Avibactam-containing antibacterial composition and application thereof

Abstract

The invention belongs to the technical field of medicines, relates to an avibactam-containing antibacterial composition and application thereof, and particularly provides application of avibactam or pharmaceutically acceptable salts thereof and carbapenem antibiotics in preparation of medicines for treating or preventing bacterial infection. The invention relates to application of carbapenem antibiotics, in particular to application of carbapenem antibiotics in preparation of medicines for treating infection caused by escherichia coli, pseudomonas aeruginosa, klebsiella pneumoniae, acinetobacter baumannii and enterobacter aerogenes, so as to solve the problem of drug resistance of carbapenem antibiotics.

Description

[0001] This application is a divisional application of the invention patent application filed on March 13, 2019, with application number 201910189632.X and invention title "Antibacterial Composition Containing Avibactam and Its Use Thereof". Technical Field

[0002] This invention relates to antibacterial compositions containing avibactam and their uses, and more specifically, to a composition containing avibactam and carbapenem antibiotics and its uses. Background Technology

[0003] Avibactam (NXL-104) is a novel β-lactamase inhibitor. Compared with classic β-lactamase inhibitors, it has the special characteristics of long-lasting effect, broad spectrum of inhibition, and no induction of β-lactamase production. Yang Fan et al. pointed out in the article "Avibactam and its compound preparation, a promising new β-lactamase inhibitor. Journal of the Third Military Medical University, 2013, 35(23):2498-2501" that avibactam can inhibit group A (including ESBLs and KPC enzymes), group C (mainly AmpC enzymes) and some group D β-lactamases (such as OXA-48), but usually cannot inhibit group B metallo-β-lactamases. The structure of avibactam is shown in Formula I.

[0004]

[0005] (Formula I) Avibactam has almost no antibacterial activity, but its activity can be effectively restored or enhanced when used in combination with cephalosporin antibiotics. The ceftazidime-avibactam combination was approved for marketing by the U.S. Food and Drug Administration in February 2015, with indications primarily including complicated intra-abdominal infections, complicated urinary tract infections, and Gram-negative bacillus infections.

[0006] Carbapenems are a class of atypical β-lactam antibiotics with a broad antibacterial spectrum and strong antibacterial activity. Due to their stability against most β-lactamases and low toxicity, they have become one of the most important antibacterial drugs for treating serious bacterial infections. Marketed varieties include imipenem, meropenem, and panipenem. Carbapenems are highly stable against group A ESBLs (Extended Spectrum Beta-Lactamases) and group C AmpC β-lactamases, but they can be hydrolyzed and inactivated by group B metallo-β-lactamases, leading to carbapenem resistance. Metallo-β-lactamases can be detected in bacteria such as *Pseudomonas aeruginosa*, *Enterobacter* spp., *Klebsiella pneumoniae*, *Acinetobacter*, *Stenotrophomonas maltophilia*, *Aeromonas hydrophila*, and *Serratia marcescens*.

[0007] In recent years, with the widespread use of carbapenem antibiotics, the problem of carbapenem antibiotic resistance has gradually become prominent and needs to be further addressed. Summary of the Invention

[0008] On one hand, the present invention provides the use of avibactam or pharmaceutically acceptable salts of it and carbapenem antibiotics in the preparation of medicaments for treating or preventing bacterial infections. In some embodiments, the bacteria include susceptible and resistant bacteria; in some embodiments, the bacteria are resistant bacteria expressing β-lactamases.

[0009] The bacteria mentioned include, but are not limited to, Staphylococcus (e.g., methicillin-resistant Staphylococcus aureus, Staphylococcus epidermidis), Enterococcus (e.g., Enterococcus faecalis, Enterococcus faecium), Stenotrophomonas maltophilia, Pseudomonas aeruginosa, Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Shigella spp., Salmonella spp., Citrobacter spp. (e.g., Citrobacter heterotrophus, Citrobacter freundii), Proteus spp., Serratia spp., Enterobacter cloacae, Acinetobacter spp. (e.g., Acinetobacter baumannii), Haemophilus influenzae, Xanthomonas aeruginosa, Pseudomonas aeruginosa, and Streptococcus spp. (e.g., Peptostreptococcus spp., Streptococcus pneumoniae, Streptococcus viridans). Streptococcus, Streptococcus pneumoniae, Moraxella spp., Escherichia coli, Actinomyces spp., Bacteroides spp. (e.g., Bacteroides fragilis, Bacteroides polymorpha, Bacteroides gigantea, Bacteroides ovalis, Bacteroides monomorpha, Bacteroides urealyticum, Bacteroides vulgaris), Prevotella spp., Fusobacterium spp., Pseudomonas aeruginosa, Neisseria meningitidis, Aeromonas hydrophila, Campylobacter jejuni, Enterobacter cloacae, Moraxella catarrhalis, Morganella morganii, Proteus mirabilis, Proteus vulgaris, Serratia marcescens, Clostridium difficile, Clostridium perfringens, Eubacterium tarda, Fusobacterium, Porphyromonas suisophilus, Propionibacterium acnes, etc.

[0010] In some specific implementations, the infection is caused by bacteria resistant to carbapenem antibiotics.

[0011] In some specific implementations, the infection is caused by one or more of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Enterobacter aerogenes, and Enterobacter cloacae.

[0012] In some specific implementations, the infection is caused by Klebsiella pneumoniae.

[0013] In some specific implementations, the infections include, but are not limited to, meningitis, urinary tract infections, gynecological infections (e.g., endometritis and pelvic inflammatory disease), skin and skin structure infections, sepsis, intra-abdominal infections, lung abscesses, chronic respiratory diseases, refractory cystitis, pyelonephritis, peritonitis, urinary tract infections, secondary infections caused by chronic respiratory diseases and gynecological adnexitis, respiratory system infections (including but not limited to pneumonia (e.g., hospital-acquired pneumonia)), urinary tract infections, reproductive system infections, biliary tract infections, intra-abdominal infections, and skin and soft tissue infections.

[0014] In some specific implementations, the infection is a lung infection, including but not limited to bacterial pneumonia (e.g., hospital-acquired pneumonia).

[0015] In some specific implementations, the infection is caused by one or more bacteria expressing one or more NDM, OXA, KPC, and GES enzymes.

[0016] In this article, the carbapenem antibiotics mentioned include, but are not limited to, meropenem, biapenem, ertapenem, imipenem, doripenem, panipenem, and faroenem.

[0017] In some specific implementation schemes, the carbapenem antibiotics described herein are selected from meropenem, biapenem, and doripenem.

[0018] In some specific implementations, the carbapenem antibiotic described herein is meropenem.

[0019] On the other hand, the present invention provides a method for treating or preventing bacterial infections, comprising administering to a patient a therapeutically effective amount of avibactam or a pharmaceutically acceptable salt thereof, and at least one carbapenem antibiotic.

[0020] The bacterial infection is caused by bacteria expressing β-lactamase. Avibactam or a pharmaceutically acceptable salt thereof may be administered simultaneously or sequentially with carbapenem antibiotics.

[0021] In some specific implementations, the avibactam or a pharmaceutically acceptable salt thereof is formulated independently and co-administered with carbapenem antibiotics.

[0022] In some specific implementations, the avibactam or its pharmaceutically acceptable salts and carbapenems are administered via intravenous infusion or injection.

[0023] In some specific embodiments, avibactam or a pharmaceutically acceptable salt thereof is administered with carbapenem antibiotics every 6 hours, 8 hours, 12 hours, or 24 hours, with each dose consisting of 40-4000 mg of carbapenem antibiotics and 10-1000 mg of avibactam. In some preferred embodiments, it is administered every 8 hours, with each dose consisting of 100-2000 mg of carbapenem antibiotics and 25-500 mg of avibactam. For patients with renal impairment, the dose may be appropriately reduced or the dosing interval extended; for example, in some specific embodiments, the dose of carbapenem antibiotics given to patients with renal impairment is halved each time, administered every 12 hours or 24 hours.

[0024] In some specific embodiments, avibactam or a pharmaceutically acceptable salt thereof is administered with meropenem every 8 or 12 hours, with each dose consisting of 200-2000 mg of meropenem and 50-500 mg of avibactam. In some preferred embodiments, 500-1000 mg of meropenem and 125-250 mg of avibactam are administered each time. In some preferred embodiments, 500 mg of meropenem and 125 mg of avibactam are administered every 8 hours.

[0025] In some specific embodiments, avibactam or a pharmaceutically acceptable salt thereof is administered with doribene every 8 or 12 hours, with each dose containing 200-1000 mg of doribene and 50-250 mg of avibactam. In some preferred embodiments, avibactam is administered every 8 hours, with each dose containing 400-800 mg of doribene and 100-200 mg of avibactam. In some preferred embodiments, avibactam is administered every 8 hours, with each dose containing 500 mg of doribene and 125 mg of avibactam.

[0026] In some specific embodiments, avibactam or a pharmaceutically acceptable salt thereof is administered with biapenem every 8 or 12 hours, with each dose containing 100-1000 mg of biapenem and 25-250 mg of avibactam. In some preferred embodiments, avibactam is administered every 12 hours, with each dose containing 200-500 mg of biapenem and 50-125 mg of avibactam. In some preferred embodiments, avibactam is administered every 12 hours, with each dose containing 300 mg of biapenem and 75 mg of avibactam.

[0027] In another aspect, the present invention provides a composition for treating or preventing bacterial infections, comprising avibactam or a pharmaceutically acceptable salt thereof, and at least one carbapenem antibiotic. The bacterial infection is caused by bacteria expressing one or more β-lactamases. The composition may be solid or liquid and is presented in pharmaceutical form, such as uncoated tablets or sugar-coated tablets, gelatin capsules, granules, suppositories, injectable formulations, ointments, creams, or gels, and is prepared according to conventional methods.

[0028] In some specific embodiments, the avibactam or a pharmaceutically acceptable salt thereof is prepared as a fixed-dose composition with carbapenem antibiotics, comprising 40-4000 mg of carbapenem antibiotics and 10-1000 mg of avibactam. In some preferred embodiments, the composition comprises 100-2000 mg of carbapenem antibiotics and 25-500 mg of avibactam.

[0029] The carbapenem antibiotics of the present invention, especially the combination of meropenem and avibactam, not only significantly reduce the development of drug resistance in susceptible bacteria, but also have significantly lower minimum inhibitory concentrations (MICs) against carbapenem-resistant Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Enterobacter aerogenes, Enterobacter cloacae, and Acinetobacter baumannii than when carbapenem antibiotics are used alone.

[0030] The carbapenem antibiotics of this invention, especially the combination of meropenem and avibactam, resulted in significantly higher survival rates in rats infected with carbapenem-resistant Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, and Acinetobacter baumannii compared to rats treated with the same dose of carbapenem antibiotics alone. The median effective dose (ED50) of the combination was significantly higher. 50 The toxicity was significantly lower than that of single-agent administration, meaning that the combination of carbapenem antibiotics and avibactam significantly improved the resistance of the aforementioned bacteria. Experimental results in healthy mice using the combination of carbapenem antibiotics and avibactam showed a significant reduction in toxicity compared to single-agent administration of carbapenem antibiotics. Detailed Implementation

[0031] The present application will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present application.

[0032] Example 1 In vitro antibacterial test of avibactam-carbapenem antibiotics 1. Experimental Materials The strain used in this experiment was obtained from a clinical isolate from Huashan Hospital of Fudan University. It was confirmed using existing technology to be a carbapenemase-producing strain (the method for confirming carbapenemase production can be found in: Pasteran. Simplified Protocol for Carba NP Test for Enhanced Detection of Carbapenemase Producers Directly from Bacterial Cultures[J]. Journal of Clinical Microbiology, 2015, 53(12): 3908-3911; the method for detecting the gene phenotype of carbapenemase production can be found in: Cheng Kuo. Evaluation of the effect of imCIM in detecting class B carbapenemases[J]. Journal of Clinical Laboratory Medicine, 2017, 35(1): 31-35; He Jianye. Analysis of drug resistance genes in carbapenem-resistant Enterobacteriaceae[J]. Laboratory Medicine, 2017, 32(11): 985-989). All other materials used in this experiment were commercially available.

[0033] 2. Experimental Methods 2.1 Preparation of antibiotic standby solution Antibiotic stock solutions (doripene, biapenem, and meropenem) with a concentration of 2560 μg / ml were serially diluted to prepare nine different concentration gradients: 128 μg / ml, 64 μg / ml, 32 μg / ml, 16 μg / ml, 8 μg / ml, 4 μg / ml, 2 μg / ml, 1 μg / ml, and 0.5 μg / ml. A blank solution with a concentration of 0 μg / ml was also prepared.

[0034] 2.2 Preparation of Avibactam Ready-to-Use Solution Avibactam stock solution with a concentration of 2560 μg / ml was serially diluted to prepare five concentration gradients of 64 μg / ml, 32 μg / ml, 16 μg / ml, 8 μg / ml, and 4 μg / ml, and a blank solution of 0 μg / ml was also prepared.

[0035] 2.3 Preparation of bacterial culture Take an appropriate amount of bacteria that has been incubated on MH agar (Mueller Hinton agar) plates for 20 hours using an inoculation needle, dilute it with physiological saline to 0.5 McFarland turbidity, and then dilute it twice with CAMHB (Mueller Hinton II Broth (Cation-Adjusted)) culture medium 10 times.

[0036] 2.4 Determination of enzyme inhibition and formulation effect by microdilution method of culture medium Take a 96-well plate, add 100 μl of bacterial culture, 50 μl of avibactam preparation solution and 50 μl of antibiotic preparation solution to each well, incubate at 37℃ for 20 h, observe the turbidity of the 96-well plate, and determine the minimum inhibitory concentration of each antibiotic against each group of bacteria under different concentrations of avibactam.

[0037] 3. Experimental Results Note: Clinical isolates without genotype labeling did not undergo genotyping.

[0038] Table 1. Doripenum-Avibactam (Escherichia coli)

[0039]

[0040]

[0041] Table 2. Biapenem-Avibactam (Escherichia coli)

[0042]

[0043]

[0044] Table 3 Meropenem-Avibactam (Escherichia coli)

[0045]

[0046]

[0047] Table 4. Doripenem-Avibactam (Acinetobacter baumannii)

[0048] Table 5. Biapenem-Avibactam (Acinetobacter baumannii)

[0049] Table 6 Meropenem-Avibactam (Acinetobacter baumannii)

[0050] Table 7 Doribenam-Avibactam (Klebsiella pneumoniae)

[0051] Table 8. Biapenem-Avibactam (Klebsiella pneumoniae)

[0052] Table 9 Meropenem-Avibactam (Klebsiella pneumoniae)

[0053]

[0054] Table 10 Doripenam-Avibactam (Pseudomonas aeruginosa)

[0055] Table 11 Biapenem-Avibactam (Pseudomonas aeruginosa)

[0056] Table 12 Meropenem-Avibactam (Pseudomonas aeruginosa)

[0057] Table 13 Doripenem-Avibactam (Enterobacter aerogenes)

[0058] Table 14 Biapenem-Avibactam (Enterobacter aerogenes)

[0059]

[0060] Table 15 Meropenem-Avibactam (Enterobacter aerogenes)

[0061]

[0062] Example 2 Avibactam-Meropenem Fractional Inhibitory Concentration (FIC) 1. Experimental Materials The strains used in this experiment were obtained from clinical isolates at Huashan Hospital, Fudan University, involving two genera and two species: 21 strains of *Klebsiella pneumoniae* (including 10 KPC-type carbapenemase-producing strains and 11 OXA-type carbapenemase-producing strains) and 2 strains of *Escherichia coli* (both NDM-type carbapenemase-producing strains). Their carbapenemase-producing strains and genotypes can be confirmed using existing techniques. All other materials used in this experiment were commercially available.

[0063] 2. Experimental Methods According to CLSI (Clinical and Laboratory Standards Institute) M07-A10 (2015 edition) recommendations, the minimum inhibitory concentrations (MICs) of meropenem and avibactam against clinical isolates were determined using the broth-microdilution method. The combined antimicrobial activity was determined using a 12×12 checkerboard dilution method combined with antimicrobial susceptibility testing.

[0064] 2.1 Preparation of antibiotic standby solution Preparation of antibacterial solutions: Meropenem and avibactam were diluted with sterile water. The concentration range of the antibacterial solutions was 128 μg / ml to 0.06 μg / ml.

[0065] 2.2 Preparation of bacterial culture The pure bacterial colonies were prepared with physiological saline to a McFarland turbidity of 0.5. The bacterial suspension was then diluted 100-fold with double-strength cationic broth to a bacterial concentration of 10. 6 CFU / mL. Then, pipette 50 μl of bacterial culture into each of the prepared 96-well microplates (containing 50 μl of antibacterial drug solution). The inoculum size is now 5 × 10⁻⁶. 5 CFU / ml.

[0066] 2.3 Cultivation Conditions Culture conditions: Ambient air, 35℃±2℃ for 16-20 hours.

[0067] 2.4 Result Judgment Partial inhibitory concentration index (FIC index = MIC) A 联合 / MIC A 单独 + MIC B 联合 / MIC B 单独 MIC A 联合 The MIC of drug A when the two drugs are combined. A 单独 This refers to the MIC (Minimum Intake) of drug A as a single agent. B 联合 The MIC of drug B when the two drugs are combined. B 单独 The MIC is the minimum effective concentration (MIC) of drug B alone. A FIC index ≤ 0.5 indicates a synergistic effect; a FIC index > 0.5~1 indicates an additive effect; a FIC index > 1~2 indicates an irrelevant effect; and a FIC index > 2 indicates an antagonistic effect.

[0068] Table 16. FIC index of meropenem-avibactam against two NDM-producing Escherichia coli strains

[0069] Table 17. FIC index of meropenem-avibactam against 10 KPC-producing Klebsiella pneumoniae strains

[0070] Table 18. FIC index of meropenem-avibactam against 11 strains of OXA-producing Klebsiella pneumoniae.

Claims

1. Use of avibactam or pharmaceutically acceptable salts and carbapenem antibiotics thereof in the preparation of medicaments for the treatment or prevention of bacterial infections, said bacteria including susceptible and resistant bacteria; preferably, said bacteria being resistant bacteria expressing β-lactamase.

2. The use according to claim 1, wherein the bacteria include Staphylococcus, Enterococcus, Stenotrophomonas maltophilia, Pseudomonas aeruginosa, Escherichia coli, Klebsiella spp., Enterobacter aerogenes, Shigella spp., Salmonella spp., Citrobacter spp., Proteus spp., Serratia spp., Enterobacter cloacae, Acinetobacter spp., Haemophilus influenzae, Xanthomonas, Pseudomonas aeruginosa, Streptococcus spp., Pneumococcus, Moraxella spp., Escherichia coli, Actinomyces spp., Bacteroides spp., Prevotella spp., Fusobacterium spp., Pseudomonas aeruginosa, Neisseria meningitidis, Aeromonas hydrophila, Campylobacter jejuni, Enterobacter cloacae, Moraxella catarrhalis, Morganella morganii, Proteus mirabilis, Proteus vulgaris, Serratia marcescens, Clostridium difficile, Clostridium perfringens, Eubacterium tarda, Fusobacterium, Porphyromonas suisophilus, and Propionibacterium acnes.

3. The use according to claim 1, wherein the infection is caused by carbapenem-resistant bacteria expressing β-lactamases, preferably, the infection is caused by one or more bacteria expressing one or more NDM, OXA, KPC, GES genotypes of β-lactamases and / or ESBLs and AmpC enzymes.

4. The use according to claim 1, wherein the infection is caused by one or more of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterobacter aerogenes, and Acinetobacter baumannii.

5. The use according to claim 1, wherein the bacterial infection is meningitis, pneumonia, urinary tract infection, gynecological infection, skin and skin structure infection, sepsis, intra-abdominal infection, lung abscess, chronic respiratory disease, refractory cystitis, pyelonephritis, peritonitis, urinary tract infection, secondary infection caused by chronic respiratory disease, and gynecological adnexitis, respiratory system infection, sepsis, urinary system infection, reproductive system infection, biliary tract infection, intra-abdominal infection, and skin and soft tissue infection; preferably, the gynecological infection is endometritis and pelvic inflammatory disease, and the respiratory system infection is pneumonia; more preferably, the pneumonia is hospital-acquired pneumonia.

6. The use according to claim 1, wherein the carbapenem antibiotic is selected from meropenem, biapenem, and doripenem.

7. A method of treating or preventing bacterial infection, comprising administering to a patient a therapeutically effective amount of avibactam or a pharmaceutically acceptable salt thereof.

8. The method of claim 7, wherein the avibactam or a pharmaceutically acceptable salt thereof is independently formulated and co-administered with a carbapenem antibiotic.

9. A composition for treating or preventing bacterial infections, comprising avibactam or a pharmaceutically acceptable salt thereof, and at least one carbapenem antibiotic.

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