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An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation
Bacterial resistance represents a major health problem worldwide and there is an urgent need to develop first-in-class compounds directed against new therapeutic targets. We previously developed a drug-discovery platform to identify new antimicrobials able to disrupt the protein–protein interaction...
Autores principales: | , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598883/ https://www.ncbi.nlm.nih.gov/pubmed/36290107 http://dx.doi.org/10.3390/antibiotics11101449 |
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author | Caputo, Alessia Sartini, Sara Levati, Elisabetta Minato, Ilaria Elisi, Gian Marco Di Stasi, Adriana Guillou, Catherine Goekjian, Peter G. Garcia, Pierre Gueyrard, David Bach, Stéphane Comte, Arnaud Ottonello, Simone Rivara, Silvia Montanini, Barbara |
author_facet | Caputo, Alessia Sartini, Sara Levati, Elisabetta Minato, Ilaria Elisi, Gian Marco Di Stasi, Adriana Guillou, Catherine Goekjian, Peter G. Garcia, Pierre Gueyrard, David Bach, Stéphane Comte, Arnaud Ottonello, Simone Rivara, Silvia Montanini, Barbara |
author_sort | Caputo, Alessia |
collection | PubMed |
description | Bacterial resistance represents a major health problem worldwide and there is an urgent need to develop first-in-class compounds directed against new therapeutic targets. We previously developed a drug-discovery platform to identify new antimicrobials able to disrupt the protein–protein interaction between the β’ subunit and the σ(70) initiation factor of bacterial RNA polymerase, which is essential for transcription. As a follow-up to such work, we have improved the discovery strategy to make it less time-consuming and more cost-effective. This involves three sequential assays, easily scalable to a high-throughput format, and a subsequent in-depth characterization only limited to hits that passed the three tests. This optimized workflow, applied to the screening of 5360 small molecules from three synthetic and natural compound libraries, led to the identification of six compounds interfering with the β’–σ(70) interaction, and thus was capable of inhibiting promoter-specific RNA transcription and bacterial growth. Upon supplementation with a permeability adjuvant, the two most potent transcription-inhibiting compounds displayed a strong antibacterial activity against Escherichia coli with minimum inhibitory concentration (MIC) values among the lowest (0.87–1.56 μM) thus far reported for β’–σ PPI inhibitors. The newly identified hit compounds share structural feature similarities with those of a pharmacophore model previously developed from known inhibitors. |
format | Online Article Text |
id | pubmed-9598883 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-95988832022-10-27 An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation Caputo, Alessia Sartini, Sara Levati, Elisabetta Minato, Ilaria Elisi, Gian Marco Di Stasi, Adriana Guillou, Catherine Goekjian, Peter G. Garcia, Pierre Gueyrard, David Bach, Stéphane Comte, Arnaud Ottonello, Simone Rivara, Silvia Montanini, Barbara Antibiotics (Basel) Article Bacterial resistance represents a major health problem worldwide and there is an urgent need to develop first-in-class compounds directed against new therapeutic targets. We previously developed a drug-discovery platform to identify new antimicrobials able to disrupt the protein–protein interaction between the β’ subunit and the σ(70) initiation factor of bacterial RNA polymerase, which is essential for transcription. As a follow-up to such work, we have improved the discovery strategy to make it less time-consuming and more cost-effective. This involves three sequential assays, easily scalable to a high-throughput format, and a subsequent in-depth characterization only limited to hits that passed the three tests. This optimized workflow, applied to the screening of 5360 small molecules from three synthetic and natural compound libraries, led to the identification of six compounds interfering with the β’–σ(70) interaction, and thus was capable of inhibiting promoter-specific RNA transcription and bacterial growth. Upon supplementation with a permeability adjuvant, the two most potent transcription-inhibiting compounds displayed a strong antibacterial activity against Escherichia coli with minimum inhibitory concentration (MIC) values among the lowest (0.87–1.56 μM) thus far reported for β’–σ PPI inhibitors. The newly identified hit compounds share structural feature similarities with those of a pharmacophore model previously developed from known inhibitors. MDPI 2022-10-21 /pmc/articles/PMC9598883/ /pubmed/36290107 http://dx.doi.org/10.3390/antibiotics11101449 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Caputo, Alessia Sartini, Sara Levati, Elisabetta Minato, Ilaria Elisi, Gian Marco Di Stasi, Adriana Guillou, Catherine Goekjian, Peter G. Garcia, Pierre Gueyrard, David Bach, Stéphane Comte, Arnaud Ottonello, Simone Rivara, Silvia Montanini, Barbara An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title | An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title_full | An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title_fullStr | An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title_full_unstemmed | An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title_short | An Optimized Workflow for the Discovery of New Antimicrobial Compounds Targeting Bacterial RNA Polymerase Complex Formation |
title_sort | optimized workflow for the discovery of new antimicrobial compounds targeting bacterial rna polymerase complex formation |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9598883/ https://www.ncbi.nlm.nih.gov/pubmed/36290107 http://dx.doi.org/10.3390/antibiotics11101449 |
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