Preparation of Artificial Ribonucleases and Their Applications to the Therapy for Cancer and AIDS

人工核糖核酸酶的制备及其在癌症和艾滋病治疗中的应用

• 批准号: 08559005
• 负责人: KOMIYAMA Makoto
• 金额: $ 2.05万
• 依托单位: Graduate School of Engineering, The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08559005/
• 关键词: RNA; Hydrolysis; Oligoamine; Artificial ribonuclease; Cancer; AIDS; Therapy; アミン

With the aims to develop new therapeutic methods for cancer and AIDS,artificial ribonucleases have been prepared, and RNA scission mechanism is investigated. It is confirmed that intramolecular acid/base cooperation is responsible for the remarkable catalysis of oligoamines for RNA hydrolysis. Novel artificial ribonucleases in which oligoamines are attached inside of DNA oligomers (which are complementary with a part of the substrate RNA) have been synthesized. These artificial enzymes cut the target RNA more precisely than the ones with the scissors at the ends of DNA oligomers, mainly because the scissors are fixed in the rigidly structured double strands. Furthermore, Conjugates of an intercalator and either glycine or iminodiacetate show effective scission of RNA.These conjugates are neutral or negatively charged, which is highly in contrast with the fact that the previously reported RNA scissors are positively charged. Thus, they can be placed exactly at the desired site, even in a strong electrostatic field induced by RNA.These artificial ribonucleases cleave only the target RNA in cells, and thus should be highly potent for the therapy of cancer and AIDS.

为了开发新的治疗癌症和艾滋病的方法，制备了人工核糖核酸酶，并研究了其切割机制。它被证实，分子内的酸/碱的合作是负责显着催化的寡胺RNA水解。已经合成了新的人工核糖核酸酶，其中寡胺附着在DNA寡聚物（与底物RNA的一部分互补）的内部。这些人工酶比在DNA寡聚体末端使用剪刀的酶更精确地切割靶RNA，主要是因为剪刀固定在刚性结构的双链中。此外，嵌入剂与甘氨酸或亚氨基二乙酸盐的偶联物均能有效地切割RNA，偶联物呈中性或带负电荷，这与先前报道的RNA剪刀带正电荷的事实形成鲜明对比。因此，即使在RNA诱导的强静电场中，它们也可以精确地放置在所需的位置。这些人工核糖核酸酶仅切割细胞中的靶RNA，因此对于癌症和艾滋病的治疗应该是非常有效的。

项目成果

Masaki Endo et al.: "Novel phosphoramidite monomer for the site-selective incorporation of a diastereochemically pure phosphoramidate to oligo-nucleotide." J.Org.Chem.61(6). 1994-2000 (1996)

Masaki Endo 等人：“用于将非对映化学纯的氨基磷酸酯位点选择性掺入寡核苷酸的新型亚磷酰胺单体。”

M.Endo, Y.Azuma, Y.Saga, A.Kuzuya, G.Kawai, and M.Komiyama: ""Molecular design for a pinpoint RAN scission. Interposition of oligoamines between two DNA oligomers."" J.Org.Chem.62(4). 846-852 (1997)

M.Endo、Y.Azuma、Y.Saga、A.Kuzuya、G.Kawai 和 M.Komiyama：“”精确 RAN 断裂的分子设计。

M.Endo et al.: "RNA hydrolysis by the cooperation of carboxylate ion and ammonium ion" J.Am.Chem.Soc.118. 5478-5479 (1996)

M.Endo 等人：“通过羧酸根离子和铵离子的合作进行 RNA 水解”J.Am.Chem.Soc.118。

八代 盛夫: "酵素に学ぶ-多核金属錯体の設計と人工ヌクレアーゼへの応用" 化学と工業. 50(7). 980-982

Morio Yashiro：“从酶中学习 - 多核金属配合物的设计及其在人工核酸酶中的应用”化学与工业 50(7)。

M.Endo, Y.Azuma, and M.Komiyama: ""Site-selective RNA hydrolysis by a novel artificial ribo-nuclease : Interposition of oligoamine between two DNA oligomers."" Nucleic Acids, Symp.Ser.35. 81-82 (1996)

M.Endo、Y.Azuma 和 M.Komiyama：“通过新型人工核糖核酸酶进行位点选择性 RNA 水解：在两个 DNA 寡聚物之间插入寡胺。”《核酸》，Symp.Ser.35。