesis of DNA conjugates"Nucleosides, Nucleotides and Nucleic Acids,. 20巻(印刷中). (2001)

Masaguki Fujii 等：“DNA 缀合物的新型固相合成”，《核苷、核苷酸和核酸》，第 20 卷（出版中）。

藤井政幸ら3名: "Synthesis and Biological Activity of DNA-NLS Peptide Conjugate"Peptide Science. 2001. 313-316 (2001)

Masayuki Fujii 等：“DNA-NLS 肽缀合物的合成和生物活性”肽科学 2001. 313-316 (2001)。

藤井政幸ら2名: "A Novel Solid Phase Synthesis of DNA Conjugates"Nucleosides, Nucleotides and Nucleic Acids. 20. 1321-1324 (2001)

Masayuki Fujii 等人：“DNA 缀合物的新型固相合成”核苷、核苷酸和核酸 20. 1321-1324 (2001)