| Project/Area Number |
07557159
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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 |
Biological pharmacy
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| Research Institution | Faculty of Pharmaceutical Sciences, Nagoya City University |
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Principal Investigator |
ONOZAKI Kikuo Fac.Pharmaceu.Sci., Nagoya City Univ., 薬学部, 教授 (20101313)
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| Co-Investigator(Kenkyū-buntansha) |
FURUTANI Taiji BioScience Inst.Dainippon Phar.Co.Ltd., 生物科学研究所, 主任研究員
KUWASHIMA Junji BioScience Inst.Dainippon Phar.Co.Ltd., 生物科学研究所, 部長
HAYASHI Hidetoshi Fac.Pharmaceu.Sci., Nagoya City Univ., 薬学部, 助教授 (80198853)
CHIBA Taku Fac.Pharmaceu.Sci., Nagoya City Univ., 薬学部, 講師 (10094385)
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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 |
¥7,500,000 (Direct Cost: ¥7,500,000)
Fiscal Year 1996: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1995: ¥5,900,000 (Direct Cost: ¥5,900,000)
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| Keywords | neoglycoprotein / cytokine / interleukin 1 / glycotechnology / arteficial glycoprotein / インターロイキン / 糖 |
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| Research Abstract |
IL-1 is potentially useful for therapy because of its antitumor effect and protective effect on animals against infection, radiation and chemotherapy. However, IL-1 exhibits serious deleterious effects. In this study we attempted to develop IL-1 with less deleterious effects by chemically introducing oligosaccharides to recombinant human IL-1alpha.
1) Tissue distribution of mannose dimer-introduced IL-1
We have previously succeeded in the synthesis of mannose dimer-introduced IL-1alpha with selective activities in vivo. In this study the effect of glycosylation on tissue distribution was examined. Mannose dimer-introduced IL-1 was labeled with ^<125>I,and then examined its tissue distribution after i.p.or i.v.injection into mice. Compared to the nonglycosylated IL-1, the glycosylated IL-1 distributed more in the liver, and less in the kidney.
2) Analysis of glycosylated site in mannose dimer-introduced IL-1
Mannose dimer-introduced IL-1 was dijested with lysylendopeptidase, and glycosylate
… More
d sites were analyzed by TOF-MASS.The conserved regions among IL-1alpha and IL-1beta were not glycosylated. However, the regions exposed to the surface of IL-1alpha molecule were glycosylated.
3) Synthesis, purification and biological activities of galactose-introduced IL-1
D-Galactose monosaccharide was coupled to IL-1alpha by the acyl azide method, and was purified by anion-exchange column chromatography. Galactose introduced per molecule of IL-1 was estimated to be 9. We examined a variety of IL-1 activities in vitro, including proliferative effect on T cells, antiproliferative effect on myeloid leukemia cells and melanoma cells, stimulatory effects on IL-6 syntheisi by melanoma cells and PGE2 synthesis by fibroblast cells. The glycosylated IL-1 exhibited reduced activities from ten to thousand times compared with nonglycosyslated IL-1 in all the activities performed in vitro. The glycosylated IL-1 exhibited reduced binding affinities to type I and II IL-1 receptors, suggesting that the reduced activities are due, at least partially, to the decrease in the binding affinities. Less
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