Overview of the Synthesis of Nucleoside Phosphates and Polyphosphates

Abstract

This overview summarizes methodology used for the synthesis of nucleoside mono-, di-, and triphosphates. Selected techniques such as the Mitsunobu reaction, displacement reactions involving nucleoside 5'-tosylates, "anion-exchange" techniques, and phosphoramidite and phosphoramidate methodologies are highlighted. The chemistry of phosphorylation is detailed with respect to advantages and limitations under various conditions. Applicability of the methods toward the synthesis of analogs such as imidophosphates, phosphorothioates, and radiolabeled nucleotides is also addressed.

Toll-Like Receptor 3–TRIF Pathway Activation by Neospora caninum RNA Enhances Infection Control in Mice

“…Quantification of the parasite burden. The parasite burdens of peritoneal cells and other sampled tissues were determined by quantitative PCR (qPCR) normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH), as previously described (31). Briefly, DNA was extracted from 50 mg of cells or tissue using SDS and proteinase K, and its content was estimated by 260/280 ratio (NanoDrop; Thermo Scientific).…”

Solid‐Phase Synthesis of Oligonucleotide 5′‐(α‐P‐Thio)triphosphates and 5′‐(α‐P‐Thio)(β,γ‐methylene)triphosphates

“…The silyl groups were then hydrolyzed with triethylammonium bicarbonate (TEAB) buffer (Scheme 1). Further activation of phosphorothioate derivative p s T 6 with diphenyl phosphoryl chloride [19] (Scheme 2) was attempted by using different solvents and by changing the reaction time (Table 1). Further activation of phosphorothioate derivative p s T 6 with diphenyl phosphoryl chloride [19] (Scheme 2) was attempted by using different solvents and by changing the reaction time (Table 1).…”

“…An aliquot of solid-supported T 6 was deprotected, and the crude was analyzed by IEX-HPLC and MS to attest the quantitative conversion into p s T 6 (Figure 2). Further activation of phosphorothioate derivative p s T 6 with diphenyl phosphoryl chloride [19] (Scheme 2) was attempted by using different solvents and by changing the reaction time ( Table 1). The resulting mixed phosphoanhydride was then substituted with an alkylammonium salt of pyrophosphate (1 m in pyridine) to give ppp s T 6 ON 2 after ammonia deprotection; the yields ranged from 8 to 32 %.…”

Phosphodeoxyribosyltransferases, Designed Enzymes for Deoxyribonucleotides Synthesis

“…Whereas chemical phosphorylation is widely used, it requires several steps of protection, deprotection, and purification (36). It is only recently that a one-pot synthesis of deoxyribonucleoside 5Ј-triphosphates without any protection on the nucleosides was reported (37).…”