A general and efficient approach to synthesize the phosphoramidites of 5'-18O labeled purine nucleosides

Abstract

5'-¹⁸O labeled RNA oligos are important probes to investigate the mechanism of 2'-O-transphosphorylation reactions. Here we describe a general and efficient synthetic approach to the phosphoramidite derivatives of 5'-¹⁸O labeled nucleosides starting from the corresponding commercially available 5'-O-DMT protected nucleosides. Using this method, we prepared 5'-¹⁸O-guanosine phosphoramidite in 8 steps (13.2% overall yield), 5'-¹⁸O-adenosine phosphoramidite in 9 steps (10.1% overall yield) and 5'-¹⁸O-2'-deoxyguanosine phosphoramidite in 6 steps (12.8% overall yield). These 5'-¹⁸O labeled phosphoramidites can be incorporated into RNA oligos by solid phase synthesis for determination of heavy atom isotope effects in RNA 2'-O-transphosphorylation reactions.

Keywords: 18O labeled RNA; 18O labeled nucleoside phosphoramidite; 2′-O-transphosphorylation; Isotopic substitution; kinetic isotope effect.