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Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action

Mycobacterium abscessus (Mab) infections are a growing menace to the health of many patients, especially those suffering from structural lung disease and cystic fibrosis. With multidrug resistance a common feature and a growing understanding of peptidoglycan synthesis in Mab, it is advantageous to i...

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Autores principales: Dousa, Khalid M., Nguyen, David C., Kurz, Sebastian G., Taracila, Magdalena A., Bethel, Christopher R., Schinabeck, William, Kreiswirth, Barry N., Brown, Sheldon T., Boom, W. Henry, Hotchkiss, Richard S., Remy, Kenneth E., Jacono, Frank J., Daley, Charles L., Holland, Steven M., Miller, Alita A., Bonomo, Robert A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8787486/
https://www.ncbi.nlm.nih.gov/pubmed/35073757
http://dx.doi.org/10.1128/mbio.03529-21
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author Dousa, Khalid M.
Nguyen, David C.
Kurz, Sebastian G.
Taracila, Magdalena A.
Bethel, Christopher R.
Schinabeck, William
Kreiswirth, Barry N.
Brown, Sheldon T.
Boom, W. Henry
Hotchkiss, Richard S.
Remy, Kenneth E.
Jacono, Frank J.
Daley, Charles L.
Holland, Steven M.
Miller, Alita A.
Bonomo, Robert A.
author_facet Dousa, Khalid M.
Nguyen, David C.
Kurz, Sebastian G.
Taracila, Magdalena A.
Bethel, Christopher R.
Schinabeck, William
Kreiswirth, Barry N.
Brown, Sheldon T.
Boom, W. Henry
Hotchkiss, Richard S.
Remy, Kenneth E.
Jacono, Frank J.
Daley, Charles L.
Holland, Steven M.
Miller, Alita A.
Bonomo, Robert A.
author_sort Dousa, Khalid M.
collection PubMed
description Mycobacterium abscessus (Mab) infections are a growing menace to the health of many patients, especially those suffering from structural lung disease and cystic fibrosis. With multidrug resistance a common feature and a growing understanding of peptidoglycan synthesis in Mab, it is advantageous to identify potent β-lactam and β-lactamase inhibitor combinations that can effectively disrupt cell wall synthesis. To improve existing therapeutic regimens to address serious Mab infections, we evaluated the ability of durlobactam (DUR), a novel diazobicyclooctane β-lactamase inhibitor to restore in vitro susceptibilities in combination with β-lactams and provide a biochemical rationale for the activity of this compound. In cell-based assays, susceptibility of Mab subsp. abscessus isolates to amoxicillin (AMOX), imipenem (IMI), and cefuroxime (CXM) was significantly enhanced with the addition of DUR. The triple drug combinations of CXM-DUR-AMOX and IMI-DUR-AMOX were most potent, with MIC ranges of ≤0.06 to 1 μg/mL and an MIC(50)/MIC(90) of ≤0.06/0.25 μg/mL, respectively. We propose a model by which this enhancement may occur, DUR potently inhibited the β-lactamase Bla(Mab) with a relative Michaelis constant (K(i) (app)) of 4 × 10(−3) ± 0.8 × 10(−3 )μM and acylation rate (k(2)/K) of 1 × 10(7) M(−1) s(−1). Timed mass spectrometry captured stable formation of carbamoyl-enzyme complexes between DUR and Ldt(Mab2-4) and Mab d,d-carboxypeptidase, potentially contributing to the intrinsic activity of DUR. Molecular modeling showed unique and favorable interactions of DUR as a Bla(Mab) inhibitor. Similarly, modeling showed how DUR might form stable Michaelis-Menten complexes with Ldt(Mab2-4) and Mab d,d-carboxypeptidase. The ability of DUR combined with amoxicillin or cefuroxime and imipenem to inactivate multiple targets such as d,d-carboxypeptidase and Ldt(Mab2,4) supports new therapeutic approaches using β-lactams in eradicating Mab.
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spelling pubmed-87874862022-02-07 Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action Dousa, Khalid M. Nguyen, David C. Kurz, Sebastian G. Taracila, Magdalena A. Bethel, Christopher R. Schinabeck, William Kreiswirth, Barry N. Brown, Sheldon T. Boom, W. Henry Hotchkiss, Richard S. Remy, Kenneth E. Jacono, Frank J. Daley, Charles L. Holland, Steven M. Miller, Alita A. Bonomo, Robert A. mBio Research Article Mycobacterium abscessus (Mab) infections are a growing menace to the health of many patients, especially those suffering from structural lung disease and cystic fibrosis. With multidrug resistance a common feature and a growing understanding of peptidoglycan synthesis in Mab, it is advantageous to identify potent β-lactam and β-lactamase inhibitor combinations that can effectively disrupt cell wall synthesis. To improve existing therapeutic regimens to address serious Mab infections, we evaluated the ability of durlobactam (DUR), a novel diazobicyclooctane β-lactamase inhibitor to restore in vitro susceptibilities in combination with β-lactams and provide a biochemical rationale for the activity of this compound. In cell-based assays, susceptibility of Mab subsp. abscessus isolates to amoxicillin (AMOX), imipenem (IMI), and cefuroxime (CXM) was significantly enhanced with the addition of DUR. The triple drug combinations of CXM-DUR-AMOX and IMI-DUR-AMOX were most potent, with MIC ranges of ≤0.06 to 1 μg/mL and an MIC(50)/MIC(90) of ≤0.06/0.25 μg/mL, respectively. We propose a model by which this enhancement may occur, DUR potently inhibited the β-lactamase Bla(Mab) with a relative Michaelis constant (K(i) (app)) of 4 × 10(−3) ± 0.8 × 10(−3 )μM and acylation rate (k(2)/K) of 1 × 10(7) M(−1) s(−1). Timed mass spectrometry captured stable formation of carbamoyl-enzyme complexes between DUR and Ldt(Mab2-4) and Mab d,d-carboxypeptidase, potentially contributing to the intrinsic activity of DUR. Molecular modeling showed unique and favorable interactions of DUR as a Bla(Mab) inhibitor. Similarly, modeling showed how DUR might form stable Michaelis-Menten complexes with Ldt(Mab2-4) and Mab d,d-carboxypeptidase. The ability of DUR combined with amoxicillin or cefuroxime and imipenem to inactivate multiple targets such as d,d-carboxypeptidase and Ldt(Mab2,4) supports new therapeutic approaches using β-lactams in eradicating Mab. American Society for Microbiology 2022-01-25 /pmc/articles/PMC8787486/ /pubmed/35073757 http://dx.doi.org/10.1128/mbio.03529-21 Text en https://doi.org/10.1128/AuthorWarrantyLicense.v1This is a work of the U.S. Government and is not subject to copyright protection in the United States. Foreign copyrights may apply.
spellingShingle Research Article
Dousa, Khalid M.
Nguyen, David C.
Kurz, Sebastian G.
Taracila, Magdalena A.
Bethel, Christopher R.
Schinabeck, William
Kreiswirth, Barry N.
Brown, Sheldon T.
Boom, W. Henry
Hotchkiss, Richard S.
Remy, Kenneth E.
Jacono, Frank J.
Daley, Charles L.
Holland, Steven M.
Miller, Alita A.
Bonomo, Robert A.
Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title_full Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title_fullStr Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title_full_unstemmed Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title_short Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
title_sort inhibiting mycobacterium abscessus cell wall synthesis: using a novel diazabicyclooctane β-lactamase inhibitor to augment β-lactam action
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8787486/
https://www.ncbi.nlm.nih.gov/pubmed/35073757
http://dx.doi.org/10.1128/mbio.03529-21
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