Nucleoside and deoxynucleoside phosphorylation in formamide solutions

Nucleosides or deoxynucleosides were converted to a number of phosphorylated nucleotide and deoxynucleotide derivatives by ammonium or alkali dihydrogen phosphates in formamide. Conversions were smaller and slower at room temperature and greater and faster at elevated temperatures. Nucleotides afforded product mixtures similar to those obtained for nucleosides under the same conditions, indicating the occurrence of transphosphorylation processes. Products of reaction at elevated temperatures were cyclic nucleotides, nucleoside monophosphates, nucleoside diphosphates and cyclic nucleotide phosphates. The relative amounts of products formed were quite temperature dependent. Cyclic nucleotides were found to be in greatest abudance for reactions run at 125° or above. Relative yields of 2, 3 and 5 nucleotides and 3 and 5 deoxynucleotides from several experiments are reported. 5-Monophosphates were generally found to be present in larger quantities than 2 or 3 monophosphates. 2-Deoxyadenosine showed a preference for phosphorylation at the 3 position. Conclusions reached from mechanistic studies are that the phosphorylations are a series of equilibrium reactions, with cyclic nucleotides being formed irreversibly.Presented in part at the 3rd Northwest and 5th Rocky Mountain Joint Regional ACS Meeting of the American Chemical Society, Salt Lake City, Utah, June, 1980.