Exploring novel amides as efflux pump inhibitors for overcoming antibiotic resistance in multidrug-resistant Pseudomonas aeruginosa

Fahad  Ullah; Maqsood  Ali; Falak  Niaz; Israr  Ali Khan; Sher  Wali Khan; Momin  Khan; Rafaqat  Ishaq; Abdul  Manan; Ying  Yu; Muhammad  Ilyas

doi:10.18502/ijm.v17i4.19271

Fahad Ullah Department of Microbiology, Institute of Pathology and Diagnostic Medicine, Khyber Medical University, Peshawar, Pakistan
Maqsood Ali Center for Stem Cells and Regenerative Medicine, Department of Orthopedic Surgery of the Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China
Falak Niaz Department of Medical Laboratory Technology, Faculty of Rehabilitation and Allied Health Sciences, Riphah International University, Malakand Campus, Malakand, Pakistan
Israr Ali Khan Department of Microbiology, Institute of Pathology and Diagnostic Medicine, Khyber Medical University, Peshawar, Pakistan
Sher Wali Khan Department of Chemistry, Women University Rawalpindi, Rawalpindi, Pakistan
Momin Khan Department of Microbiology, Institute of Pathology and Diagnostic Medicine, Khyber Medical University, Peshawar, Pakistan
Rafaqat Ishaq Department of Biochemistry & Molecular Biology, School of Medicine, University of Maryland, Baltimore, Maryland, USA
Abdul Manan Department of Medical Laboratory Technology, Faculty of Rehabilitation and Allied Health Sciences, Riphah International University, Malakand Campus, Malakand, Pakistan
Ying Yu Department of General Medicine, The 2nd Clinical Medical College, Zhejiang Chinese Medical University Hangzhou, Zhejiang Province, China
Muhammad Ilyas Department of Medical Laboratory Technology, Faculty of Rehabilitation and Allied Health Sciences, Riphah International University, Malakand Campus, Malakand, Pakistan

DOI: https://doi.org/10.18502/ijm.v17i4.19271

Keywords: Anti-bacterial agents; Checkerboard assay; Drug resistance; Efflux pump; Pseudomonas aeruginosa

Abstract

Background and Objectives: Pseudomonas aeruginosa (P. aeruginosa), a multidrug-resistant bacterium, represents a con- siderable risk in healthcare environments owing to its capacity to induce various infections. The resistance of P. aeruginosa is frequently linked to efflux pumps that actively remove antibiotics from the bacterial cell. This study investigates novel amide compounds as potential alternatives to address P. aeruginosa isolates exhibiting multidrug resistance mediated by efflux pumps.

Materials and Methods: Gram staining and biochemical assays revealed thirty-three multi-drug-resistant P. aeruginosa iso- lates from a tertiary care hospital Peshawar. After antibiotic susceptibility testing, efflux pumps were detected using Ethidium Bromide (EtBr) agar cartwheel technique and UV transilluminator. Novel amides were tested for efflux pump and anti-pseu- domonal action against efflux pump-positive isolates utilizing agar well diffusion and micro broth dilution, including synergy with ciprofloxacin and gentamicin.

Results: Three high efflux pump activity P. aeruginosa isolates were chosen using ETBr agar cartwheel technique. Novel

amides (ITC, ITD, ITE, DEP) block efflux pump, although TEM-cu is very antimicrobial. TEM-cu, DEP, ITC, and ITE have

0.19, 0.78, and 0.78 mg/ml MICs. Effectiveness against efflux pump-expressing P. aeruginosa is lowest with ITE (1.56 mg/

ml). Together with ciprofloxacin and gentamicin, TEM-cu and DEP improved antimicrobial effectiveness.

Conclusion: TEM-cu is highly effective against efflux pump-positive P. aeruginosa, while amides like ITC, ITD, ITE, and DEP block these pumps. With significant reductions, DEP and TEM-cu improve ciprofloxacin and gentamicin efficacy. This method may help overcome P. aeruginosa efflux pump-mediated resistance.