Separation of individual antiviral nucleotide prodrugs from synthetic mixtures using cross-reactivity of a molecularly imprinted stationary phase

2ʹ,3ʹ-Dideoxynucleosides (ddNs) are among the most potent of nucleoside analogues active against human immunodeficiency virus (HIV) in cell culture. d4T (2ʹ,3ʹ-dideoxy-2ʹ,3ʹdidehydrothymidine) is clinically effective acting through competitive inhibition of viral reverse transcriptase and/or incorporation and subsequent chain termination of the growing viral chain. Activation occurs via intracellular conversion to the 5ʹ-triphosphate, the kinase-mediated formation of the monophosphate being the rate-limiting step and, therefore, strategies to deliver the monophosphate have been sought. This study uses a molecularly imprinted HPLC stationary phase to separate single diastereomers for a number of nucleoside monophosphates prodrugs from synthetic mixtures. The biological activity of some individual diastereomers are unknown and a need to efficiently separate them from synthetic mixtures, has been identified. Due to cross-reactive affinity for the imprinted polymer one imprinted stationary phase was used to isolate a single diastereomer from a range of prodrug synthetic mixtures.