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Elucidation and Identification of Endogenous Benzodiazepine and Elucidation of Biological Properties of the Benzodiazepine

Research Project

Project/Area Number 01571259
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field 応用薬理学・医療系薬学
Research InstitutionTokyo University of Science

Principal Investigator

KUBOTA Kazuhiko  Science University of Tokyo, Department of Pharmaceutical Sciences, Professor, 薬学部, 教授 (30084411)

Co-Investigator(Kenkyū-buntansha) HAZATO Tadahiko  The Tokyo Metropolitan Institute of Medical Science, Department of Chemotherapy,, 化学療法研究部, 主任研究員 (60109949)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1989: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsBenzodiazepine / Receptor / Endogenous Ligand / Bovine Brain / Bovine Spinal Cord / Isolation / 牛脊随 / 精製 / 牛背髄
Research Abstract

In this study, we aimed to isolate and identify the substances that have been expected to exist in the central nervous system and bind selectively to the benzodiazepine receptor in the brain.
In order to conferm if endogenous benzodiazepine ligands isolated will bind to the benzodiazepine receptor, we obtained a fraction containing benzodiazepine receptor from the mice brain. Radioactive flunitrazepam labled with tritium was bound to the fraction, and replaced with fractions that were isolateed from the central nervous system and contained endogenous benzodiazepine ligands. Determination of endogenous benzodiazepine ligands was made through the degree of displacement of flunitrazepam by the endogenous ligands.
1)Search of endogenous ligands in the bovine brain
The bovine brain cortex was homogenized and extracted with acetone-methanol mixture. Detectable, but insufficient amount of endogenous ligands was obtained from this procedure.
2)Isolation of endogenous ligands from the bovine spinal cord.
The homogenate of the bovine spinal cord was subjected to the isolation of ligands through gel filtration and other procedures, resulting in the confirmation of endogenous benzodiazepine ligands having high affinity to benzodiazepine receptor preparation in the elutes of fraction number 47 to 49. The endogenous ligands showed their molecular weights to be less than 1, 000 and to be other substances than peptide and than diazepam, desmethyl diazepam and methy b-carboline ester. In addition, they were composed of more than 5 substances having different molecular sizes.
3)Isolation of ligands from the monkey brain and human serum
We tried to detect endogenous ligands from these sources by using similar methods as those for bovine spinal cord, but failed to isolate endogenous ligands.

Report

(3 results)
  • 1991 Final Research Report Summary
  • 1990 Annual Research Report
  • 1989 Annual Research Report
  • Research Products

    (8 results)

All Other

All Publications (8 results)

  • [Publications] K.Kubota et al.: "cholecystokinin antagonism by anthramycin, a benzodiazeine antibiotic,in the central nervous system in mice" Brain Research. 485. 62-66 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kubota el al.: "Purification and characterization of endogenous benzocliazepine from bovine spiual cord"

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 羽里 忠彦ほか: "ウシ脊髄からの内因性ベンゾジアゼピン受容体リガンド" 日本薬学会第110年会. (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kazuhiko Kubota et al.: "Cholecystokinin antagonism by anthramycin, a benzodiazepine antibiotic, in the central nervous system in mice." Brain research. 485. 62-66 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] K.Kubota: "Cholecystokinin antagonism by anthramycin,a benzodiazepine antibiotic,in the central nervous system in mice" Brain Resarch. 485. 62-66 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] K,Kubota: "Purification and characterization of endogenous benzodiazepine from cow spinal cord" Life science.

    • Related Report
      1990 Annual Research Report
  • [Publications] 羽里 忠彦: "ウシ脊随からの内因性ベンゾジアゼピン受容体リガンド分離の試み" 日本薬学会第110年会. (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 高橋、羽里、久保田ほか: "ウシ中枢神経中の内因性ベンゾジアビピンクガンドの検索" 日本薬業会第101年会.

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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