CN113173961A

CN113173961A - Method for preparing adenosine

Info

Publication number: CN113173961A
Application number: CN202010927777.8A
Authority: CN
Inventors: 陈阳; 张磊; 李佳杨; 薛百忠
Original assignee: Yuanda Life Science Liaoning Co ltd
Current assignee: Yuanda Life Science Liaoning Co ltd
Priority date: 2020-09-07
Filing date: 2020-09-07
Publication date: 2021-07-27
Anticipated expiration: 2040-09-07
Also published as: CN113173961B

Abstract

The invention provides a method for preparing adenosine, which comprises the step of carrying out hydrolysis reaction on adenosine in a protic solvent under an acid buffer system. The preparation method of the invention realizes the high-efficiency hydrolysis of adenosine by using a specific acidity buffer system as a hydrolysis condition, and the prepared product has high purity and reasonable yield.

Description

Method for preparing adenosine

Technical Field

The invention relates to the field of drug synthesis, in particular to a preparation method of adenosine.

Background

Adenosine, adenosine nucleoside, is a compound in which N-9 of adenine and C-1 of D-ribose are linked by a β -glycosidic bond, and its phosphate ester is adenylic acid. Adenosine is an endogenous nucleoside, widely present in all types of cells, and is also one of the important active ingredients in the body. Adenosine can directly enter myocardium and be phosphorylated to generate adenylic acid, and participate in myocardial energy metabolism, coronary artery vessel dilation and blood flow increase, and can be used for treating angina pectoris, myocardial infarction, arteriosclerosis, essential hypertension, and apoplexy sequela. Meanwhile, it is an important intermediate of medicine, and can be used for synthesizing Adenosine Triphosphate (ATP), adenine, adenylic acid, vidarabine and the like. Therefore, adenosine has wide market development and application prospects.

CN102675391A discloses a chemical method for synthesizing adenosine, which uses inosine as a starting material, firstly carries out acetylation reaction to generate 2 ', 3 ', 5 ' -triacetyl inosine, then generates transition state salt in saturated ammonia-methanol solution in the presence of double catalysts, and then carries out ammonolysis to generate adenosine.

CN1408720 discloses a chemical method for synthesizing adenosine, which takes hypoxanthine as a raw material, uses a chlorine reagent for chlorination in an organic alkali solution, then carries out condensation reaction with alpha, beta-1, 2,3, 5-tetraacetyl ribofuranose under the catalysis of a phosphophenol ester compound to prepare 6-chloro-9- (beta-D-2 ', 3 ', 5 ' -triacetyl ribofuranose) purine, and then carries out pressurized ammonolysis to obtain the adenosine.

There is also a related document reporting a method for chemically synthesizing adenosine, which comprises the steps of synthesizing a purine ring and then combining with a sugar ring to produce adenosine.

The methods have the defects of long process route, low yield, high cost, environmental friendliness and the like.

Therefore, the problem to be solved is to find a new method for preparing adenosine, which is simple to operate, low in cost and environment-friendly.

Disclosure of Invention

The invention overcomes the defects in the prior art and provides a preparation method of adenosine. The preparation method of the invention adopts a specific acid buffer system as a hydrolysis environment, realizes the efficient hydrolysis process of the adenylic acid shown by the following route, has proper product yield and is more suitable for industrial production.

The invention is realized by the following technical scheme:

the invention provides a method for preparing adenosine, which comprises the step of carrying out hydrolysis reaction on adenosine in a protic solvent in the presence of an acid buffer system to generate adenosine.

In the above production method, the protic solvent may be selected from one or more of an alcohol solvent, a nitrile solvent, or water; preferably, the protic solvent is water.

In the above preparation method, the acid moiety of the acidic buffer system may be selected from one or more of formate, acetate, phosphate, citrate, oxalate, phthalate or fumarate, and the cation moiety may be selected from one or more of sodium ion, potassium ion or ammonium ion.

In the above preparation method, the buffer system is selected from one or more of citric acid/sodium citrate, oxalic acid/sodium oxalate or fumaric acid/sodium fumarate systems; more preferably, the buffer system is a citric acid/sodium citrate system.

In the above production method, the pH value of the hydrolysis reaction may be 3.0 to 6.0; preferably, the pH of the hydrolysis reaction is 4.7-5.5.

In the above preparation method, the temperature of the hydrolysis reaction may be 60 ℃ to the reflux temperature of the reaction system; preferably, when water is used as the solvent, the temperature of the hydrolysis reaction is 90 ℃ to the reflux temperature; when alcohols are used as a solvent, the temperature of the hydrolysis reaction is 60 ℃ to the reflux temperature; when nitrile is used as a solvent, the temperature of the hydrolysis reaction is 80 ℃ to the reflux temperature; more preferably, the temperature of the hydrolysis reaction is the reflux temperature of the reaction system.

In the preparation method, the time of the hydrolysis reaction can be 20-60 h; preferably, the time of the hydrolysis reaction is 40-50 h.

In the above production method, the method further comprises a step of purifying the produced adenosine using alumina column chromatography and/or gel column chromatography. In one embodiment, alumina column chromatography uses an aqueous solvent as the eluent, and gel column chromatography uses an aqueous solvent as the eluent.

The invention has the following advantages and positive effects:

1. the method is used for hydrolysis under the condition of an acid buffer system, the hydrolysis process is more stable, the types of generated impurities are less, the impurity content is obviously reduced, the impurity removal burden is reduced for the subsequent refining and purification steps, and the purity of the final product is improved.

2. The invention carries out hydrolysis under the condition of an acid buffer system, realizes the high-efficiency hydrolysis of adenosine and greatly improves the yield of products.

3. The adenosine is used as the starting material, the material is cheap and has sufficient market supply, and the solvent and the reagent used in the reaction process are both conventional reagents and are cheap, so that the cost is greatly reduced compared with other methods for preparing adenosine.

4. The adenosine prepared by the method can obtain higher yield and purity no matter simple post-treatment or separation and purification by alumina or gel column chromatography.

Detailed Description

The present invention will be described in further detail with reference to specific embodiments thereof, which will be better understood from the following examples. However, it should be readily understood by those skilled in the art that the following examples are illustrative only and are not intended to limit the present invention to these specific embodiments. It will be appreciated by those skilled in the art that the present invention encompasses all modifications, alternatives, and equivalents as may be included within the scope of the claims.

EXAMPLE 1 preparation of adenosine

Adenosine acid 10.0g was weighed into a reaction flask, 75mL of purified water was added, stirring was turned on, and then an aqueous NaOH solution was slowly added dropwise thereto until the adenosine acid was completely dissolved. 10.0g of sodium citrate was added to the reaction solution, and then citric acid was slowly added thereto to adjust the pH to 4.7. Heating, refluxing for 40h, stopping heating and naturally cooling the reaction solution to room temperature. The reaction solution was concentrated to remove salt, and adenosine 4.2g was obtained in 54.5% yield and 99.5% purity.

EXAMPLE 2 preparation of adenosine

10.0g of adenylic acid was weighed into a reaction flask, 100mL of purified water was added, stirring was turned on, and then an aqueous KOH solution was slowly added dropwise thereto until all of the adenylic acid was dissolved. 10.0g of ammonium citrate was added to the reaction solution, followed by slowly adding acetic acid to adjust the pH to 6.0. Heating to 90 ℃, preserving the temperature for 60h, stopping heating and naturally cooling the reaction solution to room temperature. The reaction solution was concentrated to remove salt to obtain 5.8g of adenosine with a yield of 75.3% and a purity of 99.3%.

EXAMPLE 3 preparation of adenosine

Weighing 10.0g of adenosine, adding the adenosine into a reaction bottle, adding 60mL of methanol, starting stirring, and then slowly dropwise adding ammonia water into the reaction bottle until the adenosine is completely dissolved. 10.0g of sodium oxalate was added to the reaction solution, followed by slow addition of formic acid to adjust the pH to 3.0. Heating, refluxing for 20h, stopping heating and naturally cooling the reaction solution to room temperature. The reaction solution was subjected to concentration and desalting treatment to obtain 6.0g of adenosine, the yield was 77.9%, and the purity was 99.4%.

EXAMPLE 4 preparation of adenosine

Adenosine 10.0g was weighed into a reaction flask, 75mL of acetonitrile was added, stirring was turned on, and then an aqueous solution of NaOH was slowly added dropwise thereto until all the adenosine was dissolved. 10.0g of potassium dihydrogen phosphate was added to the reaction mixture, and then citric acid was slowly added thereto to adjust the pH to 5.2. Heating to 80 ℃, preserving the temperature for 50h, stopping heating and naturally cooling the reaction solution to room temperature. The reaction solution was treated with a gel column or an alumina column using an aqueous eluent to obtain 4.7g of adenosine, yield 61.0%, purity 99.4%.

In an embodiment of the present invention, the adenosine is prepared by a method comprising first dissolving adenosine in a protic solvent by adding an alkaline solution, and then subjecting adenosine to a hydrolysis reaction in the presence of an acidic buffer system to produce adenosine, wherein the type and concentration of the alkaline solution are used in such an amount that adenosine is dissolved in the protic solvent.

EXAMPLE 5 study of the Experimental parameters

On the basis of example 1, the relevant parameters were varied according to the following table, i.e. the same conditions as in example 1 except for the variations in the reaction conditions to be investigated, giving the following table of product yields and purities:

it can be seen that when the solvent is water, alcohol or nitrile, the yield and purity of the product are both high; when the solvent is acetone (aprotic solvent), both the yield and purity of the product are significantly reduced. When the buffer system is adopted, the yield and the purity of the product are high; when a non-buffered system is used, the yield of product is significantly reduced.

Furthermore, after studying a range of pH values from 2.0 to 10.0, it was found that the desired yield and purity could be obtained only at pH values from 3.0 to 6.0. Wherein the hydrolysis pH values of less than 3.0 and greater than 6.0 result in reduced yield and purity, and wherein hydrolysis pH values of 4.7 to 5.5 achieve high yields and purity, with hydrolysis pH values of 5.0 yielding the best results.

Further, the reaction time also has an influence on the experimental results, and an excessively short reaction time cannot give a desired yield and purity. When the reaction time is 20-60h, the ideal yield and purity can be obtained, when the reaction time is less than 20h, the yield and purity are both obviously reduced, and when the reaction time is continuously prolonged, the yield and purity are not improved any more.

Although the present invention has been described to a certain extent, it is apparent that appropriate changes in the respective conditions may be made without departing from the spirit and scope of the present invention. It is to be understood that the invention is not limited to the described embodiments, but is to be accorded the scope consistent with the claims, including equivalents of each element described.

Claims

1. A method for producing adenosine, which comprises subjecting adenosine to a hydrolysis reaction in a protic solvent in the presence of an acidic buffer system to produce adenosine.

2. The method according to claim 1, wherein the protic solvent is selected from one or more of an alcohol solvent, a nitrile solvent, or water; preferably, the protic solvent is water.

3. The method according to claim 1 or 2, wherein the hydrolysis reaction has a pH of 3.0 to 6.0.

4. The method of claim 3, wherein the hydrolysis reaction has a pH of 4.7 to 5.5.

5. A method according to any one of claims 1 to 4, characterised in that the acid moiety of the acidic buffer system is selected from one or more of formate, acetate, phosphate, citrate, oxalate, phthalate or fumarate and the cationic moiety is selected from one or more of sodium, potassium or ammonium ions.

6. A method according to any one of claims 1 to 5, wherein the buffer system is selected from one or more of citric acid/sodium citrate, oxalic acid/sodium oxalate or fumaric acid/sodium fumarate systems; more preferably, the buffer system is a citric acid/sodium citrate system.

7. The method according to any one of claims 1 to 6, wherein the temperature of the hydrolysis reaction is 60 ℃ to the reflux temperature of the reaction system; preferably, the temperature of the hydrolysis reaction is the reflux temperature of the reaction system.

8. The process of claim 7, wherein the hydrolysis reaction is carried out at a temperature of from 90 ℃ to reflux temperature with water as solvent; when alcohols are used as a solvent, the temperature of the hydrolysis reaction is 60 ℃ to the reflux temperature; the hydrolysis reaction is carried out at a temperature of from 80 ℃ to reflux temperature when nitriles are used as the solvent.

9. The process according to any one of claims 1 to 8, wherein the hydrolysis reaction time is 20 to 60 hours; preferably, the time of the hydrolysis reaction is 40-50 h.

10. The method according to any one of claims 1 to 9, wherein the method further comprises a step of purifying the adenosine produced using alumina column chromatography and/or gel column chromatography.