Synthesis of uridine-5-propylamine derivatives and their use in affinity chromatography of N-acetylglucosaminyltransferases I and II.

The C-5 substituted uridine derivatives UDP-5-propylamine (7) and UDP-GlcNAc-5-propylamine (8) were synthesized in good yields by Heck alkylation of the 5-mercuriuridines, followed by hydrogenation. The products were characterized by 1H and 13C NMR spectroscopy, electrospray mass spectrometry and UV spectrophotometry. The amines are of interest for the preparation of affinity probes for glycosyltransferases. The benzoylbenzamides of 7 and 8 show strong competitive inhibition of N-acetylglucosaminyltransferase I and II with Ki values ranging from 30 to 100 microM (without irradiation) and may be useful as active site-directed photoaffinity labels. A conjugate of 8 and Sepharose was used for affinity chromatographic purification of N-acetylglucosaminyltransferases I and II. The results indicate that this affinity gel is a stable alternative to the commonly used but unstable UDP-GlcNAc-5-Hg-thiopropyl conjugate.