1999 Fiscal Year Annual Research Report

新機能を有する核酸関連生体分子の人工創製を目指した基礎研究

Research Project

Project/Area Number	11101001
Research Institution	Nagoya University
Principal Investigator	早川芳宏名古屋大学, 大学院・人間情報学研究科, 教授 (50022702)
Co-Investigator(Kenkyū-buntansha)	関根光雄東京工業大学, 大学院・生命理工学研究科, 教授 (40111679) 牧野圭祐京都大学, エネルギー理工学研究所, 教授 (50159141)
Keywords	塩基部無保護アミダイト法 / 修飾DNAオリゴマー / アリル保護アミダイト法 / DNA-ペプチドコンジュゲート / H-ホスホネート法 / アンチセンス核酸 / 一酸化窒素による発ガン / 塩基配列認識ペプチド
Research Abstract	早川:(1)これまでの塩基部無保護アミダイト法に残存する問題点の内、出発原料である4種類の塩基部無保護5′-O-ジメトキシトリチルデオキシリボヌクレオシドの高収率合成と、インターヌクレオチド結合形成におけるアデノシンおよびシチジンにおける水酸基選択的ホスフィチル化に関して改良を加え、既存の方法をより有効なものとした。(2)この改良法によってホスホロチオエートなど、インターヌクレオチド部修飾DNAオリゴマーの高効率合成法を開発した。(3)以前報告したアリル保護アミダイト法を用いて、既存の方法では合成が困難・不可能であった、塩基に対して不安定なDNA一ペプチド共有結合性複合体(コンジュゲート)の高純度・高収率合成を達成した。関根:塩基部位無保護法の一般的合成法を確立する上で重要な、5′水酸基と塩基部のアミノ基の両方がジメトキシトリチル基によって保護された4種類のデオキシヌクレオシドを導入した固相担体の効率的合成を検討し、収率よく合成する方法を見いだした。これによって、H-ホスホネート法およびアミダイト法による塩基部位無保護法でいくつかのオリゴデオキシヌクレオチドを合成することができた。牧野:(1)ホスホロチオエート型アンチセンス核酸に関し、標的がDNAの場合にはリン酸部位の立体異性による二重鎖構造変化が安定性を支配するのに対し、RNAの場合は水和等が安定性を支配しており立体異性が大きな要因ではないことを明らかにした。(2)一酸化窒素と遺伝子中のグアノシンおよびシチジンとの反応メカニズムを明らかにし、一酸化窒素による発ガンのメカニズムを提唱した。(3)転写因子モデルとして遺伝子の特異的塩基配列を認識するペプチドの調製に成功し、ペプチド間の共同作用が認識の程度を支配することを明らかにした。

Research Products

(35 results)

All Other

All Publications (35 results)

[Publications] Sakakura, A.: "A Novel Approach to Oligodeoxyribonucleotides Bearing Phosphoric Acid Esters at the 3'-Terminals via the Phosphoramidite Method with Allyl Protection : …"Tetrahedron Lett.. 40. 4359-4362 (1999)
[Publications] Iwai, S.: "Benzimidazolium triflate-Activated Synthesis of (6-4) Photoproduct-Containing Oligonucleotides and Its Application"Nucleic Acids Res.. 27. 2299-2303 (1999)
[Publications] Kataoka, M.: "A Convenient Method for the Synthesis of N-Free 5'-O-(p, p'-Dimethoxytrityl)-2'-deoxyribonucleosides via the 5'-O-Selective Tritylation of the Parent Substances"J. Org. Chem.. 64. 6087-6089 (1999)
[Publications] Heidenhain, S. B.: "Improved Methods for the Preparation of 2'-Deoxyribonucleoside and Ribonucleoside 3'-Phosphoramidites with Allylic Protectors"Nucleosides Nucleotides. 18. 1771-1787 (1999)
[Publications] Hayakawa, Y.: "A Facile Synthesis of DNA Oligomers with Modified Backbones via the Phosphoramidite Method without Nucleoside Base Protection"Chem. Comm. Symp. Ser.. 2. 105-108 (1999)
[Publications] Kataoka, M.: "A Facile Synthesis of DNA Oligomers with Modified Backbones via the Phosphoramidite Method without Nucleoside Base Protection"Nucleic Acids Symp. Ser.. 42. 105-106 (1999)
[Publications] Kato, T.: "A Facile Method for the Oxidation of Nucleoside H-Phosphonates to the Phosphates with Bis(trimethylsilyl) Peroxide"Synlett. 1796-1798 (1999)
[Publications] Hayakawa, Y.: "Nucleotide Synthesis via Methods without Nucleoside-Base Protection"J. Pharm. Sci.. (in press).
[Publications] K. Kanaori: "Structure and stability of consecutive stereoregulated chiral phosphorothioate DNA duplex"Biochemistry. 38. 16058-16066 (1999)
[Publications] T. Suzuki: "Isolation and characterization of diazoate intermediate upon nitrous acid and nitric oxide treatment of 2'-deoxycytidine"Biochemistry. 38. 7151-7158 (1999)
[Publications] Y. Aizawa: "Stability of the dimerization domain effects the cooperative DNA binding of short peptides"Biochemistry. 38. 4008-4017 (1999)
[Publications] T. Suzuki: "Formation of 2'-deoxyoxanosine from 2'-deoxyguanosine and nitrous acid : mechanisum and intermediates"Nucleic Acids Res.. 28. 544-551 (2000)
[Publications] T. Suzuki: "Reactivity of 2'-deoxyoxanosine, a novel DNA leison"Nucleic Acids Res. Symp. Ser.. 42. 7-8 (1999)
[Publications] T. Suzuki: "Formation of 8-nitroguanine from 2ユ-deoxyguanosine by NO/NO2"Nucleic Acids Res. Symp. Ser.. 42. 155-156 (1999)
[Publications] M. Tamura: "[1, 3-Bis(pyridine-2-carboxamino)propane]-palladium(II) monohydrate and its nickel(II) analogue"Acta Cryst.. C55. 719-722 (1999)
[Publications] H. Mori: "Effects of 6-formylpterin, a xanthine oxidase inhibitor and a superoxide scavenger, on production of nitric oxide in RAW 264.7 macrophages"Biochim. Biophys. Acta. (印刷中). (2000)
[Publications] Y. Ikada: "Characterization of ultrahigh molecular weight polyethylene irradiated with g-rays and electron beams to high doses"J. Polym. Sci.. 37. 159-168 (1999)
[Publications] 牧野圭祐: "酸素ラジカル種のヒトとの関わり"日本高気圧環境学会雑誌. 33. 151-159 (1999)
[Publications] 牧野圭祐一: "酸化窒素の環境生命科学"環・京・NOW. 23. 2-7 (1999)
[Publications] 牧野圭祐: "一酸化窒素の生化学"化学と生物. 38. 14-19 (2000)
[Publications] Wada, T.: "First Synthesis of H-Phosphonate Oligonucleotides Bearing N-Unmodified Bases"Tetrahedron Lett.. 40. 915-918 (1999)
[Publications] Moriguchi, T.: "31p NMR Study on Aminoacylation of 5'-AMP and Its Analogs"J. Chem. Soc., Perkin I,. 1859-1866 (1999)
[Publications] Kawahara, S.: "Ab into and Density Functional Studies of Substiuent Effects of an A-U Base-Pair on the Stability of Hydrogen Bonding"J. Phys. Chem.(A). 103. 8516-8523 (1999)
[Publications] Tsuruoka, H.: "Selective and Facile 5'-De(thio)phosphorylation of Oligodeoxynucleotides Having a 5'-Terminal Phosphorothioate Group by Simple Thermolysis"Tetrahedron. 40. 8411-8414 (1999)
[Publications] Noguchi, Y.: "Isolation of Biuopterin-α-glucoside from Spirulina (Arthrospira) platensis and Its Physiologic Function"Mar. Biotechnol.. 1. 207-210 (1999)
[Publications] Moriguchi, T.: "Synthesis of Nucleotide Antibiotics Having N-Acyl Phosphoramidate Linkages"Nucleic Acids Symp. Ser.. 42. 15-16 (1999)
[Publications] Shoda, K.: "Synthesis and Properties of Oligonucleotides Containing 2'-O-Methyl -2-thiouridine"Nucleic Acids Symp. Ser.. 42. 161-162 (1999)
[Publications] Sekine M.: "Chemical Synthesis of Hypermethylated Capped RNA on Polymer Supports"Nucleic Acids Symp. Ser.. 15. 318 (1999)
[Publications] Sekine. M.: "Chemical Synthesis of Oligonucleotides by Use of Phenylthio Group"Current Organic Chemistry. 3. 25-66 (1999)
[Publications] Seio, K.: "Synthesis and Properties of Oligodeoxythymidylates Incorporating an Artificial Bend Motif"Helv. Chim. Acta. 83. 162-180 (2000)
[Publications] Matsuo, H.: "Efficient Synthesis of 13C, 15N Labeled RNA Containing the Cap Structure m7GpppA"J. Am. Chem. Soc.. (in press).
[Publications] Tsuruoka, H.: "Synthesis and Conformational Properties of Oligonucleotides Incorporating 2'-O-phosphorylated Ribonucleotide as Structural Motifs of Pre-tRNA Splicing Intermediates"J. Org. Chem.. (in press).
[Publications] Kawahara, S.: "The Hydrogen Bond Energy on Mismatched Base Pair Formation between Uracil Derivatives and Guanine in the Gas-Phase and the Aqueous-phase"THEOCHEM. (in press).
[Publications] Sekine, M.: "Synthesis and Properties of Oligodeoxynucleotides Incorporating a Conformationally Rigid Uridine Unit Having a Cyclic Structure at the 5'-terminal Site"J. Org. Chem.. (in press).
[Publications] Kadokura, M.: "Synthesis of 4-Thiouridine, 6-Thioinosine and 6-Thioguanosine 3', 5'-O-Bisphosphates as Donor Molecules for RNA Ligation and Their Application to…"J. Org. Chem.. (in press).

1999 Fiscal Year Annual Research Report

新機能を有する核酸関連生体分子の人工創製を目指した基礎研究

Principal Investigator

早川 芳宏 名古屋大学, 大学院・人間情報学研究科, 教授 (50022702)

Research Products

[Publications] Sakakura, A.: "A Novel Approach to Oligodeoxyribonucleotides Bearing Phosphoric Acid Esters at the 3'-Terminals via the Phosphoramidite Method with Allyl Protection : …"Tetrahedron Lett.. 40. 4359-4362 (1999)

[Publications] Iwai, S.: "Benzimidazolium triflate-Activated Synthesis of (6-4) Photoproduct-Containing Oligonucleotides and Its Application"Nucleic Acids Res.. 27. 2299-2303 (1999)

[Publications] Kataoka, M.: "A Convenient Method for the Synthesis of N-Free 5'-O-(p, p'-Dimethoxytrityl)-2'-deoxyribonucleosides via the 5'-O-Selective Tritylation of the Parent Substances"J. Org. Chem.. 64. 6087-6089 (1999)

[Publications] Heidenhain, S. B.: "Improved Methods for the Preparation of 2'-Deoxyribonucleoside and Ribonucleoside 3'-Phosphoramidites with Allylic Protectors"Nucleosides Nucleotides. 18. 1771-1787 (1999)

[Publications] Hayakawa, Y.: "A Facile Synthesis of DNA Oligomers with Modified Backbones via the Phosphoramidite Method without Nucleoside Base Protection"Chem. Comm. Symp. Ser.. 2. 105-108 (1999)

[Publications] Kataoka, M.: "A Facile Synthesis of DNA Oligomers with Modified Backbones via the Phosphoramidite Method without Nucleoside Base Protection"Nucleic Acids Symp. Ser.. 42. 105-106 (1999)

[Publications] Kato, T.: "A Facile Method for the Oxidation of Nucleoside H-Phosphonates to the Phosphates with Bis(trimethylsilyl) Peroxide"Synlett. 1796-1798 (1999)

[Publications] Hayakawa, Y.: "Nucleotide Synthesis via Methods without Nucleoside-Base Protection"J. Pharm. Sci.. (in press).

[Publications] K. Kanaori: "Structure and stability of consecutive stereoregulated chiral phosphorothioate DNA duplex"Biochemistry. 38. 16058-16066 (1999)

[Publications] T. Suzuki: "Isolation and characterization of diazoate intermediate upon nitrous acid and nitric oxide treatment of 2'-deoxycytidine"Biochemistry. 38. 7151-7158 (1999)

[Publications] Y. Aizawa: "Stability of the dimerization domain effects the cooperative DNA binding of short peptides"Biochemistry. 38. 4008-4017 (1999)

[Publications] T. Suzuki: "Formation of 2'-deoxyoxanosine from 2'-deoxyguanosine and nitrous acid : mechanisum and intermediates"Nucleic Acids Res.. 28. 544-551 (2000)

[Publications] T. Suzuki: "Reactivity of 2'-deoxyoxanosine, a novel DNA leison"Nucleic Acids Res. Symp. Ser.. 42. 7-8 (1999)

[Publications] T. Suzuki: "Formation of 8-nitroguanine from 2ユ-deoxyguanosine by NO/NO2"Nucleic Acids Res. Symp. Ser.. 42. 155-156 (1999)

[Publications] M. Tamura: "[1, 3-Bis(pyridine-2-carboxamino)propane]-palladium(II) monohydrate and its nickel(II) analogue"Acta Cryst.. C55. 719-722 (1999)

[Publications] H. Mori: "Effects of 6-formylpterin, a xanthine oxidase inhibitor and a superoxide scavenger, on production of nitric oxide in RAW 264.7 macrophages"Biochim. Biophys. Acta. (印刷中). (2000)

[Publications] Y. Ikada: "Characterization of ultrahigh molecular weight polyethylene irradiated with g-rays and electron beams to high doses"J. Polym. Sci.. 37. 159-168 (1999)

[Publications] 牧野圭祐: "酸素ラジカル種のヒトとの関わり"日本高気圧環境学会雑誌. 33. 151-159 (1999)

[Publications] 牧野圭祐一: "酸化窒素の環境生命科学"環・京・NOW. 23. 2-7 (1999)

[Publications] 牧野圭祐: "一酸化窒素の生化学"化学と生物. 38. 14-19 (2000)

[Publications] Wada, T.: "First Synthesis of H-Phosphonate Oligonucleotides Bearing N-Unmodified Bases"Tetrahedron Lett.. 40. 915-918 (1999)

[Publications] Moriguchi, T.: "31p NMR Study on Aminoacylation of 5'-AMP and Its Analogs"J. Chem. Soc., Perkin I,. 1859-1866 (1999)

[Publications] Kawahara, S.: "Ab into and Density Functional Studies of Substiuent Effects of an A-U Base-Pair on the Stability of Hydrogen Bonding"J. Phys. Chem.(A). 103. 8516-8523 (1999)

[Publications] Tsuruoka, H.: "Selective and Facile 5'-De(thio)phosphorylation of Oligodeoxynucleotides Having a 5'-Terminal Phosphorothioate Group by Simple Thermolysis"Tetrahedron. 40. 8411-8414 (1999)

[Publications] Noguchi, Y.: "Isolation of Biuopterin-α-glucoside from Spirulina (Arthrospira) platensis and Its Physiologic Function"Mar. Biotechnol.. 1. 207-210 (1999)

[Publications] Moriguchi, T.: "Synthesis of Nucleotide Antibiotics Having N-Acyl Phosphoramidate Linkages"Nucleic Acids Symp. Ser.. 42. 15-16 (1999)

[Publications] Shoda, K.: "Synthesis and Properties of Oligonucleotides Containing 2'-O-Methyl -2-thiouridine"Nucleic Acids Symp. Ser.. 42. 161-162 (1999)

[Publications] Sekine M.: "Chemical Synthesis of Hypermethylated Capped RNA on Polymer Supports"Nucleic Acids Symp. Ser.. 15. 318 (1999)

[Publications] Sekine. M.: "Chemical Synthesis of Oligonucleotides by Use of Phenylthio Group"Current Organic Chemistry. 3. 25-66 (1999)

[Publications] Seio, K.: "Synthesis and Properties of Oligodeoxythymidylates Incorporating an Artificial Bend Motif"Helv. Chim. Acta. 83. 162-180 (2000)

[Publications] Matsuo, H.: "Efficient Synthesis of 13C, 15N Labeled RNA Containing the Cap Structure m7GpppA"J. Am. Chem. Soc.. (in press).

[Publications] Tsuruoka, H.: "Synthesis and Conformational Properties of Oligonucleotides Incorporating 2'-O-phosphorylated Ribonucleotide as Structural Motifs of Pre-tRNA Splicing Intermediates"J. Org. Chem.. (in press).

[Publications] Kawahara, S.: "The Hydrogen Bond Energy on Mismatched Base Pair Formation between Uracil Derivatives and Guanine in the Gas-Phase and the Aqueous-phase"THEOCHEM. (in press).

[Publications] Sekine, M.: "Synthesis and Properties of Oligodeoxynucleotides Incorporating a Conformationally Rigid Uridine Unit Having a Cyclic Structure at the 5'-terminal Site"J. Org. Chem.. (in press).

[Publications] Kadokura, M.: "Synthesis of 4-Thiouridine, 6-Thioinosine and 6-Thioguanosine 3', 5'-O-Bisphosphates as Donor Molecules for RNA Ligation and Their Application to…"J. Org. Chem.. (in press).

早川芳宏名古屋大学, 大学院・人間情報学研究科, 教授 (50022702)