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Development of New Glycosyltransferase Inhibitors

新型糖基转移酶抑制剂的开发

基本信息

批准号：
08044129
负责人：
HATANAKA Kenichi
金额：
$ 2.82万
依托单位：
Tokyo Institute of Technology
依托单位国家：
日本
项目类别：
Grant-in-Aid for international Scientific Research
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1998
项目状态：
已结题

项目摘要

2' , 3' -Di-O-acetyluridine 5' -p-styrenesulfonate was synthesized by the reaction of 2' , 3' -di-O-acetyluridine with p-styrenesulfonyl chloride and polymerized. After removal of acetyl groups, the polymeric product was shown by NMR spectroscopy and gel permeation chromatography to be poly(uridine 5' -p-styrenesulfonate). This uridine-containing polymer was tested against the galactosyl transferase that synthesizes lactose in the presence of alpha-lactalbumin. The polymeric compound did inhibit the enzyme strongly. And this polymer inhibited also trehalose phosphate synthase which was purified from Mycobacterium.The adsorption of poly(uridine 5' -p-styrenesulfonate) on the polystyrene 96-well mutiplate was confirmed by ESCA measurements. On the poly(uridine 5' -p-styrene-sulfonate)-coated plate, adhesion of 3T3-L1 fibroblast which has galactosyl transferase on the cell surface was increased, compared with non-coated plate. And the increase in adhesive cell number was inhibited by the addition of EDTA, indicating that the cell adhesion needs metal ions. This result showed the 3T3-L1 fibroblast adhesion on the poly(uridine 5'-p-styrene-sulfonate)-coated plate may occur through the galactosyl transferase on the cell surface because this enzyme needs a divalent metal ion. On the other hand, Hem cell which does not have galactosyl transferase on the cell surface did not show the specific adhesion on the poly(uridine 5' -p-s tyrene-sulfonate)-coated plate. Moreover, it was suggested that the cell migration may occur by using the interaction betwecn galactosyl transferase on the cell surface of 3T3-L1 and poly(uridine 5'-p-styrene-sul fonate).

以2'，3'-二-O-乙酰尿苷和对苯乙烯磺酰氯为原料，经聚合反应合成了2 '，3'-二-O-乙酰尿苷5' -对苯乙烯磺酸盐。除去乙酰基后，通过NMR光谱和凝胶渗透色谱法显示聚合产物为聚（尿苷5 ′-对苯乙烯磺酸盐）。针对在α-乳白蛋白存在下合成乳糖的半乳糖基转移酶测试该含尿苷的聚合物。该聚合物确实强烈抑制酶。用ESCA方法测定了聚尿苷5' -对苯乙烯磺酸钠在聚苯乙烯96孔板上的吸附量，证实了聚尿苷5' -对苯乙烯磺酸钠对磷酸海藻糖合成酶的抑制作用。在聚尿苷5' -对苯乙烯磺酸盐包被的培养板上，具有半乳糖基转移酶的3 T3-L1成纤维细胞的粘附性比未包被的培养板增加。并且添加EDTA后粘附细胞数量的增加受到抑制，表明细胞粘附需要金属离子。该结果表明，3 T3-L1成纤维细胞在聚（尿苷5 ′-对苯乙烯磺酸酯）包被的板上的粘附可能通过细胞表面上的半乳糖基转移酶发生，因为该酶需要二价金属离子。另一方面，在细胞表面上不具有半乳糖基转移酶的Hem细胞在聚（尿苷5' -β-苯乙烯-磺酸酯）涂覆的板上不显示特异性粘附。此外，还提示3 T3-L1细胞表面的半乳糖基转移酶可能与聚尿苷5 ′-对苯乙烯磺酸酯相互作用而发生细胞迁移。