CN113292611A

CN113292611A - Cytarabine purification method

Info

Publication number: CN113292611A
Application number: CN202110712119.1A
Authority: CN
Inventors: 王淑娟; 徐昊; 李铁军; 李丹丹
Original assignee: Sinopharm A Think Pharmaceutical Co Ltd
Current assignee: Sinopharm A Think Pharmaceutical Co Ltd
Priority date: 2021-06-25
Filing date: 2021-06-25
Publication date: 2021-08-24
Anticipated expiration: 2041-06-25
Also published as: CN113292611B

Abstract

The invention relates to the technical field of medicines, and particularly relates to a method for purifying cytarabine. The method adopts alkaline substances to purify the crude cytarabine, has good batch consistency and controllable quality, and the purity can reach more than 99.9 percent. Compared with the traditional resin column purification, the method has the characteristics of simple operation, low energy consumption, short production period, low toxicity, harmlessness, green and environmental protection of the solvent and low purification cost, and is more suitable for industrial production.

Description

Cytarabine purification method

Technical Field

The invention relates to the technical field of medicines, and particularly relates to a method for purifying cytarabine.

Background

Cytarabine, chemically known as 1- β -D-arabinofuranosyl-4-amino-2 (1H) -pyrimidinone, was first synthesized in 1959 by RichardWalwick, Walden Roberts and Charles Dekker, university of California, Berkeley, division.

Cytarabine for injection was first marketed in the United states by Upjohn (Pfizer, the genus) and 6 months 1969 under the name of

The specifications on the market are 100mg, 500mg, 1g and 2g, which are the reference formulations recommended by the American orange book, but are currently on the market (for reasons of non-safety and effectiveness). HOSPIRA was approved by the FDA in 1990 to market a 20MG/ML injection, recommended as RS.

France: in 1985, PfThe marker is cytarabine for injection on the market in France and the commercial name is

The specification is 500mg, 1.0g and 2.0g are newly increased in 1987, and 100mg is newly increased in 1997.

Italy: pfizer is marketed in Italy as injectable cytarabine under the trade name Pfizer

The specification was 100mg (with 5ml solvent), 500mg (with 10ml solvent), and time to first market/re-registration was 2010.

Spain: in 1970, Pfizer marketed Spain with 100mg specification for injection, and newly increased 500mg specification in 1984.

The product is prepared by firstly approving the Japan neo-medicine company in 1971 at 4 months, wherein the product is prepared by キロサイド -injection of 20mg, and then sequentially marketing 40mg, 60mg, 100mg, 200mg, 400mg and 1g of the preparation in specification, which are all injections. Wutian 1248686is approved in 2013 by Tokayao K.K. of バファーマ, and the commercial specification is 400mg and 1g of intravenous injection.

Indications are as follows: can be used alone or in combination with other chemotherapy drugs, and is suitable for treating leukemia, especially acute myelocytic leukemia of adults or children. Can also be used for inducing and relieving acute lymphocytic leukemia, chronic myelocytic leukemia and erythroleukemia, and for treating and maintaining meningeal leukemia and other meningeal malignant tumors

The action mechanism is as follows: the product is a pyrimidine antimetabolite mainly acting on the proliferation stage of cells S, and interferes the proliferation of cells by inhibiting the synthesis of cell DNA. Cytarabine enters a human body and is converted into cytarabine triphosphate and cytarabine diphosphate after phosphorylation by kinase, the cytarabine can strongly inhibit the synthesis of DNA polymerase, and the cytarabine diphosphate can inhibit the conversion of the cytidine diphosphate into the deoxycytidine diphosphate, so that the DNA polymerization and the synthesis of cells are inhibited.

The synthetic route is as follows: the synthetic routes reported in the literature are suitable for the preparation of cytarabine starting material, but according to document 1(Zhejiang Haizhig pharmaceutical industry Co., Ltd. a method for preparing cytarabine CN1302004C.2006.01;) document 2([24.Synthesis of (C.))¹⁵N₂)[¹⁷O]Urea,(¹⁵N₂)[O₂,O^4-17O₂]Uridine and(¹⁵N₃)[O^2-17O]Cytidine, byAntonio Antael), Matthias Henz2), and Peter stratazewsk. HELVETICA CH IMICAAC TA-Vol.79 (1996)) reported that cytarabine was prepared in two parts: 1) taking an inert organic solution as a solvent, carrying out ammonolysis by adopting ammonia water and then adopting methanolic ammonia, and concentrating a reaction system to obtain a concentrated residue (crude cytarabine); 2) and purifying the residue (crude cytarabine) by adopting a cation exchange resin column, eluting by using deionized water and eluting by using dilute ammonia water, collecting product eluent, and concentrating the eluent in vacuum to obtain the crude cytarabine.

The aftertreatment of the crude cytarabine product needs resin purification, a large amount of ammonia water eluent is generated, and the ammonia water eluent needs rapid low-temperature concentration and neutralization, so that the method is time-consuming, energy-consuming and low in yield.

Therefore, the invention develops and improves the cytarabine purification process more suitable for industrial production, and finds a method suitable for the industrial purification of cytarabine, which has important practical significance for the industrial production of cytarabine.

Disclosure of Invention

In view of the above, the invention provides a method for purifying cytarabine. The cytarabine obtained by the method has the characteristics of stable character, uniform granularity, high purity and yield, high purity of over 99.9 percent, simple operation, low energy consumption, safety, environmental protection, short production period and low production cost, and is more suitable for industrial production.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a method for purifying cytarabine, which comprises the following steps:

mixing the cytarabine crude product with an alkaline substance, and stirring for reaction;

concentrating the reaction solution under reduced pressure to dryness, adding an organic solvent into the concentrate for dispersion, and filtering;

drying the obtained filter residue to obtain cytarabine;

the alkaline substance is at least one of ammonia water, triethylamine, DIPEA, sodium carbonate and potassium carbonate.

In some embodiments, the basic substance is ammonia. In some embodiments of the invention, the alkaline substance is 25-36 wt% of ammonia water, 2mol/L of potassium carbonate or 2mol/L of sodium carbonate. According to the invention, ammonia water with the concentration of 25-36 wt% and water can be diluted according to the volume ratio of 1: 1-1: 3 for use.

In some embodiments, the volume to mass ratio of the basic substance to the crude cytarabine is 5:1 in ml/g.

In some embodiments, the stirring reaction is performed at 30-40 ℃ for 10-15 hours. In some embodiments, the stirring reaction is performed at 30 ℃, 35 ℃ or 40 ℃ for 10h, 12.5h or 15 h.

In the present invention, the drying is preferably air-blast drying. Specifically, the temperature of the air-blast drying is 55-65 ℃, specifically 55 ℃, 60 ℃ or 65 ℃, and the time of the air-blast drying is 6-8 hours, specifically 6 hours, 7 hours or 8 hours.

In some embodiments, the volumetric mass ratio of the concentrate to the organic solvent is 2: 1.

In some embodiments, the solvent used for the dispersion is one or more of ethyl acetate, ethanol, and water. In some embodiments, the solvent used for the dispersion is an organic solvent, specifically ethyl acetate or ethanol.

The method for purifying the cytarabine comprises the steps of mixing a cytarabine crude product with an alkaline substance, and stirring for reaction; concentrating the reaction solution under reduced pressure to dryness, adding an organic solvent into the concentrate for dispersion, and filtering; drying the obtained filter residue to obtain cytarabine; the alkaline substance is at least one of ammonia water, triethylamine, DIPEA, sodium carbonate and potassium carbonate. The method is used for purifying the crude cytarabine, the consistency among batches is good, the quality is controllable, and the purity can reach more than 99.9 percent. Compared with the traditional resin column purification process, the purification operation is greatly simplified, resin column chromatography purification and concentration are not needed, large-scale concentration equipment is avoided, the generation of a large amount of waste liquid is reduced, the purification operation of the cytarabine raw material is simpler, the yield is higher, the production cost is reduced, the method is more suitable for industrial production, and the stable method suitable for industrial purification of cytarabine is mainly provided.

Drawings

FIG. 1 shows a liquid chromatogram of cytarabine after purification according to example 1;

FIG. 2 shows a liquid chromatogram of cytarabine after purification according to example 2;

FIG. 3 shows a liquid chromatogram of cytarabine after purification according to example 3;

FIG. 4 shows a liquid chromatogram of cytarabine after purification according to example 4;

FIG. 5 shows the liquid chromatogram of cytarabine after purification in comparative example 1.

Detailed Description

The invention provides a method for purifying cytarabine. Those skilled in the art can modify the process parameters appropriately to achieve the desired results with reference to the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention.

The test materials adopted by the invention are all common commercial products and can be purchased in the market.

The invention is further illustrated by the following examples:

example 1

Adding ammonia water (V) into a reaction kettle_{Dilute ammonia water}：m_{Cytarabine crude product}Stirring the mixture with the ratio of 5:1, adding 15kg of crude cytarabine, controlling the temperature of the system to be 30-40 ℃, and stirring the mixtureStirring for reaction for 15h, concentrating the above system under reduced pressure to dryness, and collecting solid with ethyl acetate (V)_{Ethyl acetate}：m_Cytarabine2:1), filtering, drying the solid by blowing at 65 ℃ for 6h to obtain 13.5kg of purified cytarabine with the yield of 90.0 percent, and the chromatographic detection result is shown in a figure 1.

Example 2

Adding ammonia water (V) into a reaction kettle_{Dilute ammonia water}：m_{Cytarabine crude product}Stirring the mixture at a ratio of 5:1, adding 15kg of crude cytarabine, controlling the temperature of the system to be 30-40 ℃, stirring the mixture for reaction for 15h, concentrating the system under reduced pressure until the system is dry, and adopting ethanol (V) as a solid_Ethanol：m_Cytarabine2:1), filtering, drying the solid by blowing at 55 ℃ for 8h to obtain 13.6kg of purified cytarabine with the yield of 90.7 percent, and the chromatographic detection result is shown in figure 2.

Example 3

Adding 2mol/L potassium carbonate aqueous solution (V) into a reaction kettle_{Aqueous potassium carbonate solution}：m_{Cytarabine crude product}Stirring the mixture at a ratio of 5:1, adding 10kg of crude cytarabine, controlling the temperature of the system to be 30-40 ℃, stirring the mixture for reaction for 15h, concentrating the system under reduced pressure until the system is dry, and taking ethanol/water (V) as a solid_{Ethanol/water}：m_Cytarabine2:1), filtering, drying the solid by blowing at 60 ℃ for 7h to obtain 8.1kg of purified cytarabine with the yield of 81 percent, and the chromatographic detection result is shown in figure 3.

Example 4

2mol/L sodium carbonate aqueous solution (V) is added into the reaction kettle_{Aqueous sodium carbonate solution}：m_{Cytarabine crude product}Stirring the mixture at a ratio of 5:1, adding 10kg of crude cytarabine, controlling the temperature of the system to be 30-40 ℃, stirring the mixture for reaction for 15h, concentrating the system under reduced pressure until the system is dry, and taking ethanol/water (V) as a solid_Ethanol：m_Cytarabine2:1), filtering, drying the solid by blowing at 60 ℃ for 8h to obtain 8.3kg of purified cytarabine with the yield of 83%, and the chromatographic detection result is shown in figure 4.

Comparative example 1

And (2) purifying the crude cytarabine product by using 100g of ion exchange resin, eluting with deionized water, eluting with 5% ammonia water, collecting 500mL of product eluent, concentrating under reduced pressure and vacuum to dryness to obtain the crude cytarabine product, dissolving the crude cytarabine product in water and ethanol in a refluxing manner, filtering while the product is hot, crystallizing the filtrate, filtering, washing the crystals with a small amount of ethanol, and drying the solids at 60 ℃ for 6-8h by blowing to obtain 60g of cytarabine with the yield of 60%. The results of the chromatographic measurements are shown in FIG. 5.

The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.

Claims

1. A method for purifying cytarabine, comprising:

concentrating the reaction solution under reduced pressure to dryness, adding a solvent into the concentrate for dispersion, and filtering;

drying the obtained filter residue to obtain cytarabine;

2. The method according to claim 1, wherein the basic substance is ammonia water.

3. The method according to claim 1, wherein the volume to mass ratio of the basic substance to the crude cytarabine is 5:1 in ml/g.

4. The preparation method according to claim 1, wherein the stirring reaction is performed at 30-40 ℃ for 10-15 h.

5. The method according to claim 1, wherein the stirring reaction is a stirring reaction at 35 ℃ for 12.5 hours.

6. The method of claim 1, wherein the drying is forced air drying.

7. The preparation method according to claim 4, wherein the temperature of the forced air drying is 55-65 ℃ and the time is 6-8 h.

8. The method according to claim 1, wherein the volume-to-mass ratio of the concentrate to the organic solvent is 2:1 in ml/g.

9. The method according to claim 1, wherein the solvent used for the dispersion is one or more of ethyl acetate, ethanol and water.