Preparative-scale high-performance liquid chromatographic separation and purification of 3′-azido-3′-deoxythymidine-5′-phosphate

Abstract

Crude 3′-azido-3′-deoxythymidine-5′-phosphate (AZT-P), obtained from direct phosphorylation of 3′-azido-3′-deoxythymidine (azidothymidine, AZT), was separated and purified by isocratic preparative high-performance liquid chromatography. The components in a 2.5-g load of crude AZT-P, obtained from work-up of the phosphorylation reaction, were separated in 50 min to give 1.8 g of 99.5% pure AZT-P. AZT-P was analyzed by high-performance liquid chromatography and by high-resolution nuclear magnetic resonance ( 1H, 13C, P spectroscopy. The practical and rapid preparative chromatographic method is being applied to the purification of AZT-P and other antiretroviral dideoxynucleotides, used as intermediates in the synthesis of target-directed experimental drugs for the treatment of AIDS.