Avibactam

Manufactured by AstraZeneca

Sourced in United States

Avibactam is a β-lactamase inhibitor developed by AstraZeneca. It is designed to inactivate certain types of β-lactamase enzymes that can confer resistance to β-lactam antibiotics. Avibactam is intended for use in combination with other antibacterial agents to enhance their efficacy against resistant bacterial strains.

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7 protocols using avibactam

Aztreonam-avibactam stock solution prep

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Aztreonam (Sigma-Aldrich, St Quentin Fallavier, France) and avibactam (provided as a dry powder by AstraZeneca, Macclesfield, UK) were used to prepare stock solutions of 50 mg/mL of Aztreonam in methanol/dimethyl sulphoxide (50/50, v/v) and 1 mg/mL of avibactam in sterile water. Aztreonam avibactam stock solutions were further diluted in cation-adjusted MuellereHinton broth (CAMHB).

Chauzy A., Buyck J., de Jonge B.L.M., Marchand S., Grégoire N, & Couet W. (2019). Pharmacodynamic modelling of β-lactam/β-lactamase inhibitor checkerboard data: illustration with aztreonam-avibactam. Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases, 25(4).

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Evaluation of Avibactam and Ceftazidime

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Avibactam was obtained successively from AstraZeneca (Waltham, MA, USA) and Pfizer (New York, NY, USA) and ceftazidime from AstraZeneca. Cephalothin was purchased from Sigma-Aldrich (Overijse, Belgium) and nitrocefin from Oxoid.

Frère J.M., Bogaerts P., Huang T.D., Stefanic P., Moray J., Bouillenne F, & Brans A. (2020). Interactions between Avibactam and Ceftazidime-Hydrolyzing Class D β-Lactamases. Biomolecules, 10(3), 483.

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Antibiotic Susceptibility Profiling

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MICs were determined by broth microdilution using custom frozen panels (ThermoFisher Scientific, Cleveland, OH) according to Clinical Laboratory Standards Institute (CLSI) methods (Clinical and Laboratory Standards Institute, 2015a ). MIC values were obtained in triplicate and modal values determined. β-Lactam agents tested (doubling dilution concentration range in mg/L) included ceftazidime (0.06–128), ceftaroline (0.03–64), aztreonam (0.06–64), meropenem (0.015–32) and piperacillin-tazobactam (0.06/4–128/4). Ceftazidime, ceftaroline and aztreonam were also tested at the same concentrations in the presence of a fixed concentration of 4 mg/L of Avibactam. MICs were interpreted according to current CLSI breakpoints for these compounds in the absence of the inhibitor (Clinical and Laboratory Standards Institute, 2015b ). Avibactam and ceftaroline were provided by AstraZeneca as kind gifts.

Papp-Wallace K.M., Bajaksouzian S., Abdelhamed A.M., Foster A.N., Winkler M.L., Gatta J.A., Nichols W.W., Testa R., Bonomo R.A, & Jacobs M.R. (2015). Activities of ceftazidime, ceftaroline and aztreonam alone and combined with avibactam against isogenic Escherichia coli strains expressing selected single β-lactamases. Diagnostic microbiology and infectious disease, 82(1), 65-69.

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Antimicrobial Susceptibility Testing Protocol

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MICs were determined by broth microdilution according to CLSI guidelines (Clinical and Laboratory Standards Institute, 2020 ) for the following antibiotics (potency and origin): amikacin (Amukin 500 mg/2 ml for injection, S.A. Bristol-Myers Squibb, Belgium), azithromycin (100%, SMB, Brussels, Belgium), ceftazidime (2 g for IV injection, 72.5%, PAN Pharma, Luitré, France), chloramphenicol (98%, Sigma-Aldrich), ciprofloxacin (98%, Fluka, Sigma-Aldrich), colistin sulfate salt (79.6%, Sigma-Aldrich), doxycycline hyclate (86.6%, Sigma-Aldrich), meropenem (500 mg powder for solution for injection or infusion, 92%, Hospira UK Ltd., Hurley, United Kingdom), piperacillin (94.2%, Sigma-Aldrich, Maryland Heights, MO), temocillin (84%, Eumedica Pharmaceuticals, Manage, Belgium), ticarcillin disodium salt (85.2%, Sigma-Aldrich), and tobramycin (100%, Teva, Wilrijk, Belgium). Tazobactam sodium salt (92.4%, Cubist Pharmaceuticals, Lexington, MA) and avibactam (99.6%, AstraZeneca Pharmaceuticals, Waltham, MA) were used as inhibitors for β-lactamases. Berberine (chloride hydrate, 82.1%; Sigma-Aldrich), known to attenuate the MexXY-OprM/OprA-mediated aminoglycoside resistance in P. aeruginosa (Morita et al., 2016 (link)), was used to reduce AxyXY-OprZ activity.

Chalhoub H., Kampmeier S., Kahl B.C, & Van Bambeke F. (2022). Role of Efflux in Antibiotic Resistance of Achromobacter xylosoxidans and Achromobacter insuavis Isolates From Patients With Cystic Fibrosis. Frontiers in Microbiology, 13, 762307.

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Antibiotic Interaction Study Protocol

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Chemicals were from Sigma-Aldrich, unless otherwise stated. Avibactam and ceftazidime were kindly supplied by AstraZeneca and GlaxoSmithKline, respectively.

Abboud M.I., Damblon C., Brem J., Smargiasso N., Mercuri P., Gilbert B., Rydzik A.M., Claridge T.D., Schofield C.J, & Frère J.M. (2016). Interaction of Avibactam with Class B Metallo-β-Lactamases. Antimicrobial Agents and Chemotherapy, 60(10), 5655-5662.

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Antibiotic Susceptibility Assay Protocol

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Tobramycin, ciprofloxacin, minocycline, trimethoprim, sulfamethoxazole, ceftazidime, and aztreonam were purchased from Sigma-Aldrich. Imipenem–cilastatin was obtained from its commercial source. Avibactam, batches AFCH005151 and C565/5, were kind gifts from AstraZeneca; the chemical structure is presented in Figure 2A. Nitrocefin was a kind gift from Dr. ShahriarMobashery at the University of Notre Dame in South Bend, IN, USA.

Papp-Wallace K.M., Becka S.A., Zeiser E.T., Ohuchi N., Mojica M.F., Gatta J.A., Falleni M., Tosi D., Borghi E., Winkler M.L., Wilson B.M., LiPuma J.J., Nukaga M, & Bonomo R.A. (2017). Overcoming an Extremely Drug Resistant (XDR) Pathogen: Avibactam Restores Susceptibility to Ceftazidime for Burkholderia cepacia Complex Isolates from Cystic Fibrosis Patients. ACS infectious diseases, 3(7), 502-511.

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Bocillin FL Labeling of PBPs

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Avibactam was provided by AstraZeneca Pharmaceuticals (Wilmington, NC, USA). Ceftazidime was obtained from Lab Express International (Fairfield, NJ, USA). Oxacillin and amdinocillin were from Sigma-Aldrich Canada (Oakville, ON, Canada). PBPs were labeled using the fluorescent reporter molecule Bocillin FL (Invitrogen-Molecular Probes, Eugene, OR, USA).

Asli A., Brouillette E., Krause K.M., Nichols W.W, & Malouin F. (2016). Distinctive Binding of Avibactam to Penicillin-Binding Proteins of Gram-Negative and Gram-Positive Bacteria. Antimicrobial Agents and Chemotherapy, 60(2), 752-756.

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