SIALIDASE SUBSTRATE, PRODUCTION THEREOF, METHOD FOR MEASURING SIALIDASE ACTIVITY USING THE SAME AND KIT FOR MEASURING THE ACTIVITY

摘要

NEW MATERIAL:Sodium p-nitrophenyl-N-acetyl-neuraminate expressed by formula I (Ac is acetyl). USE:A chemically stable substrate for measuring sialidase activity, good in preservation quality and capable of providing measured values of the sialidase activity with high accuracy and excellent reproducibility. PREPARATION:An N-acetyl-sialyl chloride expressed by formula II (R is acetyl) is reacted with p-nitrophenol sodium in DMF solvent and concentrated under reduced pressure and ethyl acetate and water are then added to separate the reaction solution. The separated ethyl acetate layer is subsequently concentrated. The resultant reaction product is subsequently separated by silica gel chromatography using a mixed solvent of ethyl acetate and toluene to recover a fraction containing a condensate of the sialyl chloride with the p-nitrophenol sodium. Methanol and sodium methylate are then added to carry out deacetylation and water is added to saponify the methyl ester. The resultant product is subsequently purified by using a weak acidic cation exchange resin.