CN110776520B

CN110776520B - Preparation method of cefazolin sodium impurity G

Info

Publication number: CN110776520B
Application number: CN201911048609.5A
Authority: CN
Inventors: 袁晓; 林顺权
Original assignee: Pi & Pi Technology Inc
Current assignee: Pi & Pi Technology Inc
Priority date: 2019-10-30
Filing date: 2019-10-30
Publication date: 2022-05-10
Anticipated expiration: 2039-10-30
Also published as: CN110776520A

Abstract

The invention discloses a preparation method of cefazolin sodium impurity G. According to the preparation method, the hydrolysis reaction of cefazolin sodium impurity D is adopted to obtain deacetylated cefazolin, the deacetylated cefazolin is subjected to cyclization reaction, and cefazolin sodium impurity G is obtained through separation and purification. The yield of the cefazolin sodium impurity G prepared by the method is 61.4-64.8%, the purity can reach 95%, the requirement of cefazolin sodium quality research can be met, and a technical basis is provided for improving the national quality standard of cefazolin sodium.

Description

Preparation method of cefazolin sodium impurity G

Technical Field

The invention relates to the field of pharmaceutical impurities, and in particular relates to a preparation method of cefazolin sodium impurity G.

Background

Cefazolin sodium (Cefazolin sodium), also named pioneer V, is the strongest cephalosporin of the first generation, has the advantages of strong bactericidal power, broad-spectrum antibacterial property, relative enzyme resistance, high efficiency, low toxicity, ideal pharmacokinetics and the like, has good antibacterial activity to other gram-positive cocci, and is the semisynthetic cephalosporin commonly used in clinic at present. Due to the production process and structural characteristics of antibiotics, research on related substances (impurities) is a key and difficult point in the quality control of medicines. Due to different processes, related substances of cephalosporin drugs have different sources, side reactions are introduced in the synthesis process, and starting materials and intermediates, degradation and the like are introduced, so that the quality of the drugs is reduced, and meanwhile, the impurities can cause reactions such as antibiotic allergy and the like, and even endanger life in severe cases. Therefore, the research on the cefazolin sodium impurities is particularly important, the synthesis and separation of the impurity monomers are essential to the research on the structure, toxicity and quality control of the cefazolin sodium impurities, the contrast is provided for the research on the quality of the medicine impurities, the safety limit of the impurities is controlled, and the medicine quality and the medication safety are improved.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provides the preparation method of the cefazolin sodium impurity G, which has mild reaction conditions, does not relate to ultralow temperature reaction, has few process steps and is suitable for pilot scale of laboratories.

In order to achieve the purpose, the technical scheme adopted by the invention is as follows:

the chemical structure of the cefazolin sodium impurity G in the invention is shown as the formula (I):

a preparation method of cefazolin sodium impurity G comprises the steps of hydrolyzing cefazolin sodium impurity D under an alkaline condition to obtain deacetyl cefazolin, carrying out cyclization reaction on the deacetyl cefazolin under an acidic condition, and carrying out separation and purification to obtain cefazolin sodium impurity G, wherein the specific route is as follows:

preferably, the preparation method of cefazolin sodium impurity G specifically comprises the following steps:

(1) weighing cefazolin sodium impurity D, dissolving the cefazolin sodium impurity D in water at room temperature, adjusting the pH value to be alkaline, and heating in an oil bath for hydrolysis reaction to obtain reaction liquid containing deacetyl cefazolin;

(2) and adjusting the pH value of the reaction solution containing the deacetylated cefazolin to be acidic, performing cyclization reaction under an acidic condition to obtain reaction solution containing cefazolin sodium impurity G, and finally performing separation, purification and freeze drying on the reaction solution to obtain the cefazolin sodium impurity G.

Preferably, in the step (1), the ratio of the cefazolin sodium impurity D to the water is 1g:50-100 mL.

Preferably, in the step (1), the pH is adjusted to 8 to 9 with a base.

Preferably, the base is sodium bicarbonate.

Preferably, in the step (1), the oil bath is heated at the temperature of 40-50 ℃ for 2-4 h.

Preferably, in the step (2), the pH is adjusted to 1-3 by acid, and the time of the cyclization reaction is 2-3 h.

Preferably, the acid is hydrochloric acid.

Preferably, the concentration of the hydrochloric acid is 2-4 mol/L.

Preferably, in the step (2), the final reaction solution is separated and purified by a C18 column, and then eluted with aqueous formic acid, aqueous acetonitrile acid and aqueous acetonitrile acid, respectively, and the pure fractions are diluted and then applied to a PIPI-02 column, eluted with acetonitrile, and then freeze-dried.

By optimizing the reaction conditions, the yield of the prepared cefazolin sodium impurity G is over 60 percent, and the purity is 95 percent.

Compared with the prior art, the invention has the following beneficial effects:

according to the preparation method, the hydrolysis reaction of cefazolin sodium impurity D is adopted to obtain deacetylated cefazolin, the deacetylated cefazolin is subjected to cyclization reaction, and cefazolin sodium impurity G is obtained through separation and purification. The yield of the cefazolin sodium impurity G prepared by the method is 61.4-64.8%, the purity of the cefazolin sodium impurity G reaches 95%, the requirement of cefazolin sodium quality research can be met, and a technical basis is provided for the national quality standard improvement of cefazolin sodium.

Drawings

FIG. 1 is an HPLC detection spectrum (chromatographic conditions: European pharmacopoeia 9.0) of cefazolin sodium impurity G.

FIG. 2 is an HPLC detection spectrum of a blank control (chromatographic conditions: European pharmacopoeia 9.0 edition).

FIG. 3 is an NMR spectrum of cefazolin sodium impurity G.

FIG. 4 is a carbon spectrum of the NMR detection of cefazolin sodium impurity G.

FIG. 5 is the HRMS detection mass spectrum of cefazolin sodium impurity G.

Detailed Description

To better illustrate the objects, aspects and advantages of the present invention, the present invention will be further described with reference to specific examples. It will be understood by those skilled in the art that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Example 1

The preparation method of cefazolin sodium impurity G described in this embodiment includes the following steps:

(1) weighing 2g of cefazolin sodium impurity D at room temperature, adding 100mL of water, adding sodium bicarbonate to adjust the pH to 8.5, heating in an oil bath at 50 ℃ for 4h for hydrolysis reaction to obtain reaction liquid containing deacetyl cefazolin;

(2) adding 3mol/L hydrochloric acid into the reaction solution containing deacetyl cefazolin to adjust the pH value to 2, carrying out a cyclization reaction for 3h to obtain a reaction solution containing cefazolin sodium impurity G, separating and purifying the reaction solution containing cefazolin sodium impurity G on a 70mL C18 column, eluting with 0.05% formic acid water, 1% acetonitrile acid water and 3% acetonitrile acid water respectively, diluting the pure product fraction by 2 times, loading the pure product fraction on a 70mL PIPI-02 column, eluting with 80% acetonitrile, and carrying out freeze drying to obtain cefazolin sodium impurity G, wherein the yield is 62.5%, and the purity is 95%.

Example 2

(1) weighing 2g of cefazolin sodium impurity D at room temperature, adding 100mL of water, adding sodium bicarbonate to adjust the pH to 9, heating in an oil bath at 50 ℃ for 4 hours to react, and obtaining reaction liquid containing deacetyl cefazolin;

(2) adding 4mol/L hydrochloric acid into the reaction solution containing deacetyl cefazolin to adjust the pH value to 1, carrying out cyclization reaction for 3h to obtain reaction solution containing cefazolin sodium impurity G, separating and purifying the reaction solution containing cefazolin sodium impurity G on a 70mL C18 column, and eluting with 300mL of 0.05% formic acid water, 1% acetonitrile acid water and 3% acetonitrile acid water respectively. Diluting the pure product fraction by 2 times, loading the pure product fraction on a 70mL PIPI-02 column, eluting with 80% acetonitrile, and freeze-drying to obtain cefazolin sodium impurity G with the yield of 64.8% and the purity of 95%.

Example 3

(1) weighing 2g of cefazolin sodium impurity D at room temperature, adding 160mL of water, adding sodium bicarbonate to adjust the pH to 9, and heating in an oil bath at 50 ℃ for 3 hours to react to obtain a reaction solution containing deacetyl cefazolin;

(2) adding 4mol/L hydrochloric acid into the reaction solution containing deacetyl cefazolin to adjust the pH value to 1, carrying out cyclization reaction for 2.5h to obtain reaction solution containing cefazolin sodium impurity G, separating and purifying the reaction solution containing cefazolin sodium impurity G on a 70mL C18 column, and eluting with 300mL of 0.05% formic acid water, 1% acetonitrile acid water and 3% acetonitrile acid water respectively. Diluting the pure fraction by 2 times, loading the pure fraction on a 70mL PIPI-02 column, eluting with 80% acetonitrile, and freeze-drying to obtain cefazolin sodium impurity G with yield of 62.8% and purity of 95%.

Example 4

(1) weighing 2g of cefazolin sodium impurity D at room temperature, adding 200mL of water, adding sodium bicarbonate to adjust the pH value to 8, and heating in an oil bath at 50 ℃ for 2h to react to obtain a reaction solution containing deacetyl cefazolin;

(2) adding 2mol/L hydrochloric acid into the reaction solution containing deacetyl cefazolin to adjust the pH value to 3, carrying out cyclization reaction for 2h to obtain reaction solution containing cefazolin sodium impurity G, separating and purifying the reaction solution containing cefazolin sodium impurity G on a 70mL C18 column, and eluting with 300mL of 0.05% formic acid water, 1% acetonitrile acid water and 3% acetonitrile acid water respectively. Diluting the pure product fraction by 2 times, loading the pure product fraction on a 70mL PIPI-02 column, eluting with 80% acetonitrile, and freeze-drying to obtain cefazolin sodium impurity G with the yield of 61.4% and the purity of 95%.

The HPLC detection spectrum of the cefazolin sodium impurity G prepared in the examples 1-4 is shown in figure 1, and the blank control HPLC detection spectrum is shown in figure 2. The cefazolin sodium impurity G prepared in the embodiments 1-4 is identified through MS/NMR structure, and the structure of the prepared cefazolin sodium impurity G is determined to be as shown in the formula (I):

the preparation method disclosed by the invention is mild in reaction conditions, simple in process steps, free of ultralow temperature reaction and suitable for pilot scale of laboratories. According to the invention, by optimizing reaction conditions, the yield of the prepared cefazolin sodium impurity G is 61.4-64.8%, the purity is 95%, the cefazolin sodium impurity G can be used as a reference substance of impurities in cefazolin sodium raw material or preparation detection, the accurate positioning and qualitative determination of the cefazolin sodium impurity G in the production quality control of cefazolin sodium are improved, the control of the impurities is favorably strengthened, the quality of the cefazolin sodium preparation is improved, and the safety and the effectiveness of the clinical use of the cefazolin sodium preparation are ensured.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention and not for limiting the protection scope of the present invention, and although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A preparation method of cefazolin sodium impurity G is characterized in that cefazolin sodium impurity D is hydrolyzed under alkaline conditions to obtain deacetyl cefazolin, the deacetyl cefazolin is subjected to cyclization under acidic conditions, and the cefazolin sodium impurity G is obtained through separation and purification, and the specific route is as follows:

the preparation method specifically comprises the following preparation steps:

(2) adjusting the pH value of the reaction solution containing deacetyl cefazolin to acidity, performing cyclization reaction under acidic condition to obtain reaction solution containing cefazolin sodium impurity G, and finally performing separation, purification and freeze drying on the reaction solution to obtain cefazolin sodium impurity G;

in the step (2), the final reaction solution is separated and purified by a C18 column, then is respectively eluted by formic acid water, acetonitrile acid water and acetonitrile acid water, and the pure fraction is diluted and then is loaded on a PIPI-02 column, and is eluted by acetonitrile and then is frozen and dried.

2. The method for preparing cefazolin sodium impurity G according to claim 1, wherein in step (1), the ratio of cefazolin sodium impurity D to water is 1G:50-100 mL.

3. The process for preparing cefazolin sodium impurity G according to claim 1, wherein in step (1), the pH is adjusted to 8-9 with a base.

4. The process for preparing cefazolin sodium impurity G according to claim 3, wherein the base is sodium bicarbonate.

5. The process for preparing cefazolin sodium impurity G according to claim 1, wherein in step (1), the temperature of oil bath heating is 40-50 ℃ and the time of oil bath heating is 2-4 h.

6. The process for preparing cefazolin sodium impurity G according to claim 1, wherein in step (2), pH is adjusted to 1-3 with acid, and the time of the cyclization reaction is 2-3 h.

7. The process of claim 6, wherein the acid is hydrochloric acid.

8. The process for preparing cefazolin sodium impurity G according to claim 7, wherein the concentration of hydrochloric acid is 2-4 mol/L.