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Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae.
Antimicrob Agents Chemother. 2018 06; 62(6)AA

Abstract

The imipenem-relebactam combination is in development as a potential treatment regimen for infections caused by Enterobacteriaceae possessing complex β-lactamase backgrounds. Relebactam is a β-lactamase inhibitor that possesses the diazabicyclooctane core, as in avibactam; however, the R1 side chain of relebactam also includes a piperidine ring, whereas that of avibactam is a carboxyamide. Here, we investigated the inactivation of the Klebsiella pneumoniae carbapenemase KPC-2, the most widespread class A carbapenemase, by relebactam and performed susceptibility testing with imipenem-relebactam using KPC-producing clinical isolates of Enterobacteriaceae MIC measurements using agar dilution methods revealed that all 101 clinical isolates of KPC-producing Enterobacteriaceae (K. pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, and Escherichia coli) were highly susceptible to imipenem-relebactam (MICs ≤ 2 mg/liter). Relebactam inhibited KPC-2 with a second-order onset of acylation rate constant (k2/K) value of 24,750 M-1 s-1 and demonstrated a slow off-rate constant (koff) of 0.0002 s-1 Biochemical analysis using time-based mass spectrometry to map intermediates revealed that the KPC-2-relebactam acyl-enzyme complex was stable for up to 24 h. Importantly, desulfation of relebactam was not observed using mass spectrometry. Desulfation and subsequent deacylation have been observed during the reaction of KPC-2 with avibactam. Upon molecular dynamics simulations of relebactam in the KPC-2 active site, we found that the positioning of active-site water molecules is less favorable for desulfation in the KPC-2 active site than it is in the KPC-2-avibactam complex. In the acyl complexes, the water molecules are within 2.5 to 3 Å of the avibactam sulfate; however, they are more than 5 to 6 Å from the relebactam sulfate. As a result, we propose that the KPC-2-relebactam acyl complex is more stable than the KPC-2-avibactam complex. The clinical implications of this difference are not currently known.

Authors+Show Affiliations

Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA krisztina.papp@va.gov robert.bonomo@va.gov. Department of Medicine, Case Western Reserve University, Cleveland, Ohio, USA. Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio, USA.Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA. Department of Medicine, Case Western Reserve University, Cleveland, Ohio, USA.University of Minnesota, Minneapolis, Minnesota, USA.Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA. Department of Medicine, Case Western Reserve University, Cleveland, Ohio, USA.Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA.Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA.Department of Medicine, University of North Carolina School of Medicine, Chapel Hill, North Carolina, USA.Public Health Research Institute Tuberculosis Center, New Jersey Medical School, Rutgers University, Newark, New Jersey, USA.Division of Infectious Diseases, Department of Medicine, University of Michigan, Ann Arbor, Michigan, USA.Research Service, Louis Stokes Cleveland Department of Veterans Affairs, Cleveland, Ohio, USA krisztina.papp@va.gov robert.bonomo@va.gov. Department of Medicine, Case Western Reserve University, Cleveland, Ohio, USA. Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio, USA. Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, Ohio, USA. Department of Pharmacology, Case Western Reserve University, Cleveland, Ohio, USA.

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

Language

eng

PubMed ID

29610205

Citation

Papp-Wallace, Krisztina M., et al. "Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae." Antimicrobial Agents and Chemotherapy, vol. 62, no. 6, 2018.
Papp-Wallace KM, Barnes MD, Alsop J, et al. Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae. Antimicrob Agents Chemother. 2018;62(6).
Papp-Wallace, K. M., Barnes, M. D., Alsop, J., Taracila, M. A., Bethel, C. R., Becka, S. A., van Duin, D., Kreiswirth, B. N., Kaye, K. S., & Bonomo, R. A. (2018). Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae. Antimicrobial Agents and Chemotherapy, 62(6). https://doi.org/10.1128/AAC.00174-18
Papp-Wallace KM, et al. Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae. Antimicrob Agents Chemother. 2018;62(6) PubMed PMID: 29610205.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Relebactam Is a Potent Inhibitor of the KPC-2 β-Lactamase and Restores Imipenem Susceptibility in KPC-Producing Enterobacteriaceae. AU - Papp-Wallace,Krisztina M, AU - Barnes,Melissa D, AU - Alsop,Jim, AU - Taracila,Magdalena A, AU - Bethel,Christopher R, AU - Becka,Scott A, AU - van Duin,David, AU - Kreiswirth,Barry N, AU - Kaye,Keith S, AU - Bonomo,Robert A, Y1 - 2018/05/25/ PY - 2018/01/26/received PY - 2018/03/21/accepted PY - 2018/4/4/pubmed PY - 2019/8/20/medline PY - 2018/4/4/entrez KW - carbapenemase KW - relebactam KW - β-lactamase inhibitor KW - β-lactamases KW - β-lactams JF - Antimicrobial agents and chemotherapy JO - Antimicrob Agents Chemother VL - 62 IS - 6 N2 - The imipenem-relebactam combination is in development as a potential treatment regimen for infections caused by Enterobacteriaceae possessing complex β-lactamase backgrounds. Relebactam is a β-lactamase inhibitor that possesses the diazabicyclooctane core, as in avibactam; however, the R1 side chain of relebactam also includes a piperidine ring, whereas that of avibactam is a carboxyamide. Here, we investigated the inactivation of the Klebsiella pneumoniae carbapenemase KPC-2, the most widespread class A carbapenemase, by relebactam and performed susceptibility testing with imipenem-relebactam using KPC-producing clinical isolates of Enterobacteriaceae MIC measurements using agar dilution methods revealed that all 101 clinical isolates of KPC-producing Enterobacteriaceae (K. pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Citrobacter freundii, Citrobacter koseri, and Escherichia coli) were highly susceptible to imipenem-relebactam (MICs ≤ 2 mg/liter). Relebactam inhibited KPC-2 with a second-order onset of acylation rate constant (k2/K) value of 24,750 M-1 s-1 and demonstrated a slow off-rate constant (koff) of 0.0002 s-1 Biochemical analysis using time-based mass spectrometry to map intermediates revealed that the KPC-2-relebactam acyl-enzyme complex was stable for up to 24 h. Importantly, desulfation of relebactam was not observed using mass spectrometry. Desulfation and subsequent deacylation have been observed during the reaction of KPC-2 with avibactam. Upon molecular dynamics simulations of relebactam in the KPC-2 active site, we found that the positioning of active-site water molecules is less favorable for desulfation in the KPC-2 active site than it is in the KPC-2-avibactam complex. In the acyl complexes, the water molecules are within 2.5 to 3 Å of the avibactam sulfate; however, they are more than 5 to 6 Å from the relebactam sulfate. As a result, we propose that the KPC-2-relebactam acyl complex is more stable than the KPC-2-avibactam complex. The clinical implications of this difference are not currently known. SN - 1098-6596 UR - https://www.unboundmedicine.com/medline/citation/29610205/Relebactam_Is_a_Potent_Inhibitor_of_the_KPC_2_β_Lactamase_and_Restores_Imipenem_Susceptibility_in_KPC_Producing_Enterobacteriaceae_ L2 - http://aac.asm.org/cgi/pmidlookup?view=long&pmid=29610205 DB - PRIME DP - Unbound Medicine ER -