Penems were synthesized as described in the Supporting Information Imipenem and meropenem were purchased from Carbosynth (San Diego, CA). Clavulanate was purchased from Sigma-Aldrich (St. Louis, MO).
Meropenem
Manufactured by Biosynth
Sourced in United Kingdom
Meropenem is a broad-spectrum antibiotic that belongs to the carbapenem class of antibiotics. It is used to treat a variety of bacterial infections. Meropenem works by inhibiting the synthesis of the bacterial cell wall, leading to cell death.
Automatically generated - may contain errors
Lab products found in correlation
Imipenem, by Biosynth (3 mentions)
Tigecycline, by Biosynth (2 mentions)
Amikacin, by Biosynth (2 mentions)
Clavulanate, by Merck Group (1 mentions)
Ciprofloxacin, by Biosynth (1 mentions)
Kanamycin, by Merck Group (1 mentions)
Bbl mueller hinton 2 broth, by BD (1 mentions)
Mueller hinton agar (mha), by Thermo Fisher Scientific (1 mentions)
Avibactam, by Biosynth (1 mentions)
Amoxicillin, by Merck Group (1 mentions)
Ceftazidime, by Merck Group (1 mentions)
Tazobactam, by Merck Group (1 mentions)
6 protocols using meropenem
1
Synthesis of Antimicrobial Compounds
2
Carbapenem-Resistant A. baumannii Identification
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Antimicrobial susceptibility testing for CR-AB identification was performed using broth microdilution methods according to the Clinical Laboratory Standard Institute (CLSI) guidelines [21 ]. A. baumannii isolates were grown in cation-adjusted Mueller-Hinton broth (CAMHB, Difco, BD Diagnostic Systems, USA) before adding them into 96-well plates containing antibiotics to the final concentration (approximately 105 CFU/ml). A. baumannii isolates were tested against carbapenems, including imipenem, meropenem, and doripenem (Carbosynth, UK). The microtiter plate was incubated for 16–20 h at 35±2°C. The results were evaluated based on the MIC values from the minimum concentration of drugs that showed no visible growth. Clinical and Laboratory Standards Institute (CLSI) breakpoints were used for susceptibility determination. In this study, CR-AB was defined as an isolate resistant to at least one carbapenem agent. A. baumannii ATCC 19606, obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA) was used as a quality control isolate.
3
Antibacterial Activity Evaluation Protocol
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NOSO-502 was synthesized at Nosopharm, Nîmes, France (Biosynth, ref: AC20542), ciprofloxacin (Biosynth, ref: AC58172), meropenem (Biosynth, ref: AM32026), ceftazidime-avibactam (CAZ: Biosynth, ref: AC19871; AVI: Biosynth, ref: AA158833), tigecycline (Biosynth, ref: AT10818), and amikacin (CAZ: Biosynth, ref: AA17356) were obtained from manufacturers as standard powders. The BBL Mueller-Hinton II Broth (Becton, Dickinson, ref: 212322), alone or with 1.4% wt/wt agarose (Grosseron ref A8963) and the Mueller-Hinton Agar (Oxoid, ref CM0337) were used in all experiments. When necessary, Mueller-Hinton was supplemented with kanamycin at 40 μg/mL (Sigma-Aldrich, ref: K1637).
4
Antimicrobial Compound Preparation Protocols
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The antimicrobials ceftazidime (CAZ) and amoxicillin (AMX) were sourced from Sigma Aldrich (Dorset, UK). Imipenem (IMI), meropenem (MER), avibactam (AVI) and relebactam (REL) were obtained from Carbosynth (Compton, UK). Stock solutions of all compounds were prepared in sterile dH2O, apart from AMX where DMSO was used, and stored at −20 °C, where appropriate.
5
Antibacterial Drug Susceptibility Testing
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Minimal inhibitory concentration (MIC) values of the commercially available antibacterial drugs ceftriaxone, cefepime, meropenem, levofloxacin, amikacin, and tigecycline and DFO for 55 Gram-negative isolates included in our study were determined with broth microdilution methods according to EUCAST standards (ISO 20776-1:2019). Stock solutions of ceftriaxone, cefepime, meropenem, levofloxacin, amikacin, and tigecycline (Carbosynth, Campton, UK) were prepared in accordance with the manufacturer's instructions. Water was utilized as a solvent for ceftriaxone, meropenem, levofloxacin, amikacin, and tigecycline. The solvent for cefepime was phosphate buffer (pH 6.0, 0.1 mol/L). Twofold concentrations of antibiotics and DFO ranging between 0.06 µg/mL and 64 µg/mL (from 0.06 µg/mL to 512 µg/mL for DFO) were added to microplate wells filled with cation-adjusted Mueller-Hinton broth (CAMHB). Bacterial suspension at the concentration of 5×10 CFU/mL was inoculated to the microplate wells. Inoculated microplates were incubated at 35 ±2°C for 24 h. The MIC was defined as the lowest antimicrobial drug concentration that inhibits the visible growth of the microorganism in the microdilution wells.
6
Antibiotics Susceptibility Testing Protocol
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Bacterial isolates were subjected to broth microdilution susceptibility testing, performed according to Clinical and Laboratory Standards Institute (CLSI) methods (32 ), using panels prepared in-house. MIC data for the representative panel were based on single tests; data for the challenge panel is the mode of a minimum of three experiments for the majority of BL/BLI combinations (and up to five, for some combinations). Meropenem was purchased from Carbosynth (San Diego, CA), and ceftolozane was synthesized at ACME Bioscience (Exton, PA); all other antibiotics and tazobactam were from Sigma-Aldrich. All beta-lactamase inhibitors were synthesized at Qpex Biopharma, San Diego, CA.
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