A simple and rapid synthesis of nucleotide analogues containing a phosphorothioate moiety at the terminal position of the phosphate chain

Abstract

A straightforward method for the synthesis of nucleotide analogues bearing a phosphorothioate moiety at the terminal position of the polyphosphate chain is described. Several nucleoside 5′-(2-thiodiphosphates) and 5′-(3-thiotriphosphates) were synthesized by treatment of the appropriate nucleotide imidazolide derivative with a ca. 4-fold excess of thiophosphate triethylammonium salt in DMF in the presence of zinc chloride. The HPLC reaction yields varied from 80% to 100%, in the majority of cases exceeding 90%. Separation was accomplished by Sephadex ion-exchange chromatography or reverse-phase HPLC with preparative yields of about 70%.