Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI

Abstract Staphylococcus aureus infections have emerged as a global public health threat. Two key factors—drug resistance and biofilm formation—substantially impair the efficacy of the antimicrobial treatment for S. aureus infections using conventional antibiotics. Consequently, discovering novel ant...

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Main Authors: Yuanyuan Tang, Han Deng, Zhichao Xu, Zhijian Yu, Yong Xiang, Zewen Wen, Shiqing Han, Zhong Chen, Tieying Hou
Format: Article
Language:English
Published: BMC 2025-07-01
Series:BMC Microbiology
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Online Access:https://doi.org/10.1186/s12866-025-04104-2
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author Yuanyuan Tang
Han Deng
Zhichao Xu
Zhijian Yu
Yong Xiang
Zewen Wen
Shiqing Han
Zhong Chen
Tieying Hou
author_facet Yuanyuan Tang
Han Deng
Zhichao Xu
Zhijian Yu
Yong Xiang
Zewen Wen
Shiqing Han
Zhong Chen
Tieying Hou
author_sort Yuanyuan Tang
collection DOAJ
description Abstract Staphylococcus aureus infections have emerged as a global public health threat. Two key factors—drug resistance and biofilm formation—substantially impair the efficacy of the antimicrobial treatment for S. aureus infections using conventional antibiotics. Consequently, discovering novel antimicrobial agents with potent antibacterial and antibiofilm activity has become a hotspot in recent years. Herein, the research first reported the remarkable inhibitory activity of AZD-5991, a selective Mcl-1 inhibitor, against S. aureus. The MIC50 and MIC90 values of AZD-5991 against S. aureus were 12.5 µM, and significant growth inhibition was observed at a subinhibitory concentration of 1/2 × MIC. Additionally, AZD-5991 exhibited bactericidal activity and a robust capacity for inhibiting S. aureus biofilm formation, with minimal cytotoxicity toward host cell lines. Membrane permeability assays revealed that AZD-5991 compromised S. aureus cell membrane integrity, while bacterial phospholipid components were found to neutralize the antibacterial activity of AZD-5991. Moreover, whole-genome sequencing and proteomic analysis were also applied to gain insights into the possible impact of AZD-5991 on the fatty metabolism of S. aureus. Furthermore, the antibacterial activity of AZD-5991 was remarkably declined by exogenous fatty acids linoleic acid (C18:2Δ9,12) and arachidonic acid (C20:4Δ5,8,11,14). Lastly, the biolayer interferometry assay supported the direct interaction of AZD-5991 with FabI, a key protein essential for bacterial growth and fatty acid metabolism. Conclusively, this study demonstrates that AZD-5991 inhibits S. aureus planktonic growth and biofilm formation by disrupting cell membrane integrity and targeting FabI. These findings position AZD-5991 as a promising novel antibiotic candidate for treating S. aureus infections resistant to traditional clinical antibiotics. Graphical Abstract
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spelling doaj-art-c86d97c1a3464b0fa642e5b2c7c673672025-08-20T04:01:25ZengBMCBMC Microbiology1471-21802025-07-0125111510.1186/s12866-025-04104-2Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabIYuanyuan Tang0Han Deng1Zhichao Xu2Zhijian Yu3Yong Xiang4Zewen Wen5Shiqing Han6Zhong Chen7Tieying Hou8College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech UniversityCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech UniversityCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech UniversityGuangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Affiliated Nanshan Hospital of Shenzhen UniversityMarketing Department, Guangzhou Branch Varian Medical Equipment Trading (Beijing) Corp.Guangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Affiliated Nanshan Hospital of Shenzhen UniversityCollege of Biotechnology and Pharmaceutical Engineering, Nanjing Tech UniversityGuangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Affiliated Nanshan Hospital of Shenzhen UniversityGuangdong Key Laboratory for Biomedical Measurements and Ultrasound Imaging, National-Regional Key Technology Engineering Laboratory for Medical Ultrasound, School of Biomedical Engineering, Affiliated Nanshan Hospital of Shenzhen UniversityAbstract Staphylococcus aureus infections have emerged as a global public health threat. Two key factors—drug resistance and biofilm formation—substantially impair the efficacy of the antimicrobial treatment for S. aureus infections using conventional antibiotics. Consequently, discovering novel antimicrobial agents with potent antibacterial and antibiofilm activity has become a hotspot in recent years. Herein, the research first reported the remarkable inhibitory activity of AZD-5991, a selective Mcl-1 inhibitor, against S. aureus. The MIC50 and MIC90 values of AZD-5991 against S. aureus were 12.5 µM, and significant growth inhibition was observed at a subinhibitory concentration of 1/2 × MIC. Additionally, AZD-5991 exhibited bactericidal activity and a robust capacity for inhibiting S. aureus biofilm formation, with minimal cytotoxicity toward host cell lines. Membrane permeability assays revealed that AZD-5991 compromised S. aureus cell membrane integrity, while bacterial phospholipid components were found to neutralize the antibacterial activity of AZD-5991. Moreover, whole-genome sequencing and proteomic analysis were also applied to gain insights into the possible impact of AZD-5991 on the fatty metabolism of S. aureus. Furthermore, the antibacterial activity of AZD-5991 was remarkably declined by exogenous fatty acids linoleic acid (C18:2Δ9,12) and arachidonic acid (C20:4Δ5,8,11,14). Lastly, the biolayer interferometry assay supported the direct interaction of AZD-5991 with FabI, a key protein essential for bacterial growth and fatty acid metabolism. Conclusively, this study demonstrates that AZD-5991 inhibits S. aureus planktonic growth and biofilm formation by disrupting cell membrane integrity and targeting FabI. These findings position AZD-5991 as a promising novel antibiotic candidate for treating S. aureus infections resistant to traditional clinical antibiotics. Graphical Abstracthttps://doi.org/10.1186/s12866-025-04104-2Staphylococcus aureusAntimicrobial activityAZD-5991BiofilmFabI
spellingShingle Yuanyuan Tang
Han Deng
Zhichao Xu
Zhijian Yu
Yong Xiang
Zewen Wen
Shiqing Han
Zhong Chen
Tieying Hou
Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
BMC Microbiology
Staphylococcus aureus
Antimicrobial activity
AZD-5991
Biofilm
FabI
title Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
title_full Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
title_fullStr Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
title_full_unstemmed Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
title_short Repurposing AZD-5991 for inhibiting growth and biofilm formation of Staphylococcus aureus by disrupting the cell membrane and targeting FabI
title_sort repurposing azd 5991 for inhibiting growth and biofilm formation of staphylococcus aureus by disrupting the cell membrane and targeting fabi
topic Staphylococcus aureus
Antimicrobial activity
AZD-5991
Biofilm
FabI
url https://doi.org/10.1186/s12866-025-04104-2
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