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Efficient intracellular delivery of antisense oligonucleotide and pharmaceutical factors

Research Project

Project/Area Number 08672481
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Physical pharmacy
Research InstitutionNagoya City University

Principal Investigator

YOTSUYANAGI Tosihisa  Nagoya City university, Pharmaceutical Science, Professor, 薬学部, 教授 (40080189)

Co-Investigator(Kenkyū-buntansha) HAZEMOTO Norio  Nagoya City university, Pharmaceutical Science, Associate Professor, 薬学部, 助教授 (40192273)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1996: ¥1,800,000 (Direct Cost: ¥1,800,000)
KeywordsAntisense / Oligonucleotide / DNA carrier / Oligopeptide / intracellular delivery / オリゴヌクレオチド
Research Abstract

As a carrier for intracellular delivery of an antisense and plasmid DNA,synthetic oligopeptides were examined. The oligopeptide which consisted of nine amino acids was synthesized as the carrier with a peptide synthesizer. Various peptide which consisted of lysine, leucine, cysteine, and tryptophane were synthesized and purified by the liquid chromatoqraphy which the reverse-aspect column was installed. The aptitude as the intracellular introduction carrier of the deoxyribonucleic acid with this synthetic peptide was examined by using the HeLaS3 cell by the plasmid DNA which contained the CAT gene.
The gene expression was not observed in peptide which consisted of lysine and leucine, but observed in peptide contained cysteine. The dimer was found with the mass spectrum and the gene expression ability disappeared when the disulfide linkage of the peptide was cut by the DTT processing. The CD measurement showed that peptide took the structure which consisted of two peptide chains. As a result of the gel electrophoresis, the DNA and peptide formed complex. It were confirmed the complex was taken into intracellular from the observation with confocus laser microscope. The gene expression have been observed even by cells another of the HeLaS3, such as CHO,L929, COS7 and HepG2 cells. The introduction efficiency has improved when tryptophane is introduced into peptide. It has been understood that the oligopeptide which consists of the basic amino acid and the hydrophobic amino acid with the disulfide bond has the ability to introduce the deoxyribonucleic acid into intracellular. It will be studied on reletionship between the secondary structure of peptide and the interaction with the deoxyribonucleic acid further in the future.

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (17 results)

All Other

All Publications (17 results)

  • [Publications] M.Fukaholi, S.Akazu et al: "Bacterial susceptibility of Buthyl p-Hydroxy benzoate in the presence of polyozy etylene Cetyl Ether" Biocontrol Sci. 3. 47-49 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 植田 拓也.水野 勇 ら: "ラット癌性腹膜炎におけるカルボプラチン含有αーリン酸三カルシウム細粒の腹腔内投与における基礎的検討" Drug Deliv.Sys.12. 175-179 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 田中 洵.小山 修 ら: "口腔内粘膜付着性軟膏の製剤特性:水溶性高分子-プラスチベース系" 薬剤学. 57. 8-15 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Fukaholi, Y.Takafuji, S.Akazu, H.Sato, and T.Yotsuyanagi: "Effects of Polyoxyethylene Cetyl Ether on the Cellular Uptake and Antibacterial Activity of Butyl-P-Hydroxybenzoate against Escherichia coli" Bioconfrol Sci. 3. 23〜29 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 赤毛 義実、櫨本 紀夫、竹山 廣光、水野 勇、毛利 紀章、植田 拓也、柴田 直史、四ツ柳 智久、真辺 忠夫: "Cationic liposomeを用いた癌に対する遺伝子導入の基礎的検討" Biotherapy. 12. 199〜201 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Fukahori, Y.Takafuji, S.Akazu, H.Sato and T.Yotsuyanagi: "Effects of Polyoxyethylene Cetyl Ether on the Cellular Uptake and Antibacterial activity of Buthyl p-Hydroxy-benzoate against Escherichia coli" Biocontrol Sci.3. 23-29 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Fukahori, S.Akazu, H.Sato, and T.Yotsuyanagi: "Bacterial Susceptibility of Buthyl p-Hydroxybenzoate in the Presence of Polyoxyethylene Cetyl Ether" Biocontrol Sci.3. 47-49 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Y.Akamo, N.Hazemoto, H.Takeyama, I.Mizuno, N.Mouri, T.Ueda, N.Shibata, T.Yotsuyanagi and T.Manabe: "Gene transfection of cancer using cationic liposomes" Biotherapy. 12(1). 199-201 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Ueda, I.Mizuno, T.Yotsuyanagi, T.Manabe, and S.Nakamura: "Intraperitoneal administration of alpha-tricalcium phosphate particles incorporated with carboplatin in rats bearing abdominal carcinomatosis" Drug Delivery System. 12(3). 175-179 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] J.Tanaka, O.Koyama, E.Kuwabara, M.Takahashi N.Takahashi and T.Yotsuyanagi: "Dissolution behavior and bioavailability of amphotericin B ground or lyophilized with polymers" YAKUZAIGAKU. 57(2). 79-85 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] M.Fukaholi, Y.Takafuji, et al: "Effects of polyoxyethylene Cetyl Ether on the Cellular UpAcko and Antibacterial activity of Buthyl p-Hydroxybenzoate against Escherichra coli" Biocontrol Sci.,. 3. 23-29 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] M.Fukaholi, S.Akazu, et al: "Bacterial Susceptibility of Buthyl p-Hydroxybenzoate in the presence of polyoxyacetylene Cetyl Ether" Biocontrol Sci. 3. 47-49 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 赤毛義美・櫨本紀夫, ら: "Catronic liposomeを用いた癌に対する遺伝子導入の基礎的検討" Biotherapy. 12. 199-201 (1998)

    • Related Report
      1997 Annual Research Report
  • [Publications] 植田拓也, 水野勇, ら: "ラット癌性腹膜炎におけるカルボプラチン含有αーリン酸三カルシウム細粒の腹腔内投与における基礎的検討" Drug Deliv.Sys.12. 175-179 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 田中殉, 小山修, ら: "口腔内粘膜付着性軟膏の製剤特性:水溶性高分子-プラスチベース系" 薬剤学. 57. 8-15 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] M. Fukahori et al: "Relationship between uptake of p-hydroxybenzoicacid esterby E. coli" Chom. pharm. Bull. 44. 1567-1570 (1996)

    • Related Report
      1996 Annual Research Report
  • [Publications] I. Takagi et al: "Application of alginategel as a Vehicle for liposomes I." Chom. pharm. Bull. 44. 1941-1947 (1996)

    • Related Report
      1996 Annual Research Report

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Published: 1996-04-01   Modified: 2016-04-21  

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