Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action
ABSTRACT 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 advantag...
| Main Authors: | , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2022-02-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.03529-21 |
| _version_ | 1818872270362247168 |
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| author | Khalid M. Dousa David C. Nguyen Sebastian G. Kurz Magdalena A. Taracila Christopher R. Bethel William Schinabeck Barry N. Kreiswirth Sheldon T. Brown W. Henry Boom Richard S. Hotchkiss Kenneth E. Remy Frank J. Jacono Charles L. Daley Steven M. Holland Alita A. Miller Robert A. Bonomo |
| author_facet | Khalid M. Dousa David C. Nguyen Sebastian G. Kurz Magdalena A. Taracila Christopher R. Bethel William Schinabeck Barry N. Kreiswirth Sheldon T. Brown W. Henry Boom Richard S. Hotchkiss Kenneth E. Remy Frank J. Jacono Charles L. Daley Steven M. Holland Alita A. Miller Robert A. Bonomo |
| author_sort | Khalid M. Dousa |
| collection | DOAJ |
| description | ABSTRACT 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 MIC50/MIC90 of ≤0.06/0.25 μg/mL, respectively. We propose a model by which this enhancement may occur, DUR potently inhibited the β-lactamase BlaMab with a relative Michaelis constant (Ki app) of 4 × 10−3 ± 0.8 × 10−3 μM and acylation rate (k2/K) of 1 × 107 M−1 s−1. Timed mass spectrometry captured stable formation of carbamoyl-enzyme complexes between DUR and LdtMab2-4 and Mab d,d-carboxypeptidase, potentially contributing to the intrinsic activity of DUR. Molecular modeling showed unique and favorable interactions of DUR as a BlaMab inhibitor. Similarly, modeling showed how DUR might form stable Michaelis-Menten complexes with LdtMab2-4 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 LdtMab2,4 supports new therapeutic approaches using β-lactams in eradicating Mab. IMPORTANCE Durlobactam (DUR) is a potent inhibitor of BlaMab and provides protection of amoxicillin and imipenem against hydrolysis. DUR has intrinsic activity and forms stable acyl-enzyme complexes with LdtMab2 and LdtMab4. The ability of DUR to protect amoxicillin and imipenem against BlaMab and its intrinsic activity along with the dual β-lactam target redundancy can explain the rationale behind the potent activity of this combination. |
| first_indexed | 2024-12-19T12:36:09Z |
| format | Article |
| id | doaj.art-89354fdd37e14eba94df814fcd3ea960 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-19T12:36:09Z |
| publishDate | 2022-02-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-89354fdd37e14eba94df814fcd3ea9602022-12-21T20:21:09ZengAmerican Society for MicrobiologymBio2150-75112022-02-0113110.1128/mbio.03529-21Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam ActionKhalid M. Dousa0David C. Nguyen1Sebastian G. Kurz2Magdalena A. Taracila3Christopher R. Bethel4William Schinabeck5Barry N. Kreiswirth6Sheldon T. Brown7W. Henry Boom8Richard S. Hotchkiss9Kenneth E. Remy10Frank J. Jacono11Charles L. Daley12Steven M. Holland13Alita A. Miller14Robert A. Bonomo15Louis Stokes Cleveland VA Medical Center, Case Western Reserve University, Cleveland, Ohio, USADivision of Infectious Diseases and HIV Medicine, University Hospitals Cleveland Medical Center, Cleveland, Ohio, USAMount Sinai National Jewish Health Respiratory Institute, New York City, New York, USAResearch Service, Louis Stokes Veterans Affairs Medical Center, Cleveland, Ohio, USAResearch Service, Louis Stokes Veterans Affairs Medical Center, Cleveland, Ohio, USABates College, Lewiston, Maine, USACenter for Discovery and Innovation, Hackensack Meridian Health, Nutley, New Jersey, USADepartment of Medicine, Icahn School of Medicine at Mount Sinai, New York, New York, USADivision of Infectious Diseases and HIV Medicine, University Hospitals Cleveland Medical Center, Cleveland, Ohio, USADepartment of Anesthesiology, Washington University School of Medicine, St. Louis, Missouri, USADivision of Pediatric Critical Care, UH Rainbow Babies & Children’s Hospital, Cleveland, Ohio, USALouis Stokes Cleveland VA Medical Center, Case Western Reserve University, Cleveland, Ohio, USADivision of Mycobacterial and Respiratory Infections, National Jewish Health, Denver, Colorado, USALaboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USAEntasis Therapeutics, Waltham, Massachusetts, USALouis Stokes Cleveland VA Medical Center, Case Western Reserve University, Cleveland, Ohio, USAABSTRACT 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 MIC50/MIC90 of ≤0.06/0.25 μg/mL, respectively. We propose a model by which this enhancement may occur, DUR potently inhibited the β-lactamase BlaMab with a relative Michaelis constant (Ki app) of 4 × 10−3 ± 0.8 × 10−3 μM and acylation rate (k2/K) of 1 × 107 M−1 s−1. Timed mass spectrometry captured stable formation of carbamoyl-enzyme complexes between DUR and LdtMab2-4 and Mab d,d-carboxypeptidase, potentially contributing to the intrinsic activity of DUR. Molecular modeling showed unique and favorable interactions of DUR as a BlaMab inhibitor. Similarly, modeling showed how DUR might form stable Michaelis-Menten complexes with LdtMab2-4 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 LdtMab2,4 supports new therapeutic approaches using β-lactams in eradicating Mab. IMPORTANCE Durlobactam (DUR) is a potent inhibitor of BlaMab and provides protection of amoxicillin and imipenem against hydrolysis. DUR has intrinsic activity and forms stable acyl-enzyme complexes with LdtMab2 and LdtMab4. The ability of DUR to protect amoxicillin and imipenem against BlaMab and its intrinsic activity along with the dual β-lactam target redundancy can explain the rationale behind the potent activity of this combination.https://journals.asm.org/doi/10.1128/mbio.03529-21antibiotic resistancebacteriainhibitorantibioticsdurlobactamβ-lactams |
| spellingShingle | Khalid M. Dousa David C. Nguyen Sebastian G. Kurz Magdalena A. Taracila Christopher R. Bethel William Schinabeck Barry N. Kreiswirth Sheldon T. Brown W. Henry Boom Richard S. Hotchkiss Kenneth E. Remy Frank J. Jacono Charles L. Daley Steven M. Holland Alita A. Miller Robert A. Bonomo Inhibiting Mycobacterium abscessus Cell Wall Synthesis: Using a Novel Diazabicyclooctane β-Lactamase Inhibitor To Augment β-Lactam Action mBio antibiotic resistance bacteria inhibitor antibiotics durlobactam β-lactams |
| 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 | antibiotic resistance bacteria inhibitor antibiotics durlobactam β-lactams |
| url | https://journals.asm.org/doi/10.1128/mbio.03529-21 |
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