Development of a Gene Expression Control Medicine and Analysis of the Interaction with a DNA Duplex.

• Project/Area Number: 14572012
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemical pharmacy
• Research Institution: Tokyo University of Pharmacy and Life Science
• Principal Investigator: KAWASHIMA Etsuko Tokyo University of Pharmacy and Life Science, School of Pharmacy, Associate Professor, 薬学部, 助教授 (30057343)
• Project Period (FY): 2002 – 2003
• Project Status: Completed (Fiscal Year 2003)
• Budget Amount: ¥2,700,000 (Direct Cost: ¥2,700,000); Fiscal Year 2003: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
• Keywords: Pyrrolepolyamide-nucleoside complex / Interaction of a DNA with a drug / Tm value / CD spectra / Stereoselective deuteration / (5'R)-[5'-^2H;5'-^<13>C]nucleoside / 遺伝子発現制御物質 / ポリアミド-ヌクレオシドハイブリッド / ピロールポリアミド / DNAとの相互作用 / 安定同位体標識化ヌクレオシド / [5-^2H_1;5-^<13>C]リボフラノース / 効率的合成法の開発 / 遺伝子発現制御薬 / ポリアミド修飾ヌクレオシド / 高立体選択的重水素化 / D-(5R)[5-^2H_1;5-^<13>C]リボース / {D-(5R)[5-^2H_1;5-^<13>C]リボシル}チミン

Research Abstract

Organic compounds capable of controlling gene expression by recognizing DNA sequence-specificity are expected to be viable gene therapy medicine. The nucleosides bearing pyrrolepolyamide which is minor groove binder were designed and synthesized as a lead compounds for gene therapy medicine and the analysis of DNA interaction with these compounds by Tm values and Circular Dichroism (CD) spectra was performed. In addition, the synthesis of {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}nucleoside that is useful for structural analysis of conformation in DNA was carried out. 1.Deoxyguanosine bearing pyrrolepolyamide using 3-aminopropyl linker (GAP), using aminopropyonyl linker (GBP) and adenosine bearing pyrrolepolyamide (Apy) were synthesized. 2.The investigation of affinity for DNA duplexes by Tm values and CD spectra showed that DNA included the sequence of 5'-AAATT-3' with GAP complex has potential of highly selectivity as compare with complexes of other DNA. In this result, GAP might have usable sequence selectively in gene therapy, and provide possibility to be new series of antisense or antigene drugs. 3.Highly diastereoselective synthesis of {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}thymine was established. A preparation of {(5R)-D-[5-^2H_1;5-^<13>C]ribose derivative of 1,2:5,6-di-O-isopropyliden-α-D-allofranose was successfully achieved by a ^<13>C Wittig reaction using Ph_3P^<13>CH_3I-BuLi to 5-oxoribose derivative and subsequent transformation into D-[5-^<13>C]ribose derivative with an AD reaction, selective acylation, oxidation with NaIO_4, and a stereoselective deuteride transfer reaction from Alpine-Borane-d to 5-oxo-D-[5-^<13>C]ribose derivative as the main reaction. {(5R)-D-[5-^2H_1;5-^<13>C]ribofuranosyl}thymine was synthesized in the established manner from this 5-^<13>C/^2H_1-double-labeled ribose.

Research Products

• [Publications] Kawashima, E., Umabe, K., Sekine, T.: "Synthesis of [5'-^<13>C]Ribonucleosides and 2'-Deoxy[5'-^<13>c]ribonucleosides"The Journal of Organic Chemistry. 67. 5142-5151 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kawashima, E., Itoh, D., Kamaike, K., Terui, Y., Oshima, T.: "Synthesis and Analysis of Nucleosides Bearing Pyrrolepolyamide Binding to DNA."Nucleosides, Nucleotides & Nucleic Acids.. 22. 1309-1311 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ohba, Y., Kamaike, K., Terui, Y., Oshima, T., Kawashima, E.: "Design, synthesis and analysis of antiviral nucleosides bearing pyrrolepolyamide binding to nucleic acid (II):N^2- pyrrolepolyamidopropionylguanosine"Nucleic Acids Research Supplement. 29-30 (2003)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kawashima, E., Kamaike, K.: "Synthesis of Stable-isotope (^<13>C and ^<15>N) Labeled Nucleosides and Their Applications"Mini-Reviews in Organic Chemistry.. (In press). (2004)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kawashima, E., Sekine, T., Umabe, K., Kamaike, K., Mizukoshi, T., Shimba, N., Suzuki, E., Kojima, C.: "New Sequential-Assignment Routes of Nucleic Acid NMR Signals Using A [5'-^<13>C]-Labeled DNA Dodecamer"Nucleosides, Nucleotides & Nucleic Acids.. 23. 255-262 (2004)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kawashima, E., Umabe, K., Sekine, T.: "Synthesis of [5'-^<13>C]Ribonucleosides and 2'-Deoxy[5'-^<13>c]ribonucleosides"J.Org.Chem.. 67. 5142-5151 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kawashima, E., Itoh, D., Kamaike, K., Terui, Y., Oshima, T.: "Synthesis and Analysis of Nucleosides Bearing Pyrrolepolyamide Binding to DNA."Nucleosides, Nucleotides & Nucleic Acids. 22. 1309-1311 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ohba, Y., Kamaike, K., Terui, Y., Oshima, T., Kawashima, E.: "Design, synthesis and analysis of antiviral nucleosides bearing pyrrolepolyamide binding to nucleic acid (II) : N^2-pyrrolepolyamidopropionylguanosine"Nucleic Acids Research. Supplement. 29-30 (2003)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kawashima, E., Sekine, T., Umabe, K., Kamaike, K., Mizukoshi, T., Shimba, N., Suzuki, E., Kojima, C.: "New Sequential-Assignment Routes of Nucleic Acid NMR Signals Using A [5'-^<13>C]-Labeled DNA Dodecamer"Nucleosides, Nucleotides & Nucleic Acids. 23(1&2). 255-262 (2004)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kawashima, E., Kamaike, K.: "Synthesis of Stable-isotope (^<13>C and ^<15>N) Labeled Nucleosides and Their Applications."Mini-Reviews in Organic Chemistry. in press. (2004)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kamaike, K., Namiki, T., Kawashima.E.: "6-(Levulinyloxymethyl)-3-methoxy-2-nitrobenzoyl and 2-(Levulinyl oxy-methyl)-5-methoxy-4-nitrobenzoyl Groups as Novel Base-labile Groups for 5'-Hydroxy Protection in Solid-phase Oligonucleotides Synthesi"Nucleosides, Nucleotides & Nucleic Acids. 22(4). 469-487 (2003)
• [Publications] Ohba, Y., Kamaike, K., Terui, Y., Oshima, T., Kawashima, E.: "Design, synthesis and analysis of antiviral nucleosides bearing pyrrolepolyamide binding to nucleic acid (II) : N^2-pyrrolepolyamidopropionyl"Nucleic Acids Research Supplement. 29-30 (2003)
• [Publications] Kamaike, K., Namiki, T., Kawashima, E.: "Novel Base-Labile Protecting Groups for 5'-Hydroxyl protection in Solid-Phase Oligonucleotide Synthesis"Nucleosides, Nucleotides & Nucleic Acids. 22. 1011-1015 (2003)
• [Publications] Kawashima.E., Itoh, D., Kamaike, K.Terui, Y., Oshima, T.: "Synthesis and Analysis of Nucleosides Bearing Pyrrolepolyamide Binding to DNA"Nucleosides, Nucleotides & Nucleic Acids. 22. 1309-1311 (2003)
• [Publications] Kawashima, E., Sekine, T., Umab e, K., Kamaike, K.Mizukoshi, T., Shimba, N., Suzuki, E., Kojim: "HMQC-NOESY NMR Study of a DNA Dodecamer with ^<13>C-Labeling at the 5' Positions"Nucleosides, Nucleotides & Nucleic Acids. 24. 255-262 (2004)
• [Publications] Etsuko Kawashima: "Synthesis of [5'-^<13>C] Ribonucleosides and 2'-Deoxy[5'-^<13>C] ribonucleosides"The Journal of organic chemistry. 67・15. 5142-5151 (2002)