CN110396102B

CN110396102B - Cefoxitin sodium compound pharmaceutical preparation and application thereof in prevention of infection before vaginal hysterectomy, abdominal hysterectomy and cesarean section (uterine)

Info

Publication number: CN110396102B
Application number: CN201910037285.9A
Authority: CN
Inventors: 吴丰喆; 赵春涛; 马庆双; 傅苗青
Original assignee: Guangdong Jincheng Jinsu Pharmacy Co ltd
Current assignee: Guangdong Jincheng Jinsu Pharmacy Co ltd
Priority date: 2019-01-15
Filing date: 2019-01-15
Publication date: 2021-05-04
Anticipated expiration: 2039-01-15
Also published as: CN110396102A

Abstract

The invention provides cefoxitin sodium or a composition thereof, a preparation method, a single preparation, a compound preparation and application thereof. In the cefoxitin sodium or the composition thereof, the mass content of cefoxitin is more than 93 percent. The cefoxitin sodium or the composition thereof has low impurity content and good quality stability, and is beneficial to improving the clinical curative effect and safety; the particle size range is uniform, which is beneficial to reducing the process difficulty of preparation; the cefoxitin sodium or its composition, or its preparation can be used for preventing infection before or after transvaginal hysterectomy, abdominal hysterectomy, or cesarean section.

Description

Cefoxitin sodium compound pharmaceutical preparation and application thereof in prevention of infection before vaginal hysterectomy, abdominal hysterectomy and cesarean section (uterine)

Technical Field

The invention relates to an antibiotic medicine, in particular to an application of cefoxitin sodium, a preparation and a composition thereof in preventing infection before transvaginal hysterectomy, abdominal hysterectomy and cesarean section.

Background

Cefoxitin sodium is a semisynthetic derivative of cephamycin C, the original research and development units are MERCK and Mylan, and the original research and development enterprise is ACS DOBFAR subsidiary company Antibiotics Do Brasil Ltd. BRAND-NAME (BRAND NAME): mefoxin, Mefoxin. ACS DOBFAR authorizes Jincheng medicine (Guangdong Jincheng jin Su pharmacy Co., Ltd.) to realize the qualitative and real-estate development of the original Chinese cefoxitin sodium for injection.

In recent years, cephalosporin resistant bacteria are increasing, which may be related to high activity beta lactamase induced by bacteria. The methoxyl in the structure of cefoxitin sodium has a steric hindrance effect, so that the cefoxitin sodium has higher stability on beta lactamase, has stronger activity on gram positive and negative bacteria, anaerobic bacteria or aerobic bacteria, and has wider clinical application range.

The antibacterial spectrum of cefoxitin sodium comprises Escherichia coli, pneumonia bacillus, indole positive proteus and serratia, Klebsiella, influenza bacillus, salmonella, shigella, etc.; it also has good effect on Staphylococcus and various streptococcus. The traditional Chinese medicine composition is mainly used for treating respiratory tract infection, endocarditis, peritonitis, pyelonephritis, urinary tract infection and septicemia caused by sensitive bacteria and bone, joint, skin and soft tissue infection.

The preparation of cefoxitin sodium is mainly synthesized, and the existing synthesis method has the problems of complicated steps, complex operation, high production cost, low product quality and the like. Generally, different synthesis methods and parameters lead to different impurities in the final product, thus leading to different product quality, which directly affects clinical efficacy and safety.

Although cefoxitin sodium has been widely used, as a cephalosporin product, the preparation method, the safety, the therapeutic effect and the stability of the product are always the key points of technical research and improvement. Therefore, the cefoxitin sodium has a great space for improvement.

Aiming at the problems, the inventor further improves the prior art to obtain cefoxitin sodium with higher quality, and provides a preparation of cefoxitin sodium and a new indication thereof.

Disclosure of Invention

In order to solve the above problems, the present inventors have conducted intensive studies and, as a result, have found that: by changing the synthetic reagent and the processing steps, the reaction parameter control is enhanced, the side reaction in the reaction can be well controlled, and the content of impurities in the cefoxitin sodium product is further reduced; and provides the use of cefoxitin sodium or a preparation thereof in the preparation of medicaments for treating transvaginal hysterectomy, abdominal hysterectomy, cesarean (uterine) delivery surgery and preventing infection and postoperative infection, thereby completing the invention.

The object of the present invention is to provide the following:

in a first aspect, the present invention provides cefoxitin sodium or a composition thereof, wherein cefoxitin (C) is₁₆H₁₇N₃O₇S₂) The mass content of (A) is more than 93%.

The cefoxitin sodium or the composition thereof comprises an impurity A,

the content is within 0.2 percent.

The cefoxitin sodium or the composition thereof also comprises an impurity B,

the content is within 0.3 percent;

further, the catalyst also comprises impurities C,

the content is within 0.2 percent.

In a second aspect, the present invention provides a method for preparing cefoxitin sodium or a composition thereof, comprising the steps of:

step 1: dissolving 7-aminocephalosporanic acid (7-ACA) in a reaction solvent, adding a phase transfer catalyst and an alkaline solution, dropwise adding thiophene acetyl chloride under controlled temperature, and filtering to obtain cephalothin acid after the reaction is finished;

step 2: dissolving cephalothin acid, reacting with dropwise added sodium methoxide solution, treating the reaction solution, and salifying and purifying the reaction solution and organic base to obtain 7-methoxy cephalothin acid organic base salt;

and step 3: dissolving 7-methoxy cephalothin acid organic alkali salt in methanol-water solution, dropping inorganic alkaline water solution after cooling, adjusting acid after reaction, stirring, and crystallizing to obtain deacetylated intermediate;

and 4, step 4: adding the deacetylated intermediate into dry tetrahydrofuran, cooling, adding chlorosulfonyl isocyanate for reaction, and performing post-treatment to obtain cefoxitin acid;

and 5: adding cefoxitin acid into a crystallization solvent, dropwise adding a sodium salt solution, filtering after the reaction is finished, and drying to obtain cefoxitin sodium.

Wherein, in step 1, the phase transfer catalyst comprises one of tetrabutylammonium bromide (TBAB), tetrabutylammonium chloride, dodecyltrimethylammonium chloride and tetradecyltrimethylammonium chloride, and is preferably tetrabutylammonium bromide;

the alkaline solution is sodium carbonate or sodium bicarbonate aqueous solution;

the temperature of the reaction system is-10 to 20 ℃ when the thiophene acetyl chloride is added, and preferably-5 to 5 ℃.

In the step 2, the step of the method is carried out,

the substitution reagent is selected from tert-butyl hypochlorite, N-bromosuccinimide, N-chlorosuccinimide, ethyl dichlorourethane and chlorosulfonic acid isocyanate;

after the reaction is finished, adding an inorganic salt aqueous solution into the reaction solution to separate layers, adjusting the acid of an aqueous layer, extracting, and adding an organic base into an extract.

The organic base is selected from cyclohexylamine, n-hexylamine or triethylamine, and preferably the organic base is cyclohexylamine.

In the step 3, the reaction system is cooled to below-10 ℃, preferably below-15 ℃, and more preferably below-20 ℃ after the 7-methoxy cephalothin acid organic base salt is dissolved;

in step 4, the temperature of the reaction system is reduced to below-20 ℃ after cooling, preferably to below-25 ℃;

in the step 4, the dosage of the chlorosulfonyl isocyanate is 0.2-0.35 g based on 1g of the deacetylated intermediate.

In a third aspect, the invention provides a single formulation comprising cefoxitin sodium or a composition thereof, or cefoxitin sodium prepared by the method as an active ingredient, wherein the formulation is an injection.

In a fourth aspect, the invention provides a cefoxitin sodium compound preparation, which comprises an active ingredient cefoxitin sodium and a synergist, and optionally, can also comprise pharmaceutically acceptable auxiliary materials and/or pharmaceutically active substances without incompatibility;

the synergist is preferably tazobactam sodium, sulbactam sodium or abamectin.

In a fifth aspect, the invention provides an application of the cefoxitin sodium or the composition thereof, the cefoxitin sodium prepared by the preparation method, a single cefoxitin sodium preparation or a cefoxitin sodium compound preparation in preparation of a medicament for treating infection and postoperative infection through vaginal hysterectomy, abdominal hysterectomy and cesarean (uterine) operation.

According to the cefoxitin sodium or the composition, the preparation method and the application thereof provided by the invention, the following beneficial effects are achieved:

(1) the cefoxitin sodium or the composition thereof provided by the invention has low impurity content, is beneficial to long-term storage and placement, and has good quality stability, better clinical curative effect and safety;

(2) the preparation method of cefoxitin sodium provided by the invention is simpler to operate, simpler in treatment steps, economic and reasonable, and more suitable for workshop production; the prepared cefoxitin sodium product has uniform particle size range, and is beneficial to the preparation and the split charging of subsequent preparations;

(3) the cefoxitin sodium or the composition thereof provided by the invention has the application in preparing medicines for treating vaginal hysterectomy, abdominal hysterectomy and cesarean (uterine) operation and preventing infection and postoperative infection.

Detailed Description

The features and advantages of the present invention will become more apparent and appreciated from the following detailed description of the invention.

The word "exemplary" is used exclusively herein to mean "serving as an example, embodiment, or illustration. Any embodiment described herein as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments.

The present invention is described in detail below.

The invention provides a preparation method of cefoxitin sodium or a composition thereof, which comprises the following steps:

step 2: and reacting the dissolved cephalothin acid with dropwise added sodium methoxide solution, and carrying out salt formation and purification on the reaction solution and organic base after treatment to obtain the 7-methoxy cephalothin acid organic base salt.

And step 3: dissolving 7-methoxy cephalothin acid cyclohexylamine salt in methanol-water solution, cooling, dripping inorganic alkali water solution, adjusting acid after reaction, stirring, and crystallizing to obtain deacetylated intermediate.

And 4, step 4: and adding the deacetylated intermediate into dry tetrahydrofuran, cooling, adding chlorosulfonyl isocyanate for reaction, and performing aftertreatment to obtain cefoxitin acid.

Wherein, in the step 1,

the reaction solvent is dichloromethane or ethyl acetate, and dichloromethane with better solubility to 7-aminocephalosporanic acid is preferably used as the reaction solvent.

Since 7-aminocephalosporanic acid and thiopheneacetyl chloride react in an alkaline environment, in order to improve the stability of the tetracyclic structure in 7-aminocephalosporanic acid, the alkaline solution is preferably alkalized by using sodium carbonate or sodium bicarbonate which is not very strong in alkalinity, and more preferably, the sodium carbonate or sodium bicarbonate is dissolved in water and added to the reaction system as an alkaline solution.

Based on the amount of 7-aminocephalosporanic acid, the amount of sodium carbonate or sodium bicarbonate is 1-2 times, preferably 1-1.5 times, and more preferably 1-1.3 times of the molar amount of 7-ACA.

At this time, an organic phase and an aqueous phase which are immiscible with each other are present in the reaction system, and in order to increase the contact probability of the solutes in the two phases, it is preferable to add a phase transfer catalyst to the reaction system to increase the alkalization catalytic effect of the alkaline solution on 7-aminocephalosporanic acid.

The phase transfer catalyst comprises one of tetrabutylammonium bromide (TBAB), tetrabutylammonium chloride, dodecyltrimethylammonium chloride and tetradecyltrimethylammonium chloride, and is preferably tetrabutylammonium bromide.

The addition amount of the phase transfer catalyst affects the reaction rate and the completion degree of the reaction, and the addition amount of the phase transfer catalyst is preferably 0.5 to 3% by weight, more preferably 0.5 to 2% by weight, and most preferably 0.7 to 1.5% by weight of 7-aminocephalosporanic acid.

In the step 1, after the 7-aminocephalosporanic acid is dissolved, adding an alkaline solution and a phase transfer catalyst under stirring, and cooling to ensure that the temperature of the reaction system is-10-20 ℃, preferably-5 ℃, because the thiophene acetyl chloride is an exothermic reaction in the subsequent addition, the temperature of the reaction system is easily increased. In alkaline aqueous solution, the tetracyclic structure in 7-aminocephalosporanic acid is easy to hydrolyze and open when the temperature is increased, and the stability of 7-aminocephalosporanic acid can be improved when the temperature is controlled.

In addition, the thiophene acetyl chloride is preferably added into a temperature-controlled reaction system in a dropwise manner, and multiple experiments show that the adding speed of the thiophene acetyl chloride has influence on the quality and the yield of the product, and the adding speed of the thiophene acetyl chloride should be controlled as much as possible so as to reduce the phenomenon of side reaction caused by overhigh local temperature in the reaction system.

When the local temperature of the reaction solution is too high due to too high dripping speed of the thiophene acetyl chloride, impurity A is generated in the reaction system. The process of formation of impurity a is as follows:

it was found by investigation that the formation of impurity A is mainly caused by an excessively high reaction temperature. 7-aminocephalosporanic acid and thiophene acetyl chloride generate cephalothin under the alkaline condition, and when the reaction temperature or the local temperature is too high, ester bonds of the cephalothin are hydrolyzed and then condensed into lactone to generate impurities A.

Therefore, controlling the addition rate of thiopheneacetyl chloride and the temperature of the reaction system is particularly critical to reducing the formation of impurity A. The content of the impurity A in the cefoxitin sodium of the final product can be effectively controlled within 0.1 percent, preferably within 0.08 percent through the temperature control of the ultralow temperature reaction.

Preferably, the addition amount of the thiopheneacetyl chloride is 1-1.2 times, preferably 1-1.1 times, and more preferably 1-1.05 times of the molar amount of 7-aminocephalosporanic acid.

The dripping time of the thiophene acetyl chloride is 30-90 minutes, preferably 40-60 minutes, so that the local overhigh temperature caused by the excessively fast addition of the thiophene acetyl chloride in a reaction system can be well avoided, and the generation of the impurity A is controlled.

After the thiophene acetyl chloride is dripped, the reaction is carried out for 2-5 hours, preferably 2-4 hours, and most preferably 2-3 hours in a heat preservation way.

Wherein, in the step 2,

dissolving cephalothin acid in a mixed solvent of dichloromethane and tetrahydrofuran, reacting with dropwise added sodium methoxide solution, and carrying out liquid-liquid contact to enable the reaction to be rapidly carried out.

In order to ensure the solubility of cephalothin acid, the volume usage ratio of dichloromethane and tetrahydrofuran is 5-10: 1, preferably 5-8: 1.

In order to improve the recovery rate of the product in the subsequent treatment, the use amount of the mixed solvent can be reduced; when the amount of the mixed solvent is small, the collision probability of the reaction substances is increased, and the occurrence probability of side reactions is also increased. Through research, in the step 2, each gram of cephalothin acid can be dissolved in 6-12 ml of mixed solvent, preferably 7-11 ml, and more preferably 8-10 ml.

The sodium methoxide solution should be miscible with the mixed solvent, preferably sodium methoxide is dissolved in methanol, acetonitrile, acetone, tetrahydrofuran or dichloromethane, and more preferably dissolved in methanol, so that no other solvent impurities are introduced when the methoxy group at the 7-position is generated in the reaction system. The mass percent of the sodium methoxide in the sodium methoxide solution is 25-30%, such as 30%.

In the step 2, when the reaction temperature is increased, the activity of the 7-carbon on the cephalosporin mother ring is improved, the reaction time for methoxylation of the 7-carbon can be shortened, but the temperature is increased, side reactions are caused, the impurity types are increased, and the yield and the quality of the product are reduced. Therefore, it is necessary to control the temperature of the reaction system in particular so that the reaction proceeds in an ultra-low temperature environment.

In particular, the temperature of the reaction system is reduced to-50 ℃ or lower before the addition of the sodium methoxide solution to control the reactivity of the radicals, preferably to-50 ℃ to-70 ℃. And dropwise adding a sodium methoxide solution at a controlled temperature, and stirring to fully mix the reaction substances.

And a substitution reagent is also used in the substitution reaction process, so that the 7-site carbon can be activated to improve the substitution efficiency. The substitution reagent is selected from tert-butyl hypochlorite, N-bromosuccinimide, N-chlorosuccinimide, ethyl dichlorourethane and chlorosulfonic acid isocyanate, and the tert-butyl hypochlorite with low cost and mild reaction is preferably used as the substitution reagent. Preferably, the substitution reagent is added after the sodium methoxide is added, so that the substitution efficiency of the methoxyl can be improved.

The substitution reagent is used in an amount of 1.0 to 1.3mol, preferably 1.0 to 1.2mol, for example 1.1mol, per mol of cephalothin acid.

When the substitution reagent is added into a reaction system, local concentration is instantaneously too high, so that the reactivity of reaction substances in local environment is too high, and side reaction is easily generated. Therefore, the dropping speed of the substitution agent should be appropriately controlled to reduce the local concentration in the solution at the moment of addition.

Preferably, the dropping time of the substitution reagent is 30 to 60 minutes, preferably 40 to 60 minutes, and more preferably 45 to 60 minutes. After the dropwise addition is finished, the reaction system is continuously stirred for reaction.

In the step 2, the addition amount of sodium methoxide should not be too much based on the dosage of cephalothin acid, so that the target group is fully substituted while side reactions are prevented, and the reaction yield is improved.

The amount of sodium methoxide is 1.3-1.7 mol, preferably 1.4-1.7 mol, and more preferably 1.5-1.6 mol per mol of cephalothin acid.

In order to reduce the local concentration of the solution at the moment of addition, sodium methoxide should be added dropwise into the reaction system, so as to minimize the occurrence of side reactions.

It was found that when the amount of sodium methoxide used in the reaction system or local environment is too large and the local temperature of the reaction is too high, impurity C is easily generated, and the generation process is as follows:

in the process of dropwise adding the sodium methoxide solution, the esterification reaction of cephalothin acid can be caused by overhigh local reaction temperature, and an impurity C is generated; therefore, the control of the reaction temperature is an important factor.

In addition, the over-high dropping speed of the sodium methoxide can increase the local concentration of the sodium methoxide, and the collision probability between the sodium methoxide and the cephalothin is increased, so that the content of the impurity C is increased. The dropping time of the sodium methoxide is preferably 30 to 90 minutes, and preferably 30 to 60 minutes.

Through ultralow temperature reaction and control of the addition amount and the dropping speed of sodium methoxide, the content of impurity C in the final product cefoxitin sodium can be effectively controlled within 0.1 percent, preferably within 0.08 percent.

After the addition of the reactants is complete, the reaction is stirred and the end of the reaction is monitored, preferably using a high performance liquid phase, until the reaction is complete.

Preferably, after the reaction is finished, 5-15% of sodium chloride aqueous solution and 3-7% of sodium bicarbonate aqueous solution are added into the reaction solution, so that the reaction system is layered. Preferably, the volume ratio of the sodium chloride aqueous solution to the sodium bicarbonate aqueous solution is 1-3: 1, e.g. 1: 1.

The mixed solution containing the sodium chloride aqueous solution and the sodium bicarbonate aqueous solution can be used for salifying the 7-methoxy cephalothin acid and the sodium bicarbonate in the reaction system and improving the solubility of the 7-methoxy cephalothin acid in the aqueous solution. Meanwhile, the inorganic salt contained in the water layer solution can promote the precipitation of the 7-methoxy cephalothin acid and can improve the extraction rate in the subsequent extraction treatment.

Hydrochloric acid is added to the aqueous layer to adjust the pH to 1 to 4, preferably 2 to 3, and the product in the aqueous layer is extracted with ethyl acetate, and the aqueous layer is discarded.

And then adding an organic base into the ethyl acetate layer to salify the ethyl acetate layer and the product, wherein the organic base is selected from cyclohexylamine, n-hexylamine or triethylamine, and preferably the organic base is cyclohexylamine.

Stirring, cooling, crystallizing and filtering to obtain the 7-methoxy cephalothin acid organic alkali salt.

According to the invention, through the treatments of layering, extraction, salification and the like, the extraction and purification of the 7-methoxy cephalothin acid are realized, and compared with the prior art, the method is simpler in operation, simpler in treatment steps and more suitable for workshop production.

Wherein, in the step 3,

and (3) dissolving the 7-methoxy cephalothin acid organic base salt obtained in the step (2) in a methanol-water solution, wherein the mass of methanol and water in the methanol-water solution is 1.2-1.8: 1, and preferably 1.3-1.6: 1.

Wherein the amount of the methanol-water solution is 20 to 40 times, preferably 20 to 30 times, for example 25 times of the mass of the 7-methoxy cephalothin acid organic base salt.

Because the 7-methoxy cephalothin acid structure of the 7-methoxy cephalothin acid organic base salt contains a lactam structure, the hydrolysis is easy to occur in aqueous solution at higher temperature, and particularly the hydrolysis is easy to occur under alkaline conditions.

Therefore, the methanol-water solution dissolved with the 7-methoxy cephalothin acid organic alkali salt is cooled to the temperature below minus 10 ℃, so as to reduce the hydrolytic activity of lactam; preferably to-15 deg.C, more preferably to-20 deg.C or lower, most preferably to-20 deg.C to-40 deg.C.

Preferably, an aqueous solution of an inorganic base is added to the methanol-water solution to hydrolytically remove acetyl groups from 7-methoxycephalothin acids. The aqueous solution of an inorganic base is an aqueous solution of sodium hydroxide or an aqueous solution of potassium hydroxide, and an aqueous solution of sodium hydroxide is preferably used.

Wherein the concentration of the inorganic base affects the local alkalinity of the solution at the moment of addition, and although dilution is rapidly reduced under stirring conditions, due to the high instability of the cephalosporin parent ring structure in aqueous solution, controlling the concentration of the inorganic base helps to reduce hydrolysis of lactam. However, when the concentration of the inorganic base is too low, the hydrolysis of acetyl group is not facilitated, and the volume of the reaction system is increased, which is not preferable for the precipitation of the product.

Preferably, the concentration of the inorganic base in the inorganic base aqueous solution is 1-4 mol/L, and preferably 2-3 mol/L.

Preferably, an aqueous solution of an inorganic base is added dropwise to avoid hydrolysis of the lactam due to too high a local concentration. The dropping time of the inorganic alkaline water solution is 15-90 min, preferably 45-60 min.

Monitoring the reaction in real time, preferably using a high performance liquid phase to monitor until the reaction is completed, and then heating the reaction solution to room temperature (10-30 ℃).

Adjusting the pH value of the solution to 3-4 by using inorganic acid, preferably 3-3.5; the inorganic acid is selected from any one of acetic acid, hydrochloric acid, sulfuric acid and phosphoric acid; preferably, acetic acid with weak acidity is used for adjustment, and the pH value of the solution is mildly changed to have small influence on products.

Stirring and crystallizing the solution to obtain the deacetylated intermediate.

In the hydrolysis reaction, the organic alkali salt in the organic alkali salt of the 7-methoxy cephalothin acid is remained in the mother liquor, and the deacetylated intermediate obtained by hydrolysis is further purified. Simple hydrolysis operation, low production difficulty and contribution to industrial production.

Wherein, in the step 4,

and (3) dissolving the deacetylated intermediate obtained in the step (3) in tetrahydrofuran to react with chlorosulfonyl isocyanate. Preferably, the tetrahydrofuran is subjected to dehydration and drying treatment.

The amount of the tetrahydrofuran is 5-10 ml, preferably 7-9 ml, based on 1g of the deacetylated intermediate.

The reaction activity of the chlorosulfonyl isocyanate is extremely high, and the deacetylated intermediate and chlorosulfonyl isocyanate need to be reacted at a low temperature so as to avoid a large amount of side reactions. Meanwhile, the deacetylation intermediate and chlorosulfonyl isocyanate generate heat during reaction, and the feeding speed of the chlorosulfonyl isocyanate needs to be controlled to prevent side reactions such as degradation, transesterification, ester condensation and the like caused by local overheating.

The temperature of the reaction system is preferably lowered to-20 ℃ or lower, more preferably-25 ℃ or lower. The lower the reaction temperature, the lower the reactivity, and the higher the probability of the target reaction. Researches show that when the reaction temperature is-40 to-30 ℃, more preferably-40 to-35 ℃, the reaction system has good reaction activity, the target reaction completion degree is higher, and the requirements on environment and equipment are relatively reduced.

The dosage of the chlorosulfonyl isocyanate is closely related to the reaction completion degree of the deacetylation intermediate. When the dosage of chlorosulfonyl isocyanate is too much, the activity of deacetylated intermediates is enhanced, and unnecessary side reactions are easily caused; when the dosage of chlorosulfonyl isocyanate is too low, the deacetylation intermediate is not completely reacted, so that the residual amount of the deacetylation intermediate in the product cefoxitin sodium is too high, and a residual impurity B is formed.

The residual amount of deacetylated intermediate, i.e. impurity B, is controlled as an important impurity in cefoxitin sodium; the generation source of the impurities in the cefoxitin sodium comprises intermediate residues and degradation generation.

Therefore, the appropriate dosage of chlorosulfonyl isocyanate can effectively control the residual amount of deacetylated intermediate in the product cefoxitin sodium.

Preferably, the chlorosulfonyl isocyanate is used in an amount of 0.2 to 0.35g, preferably 0.2 to 0.25g, based on 1g of the deacetylated intermediate; the addition amount can effectively control the content of the impurity B in the final product cefoxitin sodium within 0.2 percent, preferably within 0.15 percent.

The chlorosulfonyl isocyanate is preferably slowly added into the reaction system in a dropwise manner, so that the reaction process is relatively smooth and mild. The addition time of the chlorosulfonyl isocyanate is 0.5 to 2 hours, preferably 0.5 to 1 hour.

After the reaction is completed, pure water is added to quench the reaction, the reaction solution is heated to room temperature, and the reaction solution is subjected to post-treatment.

Preferably, hydrochloric acid is added into the reaction solution to adjust the pH value to 1.0-3.0, preferably to adjust the pH value to 1.5-2.0, and then the cefoxitin acid is obtained after cooling, crystallization, filtration and drying.

Wherein, in step 5,

cefoxitin acid reacts with sodium salt to generate cefoxitin sodium, and a final product with good purity, content and powder form can be obtained by recrystallization treatment in a crystallization solvent.

Researches show that products obtained by recrystallization of different crystallization solvents have great differences in purity, content and powder morphology. Generally, one or any mixture of acetone, methanol, isopropanol, tetrahydrofuran, ethyl acetate and isopropyl ether may be used as the crystallization solvent.

Through a plurality of tests, the invention particularly uses methanol, acetone or a mixed solution thereof as a crystallization solvent, so that a crystallized product has a better powder form, and the preparation and clinical use of the preparation are facilitated; and further purifying the cefoxitin sodium by recrystallization treatment to obtain precipitates with good purity content.

When a mixed solvent of methanol and acetone is used as the crystallization solvent, the volume ratio of methanol to acetone is 0.5 to 1:1, for example, 0.8: 1.

Based on 1g of cefoxitin acid, the amount of the crystallization solvent is 6-10 ml, preferably 6-8 ml, so that a good impurity removal effect can be achieved, and meanwhile, the loss of cefoxitin sodium in the crystallization process can be reduced.

Dissolving cefoxitin acid in a crystallization solvent, and adding sodium salt, wherein the sodium salt is selected from sodium acetate, sodium isooctanoate, sodium acetate trihydrate and sodium lactate, the type of the sodium salt can influence the powder form of the cefoxitin sodium during precipitation, and through multiple researches, the sodium lactate is preferably used as the sodium salt, so that the prepared cefoxitin sodium has good crystal form, and is beneficial to subsequent filtration treatment and preparation.

The sodium salt is dissolved in methanol, isopropanol or acetone, preferably in methanol, acetone or a mixed solution thereof, and then added to a solution in which cefoxitin acid is dissolved.

The molar weight of the added sodium salt is equivalent to that of cefoxitin acid, so that sodium salt residue caused by excessive addition is avoided, and instability of cefoxitin sodium/cefoxitin acid caused by excessive alkalinity in the solution is prevented.

Preferably, the mass percent of the sodium salt in the sodium salt solution is 30-40%, the sodium salt is continuously stirred in the adding process, the temperature is kept at-10-30 ℃, preferably 0-25 ℃, more preferably 10-15 ℃, and the solubility of the cefoxitin sodium can be improved.

Researches find that the dropping speed of the sodium salt obviously influences the powder state of the product. When the dropping speed of the sodium salt is too high, the phenomenon of crystal explosion is easily caused, crystal grains are instantly separated out in a short time, ordered growth cannot be carried out, and a large amount of impurities in the reaction liquid are easily mixed; also, the particle size range of the powder is too large, and the difficulty of processing the preparation is increased. Too slow a sodium salt addition rate can add unnecessary process time.

Through a large number of researches, the dropping speed of the sodium salt is controlled to be 1-4 hours, preferably 1-3 hours, and more preferably 1.5-2 hours.

In the dropwise adding process, the sodium salt and the cefoxitin acid react to release heat, and in order to avoid degradation reaction caused by overhigh local temperature, the stirring speed is kept at 50-300 revolutions per minute, preferably 150-200 revolutions per minute.

After the sodium salt is added dropwise, cooling, crystallizing and growing the crystal.

In particular, the sodium salt needs to be sterilized or disinfected before use, reducing the content of microorganisms introduced into the crystallization system.

More preferably, the crystallized solution with dissolved cefoxitin acid is decolorized, sterile filtered before the sodium salt is added to reduce the bacteria or microorganisms that may remain in the solution. The decolorization treatment can be carried out by adsorbing and decolorizing with activated carbon for injection, so that the obtained cefoxitin sodium has white color.

After the sodium salt is added, cooling the reaction system to 0-10 ℃, preferably to 2-9 ℃, and more preferably to 3-8 ℃. The cooling rate is 0.5-10 ℃/h, preferably 1-7 ℃/h, and more preferably 3-5 ℃/h.

The solubility of cefoxitin sodium can be reduced by cooling, and crystal nuclei are gradually formed. After the temperature reduction is finished, the reaction system is kept warm for 2-6 hours under stirring, so that crystal nuclei grow into crystal grains, and then the crystal grains gradually grow to the proper grain size.

The stirring speed needs to be controlled in the crystallization process, and loose crystal grains which are just formed are easily broken up when the stirring speed is too high, so that the particle size of the powder is too small; when the speed is too slow, the solute in the solution is not uniformly distributed, and the particle size range is easily overlarge.

Particularly, the stirring speed in the crystallization process is 100-150 rpm, preferably 120-140 rpm, which is beneficial to the dispersion of powder particles and the generation of crystal-form solids; the particle size distribution range can also be reduced, so that the particle size is concentrated.

The product was collected by filtration and dried in vacuo to give crystalline cefoxitin sodium.

In a preferred embodiment, the synthetic route for the preparation of cefoxitin sodium is as follows:

the preparation method of cefoxitin sodium provided by the invention is industrially feasible and low in preparation difficulty, and the prepared cefoxitin sodium is low in impurity content, high in purity, white in color and good in stability; the powder has good shape and centralized particle size range, and is helpful for reducing the difficulty of the preparation process.

Meanwhile, the preparation method of cefoxitin sodium provided by the invention is simple in synthesis steps, low in processing difficulty in the operation process and very suitable for workshop production.

The invention also provides cefoxitin sodium or a composition thereof prepared by the method, and the cefoxitin sodium or the composition thereof has high content, good purity and uniform particle size distribution. In the cefoxitin sodium or the composition thereof, the mass content of cefoxitin is more than 93 percent or the purity is more than 99.2 percent calculated on anhydrous substances.

The cefoxitin sodium or the composition thereof also contains impurity A,

the content of impurity A is controlled within 0.2%;

and an impurity B, which is a nitrogen-containing impurity,

the content of the impurity B is within 0.3 percent.

Furthermore, the cefoxitin sodium or the composition thereof also contains impurity C,

the mass content of the impurity C is controlled to be not higher than 0.2 percent.

The cefoxitin sodium has good quality stability, and after being placed under long-term test conditions (25 ℃ +/-2 ℃/65% RH +/-5% RH) for 18 months, the impurity A is preferably within 0.15%, more preferably within 0.1%, such as 0.08%. The content of impurity B is preferably 0.15% or less, more preferably 0.1% or less, for example 0.07%. The impurity C is preferably within 0.1%, more preferably within 0.08%, for example 0.06%.

The cefoxitin sodium or the composition thereof has low organic solvent residue, particularly ethyl acetate, tetrahydrofuran and methanol residue and good safety.

The invention also provides a cefoxitin sodium preparation of the cefoxitin sodium or the composition thereof prepared by the method, which is a single preparation taking the cefoxitin sodium as an active ingredient, and is preferably an injection.

In a preferred embodiment, the cefoxitin sodium preparation is packaged in a sterile manner, and the cefoxitin sodium or the composition thereof is packaged after being crushed and then is packaged in a nitrogen-filled manner.

In the preparation, cefoxitin sodium or a composition thereof is subjected to crushing treatment and then sterile subpackaging, preferably crushing until the particle size D90 is 100-300 microns, more preferably 150-200 microns, and the dissolution speed is high in use.

The invention also provides a cefoxitin sodium compound preparation which comprises an active ingredient cefoxitin sodium and a synergist, and optionally, pharmaceutically acceptable auxiliary materials and/or pharmaceutically active substances without incompatibility.

The synergist is preferably tazobactam sodium, sulbactam sodium or ababactam sodium, and more preferably tazobactam sodium.

The weight ratio of cefoxitin sodium (calculated as cefoxitin) to tazobactam sodium (calculated as tazobactam) is 2-8: 1, and preferably 4-8: 1.

In a more preferred embodiment, the weight ratio of cefoxitin sodium (calculated as cefoxitin) to tazobactam sodium (calculated as tazobactam) is 4:1, 6:1 or 8: 1.

The tazobactam sodium is irreversible beta lactamase inhibition, and when the tazobactam sodium is used together with the cefoxitin sodium, the cefoxitin sodium can be more stable to the beta lactamase, so that the antibacterial effect of the cefoxitin sodium is further improved.

The cefoxitin sodium compound preparation can be prepared by uniformly mixing active ingredients and then performing aseptic subpackaging, and can also be prepared by freeze drying.

Preferably, in the cefoxitin sodium compound preparation, active ingredients are crushed, or the particle size of the active ingredients in the prepared compound preparation is very small, so that the active ingredients can be quickly dissolved, and the clinical use requirement is met.

Because cefoxitin sodium is unstable to oxygen, nitrogen, particularly high-purity nitrogen (more than 99.999 percent) is preferably used for protection during preparation sub-packaging, the stability under long-term storage is improved by removing the oxygen in a sub-packaging bottle, the generation of oxidative degradation impurities is reduced, and the clinical curative effect and the use safety are improved.

According to the cefoxitin sodium or the composition thereof, the cefoxitin sodium preparation and the cefoxitin sodium compound preparation, the invention also provides an application of the cefoxitin sodium in preparing a medicine for preventing infection and postoperative infection through vaginal hysterectomy, abdominal hysterectomy and cesarean (uterine) delivery operations.

Examples

Example 1

Step 1: adding 500g of 7-aminocephalosporanic acid (7-ACA) and 5000ml of dichloromethane into a 10L four-neck round-bottom flask, adding a saturated aqueous solution in which 170g of sodium bicarbonate is dissolved and 5.0g of tetrabutylammonium bromide under stirring, controlling the temperature to be minus 5-0 ℃, dropwise adding 308g of thiophene acetyl chloride, and controlling the dropwise adding time to be 50-60 minutes. After reacting for 4 hours under heat preservation, filtering, washing a filter cake by dichloromethane, filtering, and drying to obtain the cephalothin acid.

Step 2: adding 5000ml of dichloromethane, 800ml of tetrahydrofuran and 600g of cephalothin acid into a 10L four-neck round-bottom flask, cooling to-60 ℃ after stirring and complete dissolution, dropwise adding 126g of methanol solution containing sodium methoxide within 60 minutes, wherein the mass concentration of the sodium methoxide is 30%, continuously stirring for 45 minutes after the addition is finished, then dropwise adding 181g of tert-butyl hypochlorite, keeping the temperature and stirring after 45 minutes of addition is finished, and adding 5% of sodium chloride aqueous solution and 5% of sodium bicarbonate aqueous solution for layering after the high-efficiency liquid phase monitoring reaction is finished; adding hydrochloric acid into the water layer to adjust the pH value to be 2.5 +/-0.5, extracting with ethyl acetate, adding cyclohexylamine into the ethyl acetate layer, stirring, crystallizing, and filtering to obtain 7-methoxy cephalothin acid cyclohexylamine salt.

And step 3: adding 500g of 7-methoxy cephalothin acid cyclohexylamine salt into 3000g of water and 4500g of methanol in a 10L four-neck round-bottom flask, cooling to-30 ℃, dropwise adding 390ml of 3.0mol/L sodium hydroxide aqueous solution within 1h, reacting for 2 hours, and monitoring by a high performance liquid phase until the reaction is complete; monitoring the high-efficiency liquid phase until the reaction is complete; heating to room temperature, adding acetic acid to adjust the pH of the system to 3.3 +/-0.2, stirring, cooling and crystallizing to obtain a deacetylated intermediate;

and 4, step 4: adding 650g of deacetylation intermediate into 5200ml of dry tetrahydrofuran, cooling to-40-37 ℃, adding 130g of chlorosulfonyl isocyanate within 45 minutes, and monitoring by a high performance liquid phase until the reaction is complete; adding 600g of pure water, heating to 20-23 ℃, stirring for 45min, dropwise adding concentrated hydrochloric acid to adjust the pH value to 1.5-2.0, cooling to 0-5 ℃, crystallizing, and filtering to obtain cefoxitin acid.

And 5: adding 520g of cefoxitin acid into 1600ml of methanol and 2000ml of acetone, and stirring and dissolving at the temperature of 10-12 ℃. Adding active carbon for decoloring and filtering; slowly dripping a sterilized methanol solution containing 140g of sodium lactate into the filtrate, wherein the mass concentration of the sodium lactate is 30 percent, and after the addition of the sodium lactate for 2 hours, the stirring speed is 175 r/m; and continuously reacting for 2 hours, cooling to 5-7 ℃ at the speed of 5 ℃/h, crystallizing, stirring at the speed of 130 rpm, filtering, washing and drying to obtain the cefoxitin sodium crystal.

Example 2

Step 1: adding 500g of 7-aminocephalosporanic acid (7-ACA) and 5000ml of dichloromethane into a 10L four-neck round-bottom flask, adding a saturated aqueous solution in which 200g of sodium bicarbonate is dissolved and 7.5g of tetrabutylammonium bromide under stirring, controlling the temperature to be between-5 and-3 ℃, dropwise adding 308g of thiophene acetyl chloride, and controlling the dropwise adding time to be between 50 and 60 minutes. After reacting for 3 hours under heat preservation, filtering, washing a filter cake by dichloromethane, filtering, and drying to obtain the cephalothin acid.

Step 2: adding 5000ml of dichloromethane, 1000ml of tetrahydrofuran and 600g of cephalothin acid into a 10L four-neck round-bottom flask, cooling to-50 ℃ after stirring and complete dissolution, dropwise adding a methanol solution containing 122.4g of sodium methoxide within 60 minutes, wherein the mass concentration of the sodium methoxide is 25%, continuously stirring for 45 minutes after the addition is finished, dropwise adding 164g of tert-butyl hypochlorite, keeping the temperature and stirring after 45 minutes of addition is finished, and adding a 5% sodium chloride aqueous solution and a 5% sodium bicarbonate aqueous solution for layering after the high-efficiency liquid-phase monitoring reaction is finished; adding hydrochloric acid into the water layer to adjust the pH value to be 2.5 +/-0.5, extracting with ethyl acetate, adding cyclohexylamine into the ethyl acetate layer, stirring, crystallizing, and filtering to obtain 7-methoxy cephalothin acid cyclohexylamine salt.

And step 3: adding 500g of 7-methoxy cephalothin acid cyclohexylamine salt into 2700g of water and 4300g of methanol in a 10L four-neck round-bottom flask, cooling to-40 ℃, dropwise adding 390ml of 3.0mol/L sodium hydroxide aqueous solution within 45min, reacting for 2 hours, and monitoring by a high performance liquid phase until the reaction is complete; monitoring the high-efficiency liquid phase until the reaction is complete; heating to room temperature, adding acetic acid to adjust the pH of the system to 3 +/-0.5, stirring, cooling and crystallizing to obtain a deacetylated intermediate;

and 4, step 4: adding 650g of deacetylation intermediate into 4600ml of dried tetrahydrofuran, cooling to-40-35 ℃, adding 148g of chlorosulfonyl isocyanate in batches within 45 minutes, and monitoring by a high performance liquid phase until the reaction is complete; adding 600g of pure water, heating to 20-23 ℃, stirring for 60min, dropwise adding concentrated hydrochloric acid to adjust the pH value to 1.5-2.0, cooling, crystallizing, and filtering to obtain cefoxitin acid.

And 5: adding 520 cefoxitin acid into 1600ml methanol and 2000ml acetone, and stirring at 10-15 ℃ to dissolve completely. Adding active carbon for decoloring and filtering; slowly dripping a degerming methanol solution containing 140g of sodium lactate into the filtrate, wherein the mass concentration of the sodium lactate is 35 percent, and after the addition is finished for 1.5 hours, the stirring speed is 150 r/m; and continuously reacting for 2 hours, cooling to 6-8 ℃ at the speed of 3 ℃/h, crystallizing, stirring at the speed of 140 rpm, filtering, washing and drying to obtain cefoxitin sodium crystals.

Example 3

Step 1: adding 500g of 7-aminocephalosporanic acid (7-ACA) and 5000ml of dichloromethane into a 10L four-neck round-bottom flask, adding a saturated aqueous solution in which 155g of sodium bicarbonate is dissolved and 3.5g of tetrabutylammonium bromide under stirring, controlling the temperature to be minus 5-0 ℃, dropwise adding 308g of thiophene acetyl chloride, and controlling the dropwise adding within 50-60 minutes to finish the dropwise adding. After reacting for 4 hours under heat preservation, filtering, washing a filter cake by dichloromethane, filtering, and drying to obtain the cephalothin acid.

Step 2: adding 5000ml of dichloromethane, 800ml of tetrahydrofuran and 600g of cephalothin acid into a 10L four-neck round-bottom flask, cooling to-70 ℃ after stirring and complete dissolution, dropwise adding a methanol solution containing 130g of sodium methoxide within 45 minutes, wherein the mass concentration of the sodium methoxide is 30%, continuously stirring for 45 minutes after the addition, then dropwise adding 196g of tert-butyl hypochlorite, keeping the temperature and stirring after 45 minutes of addition, and adding a 5% sodium chloride aqueous solution and a 7% sodium bicarbonate aqueous solution for layering after the high-efficiency liquid phase monitoring reaction is finished; adding hydrochloric acid into the water layer to adjust the pH value to be 2.5 +/-0.5, extracting with ethyl acetate, adding cyclohexylamine into the ethyl acetate layer, stirring, crystallizing, and filtering to obtain 7-methoxy cephalothin acid cyclohexylamine salt.

And step 3: adding 500g of 7-methoxy cephalothin acid cyclohexylamine salt into 3000g of water and 3900g of methanol in a 10L four-neck round-bottom flask, cooling to-20 ℃, dropwise adding 390ml of 3.0mol/L sodium hydroxide aqueous solution within 1h, reacting for 2 hours, and monitoring by a high performance liquid phase until the reaction is complete; monitoring the high-efficiency liquid phase until the reaction is complete; heating to room temperature, adding acetic acid to adjust the pH of the system to 3.3 +/-0.2, stirring, cooling and crystallizing to obtain a deacetylated intermediate;

and 4, step 4: adding 650g of deacetylation intermediate into 5700ml of dry tetrahydrofuran, cooling to-40 to-30 ℃, adding 160g of chlorosulfonyl isocyanate in batches within 45 minutes, and monitoring by a high performance liquid phase until the reaction is complete; adding 600g of pure water, heating to 20-23 ℃, stirring for 60min, dropwise adding concentrated hydrochloric acid to adjust the pH value to 1.5-2.0, cooling, crystallizing, and filtering to obtain cefoxitin acid.

And 5: adding 520 cefoxitin acid into 1600ml methanol and 2000ml acetone, and stirring at 10-15 ℃ to dissolve completely. Adding active carbon for decoloring and filtering; slowly dripping 140g of sterilized methanol solution of sodium lactate into the filtrate, wherein the mass concentration of the sodium lactate is 40 percent, and after the addition of the sodium lactate for 2 hours, the stirring speed is 2000 r/min; and continuously reacting for 2 hours, cooling to 4-8 ℃ at the speed of 4 ℃/h, crystallizing, stirring at the speed of 120 rpm, filtering, washing and drying to obtain cefoxitin sodium crystals.

Example 4

Taking the cefoxitin sodium prepared in the example 1, and crushing the cefoxitin sodium in a ball mill to ensure that the particle size D90 in the crushed cefoxitin sodium powder reaches 150 +/-10 mu m.

Example 5

Taking the cefoxitin sodium prepared in the embodiment 2, and carrying out sterile grinding to ensure that the particle size D90 in the ground cefoxitin sodium powder reaches 150 +/-10 mu m.

Example 6

Taking the cefoxitin sodium prepared in the embodiment 3, and carrying out sterile grinding to ensure that the particle size D90 in the ground cefoxitin sodium powder reaches 150 +/-10 mu m.

Example 7

Weighing cefoxitin sodium powder prepared in examples 4, 5 and 6, and adding cefoxitin acid (C) under nitrogen protection₁₆H₁₆N₃NaO₇S₂) Measuring 1.0g of the cefixime sodium preparation, sealing and packaging in a penicillin bottle to obtain the cefixime sodium preparation A, B and C.

Example 8

Weighing the cefoxitin sodium powder prepared in the embodiment 4, and preparing 100 parts of the cefoxitin sodium compound preparation according to the following formula.

The prescription is as follows:

preparing the cefoxitin sodium compound preparation E according to a conventional freeze drying technology, and aseptically packaging 1.0g of cefoxitin sodium compound preparation E in terms of cefoxitin.

Example 9

Weighing cefoxitin sodium powder prepared in the embodiment 4 according to the weight ratio of cefoxitin sodium (calculated as cefoxitin acid): 4:1 of tazobactam sodium (calculated as tazobactam acid) and 1.0g of cefoxitin are subjected to aseptic subpackage to prepare a compound preparation F.

Comparative example

Comparative example 1

As in example 1, the only difference is: the adding time of the thiophene acetyl chloride in the step 1 is 20-30 minutes; the control temperature of the reaction system is 5-10 ℃.

Comparative example 2

As in example 1, the only difference is: the concentration of sodium methoxide in the step 2 is 32 percent; the dripping time is 20-30 minutes; the temperature of the reaction system is controlled to be-40 to-30 ℃.

Comparative example 3

As in example 1, the only difference is: in the step 4, the addition amount of chlorosulfonyl isocyanate is 260g, and the dropping time is 20-30 minutes.

Examples of the experiments

Experimental example 1

The powder properties of cefoxitin sodium obtained in examples 1-3 were determined and the results were as follows:

as can be seen, the particle size of the product prepared in the examples 1-3 is uniform, the distribution range is narrow, and the particle size of the powder after the crushing treatment is relatively uniform;

as is clear from the results of the bulk density and angle of repose measurements, the products obtained in examples 4 to 6 had good flowability, and the separate packaging treatment was very facilitated after the pulverization treatment.

Experimental example 2

The residual solvent in cefoxitin sodium obtained in examples 1-3 was determined according to the guidelines of the 2015 pharmacopoeia. The results are as follows:

	ethyl acetate	Tetrahydrofuran (THF)	Methanol	Acetone (II)
					Example 1	181ppm	90ppm	186ppm	92ppm
Example 2	186ppm	92ppm	202ppm	88ppm
					Example 3	172ppm	86ppm	190ppm	80ppm

As can be seen, the organic solvent residue in examples 1 to 3 was very small, and the safety in long-term use was high.

Experimental example 3

The mass contents of cefoxitin sodium, impurities A, B and C obtained in examples 1-3 and comparative examples 1-3 were determined using high performance liquid chromatography (external standard method). The detection samples are respectively placed for 0, 6, 12 and 24 months under accelerated test conditions (30 ℃ +/-2 ℃/65% RH +/-5% RH).

The results are as follows:

it can be seen that the content of cefoxitin sodium in examples 1-3 is still stable after long-term testing, indicating that it has high stability and safety.

Example 4

The clarity of the solutions of cefoxitin sodium preparations obtained in examples 7 to 9 was measured (first method of clarity inspection method in pharmacopoeia 0902, 2015 edition: (visual method)) and insoluble fine particles (second method of insoluble fine particle inspection method in pharmacopoeia 0903, 2015 edition: microscopic counting method), and the results were as follows:

it can be seen that the solution clarity and insoluble particle detection results of the cefoxitin sodium preparation prepared in examples 7-9 are good and both meet the requirements of the existing pharmacopoeia.

Experimental example 5

The cefoxitin sodium preparations obtained in examples 7 to 9 were measured for their content (relative value of the measured amount to the labeled amount), impurity A, B and C by mass using high performance liquid chromatography (external standard method). The test samples were placed under long-term test conditions (25 ℃. + -. 2 ℃/65% RH. + -. 5% RH) for 0, 12, 24 months, respectively. Acidity and clarity of the solution were measured according to the rules of the 2015 pharmacopoeia.

The results are as follows:

therefore, after long-term tests, the content of cefoxitin sodium in the preparations A-C and F is stable, and only a small amount of cefoxitin sodium is degraded; the preparation has good long-term stability and long shelf life.

Experimental example 6

The cefoxitin sodium preparations obtained in examples 7 to 9 were measured for their content (relative value of the measured amount to the labeled amount), impurity A, B and C by mass using high performance liquid chromatography (external standard method). The detection samples are respectively placed for 1, 3 and 6 months under accelerated test conditions (40 ℃ +/-2 ℃/75% RH +/-5% RH).

The results are as follows:

therefore, after an accelerated test, the content of cefoxitin sodium in the preparation is very stable, impurities are not obviously improved, and the preparation has good stability and safety.

Experimental example 7

Endotoxin/pyrogen studies were conducted on the formulations prepared in examples 7 and 10, and it was found that the endotoxin/pyrogen levels in the formulations prepared in examples 7 and 10 were less than 0.10 EU/mg.

Experimental example 8

BALB/c female mice were randomly divided into experimental and control groups of 12 mice each. Experimental groups mice were treated as follows: administered by tail vein injection at 250mg/kg (calculated as cefoxitin) body weight before operation; the administration was 1 time. Cutting uterus at the junction of uterine horn and ovary, completely peeling off uterus, ligating the junction of cervical orifice and vagina, cutting down uterus, closing abdominal cavity, suturing skin, and sterilizing. Treatment was administered at a dose of 500mg/kg, 4 times per day after surgery. One week later, mice were sacrificed, dissected, and examined for infection.

The control mice were treated identically to the experimental groups, except that they were sequentially set as: the medicine is taken in a preoperative prevention group (no administration after operation), a postoperative treatment group (no administration before operation) and no medicine administration group (no administration before and after operation).

The administration was formulation a prepared in example 7 or formulation F prepared in example 9.

The symptom evaluation criteria were: no infection (procalcitonin PCT < 0.5 ng/ml); slight infection (0.5ng/ml < procalcitonin PCT < 2 ng/ml); severe infection (Procalcitonin PCT > 2 ng/ml).

The results were:

experimental example 9

Selecting 12 female healthy rats 200 +/-20 g for abdominal hysterectomy, and performing an operation 10min before the operation by intraperitoneally injecting the preparation A (1 g in terms of cefoxitin) prepared in the example 7 at a dose of 500 mg/kg; after operation, the cefoxitin sodium for injection is maintained for 3 days and 3 times per day, and the infection condition is observed.

Evaluation criteria:

the effect is shown: the wound starts to heal, obvious granulation grows, and purulent secretion does not exist; effective, the wound has granulation germination, red swelling, tenderness and a small amount of purulent secretion; ineffective, wound ulceration, red swelling, purulent secretion and bad smell.

According to statistics, 11 significant persons exist, 1 valid person exists, and 0 invalid person exists.

Experimental example 10

In a clinical trial using the preparation A prepared in example 7, 50 patients of 25 to 45 years old who had undergone a transvaginal hysterectomy and 50 patients of 18 to 40 years old who had undergone a caesarean section were selected for study.

Half an hour before operation, injecting the preparation A (1 g calculated as cefoxitin) into vein, and performing operation; after operation, the cefoxitin sodium for injection is used for maintaining and treating for 3 days and 3 times per day, and the infection condition is detected.

Evaluation criteria:

the effect is shown: no inflammation; effective, has slight inflammation; ineffective, severe inflammation occurs.

Statistics shows that 30 effective patients, 20 effective patients and 0 ineffective patients are in abdominal hysterectomy.

In the operation of cesarean section, 33 patients had significant effect, 17 patients had effective effect, and 0 patient had no effect. No allergic events were found during the treatment.

The invention has been described in detail with reference to specific embodiments and illustrative examples, but the description is not intended to be construed in a limiting sense. Those skilled in the art will appreciate that various equivalent substitutions, modifications or improvements may be made to the technical solution of the present invention and its embodiments without departing from the spirit and scope of the present invention, which fall within the scope of the present invention. The scope of the invention is defined by the appended claims.

Claims

1. A preparation method of cefoxitin sodium is characterized by comprising the following steps:

step 1: adding 500g of 7-aminocephalosporanic acid and 5000ml of dichloromethane into a 10L four-neck round-bottom flask, adding a saturated aqueous solution in which 170g of sodium bicarbonate is dissolved and 5.0g of tetrabutylammonium bromide under stirring, controlling the temperature to be-5-0 ℃, dropwise adding 308g of thiophene acetyl chloride, controlling the dropwise adding to be finished within 50-60 minutes, carrying out heat preservation reaction for 4 hours, filtering, washing a filter cake with dichloromethane, carrying out suction filtration, and drying to obtain cephalothin acid;

step 2: adding 5000ml of dichloromethane, 800ml of tetrahydrofuran and 600g of cephalothin acid into a 10L four-neck round-bottom flask, cooling to-60 ℃ after stirring and complete dissolution, dropwise adding 126g of methanol solution containing sodium methoxide within 60 minutes, wherein the mass concentration of the sodium methoxide is 30%, continuously stirring for 45 minutes after the addition is finished, then dropwise adding 181g of tert-butyl hypochlorite, keeping the temperature and stirring after 45 minutes of addition is finished, and adding 5% of sodium chloride aqueous solution and 5% of sodium bicarbonate aqueous solution for layering after the high-efficiency liquid phase monitoring reaction is finished; adding hydrochloric acid into the water layer to adjust the pH value to be 2.5 +/-0.5, extracting with ethyl acetate, adding cyclohexylamine into the ethyl acetate layer, stirring, crystallizing, and filtering to obtain 7-methoxy cephalothin acid cyclohexylamine salt;

and step 3: adding 500g of 7-methoxy cephalothin acid cyclohexylamine salt into 3000g of water and 4500g of methanol in a 10L four-neck round-bottom flask, cooling to-30 ℃, dropwise adding 390ml of 3.0mol/L sodium hydroxide aqueous solution within 1h, reacting for 2 hours, and monitoring by a high performance liquid phase until the reaction is complete; heating to room temperature, adding acetic acid to adjust the pH of the system to 3.3 +/-0.2, stirring, cooling and crystallizing to obtain a deacetylated intermediate;

and 4, step 4: adding 650g of deacetylation intermediate into 5200ml of dry tetrahydrofuran, cooling to-40-37 ℃, adding 130g of chlorosulfonyl isocyanate within 45 minutes, and monitoring by a high performance liquid phase until the reaction is complete; adding 600g of pure water, heating to 20-23 ℃, stirring for 45min, dropwise adding concentrated hydrochloric acid to adjust the pH value to 1.5-2.0, cooling to 0-5 ℃, crystallizing, and filtering to obtain cefoxitin acid;

and 5: adding 520g of cefoxitin acid into 1600ml of methanol and 2000ml of acetone, stirring and dissolving at the temperature of 10-12 ℃, adding activated carbon for decoloring, and filtering; slowly dripping a sterilized methanol solution containing 140g of sodium lactate into the filtrate, wherein the mass concentration of the sodium lactate is 30 percent, and after the addition of the sodium lactate for 2 hours, the stirring speed is 175 r/m; and continuously reacting for 2 hours, cooling to 5-7 ℃ at the speed of 5 ℃/h, crystallizing, stirring at the speed of 130 rpm, filtering, washing and drying to obtain the cefoxitin sodium crystal.