Azelaic Acid Exhibits Dual Antimicrobial and Quorum Sensing Inhibitory Activities Against Pathogens: In Vitro Evaluation and Molecular Docking Insights

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This in vitro study evaluated azelaic acid’s antimicrobial activity and its ability to inhibit bacterial quorum sensing (QS)-regulated virulence in reference strains and clinical isolates of Pseudomonas aeruginosa, Enterobacteriaceae, and Staphylococcus aureus, using MIC testing plus assays for QS-linked factors such as pyocyanin, elastase, alginate, and protease. Azelaic acid showed MICs of 250–1000 µg/mL, and at subinhibitory concentrations it produced dose-dependent inhibition of virulence factor production across reference and multidrug- and pan-drug-resistant clinical strains, with inhibition exceeding 90% for protease activity in several cases (p < 0.001). Molecular docking suggested azelaic acid interacts with QS-related proteins LasI, LasR, PqsD, and PqsR with moderate binding affinities and stable binding within conserved ligand-binding domains, supporting a multitarget QS-modulatory mechanism. The paper’s main limitation is that the evidence comes from in vitro assays and molecular docking rather than in vivo validation. The paper does not explicitly discuss endometriosis or adenomyosis; it was included in the corpus via a keyword match in the upstream search index.

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Abstract

The emergence of antimicrobial resistance (AMR) has driven the search for alternative therapeutic strategies, including antivirulence approaches targeting bacterial quorum sensing (QS). Azelaic acid (AzA), a naturally occurring dicarboxylic acid with known antimicrobial properties, has not previously been characterized as a QS inhibitor in Gram-negative pathogens. This study evaluated the dual antimicrobial and antivirulence activity of AzA against reference strains and clinical isolates of Pseudomonas aeruginosa, Enterobacteriaceae , and Staphylococcus aureus through in vitro assays and molecular docking analyses. Minimum inhibitory concentration (MIC) values ranged from 250 to 1000 µg/mL, with lower MICs observed in clinical isolates of E. coli and S. aureus . Subinhibitory concentrations (250, 500 and 750 µg/mL) were used to assess QS-regulated virulence factors in P. aeruginosa , including pyocyanin, elastase, alginate, and protease production. AzA exhibited a significant, dose-dependent inhibition of all evaluated virulence factors across both reference and multidrug-resistant (MDR) and pan-drug-resistant (PDR) clinical strains (p < 0.001), achieving inhibition levels exceeding 90% in several cases, particularly for protease activity. Molecular docking analyses revealed that AzA interacts with key QS-related proteins (LasI, LasR, PqsD, and PqsR), showing moderate binding affinities (−5.3 to −6.5 kcal/mol) and stable interactions within conserved ligand-binding domains. These findings suggest a multitarget modulatory mechanism affecting interconnected QS pathways. Overall, this study demonstrates, for the first time, that AzA acts as a quorum sensing inhibitor in P. aeruginosa , attenuating virulence without directly affecting bacterial growth, highlighting its potential as a promising antivirulence therapeutic strategy.
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Abstract The emergence of antimicrobial resistance (AMR) has driven the search for alternative therapeutic strategies, including antivirulence approaches targeting bacterial quorum sensing (QS). Azelaic acid (AzA), a naturally occurring dicarboxylic acid with known antimicrobial properties, has not previously been characterized as a QS inhibitor in Gram-negative pathogens. This study evaluated the dual antimicrobial and antivirulence activity of AzA against reference strains and clinical isolates of Pseudomonas aeruginosa, Enterobacteriaceae, and Staphylococcus aureus through in vitro assays and molecular docking analyses. Minimum inhibitory concentration (MIC) values ranged from 250 to 1000 µg/mL, with lower MICs observed in clinical isolates of E. coli and S. aureus. Subinhibitory concentrations (250, 500 and 750 µg/mL) were used to assess QS-regulated virulence factors in P. aeruginosa, including pyocyanin, elastase, alginate, and protease production. AzA exhibited a significant, dose-dependent inhibition of all evaluated virulence factors across both reference and multidrug-resistant (MDR) and pan-drug-resistant (PDR) clinical strains (p < 0.001), achieving inhibition levels exceeding 90% in several cases, particularly for protease activity. Molecular docking analyses revealed that AzA interacts with key QS-related proteins (LasI, LasR, PqsD, and PqsR), showing moderate binding affinities (−5.3 to −6.5 kcal/mol) and stable interactions within conserved ligand-binding domains. These findings suggest a multitarget modulatory mechanism affecting interconnected QS pathways. Overall, this study demonstrates, for the first time, that AzA acts as a quorum sensing inhibitor in P. aeruginosa, attenuating virulence without directly affecting bacterial growth, highlighting its potential as a promising antivirulence therapeutic strategy. Competing Interest Statement The authors have declared no competing interest.

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