RU2471484C2 - Method for preparing composition for injections containing sodium cevtriaxone and sodium tazobactam - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: method for preparing a composition for injections containing sodium cevtriaxone and sodium tazobactam involves the following stages: (a) suspension of raw materials, i.e. sodium cevtriaxone, sodium tazobactam, sterilised water for injections, mixed solution of ethyl acetate and isopropyl alcohol, and anhydrous ethanol in mass relation making 3-5:1:2:5:9, with volume relation of ethyl acetate to isopropyl alcohol making 1:2-4; (b) dissolution of sodium cevtriaxone and sodium tazobactam in sterilised water for injections with added activated hydrocarbon and filtration; (c) addition of the mixed solution of ethyl acetate and isopropyl alcohol to the filtrate and agitation of the mixture; addition of a seed crystal of sodium cevtriaxone to the solution for crystallisation initiation; and finally washing of the crystals in anhydrous ethanol and crystal drying; and (d) lyophilisation to form the composition for injections containing sodium cevtriaxone and sodium tazobactam.

EFFECT: composition is characterised by high uniformity, high degree of purity and safety of use.

6 cl, 2 tbl, 3 ex

Description

This application claims the priority of patent application PRC No. 200910138402.7, entitled "Method for producing an injection composition containing cevtriaxone sodium and tazobactam sodium", filed April 30, 2009 in the Patent Office of the PRC, the contents of which are fully incorporated into this description by reference.

FIELD OF TECHNOLOGY

The present invention relates to a method for producing an injection composition comprising cevtriaxone sodium and tazobactam sodium.

BACKGROUND OF THE INVENTION

Cevtriaxone sodium, which is a third generation cephalosporin, has a wide spectrum of antibacterial action. The widespread clinical use of cephalosporin antibiotics has led to the fact that, under the influence of the antibiotic, bacteria undergo genetic changes. Over the past two decades, ever new strains of drug-resistant microorganisms have been continuously identified and, in addition, cases of severe infectious lesions and infections resistant to the effects of drugs, in particular respiratory tract infections, have been identified, and thus bacterial resistance to drugs is becoming a serious problem in medical practice. For clinical use, a complex (including more than one active component) composition has been developed. Due to the difference in the specific volume of the two sterile powders, the mixture does not have sufficient homogeneity, which is an important indicator of the quality of the product. To obtain a homogeneous mixture, preparation of the mixture in liquid form is considered the best way. The regulatory document Pharmacopoeia of the People's Republic of China, Part II (Pharmacopoeia of the People’s Republic of China, part II) (2005) noted the need to determine the "transparency and color of the solution" of cevtriaxone sodium; that is, controlling the transparency and color of sterile cephalosporin products is an important aspect of their preparation. In a traditional complex composition obtained by mixing and distributing into separate packages sterile powder of cevtriaxone sodium and tazobactam sodium, the transparency and color of the sterile product affects not only the transparency of the solution of the sterile product, but also the concentration of the sterile product and the degree of coloring of the solution.

SUMMARY OF THE INVENTION

An object of the present invention is to provide a method for preparing an injection composition containing cevtriaxone sodium and tazobactam sodium, comprising mixing cevtriaxone sodium and tazobactam sodium in liquid form to form a homogeneous mixture.

To solve this problem, the present invention proposed the following technical solution, i.e. A method of obtaining an injection composition containing cevtriaxone sodium and tazobactam sodium, which comprises the following steps:

(1) establishing the ratio of the starting materials: weighing the starting materials, i.e. cevtriaxone sodium, tazobactam sodium, sterilized water for injection, a mixed solution of ethyl acetate and isopropyl alcohol and anhydrous ethanol, in a mass ratio of 3 ~ 5: 1: 2: 5: 9;

and a mixed solution of ethyl acetate and isopropyl alcohol is obtained by mixing to a homogeneous state of ethyl acetate and isopropyl alcohol in a volume ratio of 1: 2 ~ 4;

(2) dissolution and filtration: adding sterilized water for injection into the dissolution tank at a temperature of 8 ° C ~ 12 ° C, followed by the addition of cevtriaxone sodium and tazobactam sodium and stirring until they are completely dissolved; adding activated carbon in an amount of 0.2% of the total mass of liquid in the tank; mixing for 30 minutes and subsequent filtration, resulting in a filtrate;

(3) crystallization and washing: adding the filtrate to the crystallization tank; adding a mixed solution of ethyl acetate and isopropyl alcohol and thoroughly mixing the mixture; the subsequent addition of 1 ~ 5 g of seed crystals of cevtriaxone sodium to the mixed solution to initiate crystallization, growth and transformation of crystals that are incubated for at least 5 hours; subsequent triple washing of the crystals each time with a triple amount of anhydrous ethanol; and drying;

(4) lyophilization:

(4.1) pre-freezing: lowering the temperature of the product in the thermostat to -28 ° C and keeping at this temperature for 2.5 hours;

(4.2) sublimation and drying: turning on the heating system after reaching the degree of vacuum in the thermostat of less than 30 Pa, and maintaining the temperature in the range from -15 ° C to -20 ° C to perform sublimation and drying of the product;

(4.3) re-drying: raising the temperature to 20 ° C ~ 30 ° C and keeping at this temperature for 3 hours;

(4.4) purge with high purity nitrogen sterilized by filtration through a filter cartridge with a hole size of 0.22 μm;

bringing pressure to normal value; and obtaining a complex composition for injection containing cevtriaxone sodium and tazobactam sodium.

Filtering in stage (2) is performed as follows:

ultrafiltration of the mixed solution is carried out successively through a filter of titanium wire with a hole size of 0.45 μm, then through a filter cartridge with a hole size of 0.22 μm and a filter cartridge with a hole size of 0.01 μm.

The crystallization tank in step (3) is located in the aseptic (sterile) zone of class A.

The drying in stage (3) is performed as follows: apply clean hot compressed air sterilized by passing through filters; flow rate is 2 m / s; temperature is 45 ° C ~ 50 ° C; and the drying time is 4 ~ 5 hours.

The volume ratio of ethyl acetate to isopropyl alcohol when performing stage (1) is 1: 3.

The mass ratio [cevtriaxone sodium: tazobactam sodium: sterile water for injection: a mixed solution of ethyl acetate and isopropyl alcohol: anhydrous ethanol] is 3: 1: 2: 5: 9.

The present invention has several advantages: the present invention can be applied to obtain a complex composition for injection containing cevtriaxone sodium and tazobactam sodium in a ratio of 3: 1 ~ 5: 1 (cevtriaxone: tazobactam); mixing in a liquid-liquid medium may provide homogeneity of the mixture; the present invention always allows crystallization of cevtriaxone sodium and tazobactam sodium in a ratio of 3 ~ 5: 1, that is, it allows to solve the problem associated with the difficulty of maintaining in the finished product the crystallization ratio of cevtriaxone sodium to tazobactam sodium, which is 3 ~ 5: 1, which is caused by various crystallization rates of cevtriaxone sodium and tazobactam sodium due to the difference in their solubilities; at the same time, the use of the present invention can effectively increase the purity of the composition and, thus, ensure high safety of its clinical use.

DETAILED DESCRIPTION OF THE INVENTION

Below the present invention is described in more detail using graphic materials and examples of implementation, but the scope of the present invention is not limited to the above examples.

Example 1

A method of obtaining a composition for injection containing cevtriaxone sodium and tazobactam sodium, comprising the following steps:

A. Recipe Selection

Formulation 1 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 2) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 2 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 3 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 4) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 4 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 8 total 1000 vials

Recipe 5 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 6 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 10 total 1000 vials

Recipe 7 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g four anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 8 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one

sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

Recipe 9 and ratio: specification 1.0 g

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 6 anhydrous ethanol 2250 g 9 total 1000 vials

In the present example:

1. Using the formulations 1, 2 and 3, the transparency and color of mixed solutions containing various ratios of ethyl acetate and isopropyl alcohol are evaluated, and the effect of these various ratios on the yield of crystallized cevtriaxone sodium and tazobactam sodium.

2. Using formulations 4, 5, and 6, the transparency and color of mixed solutions containing various ratios of anhydrous ethanol and cevtriaxone sodium and tazobactam sodium are evaluated, and the effect of these various ratios on the yield.

3. Using formulations 7, 8, and 9, the transparency and color of mixed solutions containing various ratios of a mixed solution of ethyl acetate and isopropyl alcohol (1: 3) and cevtriaxone sodium and tazobactam sodium are evaluated, and the effect of these various ratios on the yield.

4. The homogeneity and transparency of the above formulations were compared with the homogeneity and transparency of the prior art product (in which cevtriaxone sodium and tazobactam sodium are mixed in a ratio of 3: 1, see formulations 10-1.10-2.10-3.10-4 10-5 and 10-6).

5. Results:

No. recipe exit (%) Content
riak
sone Tazo content
tank
tama The corresponding
burden
nie results those. Pharmacopoeia of China and internal control standards transparency color technical condition
Wii Farmako
Pei of China (2005) Standard
you are inward
him control one 86.5 645 202 3.2: 1 transparent <6 # (yellowish green) The color of the solution should not be darker than the color of a standard solution of degree of transparency (turbidity) No. 1. The color of the solution should not be darker than the color No. 8 of the yellow, yellowish-green or orange standard solution The color of the solution should not be darker than the color of the standard solution, degree of transparency (turbidity) No. 0.5. The color of the solution should not be darker than the color No. 6 of the yellow, yellowish-green or orange standard solution 2 85.3 643 207 3.1: 1 transparent <6 # (yellowish green) 3 85,2 650 210 3.1: 1 transparent <6 # (yellowish green) four 92.1 660 220 3: 1 transparent <5 # (yellowish green) 5 93.2 645 215 3: 1 transparent <5 # (yellowish green) 6 94.2 657 219 3: 1 transparent <5 # (yellowish green) 7 83,2 629 217 2.9: 1 transparent <7 # (yellowish green) 8 82.3 670 216 3.1: 1 transparent <7 # (yellowish green) 9 83.9 672 210 3.2: 1 transparent <7 # (yellowish green) 10-1 672 210 3.2: 1 transparent <7 # (yellowish green) 10-2 629 217 2.9: 1 transparent <7 # (yellowish green) 10-3 672 210 3.2: 1 transparent <7 # (yellowish green) 10-4 643 207 3.1: 1 transparent <7 # (yellowish green) 10-5 683 210 3.3: 1 transparent <7 # (yellowish green) 10-6 672 210 3.2: 1 transparent <7 # (yellowish green)

The experiments showed:

1. If the ratio of the mixed solution of ethyl acetate and isopropyl alcohol (1: 3) to sodium tazobactam is 1: 5, and the ratio of anhydrous ethanol to sodium tazobactam is 1: 9, the yield exceeds 93%, and both the transparency and the color of the solution (transparent, <6 # (yellowish-green)) surpass the transparency and color parameters established by the standards of the Pharmacopoeia of China (2005);

2. Based on the results of the experiments, the following formulation was selected to obtain the composition:

raw materials recipe ratio cevtriaxone sodium 894.0 g (corresponding to 750 g of cevtriaxone) 3 tazobactam sodium 294.0 g (corresponding to 250 g of tazobactam) one sterile water for injection 500 g 2 mixed solution of ethyl acetate and isopropyl alcohol (1: 3) 1250 g 5 anhydrous ethanol 2250 g 9 total 1000 vials

B. Dissolution and filtration

1. Sterilized water for injection was added to the dissolution tank, which was then cooled to 10 ° C ± 2, cevtriaxone sodium and tazobactam sodium were dissolved with stirring in the amount indicated in the formulation, 0.2% activated carbon was added with stirring, then no later than 30 minutes later filtering was performed.

2. Filtering was performed as follows: a mixed solution obtained in a tank into which activated carbon was added was passed through a sterilized pipeline using a transfer pump, sequentially passing through a filter made of titanium wire with hole sizes of 0.45 μm, a filter cartridge with hole sizes 0.22 μm and a filter cartridge with a hole size of 0.01 μm, performing ultrafiltration sterilization; then the solution was sent to a crystallization tank located in a drying chamber located in a sterile clean zone of class A; samples were taken from the filtrate and sent to the quality control department to determine such parameters as component content, endotoxin content, pH, etc.

C. Crystallization and washing of crystals

1. A mixture of ethyl acetate and isopropyl alcohol (1: 3) in an amount five times the amount of the mixture in the crystallization tank was added to the mixed solution of cevtriaxone sodium and tazobactam sodium in the crystallization tank and mixed thoroughly; then, 5 g of seed crystals of cevtriaxone sodium were added to the mixed solution to initiate crystallization and held for at least 5 hours to effect crystal growth; the product was then pumped to a sterile three-in-one drying device, then the product was washed three times with three times the amount of anhydrous ethanol and then dried.

2. Drying. For drying, clean compressed hot air was used, heated by evaporation and sterilized by filtration. The temperature at the inlet of the hot stream was set to approximately 45 ° C. The purified mixture of cevtriaxone sodium and tazobactam sodium powders was dried for approximately 4 hours to remove ethanol.

D. Lyophilization

1. Pre-freezing: the shelf was cooled to -45 ° C. The temperature of the product was lowered to -28 ° C and kept at this temperature for 2.5 hours.

2. Sublimation and drying: when the degree of vacuum in the thermostat reached a value of less than 30 Pa, the heating system was turned on and the temperature was kept below -20 ° C for sublimation and drying.

3. Re-drying: at this stage, the product temperature was kept below 30 ° C, and the re-drying time was 3 hours.

4. After lyophilization, to achieve normal pressure, the system was ventilated with high-purity nitrogen, sterilized by filtration through a filter with openings of 0.22 microns in size; the product was removed from the thermostat, thus obtaining an injection composition containing cevtriaxone sodium and tazobactam sodium.

Example 2

A method of obtaining a composition for injection containing cevtriaxone sodium and tazobactam sodium, comprising the following steps:

(1) Starting materials: respectively, starting materials were weighed, including cevtriaxone sodium, tazobactam sodium, sterilized water for injection, a mixed solution of ethyl acetate and isopropyl alcohol and anhydrous ethanol in a mass ratio of 5: 1: 2: 5: 9.

The volume ratio of ethyl acetate to isopropyl alcohol in a mixed solution of ethyl acetate and isopropyl alcohol was 1: 2;

(2) Dissolution and filtration: sterilized water for injection was added to the dissolution tank at a temperature of 8 ° C ~ 12 ° C; then cevtriaxone sodium and tazobactam sodium were added with stirring and mixed until dissolved; then added activated charcoal (0.2% by weight of the mass of liquid in the tank) and mixed for 30 minutes; then, for ultrafiltration, the mixed solution was passed through a filter of titanium wire with a hole size of 0.45 μm, a filter cartridge with a hole size of 0.22 μm and a filter cartridge with a hole size of 0.01 μm, obtaining a filtrate;

(3) Crystallization and washing: the filtrate was added to the crystallization tank, a mixed solution of ethyl acetate and isopropyl alcohol was added thereto and mixed thoroughly; then, 1 g of seed crystals of cevtriaxone sodium was added to the mixed solution to initiate crystallization and held for at least 5 hours to effect crystal growth; then the product was washed three times each time with a three-fold amount of anhydrous ethanol, and then dried as follows: pure compressed hot air sterilized by filtration was used for drying, with a flow rate of 2 m / s, a temperature of 45 ° C and a drying time of 5 hours;

(4) Lyophilization:

(4.1) Pre-freezing: the temperature of the product in the thermostat was lowered to -28 ° C and kept at this temperature for 2.5 hours;

(4.2) Sublimation and drying: when the degree of vacuum in the thermostat reached a value of less than 30 Pa, the heating system was turned on, and, for sublimation and drying, the temperature was kept below -15 ° C;

(4.3) Re-drying: the temperature was increased to 20 ° C and kept at this temperature for 3 hours;

(4.4) To achieve normal pressure, the system was ventilated with high purity nitrogen, sterilized by filtration through a 0.22 μm filter with holes, thus obtaining an injection composition containing cevtriaxone sodium and sodium tazobactam.

Example 3

A method of obtaining a composition for injection containing cevtriaxone sodium and tazobactam sodium, comprising the following steps:

(1) Starting materials: respectively, starting materials were weighed, including cevtriaxone sodium, tazobactam sodium, sterilized water for injection, a mixed solution of ethyl acetate and isopropyl alcohol and anhydrous ethanol in a mass ratio of 5: 1: 2: 5: 9.

The volume ratio of ethyl acetate to isopropyl alcohol in a mixed solution of ethyl acetate and isopropyl alcohol was 1: 4;

(2) Dissolution and filtration: sterilized water for injection was added to the dissolution tank at a temperature of 8 ° C ~ 12 ° C; then cevtriaxone sodium and tazobactam sodium were added with stirring and mixed until dissolved; then added activated charcoal (0.2% by weight of the mass of liquid in the tank) and mixed for 30 minutes; then, for ultrafiltration, the mixed solution was passed through a filter of titanium wire with a hole size of 0.45 μm, a filter cartridge with a hole size of 0.22 μm and a filter cartridge with a hole size of 0.01 μm, obtaining a filtrate;

(3) Crystallization and washing: the filtrate was added to the crystallization tank, a mixed solution of ethyl acetate and isopropyl alcohol was added thereto and mixed thoroughly; then, 3 g of seed crystals of cevtriaxone sodium was added to the mixed solution to initiate crystallization and held for at least 5 hours to effect crystal growth; then the product was washed three times each time with a three-fold amount of anhydrous ethanol and then dried as follows: pure compressed hot air sterilized by filtration was used for drying, with a flow rate of 2 m / s, a temperature of 50 ° C and a drying time of 5 hours;

(4) Lyophilization:

(4.1) Pre-freezing: the temperature of the product in the thermostat was lowered to -28 ° C and kept at this temperature for 2.5 hours;

(4.2) Sublimation and drying: when the degree of vacuum in the thermostat reached a value of less than 30 Pa, the heating system was turned on and, for sublimation and drying, the temperature was kept below -15 ° C;

(4.3) Re-drying: the temperature was increased to 20 ° C and kept at this temperature for 3 hours;

(4.4) To achieve normal pressure, the system was ventilated with high purity nitrogen, sterilized by filtration through a 0.22 μm filter with holes, thus obtaining an injection composition containing cevtriaxone sodium and sodium tazobactam.

Stability studies

Carried out stability studies of the compositions obtained by the method described above.

1. Method

Lot numbers of samples: Example 1 (080501), Example 2 (080502) and Example 3 (080503).

The studied parameters: appearance, content, transparency, relevant materials, water content, etc.

Research Method. A method for studying the content and corresponding materials in samples for 0-18 months was as follows.

Chromatography conditions and system applicability tests.

Detector: SPD-10AVP detector.

Chromatographic column: Octadecyl silane chemically bonded to silica was used as a filler.

Mobile phase: the reagent A mixture was used as the mobile phase:

water (85:15), where reagent A contained 0.015 mol / l of tetrabutylammonium phosphate (600 ml of water was added to 50 ml of a solution of 0.2 mol / l of tetrabutylammonium phosphate), methanol and acetonitrile in a ratio of 650: 250: 100, the pH was adjusted to 7.3 ± 0.1 by the addition of 0.5% phosphoric acid.

Wavelength for determination: 230 nm.

The number of theoretical plates: based on the location of the peak of cevtriaxone, the number of theoretical plates should not be less than 3000.

Definition. Approximately 100 mg of the composition obtained as described above was accurately weighed and placed in a 50 ml volumetric flask, dissolved and diluted with the mobile phase to obtain the desired dilution. The solution was shaken until a homogeneous state and 10 μl were taken, which were introduced into a liquid chromatograph; a chromatogram was recorded; appropriate amounts of cevtriaxone and tazobactam standards used as a control were selected, precisely weighed and dissolved in the mobile phase to obtain a solution containing, respectively, 1.5 mg of cevtriaxone and 0.5 mg of tazobactam (3: 1) per milliliter; the same method was used to determine the content of C ₁₈ H ₁₈ N ₈ O ₇ S ₃ and C ₁₀ H ₁₂ N ₄ O ₅ S in the test samples, using the values of the areas under the peaks and an external standard.

Procedure. A suitable amount of sample was taken and dissolved in the mobile phase to obtain a test sample solution containing approximately 2 mg of sample in a milliliter of solution; 1 ml of precisely measured was taken from this solution, which was placed in a 100 ml volumetric flask. The solution, which was then used as a control solution, was diluted with the mobile phase to obtain the desired dilution, and shaken until a homogeneous state. In accordance with the conditions of the study parameters, to clarify the measurement range so that the height of the peak of cevtriaxone was approximately 20% of the entire range of the recording device, 10 μl of the control solution were taken and injected into a liquid chromatograph; then, 10 μl of each of the solutions prepared as described above was accurately taken and, accordingly, injected into a liquid chromatograph; the chromatogram was recorded over a time of 3.5 times the retention time of cevtriaxone. If the chromatogram of the sample solution contained peaks of impurities, then the peak area of one impurity should not exceed the sum of the areas under the peaks of cevtriaxone and tazobactam in the control solution (1.0%), and the sum of the areas under the peaks of individual impurities should not be more than 4 times exceed the sum of the areas under the two main peaks of the control solution (4.0%).

2. Results

2.1 Accelerated Testing

Three batches of samples were taken (300 in each batch), which were stored at 40 ± 2 ° C and a relative humidity of 75 ± 5% for 6 months. After 1, 2, 3, 6 months, samples were taken to determine the above parameters using the methods described above; the results are presented in Table 1.

Table 1 Results of accelerated testing of samples of three batches 080501, 080502 and 080503 Vre
me Party number results Appearance Alkalinity Transparent
nost Contents water Match. materials (%) Indicated Content (%) Maxim Linen Amount Cevtriaxone Tazobactam 0 month 080501 Whitish powder 6.5 matches * 8.0 0.1 0.3 103,2 101.5 080502 Whitish powder 6.5 corresponds to 7.8 0.1 0.3 105.6 103.6 080503 Whitish powder 6.4 corresponds to 7.8 0.1 0.3 101.0 100.1 1 month 080501 Whitish powder 6.5 corresponds to 8.1 0.1 0.3 103.0 101.0 080502 Whitish powder 6.5 corresponds to 7.9 0.1 0.3 105.0 103.0 080503 Whitish powder 6.5 corresponds to 7.9 0.1 0.3 100.6 99.6 2 month 080501 Whitish powder 6.6 corresponds to 8.2 0.2 0.4 102.5 100.5 080502 Whitish powder 6.6 corresponds to 8.0 0.2 0.4 104.3 102.5 080503 Whitish powder 6.7 corresponds to 8.0 0.2 0.4 100.0 99,2 3 month 080501 Whitish powder 6.7 corresponds to 8.2 0.3 0.5 102.0 100.0 080502 Whitish powder 6.6 corresponds to 8.1 0.3 0.6 103.8 102.0 080503 Whitish powder 6.8 corresponds to 8.1 0.4 0.6 99.5 98.5 6 months 080501 Whitish powder 6.9 corresponds to 8.3 0.4 0.6 100.9 99.5 080502 Whitish powder 6.8 corresponds to 8.2 0.4 0.7 102.0 100.5 080503 Whitish powder 6.9 corresponds to 8.2 0.4 0.8 99.0 98.0 * corresponds, i.e. meets technical requirements

2.2 Long-term tests

Three batches of samples were taken (1400 in each batch), which were stored at 25 ± 2 ° C and relative humidity 60 ± 14%. After 3, 6, 9, 12, 18 and 24 months, samples were taken to determine the above parameters using the methods described above and compared with the results obtained at time 0 (0 month), making conclusions about the possibility of subsequent clinical trials. The results are presented in Table 2.

table 2 Long-term test results for samples of three batches 080501, 080502 and 080503 Time Party number results Appearance Alkalinity Sterility Transparency Water content Accordingly. materials (%) Indicated Content (%) Maximum Total Cevtriaxone Tazobactam 0 month 080501 whitish powder 6.5 acc. acc. 8.0 0.1 0.3 103.2 101.5 080502 whitish powder 6.5 acc. acc. 7.8 0.1 0.3 105.6 103.6 080503 whitish powder 6.4 acc. acc. 7.8 0.1 0.3 101.0 100.1 3 month 080501 whitish powder 6.5 acc. acc. 8.0 0.1 0.3 103.1 101,2 080502 whitish powder 6.5 acc. acc. 7.8 0.1 0.3 105.1 103,2 080503 whitish powder 6.4 acc. acc. 7.8 0.1 0.3 100.8 99.8 6 month 080501 whitish powder 6.6 acc. acc. 8.1 0.2 0.4 102.8 101.0 080502 whitish powder 6.6 acc. acc. 7.9 0.2 0.4 104.5 103.0 080503 whitish powder 6.5 acc. acc. 7.9 0.2 0.4 100.6 99.5 9 month 080501 whitish powder 6.6 acc. acc. 8.1 0.2 0.5 102.5 100.5 080502 whitish powder 6.7 acc. acc. 8.0 0.2 0.4 104.0 102.5 080503 whitish powder 6.6 acc. acc. 7.9 0.2 0.5 100.3 99,2 12 month 080501 whitish powder 6.6 acc. acc. 8.1 0.2 0.5 102,2 100.1 080502 whitish powder 6.7 acc. acc. 8.0 0.2 0.5 103.7 102,2 080503 whitish powder 6.6 acc. acc. 7.9 0.2 0.5 100.0 99.1 18 month 080501 whitish powder 6.7 acc. acc. 8.2 0.2 0.6 102.0 99.5 080502 whitish powder 6.9 acc. acc. 8.3 0.2 0.7 103,2 101.6 080503 whitish powder 6.8 acc. acc. 8.1 0.2 0.6 99.6 98.7 24 month 080501 whitish powder 6.9 acc. acc. 8.4 0.2 0.7 101.5 99,2 080502 whitish powder 7.0 acc. acc. 8.5 0.2 0.8 102.7 101.1 080503 whitish powder 6.9 acc. acc. 8.4 0.2 0.7 99,2 98.2 * acc. means "meets specifications"

3. Conclusion

The above results show that the parameters of all three batches of samples (specification 1.0 g) after storage for 24 months meet the requirements; however, significant changes in all parameters were not observed, with the exception of the “Corresponding materials” parameter, which to some extent increased compared to the value obtained at time 0 (0 month).

Hypersensitivity, hemolysis and vascular response tests with systemic administration

1. An injection composition of 150 mg / ml containing cevtriaxone sodium and tazobactam sodium (3: 1 and 5: 1) did not cause noticeable hemolysis; all results are consistent with safety requirements for administration by intravenous injection.

A 150 mg / ml injection composition containing cevtriaxone sodium and tazobactam sodium (3: 1 and 5: 1) was administered three times by intravenous injection into the rabbit’s ear vein, but no obvious response was observed; a 150 mg / ml injection composition containing cevtriaxone sodium and tazobactam sodium (3: 1 and 5: 1) was injected into the quadriceps femoris and no explicit response was also observed. Thus, the test results of the test drug for local (topical) stimulation are consistent with established requirements.

A 150 mg / ml injection composition containing cevtriaxone sodium and sodium tazobactam (3: 1 and 5: 1) was administered every second day by intramuscular injection to guinea pigs receiving a total of 3 injections each. The experimental group was examined on the 14th and 21st days after the first injection, but no symptoms of hypersensitivity were found; thus, the test results of the test drug are consistent with the requirements established for hypersensitivity tests.

2. Objective: to determine the presence of a hemolytic and agglutinating effect in the composition for injection containing cevtriaxone sodium and tazobactam sodium in a ratio of 3: 1 or 5: 1, that is, determining the safety of the drug when administered intravenously.

A response of blood vessels and muscles was observed after administration of a composition containing cevtriaxone sodium and tazobactam sodium (3: 1 and 5: 1) into the ear vein and quadriceps femoral muscle of the rabbit.

3. Determining the likelihood of a systemic allergic reaction after administration of an injection composition containing cevtriaxone sodium and tazobactam sodium (3: 1 and 5: 1).

1. Hemolytic tests

1.1 Materials

(1) Animal: One male 2.3 kg long-eared Japanese rabbit was provided by the Xi'an Medical University Experimental Animal Center, Certificate Number: ShanxiYi Dong Zi No. 08-018.

(2) Test drugs: an injection composition containing cevtriaxone sodium and tazobactam sodium, white powder, active component ratios 3: 1 and 5: 1. The composition (1.0 g / vial), including a 3: 1 ratio, contained 0.75 g of cevtriaxone and 0.25 g of tazobactam, lot number 010105; a composition (1.2 g / vial), comprising a 5: 1 ratio, contained 1.0 g of cevtriaxone and 0.2 g of tazobactam, lot number 010106. Medicines were provided by HaiKou QILI Pharmaceutical Co., LTD. Medicines were stored in a refrigerator and dissolved in sterilized water for injection before use, and then diluted to obtain a content of cevtriaxone sodium and tazobactam sodium equal to 150 mg per milliliter.

1.2 Ways

(1) Preparation of a 2% rabbit blood cell suspension: blood was collected from rabbit cardiac tissues and fibrin was separated using glass beads to obtain defibrinated blood; blood was placed in a graduated centrifuge tube into which physiological saline was added, mixed and centrifuged; the liquid above the precipitate was separated, and the liquid above the precipitate was washed with physiological saline to obtain a colorless and transparent substance; then the red blood cells were diluted with physiological saline to obtain a 2% suspension.

(2) Hemolytic tests. In 14 test tubes containing the composition for injection, including cevtriaxone sodium and tazobactam sodium, where in 7 tubes there was a composition with a ratio of cevtriaxone sodium: tazobactam sodium = 3: 1, and in 7 tubes there was a composition with a ratio of cevtriaxone sodium: tazobactam sodium = 5: 1, various liquids were added (shown in Tables 1 and 2). No sample solution was added to the sixth tube (blank control). No sample solution was added to the seventh test tube, and distilled water was added instead of physiological saline (positive hemolytic control). The contents of all tubes were carefully and uniformly mixed and incubated in an incubator, which is a water bath, at a temperature of 37 ° C, conducting observations every hour for 4 hours. If the solution turned red and transparent, this indicated the occurrence of hemolysis; if a brownish-red or red-brown flocculent precipitate appeared in the solution, this indicated the occurrence of red blood cell agglutination; if all red blood cells settled to the bottom, and the liquid above the sediment became colorless and transparent, this indicated the absence of hemolysis.

1.3 Results

The data presented in Tables 1 and 2 show the absence of hemolysis and agglutination in test tubes containing 0.1, 0.2, 0.3, 0.4 and 0.5 ml of an injection composition containing cevtriaxone sodium and tazobactam sodium (150 mg / ml, 37 ° C for 4 hours) both in a ratio of 3: 1 and in a ratio of 5: 1. The fluids above the sediment were clear and colorless, and all red blood cells settled to the bottom - this shows that the composition for injection containing cevtriaxone sodium and tazobactam sodium (150 mg / ml both in a ratio of 3: 1 and in a ratio of 5: 1 ), does not have a hemolytic effect, which is consistent with safety requirements for intravenous administration.

The description provided is a preferred embodiment of the present invention. Obviously, a person skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and such improvements and modifications are included in the scope of the present invention.

Claims (6)

1. A method of obtaining a composition for injection containing cevtriaxone sodium and tazobactam sodium, comprising the following stages:
(1) obtaining the ratio of starting materials by weighing the starting cevtriaxone sodium, tazobactam sodium, sterilized water for injection, a mixed solution of ethyl acetate and isopropyl alcohol and anhydrous ethanol in a mass ratio of 3-5: 1: 2: 5: 9;
and a mixed solution of ethyl acetate and isopropyl alcohol is obtained by mixing to a homogeneous state of ethyl acetate and isopropyl alcohol in a volume ratio of 1: 2-4;
(2) dissolution and filtration: adding sterilized water for injection to the dissolution tank at a temperature of 8-12 ° C, followed by the addition of cevtriaxone sodium and tazobactam sodium and stirring until they are completely dissolved; adding activated carbon in an amount of 0.2% of the total mass of liquid in the tank; stirring for 30 minutes and subsequent filtration, resulting in a filtrate;
(3) crystallization and washing: adding the filtrate to the crystallization tank; adding a mixed solution of ethyl acetate and isopropyl alcohol and thoroughly mixing the mixture; subsequent addition of 1-5 g of seed crystals of cevtriaxone sodium to the mixed solution to initiate crystallization, growth and transformation of crystals that are incubated for at least 5 hours; subsequent triple washing of the crystals each time with a triple amount of anhydrous ethanol, and drying;
(4) lyophilization:
(4.1) preliminary freezing: lowering the temperature of the product in the thermostat to -28 ° C and keeping at this temperature for 2.5 hours;
(4.2) sublimation and drying: turning on the heating system after reaching the degree of vacuum in the thermostat of less than 30 Pa, and maintaining the temperature in the range from -15 ° C to -20 ° C to perform sublimation and drying of the product;
(4.3) re-drying: raising the temperature to 20-30 ° C and keeping at this temperature for 3 hours;
(4.4) purge with high purity nitrogen, sterilized by filtration through a filter cartridge with a hole size of 0.22 μm, bring the pressure to normal and obtain an injection composition containing cevtriaxone sodium and sodium tazobactam. 2. The method according to claim 1, characterized in that the filtering in step (2) is performed as follows: ultrafiltration of the mixed solution through a titanium wire filter with a hole size of 0.45 μm, a filter cartridge with a hole size of 0.22 μm, and a filter cartridge with a hole size of 0.01 μm. 3. The method according to claim 1, characterized in that the crystallization tank in stage (3) is located in a clean zone of class 10000. 4. The method according to claim 1, characterized in that the drying in step (3) is performed as follows: apply clean hot compressed air sterilized by passing through filters; wherein the flow rate is 2 m / s, the temperature is 45-50 ° C, and the drying time is 4-5 hours. 5. The method according to claim 1, characterized in that the volume ratio of ethyl acetate to isopropyl alcohol in stage (1) is 1: 3. 6. The method according to claim 1, characterized in that the mass ratio [cevtriaxone sodium: tazobactam sodium: sterile water for injection: a mixed solution of ethyl acetate and isopropyl alcohol: anhydrous ethanol] is 3: 1: 2: 5: 9.