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Antibacterial effects of BB-Cl-Amidine against multidrug-resistant Gram-positive pathogens via membrane disruption

The Journal of Antibiotics volume 79, pages 30–40 (2026) Cite this article

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Abstract

The escalating prevalence of multidrug-resistant (MDR) gram-positive bacteria, including methicillin-resistant Staphylococcus aureus (MRSA) and linezolid-resistant Enterococcus faecalis, highlights the critical demand for new antibacterial agents that target resistance pathways. BB-Cl-Amidine is originally considered as a peptidyl arginine deiminase inhibitor and till now, its potential antimicrobial activity has not been explored. This study sought to evaluate the antibacterial effectiveness and underlying mechanisms of BB-Cl-Amidine against MDR gram-positive pathogens. The results showed that BB-Cl-Amidine exhibited potent antibacterial activity with minimum inhibitory concentration (MIC) values ranging from 25 μM to 50 μM against MRSA, E. faecalis and various of Gram-positive bacteria clinical isolates. At sub-MIC concentrations, BB-Cl-Amidine significantly reduced biofilm formation in both S. aureus and E. faecalis. Moreover, the increased permeability and depolarizing membrane potential of S. aureus was found by BB-Cl-Amidine. The antibacterial activity of BB-Cl-Amidine can be neutralized by cardiolipin (CL) and phosphatidylglycerol (PG). Furthermore, BB-Cl-Amidine exposure resulted in the abnormal expression of functional proteins correlated with the cell membranes and phospholipid metabolas. In summary, the potential antibacterial and anti-biofilm activities of BB-Cl-Amidine are demonstrated via membrane disruption, offering a promising scaffold for combating MDR Gram-positive infections.

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References

  1. Ahmad-Mansour N, Loubet P, Pouget C, et al. Staphylococcus aureus toxins: an update on their pathogenic properties and potential treatments[J]. Toxins. 2021;13:677.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Piewngam P, Otto M. Staphylococcus aureus colonization and strategies for decolonization. Lancet Microbe. 2024;5:e606–18.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Gagliotti C, Högberg LD, Billström H, et al. Staphylococcus aureus bloodstream infections: diverging trends of methicillin-resistant and methicillin-susceptible isolates, EU/EEA, 2005 to 2018[J]. Eurosurveillance. 2021;26:2002094.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Chen H, Song S, Cui R, Feng YW, Ge P. Global trends in staphylococcus aureus-related lower respiratory infections from 1990 to 2021: findings from the 2021 global burden of disease report. Eur J Clin Microbiol Infect Dis. 2025;44:1455–69.

  5. Liang J, Hu Y, Fu M, et al. Resistance and molecular characteristics of methicillin-resistant Staphylococcus aureus and heterogeneous vancomycin-intermediate Staphylococcus aureus[J]. Infect Drug Resist. 2023;16:379–88.

  6. Youenou B, Martins Simoes P, Tristan A, et al. Linezolid resistance: detection of the cfr (B) gene in French clinical MRSA strains[J]. J Antimicrob Chemother. 2023;78:445–9.

    Article  CAS  PubMed  Google Scholar 

  7. Garrigos C, Murillo O, Euba G, et al. Efficacy of tigecycline alone and with rifampin in foreign-body infection by methicillin-resistant Staphylococcus aureus[J]. J Infect. 2011;63:229–35.

    Article  CAS  PubMed  Google Scholar 

  8. Heidary M, Khosravi AD, Khoshnood S, et al. Daptomycin[J]. J Antimicrob Chemother. 2018;73:1–11.

    Article  CAS  PubMed  Google Scholar 

  9. Lister JL, Horswill AR. Staphylococcus aureus biofilm: recent developments in biofilm dispersal[J]. Front Cell Microb. 2014;4:178.

    Google Scholar 

  10. Willett JLE, Dale JL, Kwiatkowski LM, et al. Comparative biofilm assays using Enterococcus faecalis OG1RF identify new determinants of biofilm formation. mBio. 2021;12:e0101121.

    Article  PubMed  Google Scholar 

  11. Willett JLE, Ji MM, Dunny GM. Exploiting biofilm phenotypes for functional characterization of hypothetical genes in Enterococcus faecalis. NPJ Biofilms Microbiomes. 2019;5:23.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Vergara-Irigaray M, Maira-Litran T, Merino N, et al. Wall teichoic acids are dispensable for anchoring the PNAG exopolysaccharide to the Staphylococcus aureus cell surface[J]. Microbiology. 2008;154:865–77.

    Article  CAS  PubMed  Google Scholar 

  13. Knight JS, Subramanian V, O’Dell AA, et al. Peptidylarginine deiminase inhibition disrupts NET formation and protects against kidney, skin, and vascular disease in lupus-prone MRL/lpr mice[J]. Annals Rheum Dis. 2015;74:2199–206.

    Article  CAS  Google Scholar 

  14. Stachowicz A, Pandey R, Sundararaman N, Venkatraman V, Van Eyk JE, Fert-Bober J. Protein arginine deiminase 2 (PAD2) modulates the polarization of THP-1 macrophages to the anti-inflammatory M2 phenotype. J Inflame. 2022;19:20.

    Article  Google Scholar 

  15. Mishra N, Schwertner L, Sams K, et al. Cutting edge: protein arginine deiminases 2 and 4 regulate NLRP3 inflammasome–dependent IL-1β maturation and ASC speck formation in macrophages[J]. J Immunol. 2019;203:795–800.

    Article  CAS  PubMed  Google Scholar 

  16. Wang C, Xiong Y, Bao C, et al. Antibacterial and anti-biofilm activity of cadazolid against Staphylococcus aureus clinical isolates from China. Front Microb. 2023;14:1131178.

    Article  Google Scholar 

  17. Humphries R, Bobenchik AM, Hindler JA, Schuetz AN. Overview of changes to the clinical and laboratory standards Institute Performance Standards for Antimicrobial Susceptibility Testing, M100, 31st Edition. J Clin Microbiol. 2021;59:e0021321.

    Article  PubMed  Google Scholar 

  18. Pierce VM, Bhowmick T, Simner PJ. Guiding antimicrobial stewardship through thoughtful antimicrobial susceptibility testing and reporting strategies: an updated approach in 2023. J Clin Microbiol. 2023;61:e0007422.

    Article  PubMed  Google Scholar 

  19. Liu X, Xiong Y, Shi Y, et al. In vitro, the activities of licochalcone A against planktonic cells and biofilm of Enterococcus faecalis. Front Microbiol. 2022;13:970901.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Jacqueline C, Caillon J, Le Mabecque V, et al. In vitro activity of linezolid alone and in combination with gentamicin, vancomycin, or rifampicin against methicillin-resistant Staphylococcus aureus by time-kill curve methods. J Antimicrob Chemother. 2003;51:857–64.

    Article  CAS  PubMed  Google Scholar 

  21. Sæbø IP, Bjørås M, Franzyk H, Helgesen E, Booth JA. Optimization of the hemolysis assay for the assessment of cytotoxicity. Int J Mol Sci. 2023;24:2914.

    Article  PubMed  PubMed Central  Google Scholar 

  22. Adan A, Kiraz Y, Baran Y. Cell proliferation and cytotoxicity assays. Curr Pharm Biotechnol. 2016;17:1213–21.

    Article  CAS  PubMed  Google Scholar 

  23. Zhu Y, Xu W, Chen W, et al. Self-assembling peptide with dual function of cell penetration and antibacterial as a nano weapon to combat intracellular bacteria. Sci Adv. 2025;11:eads3844.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Duong VA, Lee H. Bottom-up proteomics: advancements in sample preparation. Int J Mol Sci. 2023;24:5350.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Chang X, Yamada R, Suzuki A, et al. Localization of peptidylarginine deiminase 4 (PADI4) and citrullinated protein in synovial tissue of rheumatoid arthritis[J]. Rheumatology. 2005;44:40–50.

    Article  CAS  PubMed  Google Scholar 

  26. Zheng J, Liu X, Xiong Y, et al. AMXT-1501 targets membrane phospholipids against Gram-positive and -negative multidrug-resistant bacteria. Emerg Microbes Infect. 2024;13:2321981.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Huang J, Xu Z, He P, et al. Repurposing TAK-285 as an antibacterial agent against multidrug-resistant Staphylococcus aureus by targeting cell membrane. Curr Microbiol. 2024;82:8.

    Article  PubMed  Google Scholar 

  28. Liu X, Xiong Y, Peng R, et al. Antibacterial activity, and mechanisms of D-3263 against Staphylococcus aureus. BMC Microbiol. 2024;24:224.

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The authors express their gratitude to Wei Guang Pan and Jie Lian from the Department of Laboratory Medicine at Shenzhen Nanshan People’s Hospital for their assistance in identifying and preserving the bacterial isolates.

Funding

This research was made possible through the following grants: National Natural Science Foundation of China (82172283); Guangdong Basic and Applied Basic Research Foundation (2022A1515110096, 2024A1515013276); Sanming Project of Medicine in Shenzhen (SMGC202305029); Shenzhen Key Medical Discipline Construction Fund (SZXK06162); and the Shenzhen Nanshan District Scientific Research Program of the People’s Republic of China (NS2024001; NS2023008; NSZD2024023; NSZD2024036; NS2022046; NS2024001Z; NS2024038).

Author information

Author notes

  1. These authors contributed equally: Zhenfeng Wang, Junhua Ma, Jintuan Lin, Congcong Li.

Authors and Affiliations

  1. Clinical Medical College, Jiamusi University, Jiamusi, Heilongjiang, China

    Zhenfeng Wang, Guiqiu Li & Ying Wei

  2. Department of Infectious Diseases and Shenzhen Key Laboratory for Endogenous Infections, Shenzhen Nanshan People’s Hospital, Shenzhen, China

    Zhenfeng Wang, Junhua Ma, Jintuan Lin, Congcong Li, Zhijian Yu & Bing Bai

  3. School of Basic Medical Sciences, Shenzhen University Medical School, Shenzhen University, Shenzhen, China

    Congcong Li

  4. Marketing Department ofGuangzhou branch ofVarian Medical Equipment Trading (Beijing) Corp, Guangzhou, China

    Yong Xiang

  5. Laboratory Medicine Center, Nanshan People’s Hospital, Shenzhen, China

    Guiqiu Li

  6. Heilongjiang Provincial Hospital, Harbin, Heilongjiang, China

    Ying Wei

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  1. Zhenfeng Wang
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Contributions

Z.W. taken part in an antimicrobial susceptibility test and growth curve experiments and drafted the manuscript. J.M. and J.L. performed the time-killing analysis. C.L. and Y.X. performed a biofilm assay and cytotoxicity assay. Y.W. taken part in the proteomics data analysis. B.B. participated in the collection and statistical analyses of the clinical isolates of S. aureus and E. faecalis coliform bacteria. Z.Y. and G.L. designed the study, took part in the analysis of the data, and provided important revisions of the manuscript for valuable intellectual content. valuable intellectual content provided important manuscript revisions. All authors have read and approved this manuscript.

Corresponding authors

Correspondence to Bing Bai, Guiqiu Li or Ying Wei.

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Conflict of interest

The authors declare no competing interests.

Ethical approval

Approval of ethics and permission for participation. All techniques were conducted in accordance with applicable norms and regulations, authorized by the Ethics Committee of Shenzhen Nanshan People’s Hospital, and in compliance with the Helsinki Declaration of 1964 and its later revisions or similar ethical standards. All experimental techniques involving human volunteers received approval from the Institutional Ethics Committee of Shenzhen Nanshan People’s Hospital. Bacterial strains were obtained in accordance with national standards for the regular clinical therapy of patients in China. Consequently, informed permission was not acquired, and a waiver of informed consent was sanctioned by the Institutional Ethics Committee of Shenzhen Nanshan People’s Hospital.

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Wang, Z., Ma, J., Lin, J. et al. Antibacterial effects of BB-Cl-Amidine against multidrug-resistant Gram-positive pathogens via membrane disruption. J Antibiot 79, 30–40 (2026). https://doi.org/10.1038/s41429-025-00869-w

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