CN110396103B

CN110396103B - Cefazolin sodium or composition thereof, preparation method and preparation thereof, and new indications of reproductive system infection

Info

Publication number: CN110396103B
Application number: CN201811183682.9A
Authority: CN
Inventors: 傅苗青; 吴丰喆; 孟宾; 周白水; 俞玉萍; 马庆双
Original assignee: Guangdong Jincheng Jinsu Pharmacy Co ltd
Current assignee: Guangdong Jincheng Jinsu Pharmacy Co ltd
Priority date: 2018-10-11
Filing date: 2018-10-11
Publication date: 2021-03-19
Anticipated expiration: 2038-10-11
Also published as: CN110396103A

Abstract

The invention provides cefazolin sodium or a composition thereof, a preparation method, a preparation and an application thereof. The preparation method provided by the invention has high reproducibility and stable and reliable production process; the prepared cefazolin sodium or the composition thereof has low impurity content, is beneficial to improving the quality of raw materials and corresponding preparations, and improves the safety and clinical treatment effect of the preparations; and has application in preparing medicine for treating genital system infection.

Description

Cefazolin sodium or composition thereof, preparation method and preparation thereof, and new indications of reproductive system infection

Technical Field

The invention relates to an antibiotic medicine, in particular to cefazolin sodium, a preparation and application thereof.

Background

Cefazolin sodium is the first generation of cephalosporin, has antibacterial effect on gram-positive bacteria and gram-negative bacteria, and is originally developed and developed as Ansfer ANCEF. It is used for treating infection of respiratory system, urinary system, soft tissue of skin, bone and joint, biliary tract, endocarditis, septicemia, and ear infection caused by sensitive bacteria.

In clinical application, cefazolin sodium causes accidental adverse reactions, such as allergic dermatitis, drug fever, nausea, vomiting, diarrhea, anaphylactic shock and the like. Although there are many factors causing adverse reactions, such as differences among human individuals, dosage, combined medication, administration mode and the like, one of the important causes of adverse reactions is impurities in cefazolin sodium, and the source of the impurities can be a synthetic intermediate or a degradation product generated in storage.

For the medicines, particularly for cephalo-type medicines, the purity of the medicines is improved, the content of impurities is reduced, and the method plays an important role in improving the safety and curative effect of the medicines. Therefore, the impurity control level is improved, the drug stability is improved, and the controllable product quality is ensured, and the control level is more and more paid more attention by pharmaceutical enterprises.

Cefazolin sodium is generally prepared by adopting a synthesis process, and different synthesis routes and control parameters may cause impurities with different levels to be generated in a product, so that cefazolin sodium preparation has poor stability and low safety.

Based on the problems in the prior art, the inventor improves the synthesis and process of cefazolin sodium, controls the impurity level, and improves the quality of cefazolin sodium so as to improve the clinical curative effect and the use safety of cefazolin sodium.

Disclosure of Invention

In order to solve the above problems, the present inventors have conducted intensive studies, and as a result, found that cefazolin sodium with a low impurity content is provided by improving a raw material synthesis process, which is advantageous for improving the quality of a raw material and a corresponding preparation, and improving the safety and clinical treatment effect of the preparation, thereby completing the present invention.

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

in a first aspect, the invention provides cefazolin sodium or a composition thereof, wherein the mass content of an active ingredient, namely cefazolin sodium, is more than 98%;

the cefazolin sodium or the composition thereof also comprises an impurity K,

the mass content of the impurity K is below 0.1 percent.

Further, the cefazolin sodium or the composition thereof also comprises an impurity N,

the mass content of the impurity N is below 0.3 percent.

Preferably, the cefazolin sodium or the composition thereof further comprises an impurity S,

the mass content of the impurity S is below 0.2 percent.

In a second aspect, the present invention provides a preparation method of cefazolin sodium, which is characterized in that the preparation method comprises the following steps:

step 1: dispersing TDA in dichloromethane, adding triethylamine, dropwise adding trimethylchlorosilane under controlled temperature, and reacting at 0-10 ℃ to obtain a dichloromethane solution of an intermediate 1;

step 2: dispersing TAA in dichloromethane, adding triethylamine, dropwise adding pivaloyl chloride under controlled temperature, and reacting at 0-10 ℃ to obtain a dichloromethane solution of an intermediate 2;

and step 3: dropwise adding the dichloromethane solution of the intermediate 2 into the dichloromethane solution of the intermediate 1, controlling the temperature to be 0-10 ℃ to react until the reaction is finished, adding water, layering, and taking a dichloromethane phase;

and 4, step 4: adding an acidic aqueous solution into the dichloromethane phase obtained in the step (3), reacting for 3 hours, and reacting until the reaction is finished; layering, adding acetone into the water phase, adjusting the pH value, filtering and drying to obtain cefazolin;

and 5: dissolving the prepared cefazolin in a mixed solvent, adding sodium salt, cooling, and crystallizing to obtain a cefazolin sodium crude product.

The method also comprises recrystallization, wherein in the recrystallization treatment, the prepared cefazolin sodium crude product is heated and dissolved in a crystallization solvent, and is filtered by a microporous filter membrane, cooled to different temperatures for growing crystals, filtered and dried to obtain cefazolin sodium pentahydrate or anhydrous cefazolin sodium.

Wherein, the crystal growth is carried out at the temperature of 30-35 ℃ for 5-10 h, and the anhydrous cefazolin sodium is obtained after filtration and drying; or the like, or, alternatively,

and cooling to 10-15 ℃, preserving heat, growing the crystal for 2-5 h, filtering, and drying to obtain cefazolin sodium pentahydrate.

In a third aspect, the invention provides a single preparation of cefazolin sodium, wherein the active ingredient is the cefazolin sodium or the composition thereof provided by the first aspect of the invention, or the cefazolin sodium prepared by the method provided by the second aspect, and the single preparation of cefazolin sodium is an injection, preferably an aseptic powder injection for injection.

In a fourth aspect, the invention provides a cefazolin sodium compound preparation, which comprises cefazolin sodium or a composition thereof, or cefazolin sodium prepared by the method provided by the second aspect of the invention, wherein the cefazolin sodium compound preparation further comprises an active ingredient beta lactamase inhibitor;

optionally, pharmaceutically acceptable adjuvants and/or pharmaceutically active substances without incompatibility are also included.

In a fifth aspect, the invention provides cefazolin sodium or a composition thereof, a single cefazolin sodium preparation, a compound cefazolin sodium preparation, or an application of cefazolin sodium prepared by the method in preparing a medicament for treating reproductive system infection.

The infection of the reproductive system comprises prostatitis, epididymitis, bartholinitis, uterine cavity infection, adnexitis and parauterine connective tissue inflammation;

especially the infections of the reproductive system including prostatitis, epididymitis, bartholinitis, uterine cavity infection, adnexitis and parauterine connective tissue inflammation caused by Escherichia coli, Proteus mirabilis, Klebsiella and enterococcus.

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

(1) the cefazolin sodium or the composition thereof provided by the invention has low impurity content, is beneficial to improving the quality of raw materials and corresponding preparations, and improves the safety and clinical treatment effect of the preparation;

(2) the preparation method of cefazolin sodium or the composition thereof improves the raw material synthesis process, so that the production method has high reproducibility and the production process is stable and reliable;

(3) the invention also provides the application of the cefazolin sodium or the composition and the preparation thereof in preparing the medicines for treating the infection of the reproductive system, in particular to the application of the medicines for treating the infection of the reproductive system, including prostatitis, epididymitis, bartholinitis, uterine cavity infection, adnexitis and parauterine connective tissue inflammation, caused by escherichia coli, proteus mirabilis, klebsiella and enterococcus.

Drawings

Figure 1 shows the dosing profile of cefazolin sodium in example 9.

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 present invention is described in detail below.

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

step 1: synthesis of intermediate 1

In the preparation method of cefazolin sodium provided by the invention, 7-amino-3 [2- (5-methyl-I, 3, 4-thiadiazole) thiomethyl ] -3-cephem-4 hydroxy acid, also called TDA, is used as a starting material. However, TDA is easily condensed with itself during the reaction to form an impurity K, as follows,

therefore, in the present invention, the carboxyl group of TDA is protected to reduce the generation of impurity K. The carboxyl protection method is various, ester bonds, amido bonds and silane bonds can be selected to protect the carboxyl, and silane substances with simple process and high reaction activity are preferably used to protect the carboxyl.

In a preferred embodiment, trimethylchlorosilane is selected to protect carboxyl groups, as shown in the following reaction scheme:

the reaction activity of the trimethylchlorosilane is very high, and the product after the reaction is easier to remove; after carboxyl is protected by trimethylchlorosilane, the protecting group can be removed under mild acidic conditions, and deprotection treatment is easy to realize, so that the process flow is simple, the treatment difficulty is low, and the synthesis process is simplified.

In the reaction process of the intermediate 1, hydrochloric acid is generated after the protection of carboxyl by trimethylchlorosilane, and in order to improve the reaction yield, alkali is added into a reaction system to promote the reaction to proceed towards the direction of generating the intermediate 1.

In order to ensure the stability of TDA, a weak base is preferably added into the reaction system, and triethylamine which is miscible with dichloromethane is preferably added to remove hydrochloric acid continuously generated in the reaction system.

In a preferred embodiment, an excessive amount of triethylamine is added into the reaction system, so that the triethylamine is prevented from being added again in the subsequent step 3, and the treatment process is simplified.

The triethylamine is added in an amount of 1.5 to 4 times, preferably 2 to 3.5 times, and most preferably 2 to 3 times the molar equivalent of trimethylchlorosilane.

The exothermic reaction of the trimethylchlorosilane can cause the temperature of a reaction system to rise, and the reaction activity of the trimethylchlorosilane is improved, so that the reaction is violent, and potential safety hazards are generated. Meanwhile, when the local temperature is too high in the reaction, the generation of the impurity K is increased, and the mass content of the impurity K in the prepared cefazolin sodium is increased.

Preferably, TDA is dispersed in dichloromethane, triethylamine is added, and the temperature of a reaction system is reduced to below-5 ℃ under stirring, more preferably to between-5 and-10 ℃; and dropwise adding trimethylchlorosilane into the reaction system. The reaction temperature was monitored so that the temperature of the reaction system did not exceed 5 ℃ during the dropwise addition of trimethylchlorosilane.

The trimethylchlorosilane is added in the same amount as the molar equivalent of TDA or in a slight excess amount to improve the yield.

After the dropwise addition is finished, in order to improve the reaction rate and shorten the reaction time, the reaction is carried out for 2-3h at the temperature of 0-10 ℃ and preferably 3-7 ℃ to obtain a dichloromethane solution of the intermediate 1.

The generation of the impurity K is obviously reduced by protecting the carboxyl of the TDA and controlling the reaction temperature.

Step 2: synthesis of intermediate 2

The invention uses Tetrazole Acetic Acid (TAA) and pivaloyl chloride to prepare an intermediate 2, and the pivaloyl chloride generates hydrochloric acid after reaction. In order to increase the reaction yield, a base is added to the reaction system to promote the reaction toward the production of intermediate 2.

Preferably, the same weak base as that in step 1, triethylamine, is added to the reaction system to remove hydrochloric acid continuously generated in the reaction system. In a preferred embodiment, the molar equivalent of triethylamine added to the reaction system is substantially equivalent to pivaloyl chloride.

The pivaloyl chloride is added in the same amount as or in a slight excess of the molar equivalents of TAA to improve the yield.

Because the pivaloyl chloride has high reaction activity, the reaction activity of the pivaloyl chloride is further improved by heat release after the pivaloyl chloride reacts with TAA, the temperature of a reaction system is easily increased sharply in a short time, and the operation safety is influenced.

Preferably, TAA is dispersed in dichloromethane, triethylamine is added, and the temperature is reduced to below-5 ℃ under stirring, more preferably to between-5 and-10 ℃; and dropwise adding pivaloyl chloride into the reaction system, and monitoring the reaction temperature to ensure that the temperature of the reaction system is not more than 5 ℃, preferably not more than 3 ℃ in the process of dropwise adding pivaloyl chloride.

After the dropwise addition is finished, in order to improve the reaction rate and shorten the reaction time, the reaction is carried out for 1-2 hours at 0-10 ℃ and preferably at 0-5 ℃ to obtain a dichloromethane solution of the intermediate 2.

The synthesis scheme of the intermediate 2 is as follows:

and step 3: synthesis of intermediate 3

Intermediate 3 was prepared using intermediate 1 prepared in step 1 and intermediate 2 prepared in step 2. Preferably, the reaction is carried out by adding dropwise a dichloromethane solution of the intermediate 2 prepared in the step 2 to a dichloromethane solution of the intermediate 1 prepared in the step 1.

This is because intermediate 2 has an activated acid anhydride group, and when intermediate 2 is added dropwise to intermediate 1, the activated amino group in intermediate 1 preferably attacks the carbonyl group of TAA to produce intermediate 3. If intermediate 1 is added dropwise to intermediate 2, this results in the amino group simultaneously attacking the pentanoic carbonyl group and producing the impurity N in the subsequent step 4.

In addition, because the reaction solution of the intermediate 1 contains excessive triethylamine, carboxylic acid groups are generated after the acid anhydride groups of the intermediate 1 and the intermediate 2 react, and the forward progress of the reaction is influenced. The reaction can be smoothly carried out forward by adding the solution of the intermediate 2 to the solution of the intermediate 1 containing an excess of triethylamine to produce the intermediate 3, without adding a weak base such as triethylamine to the solution to promote the reaction toward the production of the intermediate 3.

The synthesis scheme for intermediate 3 is as follows:

the acid anhydride group has high activity, and generates a large amount of heat during decomposition, and the temperature of the reaction solution can be increased in a short time, thereby further increasing the activity of the acid anhydride group in the intermediate 2. When the activity of intermediate 2 is increased, the activity of both carbonyl carbons is increased, resulting in an increase in impurity N.

Therefore, it is necessary to control the temperature at which the intermediate 3 is reacted.

And the reaction temperature is controlled to be-10-0 ℃ and preferably-5-0 ℃ during dropwise adding, so that the reaction activity of the anhydride is reduced, and the occurrence of side reactions is reduced. After the dropwise addition is finished, in order to improve the reaction rate and shorten the reaction time, the reaction is continued for 2-4 hours at 0-10 ℃, preferably 3-8 ℃, so that the reaction yield is improved.

Optionally, water may be added to the reaction system after the reaction is completed to separate layers, and a dichloromethane phase is left. The intermediate 3 can be primarily extracted and purified by dissolving the water-soluble substance in the water layer, thereby reducing the content of water-soluble impurities therein.

Preferably, the addition amount of water is 0.5-1 time of the reaction volume in the step 3.

And 4, step 4: synthesizing cefazolin;

and (3) adding an acidic aqueous solution into the reaction liquid in the step (3) or the reserved dichloromethane phase to deprotect the carboxyl group. The acidic component in the acidic aqueous solution can be any or combination of sulfuric acid, tartaric acid, citric acid and hydrochloric acid, and is preferably tartaric acid with mild acidity; more preferably, the mass fraction of the acidic component is 15-25%.

Preferably, the volume of the acidic aqueous solution added is substantially the same as the volume of the reaction solution or the remaining dichloromethane phase, so that the molar equivalent of the acidic component added is in excess with respect to the molar equivalent of the intermediate 3 to sufficiently deprotect the carboxyl group of the intermediate 3.

Preferably, the volume of the acidic aqueous solution is 0.8-1.2 times of the volume of the reaction solution, more preferably 0.9-1.1 times, and the generated cefazolin can be fully dissolved; and, the amount of organic solvent used is reduced in the subsequent treatment.

In order to improve the reaction rate of deprotection of carboxyl, the reaction is preferably carried out for 3-5h at 30-40 ℃, after the reaction is finished, layering is carried out, a water phase is taken, and the cefazolin is dissolved in the water phase.

Adding acetone into the water phase, and adjusting the pH value to 1.0-3.0, preferably 1.0-2.0; and carrying out heat preservation and stirring reaction for 2 hours at the temperature of 0-10 ℃, carrying out suction filtration, and drying to obtain the cefazolin. The following were used:

the pH of the solution is preferably adjusted by using a hydrochloric acid solution, and the solubility of the hydrochloric acid solution is preferably 10-20% by volume, for example, 15% by volume.

Researches show that a small amount of amino in the anhydride group in the intermediate 2 and the amino in the intermediate 1 attack pivalate carbonyl in the reaction process, and the generated impurity is deprotected by tartaric acid to generate the impurity N.

The generation of impurity N is schematically shown below:

the impurity N has a structure different from that of cefazolin, the cefazolin contains a tetrazole group, and a corresponding part in the impurity N is a trimethyl group. Researches find that based on the fact that the solubility of the impurity N and the solubility of the cefazolin in the acetone solvent are different, acetone is added into the water phase, and the proportion of the mixed solution is adjusted to increase the solubility of the impurity N, reduce the solubility of the cefazolin in the mixed solvent, reduce the content of the impurity N and improve the yield of the cefazolin.

The volume ratio of the added volume of the acetone to the volume of the water phase is 1; 1 to 5, preferably 1:1.2 to 1.5, and more preferably 1:1.2 to 1.4.

Through multiple tests, the hydrochloric acid solution is dropwise added into the mixed solution added with the acetone to adjust the pH value to 1.0-1.5, so that a large amount of cefazolin can be separated out, and the yield is improved; moreover, most of impurities, particularly N, are dissolved in the mother liquor, so that the content of the N impurities is effectively reduced.

And 5: preparing a crude product of cefazolin sodium;

and (4) alkalizing the cefazolin prepared in the step (4) by using sodium salt, and generating cefazolin sodium from the cefazolin. In the process of preparing sodium salt from cefazolin, impurities S can be generated by hydrolysis, and the impurities S are both starting raw materials TDA and degradation impurities. Through the process research, we find that the content of the impurity S can be well controlled by controlling the salification reaction reagent and the temperature.

In the salt-forming reaction, the cefazolin is dissolved in a mixed solvent, preferably a mixed solvent with the volume ratio of water to acetone of 1: 1-3, organic impurities in the cefazolin which are not completely removed in the steps can be dissolved in acetone, and the cefazolin and the generated product of cefazolin sodium are further purified.

The sodium salt is selected from sodium bicarbonate, sodium carbonate and sodium lactate, and sodium bicarbonate with weak alkalinity and good water solubility is preferably used. Adding sodium salt into the reaction solution, and continuously stirring for reaction for 0.5-1.5 h; in order to improve the reaction rate, the reaction is preferably carried out for 1 to 1.5 hours at the temperature of between 30 and 40 ℃, so that the cefazolin can be fully reacted into cefazolin sodium, and the content of the impurity S is greatly reduced.

Optionally, before the sodium salt is added, decoloring treatment can be performed, preferably decoloring treatment is performed by using activated carbon, pyrogen in the solution can be adsorbed, and the clinical safety of the product of cefazolin sodium is improved.

When in decolorization, the adding amount of the active carbon is 1-2g/L, which can meet the requirements of decolorization and pyrogen removal treatment.

After the sodium salt is added for reaction, stirring and growing crystals for 3-6 h under heat preservation, cooling the temperature of the reaction system to 0-10 ℃, preferably 0-5 ℃, and crystallizing to obtain a cefazolin sodium crude product.

Step 6: recrystallizing the cefazolin sodium crude product;

and (5) recrystallizing the cefazolin sodium crude product obtained in the step (5), so that the purity of the product is further improved, and the content of impurities is reduced.

The crystallization solvent is used for recrystallization, and the water-containing mixed solvent is used as the crystallization solvent, so that organic impurities with good water solubility can be dissolved in the crystallization mother liquor, and a large amount of precipitation of cefazolin sodium is ensured.

The crystallization solvent is a mixed solvent of water for injection and one or more of ethanol, isopropanol, methanol, acetonitrile and acetone, and preferably an isopropanol aqueous solution or an acetone aqueous solution is used as the crystallization solvent. In the crystallization solvent, the impurity removal capacity of recrystallization is strong, and the degradation of cefazolin sodium is not caused.

In the crystallization solvent, the volume ratio of the injection water to the acetone/isopropanol is 1: 1-3, preferably 1: 1.5-2.5.

In a preferred embodiment, a mixed solution of isopropanol/water in a volume ratio of 2 may be used as the crystallization solvent, as shown below:

and (3) dissolving the crude product of cefazolin sodium prepared in the step (5) in a crystallization solvent, heating to improve the solubility of the cefazolin sodium in the mixed solvent, and reducing the dosage of the solvent. However, the cefazolin sodium is unstable in an aqueous solution and has poorer stability when being heated, and multiple experiments show that the content of impurities S in the degradation product of the cefazolin sodium can be reduced by preferably heating a recrystallization solvent to 40-50 ℃, filtering and sterilizing the recrystallization solvent through a microporous filter membrane while the recrystallization solvent is hot and then cooling the recrystallization solvent.

During recrystallization, the cooling rate and the stirring rate can affect various indexes such as purity, form, particle size and the like of a precipitated product, and directly affect the quality of crystallized cefazolin sodium, so that the quality stability of a preparation prepared from the cefazolin sodium is poor, and the industrial production treatment difficulty is high.

When the cooling rate is too fast, the product is separated out fast, and is easy to embed and mix with impurities in the mother liquor, so that the impurity content of the product is too high; when the cooling rate is too slow, the precipitation time is too long, and the crystallization efficiency is low. Preferably, the cooling rate is 5-10 ℃/h, preferably 6-8 ℃/h, the solution in which the cefazolin sodium is dissolved is cooled to 0-5 ℃, and the solution is subjected to heat preservation and crystallization.

During crystallization, the stirring speed is controlled to be 60-100 r/min (revolutions per minute), preferably 60-80 r/min, so that the dispersion of powder particles is facilitated, and the powder particles are not easy to aggregate; meanwhile, collision among powder particles is not excessive and violent, so that the range of particle size distribution can be reduced, the particle sizes are concentrated, and preparation of the preparation is facilitated.

Wherein, when the temperature is reduced to 30-35 ℃, the crystal is kept for 5-10 h, and then the anhydrous cefazolin sodium can be obtained by filtering;

and when the temperature is reduced to 10-15 ℃, keeping the temperature and growing the crystal for 2-5 h, and filtering to obtain the cefazolin sodium pentahydrate.

And drying the filter cake for 5-10 hours at the temperature of 30-40 ℃ and the vacuum degree of-0.09 MPa to-0.1 MPa to respectively obtain dry anhydrous cefazolin sodium and pentahydrate cefazolin sodium.

The cefazolin sodium provided by the invention can be anhydrous cefazolin sodium, and can also contain cefazolin sodium pentahydrate.

Meanwhile, the invention also provides cefazolin sodium or a composition thereof prepared by the method, and the cefazolin sodium or the composition thereof has low powder chroma and uniform particle size. Wherein, in the cefazolin sodium or the composition thereof, the mass content of the cefazolin sodium is more than 98 percent, preferably more than 99 percent calculated by anhydrous substance.

The cefazolin sodium or the composition thereof also contains an impurity K, and the content of the impurity K is within 0.1%, preferably within 0.08%.

The content of the impurity N is 0.3% or less, preferably 0.2% or less, and most preferably 0.15% or less.

More preferably, the cefazolin sodium or the composition thereof further contains impurity S, and the content of the impurity S is within 0.2%, preferably within 0.15%, more preferably within 0.1%.

Further, the cefazolin sodium or the composition thereof also contains polymer impurities, and the mass content of the polymer impurities is not higher than 0.1% or not higher than 0.15%. The polymer is generally the cause of antibiotic allergy, and the control of the content of the polymer is very helpful to reduce the sensitization rate and improve the clinical safety.

The invention provides a cefazolin sodium preparation of cefazolin sodium or a composition thereof prepared by the method, which is a single injection prepared by taking cefazolin sodium as a main active ingredient.

The cefazolin sodium preparation is preferably packaged in an aseptic mode, and the cefazolin sodium or the composition thereof is packaged after being crushed and sealed.

The cefazolin sodium preparation can also be a freeze-dried preparation and is prepared by using conventional technology and auxiliary materials in the prior art.

The invention also provides a cefazolin sodium compound preparation of cefazolin sodium prepared by using the cefazolin sodium or the composition thereof or the method, wherein the cefazolin sodium compound preparation comprises active ingredients of cefazolin sodium and beta lactamase inhibition, and optionally, the cefazolin sodium compound preparation further comprises pharmaceutically acceptable auxiliary materials and/or pharmaceutically active substances without incompatibility.

The beta lactamase inhibitor is preferably tazobactam sodium, sulbactam sodium or ababactam sodium, and is more preferably tazobactam sodium.

In a more preferred embodiment, the cefazolin sodium (in the form of cefazolin C)₁₄H₁₄N₈O₄S₃Calculated) and sulbactam sodium (calculated as sulbactam C)₈H₁₁NO₅S) is 1-4: 1, such as 1:1.

The cefazolin sodium is extremely unstable to beta lactamase, and the inhibition of the beta lactamase can inhibit the beta lactamase and reduce the degradation of the cefazolin sodium, thereby improving the antibacterial action of the cefazolin sodium.

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

Furthermore, the single preparation and the compound preparation are subjected to nitrogen filling treatment after being subpackaged, so that the stability under long-term storage is improved, the generation of oxidative degradation impurities is reduced, and the clinical curative effect and the use safety are improved.

According to the cefazolin sodium or the composition thereof, the single preparation of the cefazolin sodium and the compound preparation of the cefazolin sodium, the invention also provides an application in preparing the medicines for treating the infection of the reproductive system.

Further, the infection of the reproductive system comprises prostatitis, epididymitis, bartholinitis, uterine cavity infection, adnexitis and parauterine connective tissue inflammation; particularly infections of the reproductive system including those caused by Escherichia coli, Proteus mirabilis, Klebsiella and enterococcus.

Examples

Example 1

Adding 1033g of TDA, 5000ml of dichloromethane and 667g of triethylamine into a reaction bottle, starting stirring, cooling a reaction system to-6 ℃, dropwise adding 358g of trimethylchlorosilane into the reaction system, heating to 3-5 ℃ after dropwise adding is finished, and keeping the temperature for 2 hours to obtain a dichloromethane solution of an intermediate 1;

adding 384g of TAA and 2000ml of dichloromethane into a reaction bottle, adding 333g of triethylamine, cooling to-10 ℃ under stirring, dropwise adding 3980g of pivaloyl chloride into the reaction system, controlling the temperature to be 0-2 ℃, and after dropwise adding, keeping the temperature at 3-5 ℃ for reaction for 1.5h to obtain a dichloromethane solution of the intermediate 2.

Dropwise adding the dichloromethane solution of the intermediate 2 into the dichloromethane solution of the intermediate 1, controlling the dropwise adding temperature to be-5 to-2 ℃, reacting for 2 hours at 3 to 5 ℃ after the dropwise adding is finished, adding 5000ml of water after the reaction is finished, layering, and reserving a dichloromethane phase;

adding 6750g of 20% tartaric acid aqueous solution into the retained dichloromethane phase, reacting for 3h at 35 ℃, layering after the reaction is finished, adding 5000ml of acetone into the water phase, adjusting the pH value to 1.0 by using 15% hydrochloric acid, stirring for 2h at 0-5 ℃, performing suction filtration and drying to obtain 1167g of cefazolin, wherein the yield is as follows: 85.6 percent;

adding 909g of cefazolin into a reaction bottle, adding 2000ml of water and 3000ml of acetone, heating to 35 ℃, adding 10g of activated carbon for decoloring for 30 minutes after the solution is clarified, and filtering by a microporous filter membrane; adding 252g of sodium bicarbonate, stirring at 30-35 ℃ for crystal growth for 5h, continuously cooling to 0-3 ℃, and crystallizing for 2h to obtain 883g of crude cefazolin sodium product with yield of 92.7%;

adding 800g of cefazolin sodium crude product into a reaction bottle, adding 1500ml of water for injection and 3000ml of acetone, heating to 45 ℃, adding medicinal activated carbon for injection to decolor for 30 minutes after the solution is clarified, and filtering with a 0.2-micron microporous filter membrane to remove the activated carbon; cooling to 0-3 ℃ at the speed of 6 ℃/h, stirring at the speed of 65r/min, cooling to 30-35 ℃, keeping the temperature and growing the crystals for 8h, filtering, and drying a filter cake for 6h under the conditions of the temperature of 35 ℃ and the vacuum degree of-0.05 MPa to obtain an anhydrous cefazolin sodium fine product 748g, wherein the yield is as follows: 93.5 percent.

Example 2

The method is the same as the synthesis method of the embodiment 1, and is different in that 1500ml of injection water and 3000ml of isopropanol are used as crystallization solvents in recrystallization treatment, the temperature is reduced to 10-15 ℃, crystal growth is carried out for 3 hours, so that 884g of fine cefazolin sodium pentahydrate is obtained, and the yield is as follows: 93.0 percent.

Example 3

The same synthesis as in example 1, except that in the preparation of cefazolin, 20% hydrochloric acid solution was used to adjust the pH to 1.5.

Example 4

And (2) crushing the cefazolin sodium prepared in the example 1 to 35-45 micrometers of D90, and filling nitrogen into 1g of cefazolin sodium according to the amount of the cefazolin, and subpackaging to obtain the single preparation of the cefazolin sodium.

Example 5

Crushing the cefazolin sodium prepared in the example 1 to D90 of 35-45 microns; the medicinal sulbactam sodium obtained from any commercial manufacturer is crushed to D90 of 35-50 microns.

The mass ratio of cefazolin to sulbactam is 1:1, and after the cefazolin and sulbactam are uniformly mixed, the cefazolin sodium compound preparation is obtained by filling nitrogen in a dosage of 1g of cefazolin and subpackaging.

Example 6

The cefazolin sodium prepared in example 1 is crushed to D90 of 35-45 micrometers, and the medicinal abamectin sodium purchased from any commercial manufacturer is crushed to D90 of 35-50 micrometers.

100 parts of cefazolin sodium compound preparation are prepared according to the following formula:

and after uniformly mixing, subpackaging 1.0g of cefazolin to obtain the cefazolin sodium compound preparation. )

Comparative example

Comparative example 1

The synthesis method is the same as that of example 1, except that: cefazolin was produced directly using TDA with intermediate 2 as follows:

and (2) dropwise adding the dichloromethane solution of TDA into the dichloromethane solution of the intermediate 2, controlling the dropwise adding temperature to be-5 to-2 ℃, reacting for 2 hours at 3 to 5 ℃ after the dropwise adding is finished, adding 5000ml of water after the reaction is finished, layering, and reserving a dichloromethane phase.

Comparative example 2

The synthesis method is the same as that of example 1, except that: in the process of preparing cefazolin, acetone is not added into the water phase, and 15% hydrochloric acid solution is directly used for adjusting and adjusting the pH value to 3.

Comparative example 3

The synthesis method is the same as that of example 1, except that: in the preparation of the sodium salt of cefazolin, sodium hydroxide is used as the sodium salt.

Comparative example 4

The synthesis method is the same as that of example 1, except that: when the crude product of cefazolin sodium is recrystallized, the temperature is reduced at the speed of 20 ℃/h, and the stirring speed is 170 r/min.

Examples of the experiments

Experimental example 1

The clarity of the solutions of cefazolin sodium obtained in examples 1 to 6 (first method of the clarity inspection method in pharmacopoeia 0902, 2015 edition: visual method) and insoluble particles (second method of the insoluble particles inspection method in pharmacopoeia 0903, 2015 edition: microscopic counting method) were measured, and the results were as follows:

it can be seen that the clarity of the solution of cefazolin sodium prepared in examples 1-6 and the detection result of insoluble particles are good, and both meet the requirements of the existing pharmacopoeia.

Experimental example 2

The particle size (μm) of the cefazolin sodium prepared in examples 1-3 and comparative example 4 was measured, and the results were as follows:

as can be seen, the particle size of the product prepared in the examples 1-3 is more uniform and fine, the distribution range is narrower, and the product is very favorable for subpackage treatment after crushing treatment.

Experimental example 3

The residual solvents of the cefazolin sodium crude drugs prepared in examples 1-3 were determined by gas phase external standard method, and the results were as follows:

	methylene dichloride	Isopropanol (I-propanol)	Acetone (II)
				Example 1	/	/	80ppm
Example 2	/	92ppm	/
				Example 3	/	/	87ppm

"/" indicates no detection.

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

Experimental example 4

The mass contents of cefazolin sodium (calculated as anhydrous cefazolin sodium), impurity K, N, S and polymer obtained in examples 1-3 and comparative examples 1-3 were determined by 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 cefazolin sodium in examples 1-3 remained stable after long-term testing. The small increase in impurities, especially polymer impurities, indicates a higher stability and therefore a higher safety.

Experimental example 5

The cefazolin sodium preparations obtained in examples 4-6 were measured for their preparation content (relative value of the measured amount to the labeled amount), impurity K, N, S and polymer mass content (calculated as anhydrous cefazolin sodium) by high performance liquid chromatography (external standard method). The test samples were placed under long-term test conditions (30 ℃. + -. 2 ℃/65% RH. + -. 5% RH) for 0, 12, 24 months, respectively.

Acidity was measured according to the 2015 th pharmacopoeia method.

After long-term tests, the cefazolin sodium preparations prepared in examples 4 to 6 have good long-term stability, stable active ingredient content, little degradation, high clinical efficacy and high safety.

Experimental example 6

Healthy adult male SD rats, 8 in each group, were inserted into the dorsal lobe of the prostate of each rat, and infected bacteria (respectively using Escherichia coli, Proteus mirabilis, and Klebsiella) were injected for molding to prepare a prostatitis model animal. The test group used the single formulation prepared in example 4, and the administration was carried out by tail-intraperitoneal injection at a dose of 0.2g/kg, with the administration frequency of 2 times/day, and the treatment period of 2 days; control group, no drug treatment. A normal group was also set, healthy rats, and no drug treatment was given.

The rats were sacrificed and the prostate was dissected and examined for light microscopy pathology.

The results show that the inflammatory symptoms are significantly better; no allergic symptoms were observed during the treatment.

Experimental example 7

Healthy adult male SD rats are selected, each group comprises 8 animals, infectious bacteria (Escherichia coli, Proteus mirabilis and Klebsiella) are directly injected into the heads of the epididymis on one side of the rats respectively for molding, and epididymitis model animals are prepared. The test group used the preparation prepared in example 5, and the administration frequency was 2 times/day, and the treatment period was 3 days, with a dose of 0.1g/kg administered by tail intraperitoneal injection; control group, no drug treatment. A normal group was also set, healthy rats, and no drug treatment was given.

The rat was sacrificed and epididymal tail tissue was removed for microscopic pathological observation.

Experimental example 8

12 healthy male beagle dogs with the weight of 8-10 kg are selected and divided into a test group and a control group, and 6 dogs are selected.

Establishing a reproductive system infection model: beagle dogs were anesthetized and a germ line was injected with proteus mirabilis to obtain a reproductive system infection model.

Test groups: selecting male beagle dogs with reproductive infection to inject the cefazolin sodium compound preparation prepared in the example 5 into the veins, wherein the dosage is 80mg/kg, the cefazolin sodium compound preparation is taken twice a day, the administration is continuously carried out for 5 days, and the vas deferens infection condition is detected after 5 days.

Control group: selecting a male beagle dog with reproductive infection, carrying out no treatment, and detecting the infection condition of the vas deferens after 5 days.

Evaluation criteria:

and (3) curing: the symptoms and signs disappeared.

The method has the following advantages: symptoms and signs were partially reduced.

And (4) invalidation: there was no reduction or an increase in symptoms.

The results of the pharmacological studies on beagle dogs are shown in the following table:

group of	Severe inflammation	Slight inflammation	Without inflammation
				Test group	0	1	5
Control group	4	2	0

Experimental example 9

Clinical trials were conducted using the formulation prepared in example 4, and 100 patients with bartholinitis, uterine cavity infection, adnexitis, and parauterine connective tissue inflammation, each of which was 25-50 years old and 60-75kg in weight, were selected for study.

The treatment method comprises the following steps: intravenous drip, 1.0g dose/time, 2 times daily.

The treatment period is as follows: and 7 days.

Evaluation criteria:

and (3) curing: the symptoms and signs disappear, or the calcitonin PCT is less than 0.25 ng/ml.

The method has the following advantages: partial reduction of symptoms and signs, or calcitonin 0.25 < PCT < 0.5 ng/ml.

And (4) invalidation: no reduction or an exacerbation of symptoms, or calcitonin PCT > 0.5 ng/ml. The treatment results are as follows:

and (3) drawing blood for sample reservation, detecting the blood concentration of the cefazolin sodium in the blood sample, calculating the average value of all subjects, and drawing a pharmaceutical time curve. The results are shown in figure 1, when the cefazolin sodium pharmaceutical preparation is used for treating the infection of the reproductive system, the maximum concentration reaches 185.1 mu g/mL, and the half-life period of the drug in blood plasma is about 116 min.

Statistically, the treatment results are shown in the following table:

kind of disease	Cure of disease	Is effective	Invalidation
				Bartholinitis	82	18	0
Infection of uterine cavity	87	12	1
				Adnexitis of uterus	86	13	1
Periuterine connective tissue inflammation	84	16	0

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 cefazolin sodium is characterized in that the mass content of an active ingredient cefazolin sodium in the cefazolin sodium is more than 98%;

the cefazolin sodium also comprises an impurity K,

the mass content of the impurity K is below 0.1 percent;

the impurity N is a nitrogen-containing impurity,

the mass content of the impurity N is below 0.3 percent;

the impurities of the S are removed from the slurry,

the mass content of the impurity S is below 0.2 percent;

the method for preparing cefazolin sodium comprises the following steps:

step 2: dispersing TAA in dichloromethane, adding triethylamine, cooling to below-5 ℃ under stirring, controlling the temperature to be dropwise added with pivaloyl chloride, reacting at the temperature of 0-10 ℃ for 1-2 h at the temperature of not more than 5 ℃ in the process of dropwise adding the pivaloyl chloride to obtain a dichloromethane solution of an intermediate 2;

and 5: dissolving the prepared cefazolin in a mixed solvent, adding sodium salt, reacting for 1-1.5h at 30-40 ℃, cooling, and crystallizing to obtain a cefazolin sodium crude product; the mixed solvent is a mixed solution of water and acetone in a volume ratio of 1: 1-3; the sodium salt is sodium bicarbonate;

the preparation method also comprises the step of recrystallization,

wherein, in the recrystallization treatment, the prepared cefazolin sodium crude product is heated and dissolved in a crystallization solvent, filtered by a microporous filter membrane, respectively cooled to different temperatures for growing crystals, filtered and dried to obtain cefazolin sodium pentahydrate or anhydrous cefazolin sodium;

and controlling the stirring speed to be 60-100 r/min during crystallization.

2. The preparation method according to claim 1, wherein after the cefazolin sodium is heated and dissolved, the temperature is reduced to 30-35 ℃, the crystal is grown for 5-10 hours under the condition of heat preservation, and the anhydrous cefazolin sodium is obtained after filtration and drying; or the like, or, alternatively,

heating and dissolving cefazolin sodium, cooling to 10-15 ℃, keeping the temperature and growing the crystal for 2-5 h, filtering, and drying to obtain cefazolin sodium pentahydrate.