SU504784A1 - The method of obtaining ribonucleoside-5 monophosphates - Google Patents

Description

one

This invention relates to an improved process for the preparation of compounds which are used in biochemistry.

A known method for preparing ribonucleoside 5-monophosphates is phosphorylated corresponding ribonucleosides with preliminary protection of the 2, 3-cis-diol hydroxyl groups of ribonucleosides not involved in the formation of phosphodiester bonds. For this purpose, 2, 3-O-alklidene-nucleosides are used.

However, the known method has a number of significant drawbacks, the main of which is the difficulty of removing the protective group carried out under severe conditions leading to the preparation of noboche substances. As a result, additional purification of the target product is required.

The proposed method makes it possible to simplify the technology for the preparation of the target product due to the use of puleoside arnbornate esters as intermediate compounds.

The described method for preparing the ribonucleoside-5-monophosphates is to that ribonucleosides treated an arylboronic acid in nol polar solvents, e.g. dnoksane, pyridine or dnmetilformamide then obtained arnlbornye esters is subjected to phosphorylation | shyu, removal zaschitNOI group with water and the desired product recovery known method .

As a rule, P-difennl-P-morpholidopyrophosphochloride in dioxane or p-cyaethyl phosphate or 2-cyanoethyl phosphate in pyridine is used as phosphorylating agents.

Example 1. To a solution of 5.5 g of anhydrous diphenylphosphoric acid in 40 ml of dioxane, 4.5 g of morpholidophosphodichloride and 3.2 ml of 2,6-lutidine are added with stirring. After 15 minutes, adenosine phenylborne efnr in 12 ml of dioxane is added (obtained by heating a mixture of 6.2 g of adenosine and 3.06 g of ph | enylboric acid in donoxane; it can be isolated in analytical form in a yield of 95% by subsequent evaporation of dioxane and adding 10 -fold amount of sulfuric ether) and the mixture is left under stirring at 18-20 ° C for 48 hours. In these conditions, the reaction is accompanied by a quantitative and selective phosphorylated nervous hydroxyl group of nucleosides (chromatographic control of the sample after preliminary hydrolysis in the system: ethanol — 0.5L1 ammonium acetate, saturated with brown 2: 2). Then 300 ml of water (pI solution 1.6-2.0) H are stirred for 4 hours at 30-40 ° C until complete hydrolysis of the P-N and P-CI bonds formed during phosphorylation of adenosine-5 phosphomorphyl chloride. (Analysis t. C.

chromatography in the system: acetone - / g-butanol - H2O 8: 1: 1). The resulting solution is neutralized with a 3% ammonia solution to a pH of 6.0 and placed at 5-8 ° C on a cooled column with KU-2 (H +) resin (6.0X X 5.30). The column is washed with water from impurities of phosphoric, diphenylphosphoric n phenylboronic acids. Adenosine-5-monophosphate is diluted in the form of ammonium salt with 1% ammonia solution. 75% yield (spectrophotometric measurement). The ammonium salt solution is evaporated to a concentration of nucleotide material of 15–20% and acidified with 10% hydrochloric acid to a pH of 1.8. The precipitate formed during cooling is filtered off, washed with water, alcohol, dried in vacuum at 40-45 ° C. Get about 3 g (58-60%) of adenosine 5-monophosphate; content of the main substance is 98.5-100%; impurities of the original nucleoside and isomeric nucleoside mono- and diphosphatone are absent {chromatographic and electrophoretic control); impurity of PO4 and C1-ions does not exceed 0.1%.

Example 2. To a solution of 1.25 g of diphenylphosphoric acid in 10 ml of dioxane was added 1.03 g of morpholidophosphodichloride and 0.7 ml of 2,6 lutidine. After 15 minutes, 1.3 g / g-tolylboronic ester of adenosine was added (obtained from 1.04 g of adenosine and 0.54 g of p-tolylboronic acid, similarly to phenylboronic acid ester; the yield was 91% in pure form) and left with stirring and 18-20 ° C for 48 hours (control of the end of the chromatographic reaction). Treatment of the reaction product was the same as Example 1. The yield of adenosine-5-monophosphate was 55%. Content of the main substance is 98.5%.

Claims (1)

Invention Formula A method for preparing ribonucleoside-5-myophosphates by phosphorylation of corresponding riboucleosides with preliminary protection of hydroxyl groups and subsequent isolation of the target product by a known method, characterized in that, in order to simplify the process, ribonucleosides are treated with arylboronic acid in polar solvents, for example dioxane, pyridine, or dimethylformamide after which the resulting arylborne esters are phosphorylated, followed by removal of the protective group by treatment with water.