Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections

<b>Background/Objectives</b>: <i>Mycobacterium abscessus</i> (MAB) is a highly resilient pathogen that causes difficult-to-treat pulmonary infections, particularly in individuals with cystic fibrosis (CF) and other underlying conditions. Its ability to form robust biofilms wi...

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Main Authors: Elizaveta Dzalamidze, Mylene Gorzynski, Rebecca Vande Voorde, Dylan Nelson, Lia Danelishvili
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Pharmaceuticals
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Online Access:https://www.mdpi.com/1424-8247/18/2/225
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author Elizaveta Dzalamidze
Mylene Gorzynski
Rebecca Vande Voorde
Dylan Nelson
Lia Danelishvili
author_facet Elizaveta Dzalamidze
Mylene Gorzynski
Rebecca Vande Voorde
Dylan Nelson
Lia Danelishvili
author_sort Elizaveta Dzalamidze
collection DOAJ
description <b>Background/Objectives</b>: <i>Mycobacterium abscessus</i> (MAB) is a highly resilient pathogen that causes difficult-to-treat pulmonary infections, particularly in individuals with cystic fibrosis (CF) and other underlying conditions. Its ability to form robust biofilms within the CF lung environment is a major factor contributing to its resistance to antibiotics and evasion of the host immune response, making conventional treatments largely ineffective. These biofilms, encased in an extracellular matrix, enhance drug tolerance and facilitate metabolic adaptations in hypoxic conditions, driving the bacteria into a persistent, non-replicative state that further exacerbates antimicrobial resistance. Treatment options remain limited, with multidrug regimens showing low success rates, highlighting the urgent need for more effective therapeutic strategies. <b>Methods</b>: In this study, we employed artificial sputum media to simulate the CF lung environment and conducted high-throughput screening of 24,000 compounds from diverse chemical libraries to identify inhibitors of MAB biofilm formation, using the Crystal Violet (CV) assay. <b>Results</b>: The screen established 17 hits with ≥30% biofilm inhibitory activity in mycobacteria. Six of these compounds inhibited MAB biofilm formation by over 60%, disrupted established biofilms by ≥40%, and significantly impaired bacterial viability within the biofilms, as confirmed by reduced CFU counts. In conformational assays, select compounds showed potent inhibitory activity in biofilms formed by clinical isolates of both MAB and <i>Mycobacterium avium</i> subsp. <i>hominissuis</i> (MAH). Key compounds, including ethacridine, phenothiazine, and fluorene derivatives, demonstrated potent activity against pre- and post-biofilm conditions, enhanced antibiotic efficacy, and reduced intracellular bacterial loads in macrophages. <b>Conclusions</b>: This study results underscore the potential of these compounds to target biofilm-associated resistance mechanisms, making them valuable candidates for use as adjuncts to existing therapies. These findings also emphasize the need for further investigations, including the initiation of a medicinal chemistry campaign to leverage structure–activity relationship studies and optimize the biological activity of these underexplored class of compounds against nontuberculous mycobacterial (NTM) strains.
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spelling doaj-art-8fb7b52f822c4a33a994087a2aca475f2025-08-20T02:45:01ZengMDPI AGPharmaceuticals1424-82472025-02-0118222510.3390/ph18020225Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> InfectionsElizaveta Dzalamidze0Mylene Gorzynski1Rebecca Vande Voorde2Dylan Nelson3Lia Danelishvili4Department of Biomedical Sciences, Carlson College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USADepartment of Biomedical Sciences, Carlson College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USADepartment of Biomedical Sciences, Carlson College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USADepartment of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, OR 97331, USADepartment of Biomedical Sciences, Carlson College of Veterinary Medicine, Oregon State University, Corvallis, OR 97331, USA<b>Background/Objectives</b>: <i>Mycobacterium abscessus</i> (MAB) is a highly resilient pathogen that causes difficult-to-treat pulmonary infections, particularly in individuals with cystic fibrosis (CF) and other underlying conditions. Its ability to form robust biofilms within the CF lung environment is a major factor contributing to its resistance to antibiotics and evasion of the host immune response, making conventional treatments largely ineffective. These biofilms, encased in an extracellular matrix, enhance drug tolerance and facilitate metabolic adaptations in hypoxic conditions, driving the bacteria into a persistent, non-replicative state that further exacerbates antimicrobial resistance. Treatment options remain limited, with multidrug regimens showing low success rates, highlighting the urgent need for more effective therapeutic strategies. <b>Methods</b>: In this study, we employed artificial sputum media to simulate the CF lung environment and conducted high-throughput screening of 24,000 compounds from diverse chemical libraries to identify inhibitors of MAB biofilm formation, using the Crystal Violet (CV) assay. <b>Results</b>: The screen established 17 hits with ≥30% biofilm inhibitory activity in mycobacteria. Six of these compounds inhibited MAB biofilm formation by over 60%, disrupted established biofilms by ≥40%, and significantly impaired bacterial viability within the biofilms, as confirmed by reduced CFU counts. In conformational assays, select compounds showed potent inhibitory activity in biofilms formed by clinical isolates of both MAB and <i>Mycobacterium avium</i> subsp. <i>hominissuis</i> (MAH). Key compounds, including ethacridine, phenothiazine, and fluorene derivatives, demonstrated potent activity against pre- and post-biofilm conditions, enhanced antibiotic efficacy, and reduced intracellular bacterial loads in macrophages. <b>Conclusions</b>: This study results underscore the potential of these compounds to target biofilm-associated resistance mechanisms, making them valuable candidates for use as adjuncts to existing therapies. These findings also emphasize the need for further investigations, including the initiation of a medicinal chemistry campaign to leverage structure–activity relationship studies and optimize the biological activity of these underexplored class of compounds against nontuberculous mycobacterial (NTM) strains.https://www.mdpi.com/1424-8247/18/2/225<i>Mycobacterium abscessus</i>nontuberculous mycobacteriaNTMhigh-throughput screeningsmall moleculesbiofilm
spellingShingle Elizaveta Dzalamidze
Mylene Gorzynski
Rebecca Vande Voorde
Dylan Nelson
Lia Danelishvili
Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
Pharmaceuticals
<i>Mycobacterium abscessus</i>
nontuberculous mycobacteria
NTM
high-throughput screening
small molecules
biofilm
title Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
title_full Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
title_fullStr Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
title_full_unstemmed Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
title_short Discovery of Biofilm-Inhibiting Compounds to Enhance Antibiotic Effectiveness Against <i>M. abscessus</i> Infections
title_sort discovery of biofilm inhibiting compounds to enhance antibiotic effectiveness against i m abscessus i infections
topic <i>Mycobacterium abscessus</i>
nontuberculous mycobacteria
NTM
high-throughput screening
small molecules
biofilm
url https://www.mdpi.com/1424-8247/18/2/225
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