In silico identification and experimental validation of hits active against KPC-2 β-lactamase.
Bacterial resistance has become a worldwide concern, particularly after the emergence of resistant strains overproducing carbapenemases. Among these, the KPC-2 carbapenemase represents a significant clinical challenge, being characterized by a broad substrate spectrum that includes aminothiazoleoxim...
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doaj-52763062b4fa47ef8e491ae03bbcd7af2021-03-03T21:05:02ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011311e020324110.1371/journal.pone.0203241In silico identification and experimental validation of hits active against KPC-2 β-lactamase.Raphael KleinPasquale LincianoGiuseppe CelenzaPierangelo BellioSofia PapaioannouJesus BlazquezLaura CendronRuth BrenkDonatella TondiBacterial resistance has become a worldwide concern, particularly after the emergence of resistant strains overproducing carbapenemases. Among these, the KPC-2 carbapenemase represents a significant clinical challenge, being characterized by a broad substrate spectrum that includes aminothiazoleoxime and cephalosporins such as cefotaxime. Moreover, strains harboring KPC-type β-lactamases are often reported as resistant to available β-lactamase inhibitors (clavulanic acid, tazobactam and sulbactam). Therefore, the identification of novel non β-lactam KPC-2 inhibitors is strongly necessary to maintain treatment options. This study explored novel, non-covalent inhibitors active against KPC-2, as putative hit candidates. We performed a structure-based in silico screening of commercially available compounds for non-β-lactam KPC-2 inhibitors. Thirty-two commercially available high-scoring, fragment-like hits were selected for in vitro validation and their activity and mechanism of action vs the target was experimentally evaluated using recombinant KPC-2. N-(3-(1H-tetrazol-5-yl)phenyl)-3-fluorobenzamide (11a), in light of its ligand efficiency (LE = 0.28 kcal/mol/non-hydrogen atom) and chemistry, was selected as hit to be directed to chemical optimization to improve potency vs the enzyme and explore structural requirement for inhibition in KPC-2 binding site. Further, the compounds were evaluated against clinical strains overexpressing KPC-2 and the most promising compound reduced the MIC of the β-lactam antibiotic meropenem by four-fold.https://doi.org/10.1371/journal.pone.0203241 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Raphael Klein Pasquale Linciano Giuseppe Celenza Pierangelo Bellio Sofia Papaioannou Jesus Blazquez Laura Cendron Ruth Brenk Donatella Tondi |
spellingShingle |
Raphael Klein Pasquale Linciano Giuseppe Celenza Pierangelo Bellio Sofia Papaioannou Jesus Blazquez Laura Cendron Ruth Brenk Donatella Tondi In silico identification and experimental validation of hits active against KPC-2 β-lactamase. PLoS ONE |
author_facet |
Raphael Klein Pasquale Linciano Giuseppe Celenza Pierangelo Bellio Sofia Papaioannou Jesus Blazquez Laura Cendron Ruth Brenk Donatella Tondi |
author_sort |
Raphael Klein |
title |
In silico identification and experimental validation of hits active against KPC-2 β-lactamase. |
title_short |
In silico identification and experimental validation of hits active against KPC-2 β-lactamase. |
title_full |
In silico identification and experimental validation of hits active against KPC-2 β-lactamase. |
title_fullStr |
In silico identification and experimental validation of hits active against KPC-2 β-lactamase. |
title_full_unstemmed |
In silico identification and experimental validation of hits active against KPC-2 β-lactamase. |
title_sort |
in silico identification and experimental validation of hits active against kpc-2 β-lactamase. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2018-01-01 |
description |
Bacterial resistance has become a worldwide concern, particularly after the emergence of resistant strains overproducing carbapenemases. Among these, the KPC-2 carbapenemase represents a significant clinical challenge, being characterized by a broad substrate spectrum that includes aminothiazoleoxime and cephalosporins such as cefotaxime. Moreover, strains harboring KPC-type β-lactamases are often reported as resistant to available β-lactamase inhibitors (clavulanic acid, tazobactam and sulbactam). Therefore, the identification of novel non β-lactam KPC-2 inhibitors is strongly necessary to maintain treatment options. This study explored novel, non-covalent inhibitors active against KPC-2, as putative hit candidates. We performed a structure-based in silico screening of commercially available compounds for non-β-lactam KPC-2 inhibitors. Thirty-two commercially available high-scoring, fragment-like hits were selected for in vitro validation and their activity and mechanism of action vs the target was experimentally evaluated using recombinant KPC-2. N-(3-(1H-tetrazol-5-yl)phenyl)-3-fluorobenzamide (11a), in light of its ligand efficiency (LE = 0.28 kcal/mol/non-hydrogen atom) and chemistry, was selected as hit to be directed to chemical optimization to improve potency vs the enzyme and explore structural requirement for inhibition in KPC-2 binding site. Further, the compounds were evaluated against clinical strains overexpressing KPC-2 and the most promising compound reduced the MIC of the β-lactam antibiotic meropenem by four-fold. |
url |
https://doi.org/10.1371/journal.pone.0203241 |
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