Design and Syntheses of Biologically Functional Molecules Based on the Mechanisms of Life

基于生命机制的生物功能分子的设计与合成

• 批准号: 11470469
• 负责人: IMANISHI Takeshi
• 金额: $ 7.55万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 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/en/grant/KAKENHI-PROJECT-11470469/
• 关键词: Nucleoside Analogs; Oligonucleotides; Antisence; Pyridoxal model compounds; unnatural amino acids; Chemiluminescence; Cationic liposome; Gene transfection; オリゴヌクレオチド類縁体; オリゴヌクレオシド; 非天然型アミノ酸

We studied the four subjects and obtained the results shown below:(1) We have succeeded to synthesize 2'-0,4'-C-methylene bridged nucleic acid analogs (BNAs) and oligonucleotides containing the BNA (BNA-ODNs). The BNA-ODNs were found to have more effective duplex and triplex forming ability than natural ODNs and modified ODNs ever reported. Comparison of various properties of BNA-ODNs with those of natural ODNs and S-ODNs showed that the BNA-ODNs have an excellent antisense ability.(2). We have designed and synthesized novel chiral vitamin B_6 model compounds having a chiral ansa-structure and/or a chiral ionophore structure, which were found to be effective for asymmetric α-alkylation ofα-amino esters and petides at the N-terminal position. Catalytic β-replacement reaction of a Ser derivative with thiols was also successfully achieved by employing the pyridoxal model compounds.(3) We have synthesized fire-fly luciferin analogs having a heterocyclic structure in place of the benzothiazole structure of the fire-fly luciferin molecule, and studied their bioluminescent and chemiluminescent activities. As a result, it was found that the benzimidazole derivative shows effective light-emmisio by treatment with luciferase, while the benzoxazole derivative is effective for chemiluminescent induced by oxidative treatment.(4) We have synthesized cationic lipid having a symmetrical structure, and studied gene transfection activity of liposome prepared from the lipid. As a consequence, it was found that the liposome composed of the symmetrical lipid, which was more easily prepared than that having an unsymmetrical structure, is low toxic and effective for gene transfection.

（1）成功地合成了2 ′-0，4 ′-C-亚甲基桥联核酸类似物（BNAs）和含有BNAs的寡核苷酸（BNA-ODNs）。BNA-ODNs具有比天然ODNs和修饰ODNs更强的双链和三链形成能力。与天然ODNs和S-ODNs的各种性质比较表明，BNA-ODNs具有良好的反义能力。（二）、我们设计并合成了具有手性柄结构和/或手性离子载体结构的新型手性维生素B_6模型化合物，发现它们对α-氨基酯和肽的N-末端不对称α-烷基化反应有效。以吡哆醛为模型化合物，成功地实现了丝氨酸衍生物与硫醇的催化β-取代反应。(3)我们合成了具有杂环结构的萤火虫荧光素类似物，取代了萤火虫荧光素分子中的苯并噻唑结构，并研究了它们的生物发光和荧光活性。结果发现，苯并咪唑衍生物通过用荧光素酶处理显示出有效的发光，而苯并恶唑衍生物对于由氧化处理诱导的荧光是有效的。(4)我们合成了具有对称结构的阳离子脂质体，并研究了由该脂质体制备的脂质体的基因转染活性。结果发现，由对称脂质组成的脂质体比具有不对称结构的脂质体更容易制备，其毒性低，并且对基因转染有效。

项目成果

S. Obika, W. Yu, A. Shimoyama, T. Uneda, T. Minami, K. Miyashita, T. Doi and T. Imanishi: "Properties of cationic liposomes composed of cationic lipid YKS-220 having an ester linkage: adequate stability, high transfection efficiency, and low cytotoxicity"

S. Obika、W. Yu、A. Shimoyama、T. Uneda、T. Minami、K. Miyashita、T. Doi 和 T. Imanishi：“由具有酯键的阳离子脂质 YKS-220 组成的阳离子脂质体的特性：足够

S.Obika, T.Imanishi et al.: "Synthesis and conformation of 3', 4'-BNA monomers, 3'-0, 4'-C-methyleneribonucleosides"Tetrahedron. (印刷中). (2002)

S.Obika、T.Imanishi 等人：“3、4-BNA 单体、3-0、4-C-亚甲基核糖核苷的合成和构象”Tetrahedron（印刷中）。

K. Miyashita, H. Miyabe, K. Tai, H. Iwaki and T. Imanishi: "Asymmetric a-alkylation of α-amino esters using pyridoxal derivatives having a chiral ansa-structure and a chiral ionophore function: a novel example of double asymmetric induction"Tetrahedron. 5

K. Miyashita、H. Miyabe、K. Tai、H. Iwaki 和 T. Imanishi：“使用具有手性 ansa 结构和手性离子载体功能的吡哆醛衍生物对 α-氨基酯进行不对称 α-烷基化：双不对称感应“四面体.5”

S. Obika, Y. Hari, H. Inohara and T. Imanishi: "2,4'-BNA bearing unnatural nucleobases: Toward the expansion of the target sequence of double-stranded DNA in triplex formation"Nucleic Acids Res. Suppl.. 1. 171-172 (2001)

S. Obika、Y. Hari、H. Inohara 和 T. Imanishi：“带有非天然核碱基的 2,4-BNA：以三链体形式扩展双链 DNA 的靶序列”《核酸研究》。

K.Miyashita, T.Imanishi et al.: "Oxiranes having an acridane structure as a novel chemiluminescent precursor: synthesis and chemiluminescent studies"Tetrahedron. 57・16. 3361-3367 (2001)

K. Miyashita、T. Imanishi 等：“具有吖啶结构的环氧乙烷作为新型化学发光前体：合成和化学发光研究”Tetrahedron 57・16 (2001)。