Cefozopran sodium for injection and a method for preparing the same

Abstract

The present application provides a kind of cefozopran sodium for injection and its preparation method, the preparation is sterile freeze-dried powder injection, including cefozopran sodium 100 parts, freeze-dried skeleton support agent 15~40 parts, stabilizer 3~12 parts, protective agent 10~25 parts and pH adjusting agent by weight parts;Freeze-dried skeleton support agent is compounded by mannitol, low molecular dextran 40, xylitol, the protective agent is injection grade trehalose, low-temperature liquid preparation, sterile filtration, freeze-drying and full pressure plug, cover preparation are made.The present application is synergized by trehalose forming glassy matrix and multiple stabilization system, and the preparation has good stability, loose and complete appearance, rapid reconstitution, high content, and is suitable for industrial production.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical preparation technology, and in particular to a cefazolin sodium for injection and its preparation method. Background Technology

[0002] Cefazolin sodium is a third-generation semi-synthetic cephalosporin antibiotic that exerts its antibacterial effect by inhibiting the biosynthesis of bacterial cell wall peptidoglycan. It has broad-spectrum antibacterial activity against both Gram-positive and Gram-negative bacteria and is widely used clinically to treat various infectious diseases caused by susceptible bacteria, such as lower respiratory tract infections, urinary tract infections, abdominal infections, pelvic infections, sepsis, and skin and soft tissue infections.

[0003] However, cefazolin sodium contains a β-lactam ring in its molecular structure, making it sensitive to moisture, temperature, light, and acid / alkaline environments. In aqueous solutions, it is prone to hydrolysis and ring-opening degradation, generating cephalosporin polymers and related impurities. This not only reduces efficacy but may also trigger adverse reactions such as allergies. Currently, most commercially available cefazolin sodium for injection is aseptically packaged as a powder for injection, meaning the active pharmaceutical ingredient is directly repackaged.

[0004] Chinese patent CN101843589B, "Cefazolin Sodium Composition for Injection: Sterile Powder," points out that existing cefazolin sodium raw materials generally suffer from problems such as low purity, poor clarity after dissolution, and poor stability in aqueous solutions. Furthermore, cefazolin sodium is a crystalline powder and exhibits polymorphism, with different crystal forms showing significant differences in solubility, dissolution rate, and bioavailability, which are important factors affecting drug quality and clinical efficacy. Although this patent improves the crystal form by introducing sodium benzoate, it still does not solve problems such as slow reconstitution rate of the finished product, easy adhesion to the bottle wall, and poor long-term storage stability.

[0005] Chinese patent CN101780052B, "A Cefazolin Sodium Liposome Injection," uses a lyophilized liposome formulation to encapsulate the drug in a liposome matrix composed of phospholipids and cholesterol to improve encapsulation efficiency and stability. However, liposome formulations are complex in composition, have high costs, and require complicated preparation processes. Furthermore, problems such as lyophilized cake collapse, shrinkage, and drug degradation can easily occur during the lyophilization process, which is not conducive to industrial production.

[0006] Chinese patent CN105622635A, "A Novel Crystal Form of Cefazolin Sodium with Reduced Allergic Reactions and Its Formulation," reduces impurities and allergic reactions through a specific crystal form. However, its approach focuses on improving the crystal form of the active pharmaceutical ingredient. The preparation requires the introduction of organic solvents such as isopropanol and tetrahydrofuran for recrystallization, and the formulation is still a conventional aseptically packaged powder for injection. It fails to address the long-term stability and reconstitution issues of the finished product from the perspectives of formulation and freeze-drying process.

[0007] In summary, existing technologies have improved cefazolin sodium from different angles, such as crystal form modification and liposome encapsulation, but none of them have been able to solve the problems of slow reconstitution, poor appearance, and insufficient long-term storage stability of the finished product while taking into account the simplicity of the process and controllable cost. Summary of the Invention

[0008] In view of this, the present invention proposes a cefazolin sodium for injection and its preparation method to solve the above problems.

[0009] The technical solution of the present invention is implemented as follows: a cefazolin sodium for injection, comprising, by weight, 100 parts of cefazolin sodium, 15-40 parts of lyophilized matrix support agent, 3-12 parts of stabilizer, 10-25 parts of protectant and pH adjuster. The freeze-dried skeleton support agent is a compound of mannitol, low molecular weight dextran 40 and xylitol; The pH adjuster is a citrate-sodium citrate buffer, used to make the pH value of the finished product after reconstitution with water 6.0~6.5.

[0010] Furthermore, in the freeze-dried skeleton support agent, the weight ratio of mannitol, low molecular weight dextran 40, and xylitol is 1:(0.2~0.8):(0.02~0.08).

[0011] Furthermore, the stabilizer is selected from one or more of glycine, arginine, histidine, and proline.

[0012] Furthermore, the protective agent is injection-grade trehalose.

[0013] Furthermore, in the citric acid-sodium citrate buffer, the molar ratio of citric acid to sodium citrate is 1:(3~5).

[0014] Furthermore, it also includes an antioxidant and a metal chelating agent, wherein the antioxidant is vitamin C and the metal chelating agent is sodium edetate; by weight, vitamin C is 0.05~0.5 parts and sodium edetate is 0.01~0.1 parts.

[0015] The preparation method of the above-mentioned cefazolin sodium for injection includes the following steps: S1. Under light-protected conditions, dissolve sodium edetate in water for injection at 2-8℃, then add cefazolin sodium and a protective agent and stir with ultrasound to aid dissolution. After adding vitamin C, add a pH adjuster to adjust the pH of the solution to 6.0-6.5. Add a lyophilized matrix support agent and a stabilizer and mix well to obtain the drug solution. S2. Nitrogen gas is introduced into the drug solution and preparation container to replace the headspace residual oxygen content to ≤1.0%. Then, the drug solution is aseptically filtered through a 0.22μm microporous membrane, filled into vials and partially stoppered. S3. Place the filled vials in a freeze dryer, and then pre-freeze, anneal, cool, sublime dry, desorption dry, and vacuum anneal until the moisture content is ≤1.5%. S4. After the desorption drying is completed, nitrogen gas is introduced into the freeze dryer under vacuum until the headspace residual oxygen content is ≤1.0%, and then the freeze dryer is fully plugged and capped.

[0016] Furthermore, the parameters for ultrasonic-assisted dissolution in step S1 are: ultrasonic frequency 20~40kHz, power 100~300W, dissolution temperature 2~6℃, dissolution time 1~3 hours, and stirring speed 100~300r / min.

[0017] Furthermore, the specific parameters of the freeze-drying process in step S3 are as follows: Pre-freezing: Lower the product temperature to -40 to -45°C at a rate of 0.5 to 2°C / min and hold for 2 to 4 hours; Annealing treatment: Then anneal at a rate of 0.1~0.5℃ / min to -20~-15℃ for 1~3 hours; Further cooling: Cool to -45~-50℃ and maintain for 1~2 hours; Sublimation drying: Sublimate drying at -20~-5℃ and vacuum degree 10~30Pa for 8~16 hours; Desorption drying: Desorption drying at 25~35℃ and vacuum degree ≤10Pa for 4~8 hours; Vacuum post-annealing: Hold at 30~35℃ and vacuum degree ≤10Pa for 1~2 hours.

[0018] Compared with the prior art, the beneficial effects of the present invention are: The total excipient content of this invention is controlled at 28% to 78% of the active pharmaceutical ingredient (API). The total package weight of a 1g specification product is approximately 1.3 to 1.8g, which is compatible with the 10mL controlled vial packaging limit. The package weight is moderate and convenient for clinical reconstitution. The stabilizer is an injection-grade excipient, with a dosage of 3% to 12% of the API, which is within the safe dosage range for injection excipients.

[0019] This invention utilizes injectable trehalose as a glassy preservative. During freeze-drying, trehalose forms a continuous amorphous glassy matrix, which binds cefazolin sodium molecules through numerous intermolecular hydrogen bonds, reducing the mobility of β-lactam ring molecules, inhibiting drug hydrolysis and ring-opening, and decreasing the formation of polymers and related substances. Combined with stabilizers, vitamin C antioxidants, calcium sodium edetate metal chelating agents, and a citrate-sodium citrate buffer system, a multi-stabilizing system is formed. After 6 months of accelerated testing at 40℃±2℃ and 75%RH±5%, the polymer content of the finished product is ≤0.13%, and the total amount of related substances is ≤0.97%, demonstrating significantly better stability than commercially available aseptically packaged powder injections. This invention uses mannitol and low molecular weight dextran 40 in combination, xylitol as a freeze-drying framework support, and introduces annealing treatment during the freeze-drying process. The resulting product is white, loose, intact, without collapse or powder shedding, with a moisture content ≤1.5%, and a reconstitution time shortened to 14-19 seconds, significantly improving the convenience of clinical use.

[0020] This invention employs low-temperature, light-protected solution preparation, nitrogen purging, and vacuum nitrogen-filled plugging processes to control residual oxygen levels and light exposure throughout the process, further inhibiting drug oxidative degradation and achieving a finished product purity of over 99%. The preparation method of this invention is stable, operates under mild conditions, and is simple to use; all raw materials and excipients are readily available, making it suitable for industrial production. Detailed Implementation

[0021] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0022] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0023] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0024] Raw material description: Low molecular weight dextran 40, injection grade solid powder, weight average molecular weight 32,000~42,000; Trehalose: Injection-grade trehalose dihydrate; Cefazolin sodium: meets the quality standards of the Chinese Pharmacopoeia, with a purity of ≥99.0%.

[0025] Example 1 The sterile lyophilized powder for injection of cefazolin sodium in this embodiment, calculated per 1000 vials, has the following formulation: Cefazolin sodium 1000g; 200g of freeze-dried skeletal support agent, in which the weight ratio of mannitol, low molecular weight dextran 40 and xylitol is 1:0.5:0.05; Stabilizer (glycine) 50g; Protectant (trehalose) 200g; Appropriate amount of citric acid-sodium citrate buffer (molar ratio of citric acid to sodium citrate 1:3); Vitamin C 2g; Sodium calcium edetate 0.5g; Add water for injection to a total of 10L.

[0026] Preparation method: S1. Under light-protected conditions, 8L of water for injection was cooled to 4°C, and sodium calcium edetate was added and stirred until completely dissolved. Cefazolin sodium and trehalose were added, and the mixture was sonicated at 30kHz, 200W, and 4°C, and stirred at 200r / min for 3 hours to aid dissolution. After dissolution, vitamin C was added and stirred until dissolved. The pH of the solution was adjusted to 6.2 using citrate-sodium citrate buffer. The lyophilized matrix support and glycine were added and stirred until completely dissolved. Water for injection was added to bring the volume to 10L to obtain the drug solution.

[0027] S2. Purge the drug solution and preparation container with nitrogen until the headspace residual oxygen content is ≤1.0%. Then, sterilely filter the drug solution through a 0.22μm microporous membrane. Aseptically fill the filtered drug solution into 15mL controlled vials, each containing 10mL of solution, and add a 1 / 2 butyl rubber stopper.

[0028] S3. Programmed freeze drying: Place the filled vials in a freeze dryer and dry them according to the following program: Pre-freezing: Cool to -42°C at a rate of 1°C / min and hold for 3 hours; Annealing treatment: Heat to -18℃ at a rate of 0.3℃ / min and hold for 2 hours; Further cooling: Cool to -48℃ at a rate of 1℃ / min and hold for 1.5 hours; Sublimation drying: Separator temperature -10℃, vacuum degree 20Pa, sublimation drying for 12 hours; Desorption drying: partition temperature 30℃, vacuum degree 8Pa, desorption drying for 6 hours; Vacuum post-annealing: Hold at 30℃ and vacuum degree ≤10Pa for 1.5 hours, and control the moisture content of the product at the drying endpoint to ≤1.5%.

[0029] S4. Vacuum Nitrogen Filling and Capping: After the analytical drying is completed, high-purity nitrogen is filled into the freeze-drying chamber under vacuum until the residual oxygen content in the headspace of the vial is ≤1.0%. Then, the rubber stopper is fully pressed, and after exiting the chamber, an aluminum-plastic composite cap is rolled to obtain the finished product.

[0030] Example 2 The sterile lyophilized powder for injection of cefazolin sodium in this embodiment, calculated per 1000 vials, has the following formulation: Cefazolin sodium 1000g; 150g of freeze-dried scaffold support agent, wherein the weight ratio of mannitol, low molecular weight dextran 40 and xylitol is 1:0.8:0.08; Stabilizer (arginine) 30g; Protectant (trehalose) 100g; Appropriate amount of citric acid-sodium citrate buffer (molar ratio of citric acid to sodium citrate 1:3); Vitamin C 1.5g; Sodium calcium edetate 0.3g; Add water for injection to a total of 10L.

[0031] Preparation method: S1. Dissolve at 2°C, stir at 300 rpm, and dissolve for 1 hour; adjust the pH of the solution to 6.0 with a buffer; the remaining preparation steps are the same as in Example 1.

[0032] S2, the operation is the same as in Example 1.

[0033] S3, Freeze-drying process parameters: Pre-freezing: Cool to -40℃ at a rate of 0.5℃ / min and hold for 4 hours; Annealing treatment: Heat to -20℃ at a rate of 0.1℃ / min and hold for 3 hours; Further cooling: Cool to -45℃ at a rate of 0.5℃ / min and hold for 2 hours; Sublimation drying: shelf temperature -5℃, vacuum degree 30Pa, sublimation drying for 16 hours; Desorption drying: partition temperature 30℃, vacuum degree 10Pa, desorption drying for 8 hours; Vacuum post-annealing: Hold at 30℃ and vacuum degree ≤10Pa for 1 hour.

[0034] S4. Headspace residual oxygen content after nitrogen filling ≤1.0%, full-pressure plug, crimped cap.

[0035] Example 3 The sterile lyophilized powder for injection of cefazolin sodium in this embodiment, calculated per 1000 vials, has the following formulation: Cefazolin sodium 1000g; 400g of freeze-dried skeletal support, wherein the weight ratio of mannitol, low molecular weight dextran 40 and xylitol is 1:0.2:0.02; Stabilizer (histidine) 120g; Protectant (trehalose) 250g; Appropriate amount of citric acid-sodium citrate buffer (molar ratio of citric acid to sodium citrate 1:5); Vitamin C 3g; Sodium calcium edetate 1g; Add water for injection to a total of 10L.

[0036] Preparation method: S1. Dissolve at 6°C, stir at 100 r / min, and dissolve for 3 hours; adjust the pH of the solution to 6.5 with a buffer; the remaining preparation procedures are the same as in Example 1.

[0037] S2, the operation is the same as in Example 1.

[0038] S3, Freeze-drying process parameters: Pre-freezing: Cool to -45°C at a rate of 2°C / min and hold for 2 hours; Annealing treatment: Heat to -15℃ at a rate of 0.5℃ / min and hold for 1 hour; Further cooling: Cool to -50℃ at a rate of 2℃ / min and hold for 1 hour; Sublimation drying: partition temperature -20℃, vacuum degree 10Pa, sublimation drying for 8 hours; Desorption drying: partition temperature 35℃, vacuum degree 5Pa, desorption drying for 4 hours; Vacuum post-annealing: Hold at 35℃ and vacuum degree ≤10Pa for 1 hour.

[0039] S4. Headspace residual oxygen content after nitrogen filling ≤0.5%, full-pressure plug, crimped cap.

[0040] Comparative Example 1 Commercially available general-purpose cefazolin sodium for injection (sterile packaged powder for injection, containing only cefazolin sodium raw material, without additional excipients).

[0041] Comparative Example 2 The formulation of this comparative example is basically the same as that of Example 1, except that no protective agent is added, while the other components and their amounts remain unchanged. The preparation method is the same as that of Example 1.

[0042] Comparative Example 3 The formulation of this comparative example is basically the same as that of Example 1, except that glycine stabilizer is not added, while the other components and their amounts remain unchanged. The preparation method is the same as that of Example 1.

[0043] Comparative Example 4 The preparation method of this comparative example is basically the same as that of Example 1, except that the freeze-drying process does not involve annealing. After pre-freezing, it directly enters the sublimation drying stage, while the other parameters remain unchanged.

[0044] Comparative Example 5 For 1000 bottles, the following ingredients are used: 1000g of cefazolin sodium; 200g of lyophilized skeleton support agent; and a weight ratio of mannitol to low molecular weight dextran 40 of 2:1. Xylitol is not added. The stabilizers, protectants, pH adjusters, other excipients, and preparation process are completely consistent with those in Example 1.

[0045] Experimental Example 1: Basic Quality Inspection of Finished Products The finished products of each embodiment and comparative example were subjected to quality testing, and the testing methods are as follows: 1. Appearance: Visually observe the shape and color of the freeze-dried cake, and record whether there is any collapse, shrinkage, or powder coating on the bottle wall.

[0046] 2. Reconstitution time: Take one sample vial, add 10 mL of 0.9% sodium chloride injection at 25℃, shake gently, and record the time from the addition of the solvent until the solution is completely clear and free of visible particles.

[0047] 3. Moisture content: determined by Karl Fischer method.

[0048] 4. pH value: Take the finished product, redissolve it in water to a solution of 0.1 g / mL, and test it according to the pH determination method in the General Rules of Chinese Pharmacopoeia.

[0049] 5. Content: Determined by high performance liquid chromatography.

[0050] The chromatographic conditions were as follows: the column was an octadecylsilane-bonded silica column (C18 column, 250 mm × 4.6 mm, 5 μm); the column temperature was 30 ℃; the mobile phase was pH 7.0 phosphate buffer-acetonitrile (85:15, v / v), with isocratic elution; the flow rate was 1.0 mL / min; the detection wavelength was 254 nm; and the injection volume was 20 μL.

[0051] Assay: Take an appropriate amount of this product, dissolve it in the mobile phase, and quantitatively dilute it to prepare a test solution containing 0.1 mg of cefazolin per 1 mL; separately take an appropriate amount of cefazolin reference standard, dissolve it in the mobile phase, and quantitatively dilute it to prepare a reference solution of the same concentration. Accurately measure 20 μL each of the test solution and the reference solution, and inject them separately into the liquid chromatograph. Calculate the cefazolin sodium content based on the peak area using the external standard method.

[0052] 6. Sterility and endotoxins: Sterility testing shall be performed in accordance with Chinese Pharmacopoeia General Chapter 1101, and bacterial endotoxin testing shall be performed in accordance with Chinese Pharmacopoeia General Chapter 1143, with the limit being less than 0.1 EU of endotoxin per 1 mg of cefazolin sodium.

[0053] 7. Content uniformity: Performed in accordance with the method for checking the content uniformity of injections in Chinese Pharmacopoeia General Chapter 0102.

[0054] The test results are as follows: Table 1. Basic Quality Inspection Results of Finished Products

[0055] Note: The sterility, bacterial endotoxin, and fill volume variation test results of all examples and comparative samples comply with the relevant provisions of the Chinese Pharmacopoeia for injections.

[0056] As shown in Table 1, the freeze-dried products of this invention exhibit good shape retention and reconstitution time (≤20 seconds), significantly faster than commercially available powder injections. The pH value of the finished product after reconstitution with water is within the range of 6.0-6.5, meeting the Chinese Pharmacopoeia's requirements for cefazolin sodium injection (6.0-8.0). All quality indicators meet the requirements of the Chinese Pharmacopoeia for injections. Comparative Examples 2-4, lacking trehalose as a glassy protective agent, stabilizer, or annealing process, showed varying degrees of decrease in shape retention and reconstitution performance.

[0057] Experimental Example 2: Accelerated Stability Study The samples from each embodiment and comparative example were subjected to accelerated testing at a temperature of 40℃±2℃ and a relative humidity of 75%RH±5% for 6 months. Samples were taken at 0, 3, and 6 months to detect the cefazolin polymer content and total related substances. Detection method: 1. Cefazolin polymers: Size exclusion chromatography was used with dextran gel G-10 as the packing material, and the polymers were determined according to the method for determining cephalosporin antibiotic polymers in the Chinese Pharmacopoeia.

[0058] 2. Related substances: The total amount of related substances was calculated by reversed-phase high-performance liquid chromatography with gradient elution and self-comparison method.

[0059] The stability test results are shown in Table 2.

[0060] Table 2. Results of accelerated stability testing after 6 months

[0061] As shown in Table 2, the initial polymer and related substance contents of the embodiments of the present invention are low, and the impurity growth rate is slow after 6 months of accelerated processing, indicating better stability than commercially available ordinary powder injections. In contrast, the comparative examples, lacking corresponding technical features, all showed significantly increased impurity growth rates, demonstrating that the multiple stabilizing system and process optimization of the present invention synergistically exert excellent stabilizing effects.

[0062] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A cefazolin sodium for injection, which is a sterile lyophilized powder for injection, characterized in that, By weight, it includes 100 parts of cefazolin sodium, 15-40 parts of lyophilized skeleton support agent, 3-12 parts of stabilizer, 10-25 parts of protectant and pH adjuster; The freeze-dried skeleton support agent is a compound of mannitol, low molecular weight dextran 40 and xylitol; The pH adjuster is a citrate-sodium citrate buffer, used to make the pH value of the finished product after reconstitution with water 6.0~6.

5.

2. The cefazolin sodium for injection as described in claim 1, characterized in that, In the freeze-dried skeleton support, the weight ratio of mannitol, low molecular weight dextran 40, and xylitol is 1:(0.2~0.8):(0.02~0.08).

3. The cefazolin sodium for injection as described in claim 1, characterized in that, The stabilizer is selected from one or more of glycine, arginine, histidine, and proline.

4. The cefazolin sodium for injection as described in claim 1, characterized in that, The protective agent is injectable trehalose.

5. The cefazolin sodium for injection as described in claim 1, characterized in that, In the citric acid-sodium citrate buffer, the molar ratio of citric acid to sodium citrate is 1:(3~5).

6. The cefazolin sodium for injection as described in claim 1, characterized in that, It also includes antioxidants and metal chelating agents, wherein the antioxidant is vitamin C and the metal chelating agent is sodium edetate; by weight, vitamin C is 0.05-0.5 parts and sodium edetate is 0.01-0.1 parts.

7. The method for preparing cefazolin sodium for injection according to any one of claims 1 to 6, characterized in that, Includes the following steps: S1. Under light-protected conditions, dissolve sodium edetate in water for injection at 2-8℃, then add cefazolin sodium and a protective agent and stir with ultrasound to aid dissolution. After adding vitamin C, add a pH adjuster to adjust the pH of the solution to 6.0-6.

5. Add a lyophilized matrix support agent and a stabilizer and mix well to obtain the drug solution. S2. Nitrogen gas is introduced into the drug solution and preparation container to replace the headspace residual oxygen content to ≤1.0%. Then, the drug solution is aseptically filtered through a microporous filter membrane, filled into vials and partially stoppered. S3. Place the filled vials in a freeze dryer, and then pre-freeze, anneal, cool, sublime dry, desorption dry, and vacuum anneal until the moisture content is ≤1.5%. S4. After the desorption drying is completed, nitrogen gas is introduced into the freeze dryer under vacuum until the headspace residual oxygen content is ≤1.0%, and then the freeze dryer is fully plugged and capped.

8. The preparation method according to claim 7, characterized in that, The parameters for ultrasonic-assisted dissolution in step S1 are: ultrasonic frequency 20~40kHz, power 100~300W, dissolution temperature 2~6℃, dissolution time 1~3 hours, and stirring speed 100~300r / min.

9. The preparation method according to claim 7, characterized in that, The specific parameters of the freeze-drying process in step S3 are as follows: Pre-freezing: Lower the product temperature to -40 to -45°C at a rate of 0.5 to 2°C / min and hold for 2 to 4 hours; Annealing treatment: Then anneal at a rate of 0.1~0.5℃ / min to -20~-15℃ for 1~3 hours; Further cooling: Cool to -45~-50℃ and maintain for 1~2 hours; Sublimation drying: Sublimate drying at -20~-5℃ and vacuum degree 10~30Pa for 8~16 hours; Desorption drying: Desorption drying at 25~35℃ and vacuum degree ≤10Pa for 4~8 hours; Vacuum post-annealing: Hold at 30~35℃ and vacuum degree ≤10Pa for 1~2 hours.

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