Skip to main content

A study on the antibacterial activity and antimicrobial resistance of pyridinium cationic pillar[5]arene against Staphylococcus aureus and Escherichia coli


An increasing number of infections caused by multidrug-resistant (MDR) Staphylococcus aureusS. aureus) and Escherichia coliE. coli) have severely affected human society. Thus, it is essential to develop an alternative type of antibacterial agents that has a different bacterial resistance mechanism from that of traditional antibiotics. After the synthesis and structural characterization of a cationic pillar[5]arene with pyridinium groups (PP5), the antibacterial and antibiofilm activities as well as its microbial resistance were systematically investigated. In-depth evaluation of biological studies revealed that PP5 was an active antibacterial agent, with surprising antibiofilm formation ability against E. coli and S. aureus. From the results of differential scanning calorimetry and transmission electron microscopy, it was concluded that the microbicidal activity of PP5 was due to the physical disruption of the pathogen’s membrane and the subsequent leakage of cytoplasmic components, which could greatly reduce the rapid generation of resistance. It was presented that the easily available PP5 has high activity to inhibit Gram-positive and Gram-negative bacteria and/or their biofilms with low cytotoxicity. This pillar[5]arene derivative can be used as a good candidate for controlling drug-resistant pathogenic bacterial infections and treating MDR bacteria.

This is a preview of subscription content, access via your institution.

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5





Colony-forming units


Confocal laser scanning microscope


Crystal violet


Differential scanning calorimetry


Dimethyl sulfoxide

E. coli:

Escherichia coli


Large unilamellar liposomes


Luria Broth


Methicillin-resistant Staphylococcus aureus


Minimum bactericidal concentration


Minimum inhibitory concentration


Polymyxin B




Multidrug-resistant Staphylococcus aureus




Polydispersity index


Propidium iodide

P. aeruginosa:

Pseudomonas aeruginosa PAO1


Pyridine-based pillar[5]arene


Standard deviations

S. aureus:

Staphylococcus aureus


Supporting information




Transmission electron microscopy


Trimethylamine based pillar[5]arene


Tryptic soy broth


  • Allison KR, Brynildsen MP, Collins JJ (2011) Metabolite–enabled eradication of bacterial persisters by aminoglycosides. Nature 473:216–220

    CAS  Article  Google Scholar 

  • Barbosa M, Costa F, Monteiro C, Duarte F, Martins MCL, Gomes P (2019) Antimicrobial coatings prepared from Dhvar–5–click–grafted chitosan powders. Acta Biomater 84:242–256

    CAS  Article  Google Scholar 

  • Benhamou RI, Shaul P, Herzog IM, Fridman M (2015) Di–N–methylation of anti–gram–positive aminoglycoside–derived membrane disruptors improves antimicrobial potency and broadens spectrum to gram–negative bacteria. Angew Chem Int Ed 54:13617–13621

    CAS  Article  Google Scholar 

  • Chanawanno K, Chantrapromma S, Anantapong T, Kanjana-Opas A, Fun HK (2010) Synthesis, structure and in vitro antibacterial activities of new hybrid disinfectants quaternary ammonium compounds: pyridinium and quinolinium stilbene benzenesulfonates. Eur J Med Chem 45:4199–4208

    CAS  Article  Google Scholar 

  • Conlon BP, Nakayasu ES, Fleck LE, LaFleur MD, Isabella VM, Coleman K, Leonard SN, Smith RD, Adkin JN, Lewis K (2013) Activated ClpP kills persisters and eradicates a chronic biofilm infection. Nature 503:365–370

    CAS  Article  Google Scholar 

  • Gao L, Li M, Ehrmann S, Tu Z, Haag R (2019) Positively charged nanoaggregates based on zwitterionic pillar[5]arene that combat planktonic bacteria and disrupt biofilms. Angew Chem Int Ed 58:3645–3649

    CAS  Article  Google Scholar 

  • Gao LY, Wang HJ, Zheng B, Huang FH (2021) Combating antibiotic resistance: current strategies for the discovery of novel antibacterial materials based on macrocycle supramolecular chemistry. Giant 7:100066

    CAS  Article  Google Scholar 

  • Gatadi S, Gour J, Shukla M, Kaul G, Dasgupta A, Madhavi YV, Chopra S, Nanduri S (2019) Synthesis and evaluation of new 4–oxoquinazolin–3(4H)–yl) benzoic acid and benzamide derivatives as potent antibacterial agents effective against multidrug resistant Staphylococcus aureus. Bioorg Chem 83:569–579

    CAS  Article  Google Scholar 

  • Guo S, Huang Q, Chen Y, Wei J, Zheng J, Wang L, Wang Y, Wang R (2021) Synthesis and bioactivity of guanidinium–functionalized pillar[5]arene as a biofilm disruptor. Angew Chem Int Ed 60:618–623

    CAS  Article  Google Scholar 

  • He L, Pei H, Zhang C, Shao M, Li D, Tang M, Wang T, Chen X, Xiang M, Chen L (2018) Design, synthesis and biological evaluation of 7H–pyrrolo[2,3–d]pyrimidin–4–amine derivatives as selective Btk inhibitors with improved pharmacokinetic properties for the treatment of rheumatoid arthritis. Eur J Med Chem 145:96–112

    CAS  Article  Google Scholar 

  • Joseph R, Kaizerman D, Herzog IM, Hadar M, Cohen Y (2016a) Phosphonium pillar[5]arenes as a new class of efficient biofilm inhibitors: importance of charge cooperativity and the pillar platform. Chem Commun 52:10656–10659

    CAS  Article  Google Scholar 

  • Joseph R, Naugolny A, Feldman M, Herzog IM, Fridman M, Cohen Y (2016b) Cationic pillararenes potently inhibit biofilm formation without affecting bacterial growth and viability. J Am Chem Soc 138:754–757

    CAS  Article  Google Scholar 

  • Kaizerman-Kane D, Hadar M, Joseph R, Logviniuk D, Zafrani Y, Fridman M, Cohen Y (2021) Design guidelines for cationic pillar[n]arenes that prevent biofilm formation by Gram–positive pathogens. ACS Infect Dis 7:579–585

    CAS  Article  Google Scholar 

  • Kang HK, Seo CH, Luchian T, Park Y (2018) Pse–T2, an antimicrobial peptide with high–level, broad spectrum antimicrobial potency and skin biocompatibility against multidrug–resistant Pseudomonas aeruginosa infection. Antimicrob Agents Chemother 62:e01493-e1518

    PubMed  PubMed Central  Google Scholar 

  • Le P, Kunold E, Macsics R, Rox K, Jennings MC, Ugur I, Reinecke M, Chaves-Moreno D, Hackl MW, Fetzer C, Mandl FAM, Lehmann J, Korotkov VS, Hacker SM, Kuster B, Antes I, Pieper DH, Rohde M, Wuest WM, Medina E, Sieber SA (2020) Repurposing human kinase inhibitors to create an antibiotic active against drug–resistant Staphylococcus aureus, persisters and biofilms. Nat Chem 12:145–158

    CAS  Article  Google Scholar 

  • Liu YH, Jiang Y, Zhu JL, Huang JY, Zhang HJ (2019) Inhibition of bacterial adhesion and biofilm formation of sulfonated chitosan against Pseudomonas aeruginosa. Carbohyd Polym 206:412–419

    CAS  Article  Google Scholar 

  • O’Brien–Simpson NM, Li FW, Pantarat N, Hossain MA, Separovic F, Wade JD, Reynolds EC (2017) Fluorescent ion efflux screening assay for determining membrane–active peptides. Aust J Chem 70:220–228

    Article  Google Scholar 

  • Ogoshi A, Kanai S, Fujinami S, Yamagishi T, Nakamoto Y (2008) para-bridged symmetrical pillar[5]arenes: their lewis acid catalyzed synthesis and host–guest property. J Am Chem Soc 130:5022–5023

    CAS  Article  Google Scholar 

  • Ogoshi T, Yamagishi T, Nakamoto Y (2016) Pillar-shaped macrocyclic hosts pillar[n]arenes: new key players for supramolecular chemistry. Chem Rev 116:7937–8002

    CAS  Article  Google Scholar 

  • Pernak J, Kalewska J, Ksycififiska H, Cybulski J (2001) Synthesis and anti-microbial activities of some pyridinium salts with alkoxymethyl hydrophobic group. Eur J Med Chem 36:899–907

    CAS  Article  Google Scholar 

  • Petrillo AD, Fais A, Pintus F, Santos-Buelga C, González-Paramás AM, Piras V, Orrù G, Mameli A, Tramontano E, Frau A (2017) Broad–range potential of Asphodelusmicrocarpus leaves extract for drug development. BMC Microbiol 17:159

    Article  Google Scholar 

  • Si W, Li Z, Hou JL (2014) Voltage–driven reversible insertion into and leaving from a lipid bilayer: tuning transmembrane transport of artificial channels. Angew Chem Int Ed 126:4666–4669

    Article  Google Scholar 

  • Si F, Treut GL, Sauls JT, Vadia S, Levin PA, Jun S (2019) Mechanistic origin of cell-size control and homeostasis in bacteria. Curr Biol 29(2019):1760–1770

    CAS  Article  Google Scholar 

  • Singh PK, Parsek MR, Greenberg EP, Welsh MJ (2002) A component of innate immunity prevents bacterial biofilm development. Nature 417:552–555

    CAS  Article  Google Scholar 

  • Stockert JC, Blázquez-Castro A, Caete M, Horobin RW, Villanueva Á (2012) MTT assay for cell viability: intracellular localization of the formazan product is in lipid droplets. Acta Histochem 114:785–796

    CAS  Article  Google Scholar 

  • Sun ZM, Shi CG, Wang XY, Fang Q, Huang JY (2017) Synthesis, characterization, and antimicrobial activities of sulfonated chitosan. Carbohyd Polym 155:321–328

    CAS  Article  Google Scholar 

  • Tan H, Peng Z, Li Q, Xu X, Guo S, Tang T (2012) The use of quaternised chitosan–loaded PMMA to inhibit biofilm formation and downregulate the virulence–associated gene expression of antibiotic–resistant Staphylococcus. Biomaterial 33:365–377

    CAS  Article  Google Scholar 

  • Xu LC, Siedlecki CA (2022) Submicron topography design for controlling staphylococcal bacterial adhesion and biofilm formation. J Biomed Mater Res a ASAP.

    Article  Google Scholar 

  • Xue M, Yang Y, Chi XD, Zhang ZB, Huang FH (2012) Pillararenes, a new class of macrocycles for supramolecular chemistry. Accounts Chem Res 45:1294–1308

    CAS  Article  Google Scholar 

  • Yan S, Chen S, Gou X, Yang J, An J, Yang Y (2019) Biodegradable supramolecular materials based on cationic polyaspartamides and pillar[5]arene for targeting gram–positive bacteria and mitigating antimicrobial resistance. Adv Funct Mater 29:1904683

    Article  Google Scholar 

  • Yang H, Jin LQ, Zhao DQ, Lian ZF, Appu M, Huang JY, Zhang ZB (2021) Antibacterial and antibiofilm formation activities of pyridinium–based cationic pillar[5]arene against Pseudomonas aeruginosa. J Agr Food Chem 69:4276–4283

    CAS  Article  Google Scholar 

  • Zhang M, Zhu PP, Xin P, Si W, Li ZT, Hou JL (2017) Inside back cover: synthetic channel specifically inserts into the lipid bilayer of Gram–positive bacteria but not that of mammalian erythrocytes. Angew Chem Int Ed 56:2999–3003

    CAS  Article  Google Scholar 

  • Zhou C, Chia GWN, Ho JCS, Seviour T, Sailov T, Liedberg B, Kjelleberg S, Hinks J, Bazan GC (2018) Informed molecular design of conjugated oligoelectrolytes to increase cell affinity and antimicrobial activity. Angew Chem Int Ed 57:8069–8072

    CAS  Article  Google Scholar 

Download references


This work was supported by the Natural Science Foundation of Zhejiang Province of China (LY20B040001), the Project of Public Welfare Technology of Zhejiang Province of China (LGN19C200008), and the Science & Technology Innovation Program of Zhejiang Province (2018R52051).

Author information

Authors and Affiliations



Hua Yang and Dengqi Zhao conducted the lab work, data analysis, and drafted the manuscript. Lijun Yang contributed partly to the lab work. Shuping Wang conducted the MTT. Jianying Huang, Zibin Zhang, and Shijun Li conceived of the study, performed data review, and contributed to the manuscript writing. All authors have read and approved the final manuscript.

Corresponding authors

Correspondence to Jianying Huang or Zibin Zhang.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 9614 KB)

Supplementary file2 (DOCX 17 KB)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Yang, H., Zhao, D., Wang, S. et al. A study on the antibacterial activity and antimicrobial resistance of pyridinium cationic pillar[5]arene against Staphylococcus aureus and Escherichia coli. Int Microbiol (2022).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI:


  • Pillar[5]arenes
  • Pyridinium
  • Biofilm
  • Antibacterial agents
  • Drug-resistant bacteria
  • Liposome