Phosphorylation of D-Glucose Derivatives with Inorganic Monoimido-cyclo-triphosphate

Abstract

Sodium cyclo-triphosphate, Na 3 P 3 O 9 (P 3m ), is a simple and efficient phosphorylating agent. Previously, we reported that alkylamines, 1) aminoalcohols, 2) and carbohydrates [3][4][5][6][7][8] are readily phosphorylated with P 3m to give their triphosphate derivatives. In particular, the reaction of D-glucose with P 3m in aqueous solution afforded 1-O-triphospho-b-D-glucose stereoselectively in good yield (47%) in a one-step process without protection of hydroxyl groups. [3][4][5] This reaction was found to be applicable to other monosaccharides and oligosaccharides. [6][7][8] Unfortunately, the products formed from carbohydrates are easily decomposed to monophosphate derivatives.Imido-cyclo-triphosphates, Na 3 P 3 O 9Ϫn (NH) n , have been known since 1962. 9) Compared with the P-O-P linkage, the P-NH-P linkage is stable and difficult to hydrolyze. 9) We therefore explored the use of monoimido-cyclo-triphosphate (MCTP) for the phosphorylation of biologically important compounds. The structure of MCTP is shown in Chart 1: the six-membered ring is composed of one P-NH-P and two P-O-P linkages. We recently demonstrated that the phosphorylation of methylamine 10) and amino acids 11) proceeded with MCTP. More recently, we reported that cyclodextrins reacted with MCTP to form 2-O-diphosphoramidophosphonocyclodextrins regioselectively in aqueous solution.12) Organic compounds containing an amino or a hydroxyl group were easily phosphorylated by MCTP. Kamasaka et al. 13) have reported that phosphoryl oligosaccharides had the ability to form a soluble complex with calcium and had an inhibitory effect on the formation of calciumphosphate precipitate. Moreover, phosphoryl oligosaccharides effectively enhanced remineralization of enamel and dentin lesion. From the point of chelating of calcium by them, triphosphoryl saccharides lead to an advantageous food ingredient as a soluble calcium. This ability would be improved by diphosphoramidophosphate ester because it was not hydrolyzed to the imidophosphate ester and diphosphate.In the present work, we first studied the reaction of D-glucose with MCTP in aqueous solution, and then investigated the phosphorylation of D-glucose derivatives by MCTP in order to develop a one-step synthesis of phosphorylated monosaccharides. Results and DiscussionPhosphorylation of D-Glucose (1), D-Glucuronic Acid (2) and 2-Deoxy-D-glucose (3) with MCTP D-Glucose (1) and its derivatives used in the present study are shown in Table 1. Phosphorylation was carried out essentially according to the previously described method.10-12) Figure 1 shows HPLC profiles for the reaction mixture of D-glucose (1) (1.5 M) and MCTP (0.3 M) incubated at pH 12 and 25°C. A peak attributed to the phosphorylated product (4) appeared at a retention time of about 20 min. The other chromatographic peaks were assigned to MCTP and its hydrolytic product (monoimidotriphosphate, MTP), respectively.To identify the product 4,

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

“…At pH 12, P 3m and MCTP are easily attacked by nucleophilic reagents such as amines, 16,17) amino acids 18,19) and D-glucose derivatives. 14,15) In the present study, the lone electron pair on the hydroxyl group of the 5Ј-deoxy-b-D-ribofuranose unit nucleophilically attacks a phosphorus atom of P 3m or MCTP, cleaving its six-membered ring. The reaction of nucleosides or nucleotides with P 3m gave triphosphate derivatives of nucleosides and nucleotides, which were immediately hydrolyzed to give nucleoside 2Ј-monophosphate and nucleoside 3Ј-monophosphate or nucleoside 2Ј,5Ј-diphosphate and nucleoside 3Ј,5Ј-diphosphate via a 2Ј,3Ј-cyclic monophosphate derivative.…”

“…15) This product shows a characteristic P a signal around 1 ppm in the 31 P-NMR spectrum. In the 1 H decoupled-31 P-NMR spectrum (Fig.…”

Phosphorylation of Nucleosides and Nucleotides with Inorganic Monoimido-cyclo-Triphosphate

“…More recently, we reported that D-glucose, D-glucuronic acid and 2-deoxy-D-glucose reacted with MCTP to form 1-O-diphosphoramidophosphono-b-D-aldoses stereoselectively. 22) Organic compounds containing amino or hydroxyl group were easily phosphorylated by MCTP. We also reported that D-glucose and gluco-oligosaccharides reacted with DCTP.…”

“…method. [20][21][22] HPLC analysis served as a tool for evaluating the yields of products from their peak area. Figure 3 shows HPLC profiles for the reaction mixture of adenosine (1) (0.1 M) and MCTP (0.4 M) incubated at pH 12 and 40°C.…”

“…In the 31 P-NMR spectra, the peak at 0.3 ppm was assigned to P a of 5, and the peak at 1.1 ppm to P a of 6. A previous study indicated that the phosphorylation products of Dglucose derivatives 22) with MCTP are diphosphoramidophosphono-D-aldoses with an -O-P a -NH-P b -bond. These products show a characteristic P a signal at around 0 ppm in their 31 P-NMR spectra.…”