Antibacterial composition of cefoperazone and avibactam and application thereof

Abstract

The invention belongs to the technical field of medicine, relates to an antibacterial composition containing cefoperazone and avibactam and application of the antibacterial composition, and particularly provides avibactam or pharmaceutically acceptable salt and solvate of avibactam and cefoperazone or pharmaceutically acceptable salt and solvate of cefoperazone. The invention relates to application of cefoperazone in preparation of drugs for treating or preventing bacterial infection, in particular to application of cefoperazone in preparation of drugs for treating infection caused by klebsiella pneumoniae and enterobacter cloacae, so as to solve the problem that drug resistance and clinical requirements of cefoperazone and other antibiotic products cannot be met.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular, relates to an antibacterial composition containing cefoperazone and avibactam and application thereof. Background Art

[0002] Cefoperazone is a third-generation cephalosporin antibiotic commonly used in clinical practice. Its sodium salt is the main medicinal form, and its structure is shown in formula (1). Cefoperazone sodium can be used to treat the following infections caused by sensitive bacteria: upper and lower respiratory tract infections; peritonitis, cholecystitis, cholangitis and other intra-abdominal infections; sepsis; meningitis; skin and soft tissue infections; pelvic inflammatory disease, endometritis, gonorrhea and other reproductive tract infections; upper and lower urinary tract infections; bone and joint infections. It can be used to prevent postoperative infections in patients undergoing abdominal, gynecological, cardiovascular and orthopedic surgeries.

[0003] It has good antibacterial effect on Escherichia coli, Klebsiella, Proteus, Salmonella typhi, Shigella, Citrobacter and other Enterobacteriaceae and Pseudomonas aeruginosa, but has poor effect on Enterobacter aerogenes, Enterobacter cloacae, Salmonella typhimurium and Acinetobacter. Haemophilus influenzae, Neisseria gonorrhoeae and Neisseria meningitidis are highly sensitive to this product. This product also has good effect on various groups of streptococci and pneumococci, and has moderate effect on Staphylococcus aureus (methicillin-sensitive strains). Cefoperazone has good effect on most Gram-positive anaerobes and some Gram-negative anaerobes.

[0004] After years of use, many pathogens have developed resistance to cefoperazone, such as Enterococcus and Bacteroides fragilis. In order to solve the problem of drug resistance, a compound preparation of cefoperazone was developed, which was formulated with β-lactamase inhibitors and cefoperazone. The compound products currently on the market are mainly cefoperazone sulbactam and cefoperazone tazobactam. The main disadvantages of sulbactam and tazobactam are that the inhibition enzyme spectrum is narrow and can only inhibit some class A enzymes. In fact, various pathogenic bacteria produce four types of β-lactamases, A, B, C, and D. In the past 10 years, no new cefoperazone compound preparations for medicinal use have been reported. Therefore, it is necessary to develop new compound preparations to meet clinical needs.

[0005] Avibactam is a known new type of β-lactamase inhibitor, with a structure as shown in formula (2). It is significantly different from previous enzyme inhibitors, mainly in that: the chemical structure does not contain the traditional β-lactam four-membered ring, and the mechanism of action is to form a conjugate with the enzyme through a covalent bond. The binding is reversible, has a long-term effect, has a broad enzyme inhibition spectrum, and will not induce the production of new enzymes.

[0006] Chinese Patent No. 03802878.6, filed in 2003, discloses that β-lactam antibiotics can be used in combination with avibactam. The patent mentions dozens of β-lactam antibiotics including penicillins, carbapenems, cephalosporins, etc., but no detailed research has been conducted.

[0007] Although β-lactam antibiotics have similar mechanisms of action, there are hundreds of types of them. Different specific drugs often have different pharmacodynamic properties, and their effects on different bacteria are particularly difficult to predict. To be suitable for clinical use, very specific research must be conducted on a particular drug and a suitable dosing regimen must be developed. Summary of the invention

[0008] The present invention provides use of a cefoperazone component and an avibactam component in preparing a drug for treating or preventing bacterial infection. The cefoperazone component comprises cefoperazone having a structure represented by formula (1) or a pharmaceutically acceptable salt or solvate thereof, and the avibactam component comprises avibactam having a structure represented by formula (2) or a pharmaceutically acceptable salt or solvate thereof. ; The mass ratio of the active ingredient calculated as cefoperazone of formula (1) to avibactam of formula (2) is 50:1 to 1:50.

[0009] In some embodiments, the bacterium is a bacterium expressing β-lactamase, a bacterium expressing one or more β-lactamases. Further, according to the Ambler classification, the β-lactamase is a Class A, Class B, or Class C enzyme. Further, the genotype of the β-lactamase includes TEM, SHV, CTX-M, CepA, KPC, AmpC, ACT, CMY, and OXA.

[0010] The mass ratio of the active ingredient calculated as cefoperazone to the active ingredient calculated as avibactam is 50:1 to 1:50, which has antibacterial activity. In some specific embodiments, the mass ratio has good antibacterial activity when it is 15:1 to 1:10; further, the mass ratio has better antibacterial activity when it is 2:1 to 1:10.

[0011] The bacteria include but are not limited to Pseudomonas (Pseudomonas aeruginosa, Pseudomonas cepacia, Pseudomonas parvum), Xanthomonas, Legionella, Alcaligenes, Brucella (sheep, abortion, swine Brucella), Bordetella, Flavobacterium, Acinetobacter (e.g., cephalosporin-sensitive or -resistant Acinetobacter baumannii, Burkholderia cepacia, Alcaligenes, Stenotrophomonas maltophilia), Moraxella, Bulahanella subgenus, Neisseria (meningococci, gonococci), Escherichia, Citrobacter (e.g., Citrobacter heteromorphis, Citrobacter freundii), Salmonella (Salmonella typhi, Salmonella paratyphi, Choleraesuis, Enteritidis), Shigella (Shigella dysenteriae, Shigella flexneri, Shigella sonnei), Klebsiella (Pneumoniae, Acidogenic bacilli , Oxytocin), Enterobacter (cloacae, Enterobacter aerogenes), Serratia (bizarre, common Proteus), Proteus, Morganella, Yersinia, Vibrio, Haemophilus (influenza, parainfluenza, Haemophilus influenzae β-lactamase (-), Haemophilus influenzae β-lactamase (+)), Pasteurella, Micrococcus, Staphylococcus (Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus), Streptococcus (Streptococcus pyogenes, Group B Streptococcus, Streptococcus pneumoniae, Streptococcus viridans), Enterococcus (Enterococcus faecalis, Enterococcus faecium), Diplococcus, Peptococcus, Bacillus, Fusobacterium (Clostridium tetani, Clostridium perfringens, Clostridium botulinum, Clostridium difficile, Clostridium septicum), Actinomycetes, Bacteroides. Bacteroides spp. (e.g., Bacteroides fragilis, Bacteroides thetaiotaomicron, Bacteroides gibbsii, Bacteroides ovatus, Bacteroides monomorpha, Bacteroides urealyticum, Bacteroides vulgaris).

[0012] The bacterial infections treated include, but are not limited to, bacterial infections including meningitis, urinary tract infections (e.g., pyelonephritis, refractory cystitis), gynecological infections (e.g., endometritis, pelvic inflammatory disease, adnexitis), skin and soft tissue infections, sepsis, intra-abdominal infections, lung abscesses, chronic respiratory diseases, peritonitis, secondary infections caused by chronic respiratory diseases, respiratory system infections (including, but not limited to, pneumonia (e.g., hospital-acquired pneumonia)), sepsis, reproductive system infections, biliary tract infections, bone infections, and joint infections; the infections prevented include abdominal, gynecological, cardiovascular, and post-orthopedic surgical infections.

[0013] The infection may be caused by resistant bacteria, non-resistant bacteria, or a mixture of resistant and non-resistant bacteria. The infection may be caused by β-lactamase-producing bacteria, non-β-lactamase-producing bacteria, or a mixture of β-lactamase-producing bacteria and non-β-lactamase-producing bacteria.

[0014] The infection may be caused by bacteria resistant to cephalosporin antibiotics. The infection may be caused by one or two of Klebsiella pneumoniae and Enterobacter cloacae.

[0015] Specifically, the infection may be caused by one or more bacteria expressing one or more TEM, SHV, CTX-M, CepA, KPC, AmpC, ACT, CMY, OXA enzymes.

[0016] The pharmaceutically acceptable salt of avibactam may be avibactam sodium salt.

[0017] The pharmaceutically acceptable salt of cefoperazone may be cefoperazone sodium salt.

[0018] In another aspect, the present invention provides a method for treating or preventing bacterial infection, comprising administering a therapeutically effective amount of a cefoperazone component and an avibactam component to a patient.

[0019] The avibactam or its pharmaceutically acceptable salt or solvate and cefoperazone or its pharmaceutically acceptable salt or solvate are independently formulated and co-administered.

[0020] In another aspect, the present invention provides a composition for treating or preventing bacterial infection, comprising a cefoperazone component and an avibactam component; wherein the mass ratio of the active ingredient calculated as cefoperazone to the active ingredient calculated as avibactam is 50:1 to 1:50, preferably 15:1 to 1:10.

[0021] The composition can be solid or liquid, and is presented in a pharmaceutical form, such as plain tablets or sugar-coated tablets, gelatin capsules, granules, suppositories, injection preparations, ointments, creams, gels, and is prepared according to conventional methods. In the case of injection preparations, the composition can be dry mixed powder injections and freeze-dried powder injections.

[0022] The injection of the avibactam component and the cefoperazone component can be administered by intravenous drip or intravenous injection.

[0023] In vitro antibacterial experiments show that the antibacterial activity of cefoperazone and avibactam in combination according to the technical solution of the present invention, especially in combination according to the mass ratio of the present invention, is significantly stronger than that of cefoperazone alone and cefoperazone-sulbactam combination.

[0024] The dosages and ratios of cefoperazone and avibactam involved in the specification and examples of this application are the dosages and ratios after conversion to active ingredients. DETAILED DESCRIPTION

[0025] The present application is further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present application and are not used to limit the scope of protection of the present application.

[0026] Example 1 Study on the antibacterial activity of cefoperazone-avibactam in multiple genotypes 1. Antimicrobial drugs Cefoperazone (using cefoperazone sodium), avibactam (using avibactam sodium), and sulbactam.

[0027] 2. Test bacteria Clinically isolated strains producing CTX-M, KPC, AmpC, ACT, and β-lactamase genotypes and purchased OXA standard strains.

[0028] 3. Experimental plan According to the CLSI drug sensitivity test standards of the United States Committee for Clinical Laboratory Standards, the minimum inhibitory concentration (MIC) of cefoperazone, avibactam and other antibiotics against clinical isolates was determined by broth microdilution method.

[0029] 3.1 Preparation of bacterial culture Take a fresh nutrient agar slant of the test strain, inoculate 2 mL of CAMHB medium, and culture at 37°C for 6 h. Compare the turbidity with a No. 0.5 McFarland turbidimeter tube, dilute appropriately, and adjust the concentration to about (2-4) × 10 5 CFU / mL is reserved.

[0030] 3.2 Drug dissolution and dilution .

[0031] Take about 5 mg of cefoperazone, avibactam, and sulbactam, add sterile water and dissolve into a solution of 1280 μg / mL.

[0032] A 1280 μg / mL solution of cefoperazone and a 1280 μg / mL solution of avibactam were taken out and mixed evenly in a ratio of 1:1 to serve as the mother solution of Group 1.

[0033] Pipette a 1280 μg / mL solution of cefoperazone and a 160 μg / mL solution of avibactam (1280 μg / mL diluted to 160 μg / mL) and mix them evenly in a 1:1 ratio to serve as the mother solution for Group 2.

[0034] A 1280 μg / mL solution of cefoperazone was diluted to 640 μg / mL as the stock solution for Group 3.

[0035] A 1280 μg / mL solution of cefoperazone and a 1280 μg / mL solution of sulbactam were taken out and mixed evenly in a ratio of 1:1 to serve as the mother solution of Group 4.

[0036] Take an appropriate amount of the mother solution and dilute it step by step with CAMHB medium to obtain solutions with concentrations of 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.12, and 0.06 μg / mL (calculated as cefoperazone) in each group. The drug dilution method for each group is shown in Table 2.

[0037] .

[0038] 3.3 Inoculation and cultivation 100 μL of the above-mentioned drug solutions of different concentrations were respectively pipetted into a sterile 96-well plate, and 100 μL of the bacterial suspension was added to each well. The bacterial concentration was approximately (1-2)×10 5 CFU / mL.

[0039] Set up a positive control well: 100 μL bacterial solution + 100 μL culture medium; negative control well: 200 μL culture medium. Incubate at 37℃ for 16-20 h, take out and observe the results, and record the growth status.

[0040] 3.4 Criteria The results were observed with the naked eye, and the concentration that inhibited the visible growth of bacteria was recorded as the minimum inhibitory concentration (MIC) of the compound. The results are summarized in Table 3.

[0041] ; Note: The MIC values ​​of cefoperazone / avibactam combinations in the table are shown as cefoperazone concentrations in µg / mL.

[0042] According to the test results, the activity of the combined administration of cefoperazone and avibactam is significantly higher than that of cefoperazone alone and cefoperazone and sulbactam combined. Background technology It is known that cefoperazone has poor inhibitory activity against Enterobacter cloacae. Group 1 of Example 1 of the present invention has good inhibitory activity against enzyme-producing Enterobacter cloacae, which is a significant improvement over the prior art.

[0043] Example 2 Study on the antibacterial activity of cefoperazone-avibactam in multiple ratios 1. Antimicrobial drugs Cefoperazone and avibactam sodium were prepared at different concentration ratios.

[0044] 2. Test bacteria KPC-2 genotype Klebsiella pneumoniae and AmpC genotype Enterobacter cloacae isolated clinically.

[0045] 3.1 Preparation of bacterial culture Take a fresh nutrient agar slant of the test strain, inoculate 2 mL of CAMHB medium, and culture at 37°C for 6 h. Compare the turbidity with a No. 0.5 McFarland turbidimeter tube, dilute appropriately, and adjust the concentration to about (2-4) × 10 5 CFU / mL is reserved.

[0046] 3.2 Drug dissolution and dilution .

[0047] Take about 5 mg of cefoperazone and avibactam and add sterile water to dissolve into solutions with a concentration of 1280 μg / mL.

[0048] The above 1280 μg / mL compound solution was diluted with CAMHB medium to obtain compound solutions of various concentrations, as shown in Table 5.

[0049] .

[0050] Solutions of different concentrations were mixed to obtain mother solutions of each group of compounds, as shown in Table 6.

[0051] .

[0052] Take appropriate amount of mother solution of each group and dilute it step by step with CAMHB medium to obtain solutions with concentrations of 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.12, and 0.06 μg / mL (calculated as cefoperazone) in each group. The dilution method of each group of drugs is shown in Table 7.

[0053] .

[0054] 3.3 Inoculation and cultivation 100 μL of the above-mentioned drug solutions of different concentrations were respectively pipetted into a sterile 96-well plate, and 100 μL of the bacterial suspension was added to each well. The bacterial concentration was approximately (1-2)×10 5 CFU / mL.

[0055] Set up a positive control well: 100 μL bacterial solution + 100 μL culture medium; negative control well: 200 μL culture medium. Incubate at 37℃ for 16-20 h, take out and observe the results, and record the growth status.

[0056] 3.4 Criteria The results were observed with the naked eye, and the concentration that inhibited the visible growth of bacteria was recorded as the minimum inhibitory concentration (MIC) of the compound. The results are summarized in Table 8.

[0057] .

[0058] .

[0059] In summary, the activity of the combined administration of cefoperazone and avibactam is significantly higher than that of cefoperazone alone and cefoperazone and sulbactam together. According to the relevant background technology, the inhibitory activity of cefoperazone alone against Enterobacter cloacae is poor, and Group 1 of Example 1 of the present invention has good inhibitory activity against enzyme-producing Enterobacter cloacae, which is a significant improvement over the prior art.

[0060] According to the data in Tables 8 and 9, as the proportion of Avitam increases, the MIC value tends to decrease, proving that the antibacterial activity is increasing.

[0061] The preferred specific embodiments of the present invention are described in detail above. It should be understood that ordinary technicians in the field can make many modifications and changes based on the concept of the present invention without creative work. Therefore, all technical solutions that can be obtained by technicians in the technical field based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art should be within the scope of protection determined by the claims.

Claims

1. Use of a cefoperazone component and an avibactam component in the preparation of a drug for treating or preventing bacterial infection, wherein the cefoperazone component comprises cefoperazone of the structure shown in formula (1) or a pharmaceutically acceptable salt or solvate thereof, and the avibactam component comprises avibactam of the structure shown in formula (2) or a pharmaceutically acceptable salt or solvate thereof, ； It is characterized in that The mass ratio of cefoperazone of formula (1) to avibactam of formula (2) is 50:1 to 1:

50.

2. The use according to claim 1, characterized in that The bacteria are bacteria that express one or more β-lactamases.

3. The use according to claim 2, characterized in that The genotypes of β-lactamase expressed by the bacteria include TEM, SHV, CTX-M, CepA, KPC, AmpC, ACT, CMY, and OXA.

4. The use according to claim 1, characterized in that The bacteria include Pseudomonas, Xanthomonas, Legionella, Alcaligenes, Brucella, Bordetella, Flavobacterium, Acinetobacter, Moraxella, Bulahanella, Neisseria, Escherichia, Citrobacter, Salmonella, Shigella, Klebsiella, Enterobacter, Serratia, Proteus, Morganella, Yersinia, Vibrio, Haemophilus, Pasteurella, Micrococcus, Staphylococcus, Streptococcus, Enterococcus, Diplococcus, Peptococcus, Bacillus, Fusobacterium, Actinomycetes, and Bacteroides.

5. The use according to claim 1, characterized in that The bacterial infections treated include meningitis, urinary tract infection, gynecological infection, skin and soft tissue infection, sepsis, intra-abdominal infection, lung abscess, chronic respiratory disease, peritonitis, secondary infection caused by chronic respiratory disease, respiratory system infection, sepsis, reproductive system infection, biliary tract infection, bone infection, and joint infection; the bacterial infections prevented include abdominal, gynecological, cardiovascular, and post-orthopedic surgical infections.

6. The use according to claim 1, characterized in that The pharmaceutically acceptable salt of cefoperazone is cefoperazone sodium.

7. The use according to claim 1, characterized in that The pharmaceutically acceptable salt of avibactam is avibactam sodium.

8. The use according to claim 1, characterized in that The mass ratio of cefoperazone of formula (1) to avibactam of formula (2) is 15:1 to 1:

10.

9. A method for treating or preventing bacterial infection, characterized in that The method comprises administering to a patient a therapeutically effective amount of the cefoperazone component and the avibactam component according to any one of claims 1 to 8.

10. A composition for treating or preventing bacterial infection, characterized in that: The composition comprises the cefoperazone-like component and the avibactam-like component as claimed in any one of claims 1 to 8.

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