Phosphorylation of Nucleosides with Sodium cyclo-Triphosphate

Abstract

Phosphorylation of nucleosides with sodium cyclo-triphosphate (P3m) was studied under various conditions (mixing ratio of P3m to nucleoside, pH, reaction temperature and time). 1) Adenosine, cytidine, guanosine, and uridine were easily phosphorylated by P3m to form selectively nucleoside 2′-monophosphates, 3′-monophosphates, and 2′,3′-cyclic monophosphates. On the other hand, 2′-deoxyadenosine, 3′-deoxyadenosine, 2′-deoxycytidine, and thymidine could not be phosphorylated by P3m. 2) The phosphorylation of adenosine, cytidine, guanosine, and uridine varied strongly, depending on the reaction conditions; mixing ratio, pH, reaction temperature and time. Under conditions of a high mixing ratio of P3m to nucleoside (5:1–10:1), high pH (12–14), high temperature (70–100 °C) for a short time (1–2d) or low temperature (room temperature) for a long time (70–150 d), nucleoside 2′- and 3′-monophosphate could be obtained in high yield (about 50–100%). 3) At the initial stage of the phosphorylation reactions of adenosine, cytidine, and guanosine with P3m, nucleoside 2′,3′-cyclic monophosphates were formed, though a small amount (about 1–3%), but in the course of the reaction for a long time, they were hydrolyzed to nucleoside 2′- and 3′-monophosphates. 4) 2′-deoxyadenosine, 3′-deoxyadenosine, 2′-deoxycytidine, and thymidine could not be phosphorylated by P3m. This means that the presence of hydroxyl groups at the 2′- and 3′-positions of the ribose was necessary for the phosphorylation of nucleosides with P3m. 5) Phosphorylation of nucleosides with short-chain phosphates (ortho-, pyro-, and triphosphate) could not be observed at all. 6) The mechanism of formation of nucleoside 2′-monophosphates, 3′-monophosphates, and 2′,3′-cyclic monophosphates in the phosphorylation of nucleosides with P3m was discussed.

Phosphorylation of 5'-Deoxy-5-fluorouridine with Inorganic Phosphorylating Agents

“…We succeeded in the phosphorylation of nucleosides and nucleotides using inorganic phosphorylating agents such as cyclo-triphosphate 9,10) and monoimido-cyclo-triphosphate.…”

“…1, P 3m and MCTP have six-membered rings composed of three and two P-O-P linkages, respectively. In the phosphorylation of nucleosides 9) and nucleotides 10) by P 3m , the 2Ј-and 3Ј-OH groups of the b-D-ribofuranosyl unit on the nucleosides and nucleotides were selectively phosphorylated with the yield of 80-97% without protection of the other hydroxyl groups. In the reaction of nucleoside or nucleotides and P 3m , the main products were 3Ј-monophosphate or 3Ј,5Ј-diphosphate derivatives.…”

Phosphonylation of L-Dopa With Sodium Diphosphonate in Aqueous Solution

“…Sodium cyclo-triphosphate (P 3m ), Na 3 P 3 O 9 , which is simple inorganic condensed phosphate having a six-membered ring, is a very attractive phosphorylation reagent for synthesizing biologically important compounds in one step in aqueous solution [1][2][3][4] . The phosphorylation with P 3m proceeded under alkaline condition such as pH 12.…”

Phosphorylation of Saccharides by Inorganic Cyclo-Triphosphate in Aqueous Solution

“…pH 12, P 3m is easily attacked by nucleophilic ents such as amine 38 , nucleoside 39 , amino [40][41][42] , phenols 43 , and aminoalcohol 44,45 . The lone tron pair on the anomeric hydroxyl group of cing β-D-glucopyranose nucleophilically attacks osphorus atom on P 3m to open its six-membered (SCHENE 1).…”