Development of New Glycosyltransferase Inhibitors

以2‘，3’-二-O-乙酰尿苷和对苯磺酰氯为原料合成了2‘，3’-二-O-乙酰尿苷5‘-对苯磺酸酯。去除乙酰基后的产物经核磁共振波谱和凝胶渗透色谱鉴定为聚5‘-对苯磺酸尿苷。这种含有尿苷的聚合物被用来对抗在α-乳清蛋白存在下合成乳糖的半乳糖转移酶。聚合物确实对酶有很强的抑制作用。该聚合物对从分枝杆菌中分离纯化的磷酸海藻糖合成酶也有抑制作用。ESCA测试证实了聚尿苷5‘-对苯磺酸盐在聚苯乙烯96孔板上的吸附。含半乳糖基转移酶的3T3-L1成纤维细胞在聚尿苷5‘-对苯磺酸包被板上的黏附力较未包被板增加。EDTA的加入抑制了细胞黏附数量的增加，说明细胞的黏附需要金属离子。这一结果表明，3T3-L1成纤维细胞与聚尿苷5‘-对苯磺酸涂层平板的黏附可能是通过细胞表面的半乳糖转移酶发生的，因为该酶需要一个二价金属离子。而不含半乳糖基转移酶的HEM细胞在聚尿苷5‘-对-S酪磺酸盐包被的平板上没有表现出特异性的黏附。此外，还认为3T3-L1细胞表面的半乳糖基转移酶与聚尿苷5‘-对苯磺酸之间的相互作用可能是细胞迁移的机制。

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).