| Project/Area Number |
07557014
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
General medical chemistry
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| Research Institution | The University of Tokushima |
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Principal Investigator |
YAMAMOTO Shozo Department of Biochemistry, The University of Tokushima, School of Medicine Professor, 医学部, 教授 (50025607)
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| Co-Investigator(Kenkyū-buntansha) |
KATORI Makoto Kitasato University School of Medicine Professor, 医学部, 教授 (50050365)
HIGUCHI Shohei Taisho Pharmaceutical Company, Development and Research Institute Division Head, 開発研究所, 室長
HARADA Yoshiteru Kitasato University School of Medicine Professor, 医学部, 教授 (20050677)
TANABE Tadashi National Cardiovasculal Disease Center, Research Institute Division Head, 研究所, 室長 (60025624)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥15,500,000 (Direct Cost: ¥15,500,000)
Fiscal Year 1996: ¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1995: ¥9,000,000 (Direct Cost: ¥9,000,000)
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| Keywords | Antiinflammatory drug / Inflammation / Prostaglandin / Cyclooxygenase / Isozyme / Glucocorticoid / Steroid / Enzyme inhibitors / シクロオキシゲナーゼ-2 / トロンボキサン / ウエスタンブロット法 / ノーザンブロット法 |
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| Research Abstract |
Fatty acid cyclooxygenase (COX) initiates the synthesis of bioactive prostaglandins and thromboxanes, and has been considered as a target of anti-inflammatory drugs. The enzyme had been found in vesicular gland, platelets, stomach and various other animal tissues. In 1991 an isozyme of the enzyme (COX-2) was discovered, and found to be an inducible enzyme. Since COX-2 is induced rapidly and transiently by LPS and proinflammatory cytokines, more attentions have been directed to COX-2 rather than COX-1 in the development of anti-inflammatory drugs. The purpase of this project is to develop and establish practicallly convenient assay methods of the protein and mRNA of COX isozymes and the in vivo effect of anti-inflammatory drugs.
1) Antibodies were raised with a COX-2 fragment with a specific sequence. COX-1 selective antibodies were found among monoclonal antibodies previously raised against the enzymes of bovine vesicular gland and human platelets. These antibodies for COX-1 and COX-2 could be applied to Western blottings of the enzymes.
2) Amplification of mRNAs for COX-1 and COX-2 by RT-PCR was attempted to develop sensitive and convenient assays. Non-radioactive probes by the use of a fluorescent label are being prepared.
3) For the assessment of in vivo effects of drugs, diseases models were investigated. Rat pleurisy induced by carrageenin was used for COX-2 studies. Non-treated (for COX-1) and LPS-treated (for COX-2) rats received arachidonic acid intravenously, and the changes of arachidonate metabolites in the blood were determined quantitatively upon administration of drugs.
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