skip to main content


Title: Synthetic ionophores as non-resistant antibiotic adjuvants
Antimicrobial resistance is a world-wide health care crisis. New antimicrobials must both exhibit potency and thwart the ability of bacteria to develop resistance to them. We report the use of synthetic ionophores as a new approach to developing non-resistant antimicrobials and adjuvants. Most studies involving amphiphilic antimicrobials have focused on either developing synthetic amphiphiles that show ion transport, or developing non-cytotoxic analogs of such peptidic amphiphiles as colistin. We have rationally designed, prepared, and evaluated crown ether-based synthetic ionophores (‘hydraphiles’) that show selective ion transport through bilayer membranes and are toxic to bacteria. We report here that hydraphiles exhibit a broad range of antimicrobial properties and that they function as adjuvants in concert with FDA-approved antibiotics against multi-drug resistant (MDR) bacteria. Studies described herein demonstrate that benzyl C 14 hydraphile (BC 14 H) shows high efficacy as an antimicrobial. BC 14 H, at sub-MIC concentrations, forms aggregates of ∼200 nm that interact with the surface of bacteria. Surface-active BC 14 H then localizes in the bacterial membranes, which increases their permeability. As a result, antibiotic influx into the bacterial cytosol increases in the presence of BC n Hs. Efflux pump inhibition and accumulation of substrate was also observed, likely due to disruption of the cation gradient. As a result, BC 14 H recovers the activity of norfloxacin by 128-fold against resistant Staphylococcus aureus . BC 14 H shows extremely low resistance development and is less cytotoxic than colistin. Overall, synthetic ionophores represent a new scaffold for developing efficient and non-resistant antimicrobial-adjuvants.  more » « less
Award ID(s):
1710594
NSF-PAR ID:
10249494
Author(s) / Creator(s):
; ; ; ; ;
Date Published:
Journal Name:
RSC Advances
Volume:
9
Issue:
4
ISSN:
2046-2069
Page Range / eLocation ID:
2217 to 2230
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. The threat of antibiotic resistance warrants the discovery of agents with novel antimicrobial mechanisms. Antimicrobial peptides (AMPs) directly disrupting bacterial membranes may overcome resistance to traditional antibiotics. AMP development for clinical use has been mostly limited to topical application to date. We developed a rational framework for systematically addressing this challenge using libraries composed of 86 novel Trp- and Arg-rich engineered peptides tested against clinical strains of the most common multidrug-resistant bacteria known as ESKAPE pathogens. Structure-function correlations revealed minimum lengths (as low as 16 residues) and Trp positioning for maximum antibacterial potency with mean minimum inhibitory concentration (MIC) of 2–4 μM and corresponding negligible toxicity to mammalian cells. Twelve peptides were selected based on broad-spectrum activity against both gram-negative and -positive bacteria and <25% toxicity to mammalian cells at maximum test concentrations. Most of the selected PAX remained active against the colistin-resistant clinical strains. Of the selected peptides, the shortest (the 16-residue E35) was further investigated for antibacterial mechanism and proof-of-concept in vivo efficacy. E35 killed an extensively-resistant isolate of Pseudomonas aeruginosa (PA239 from the CDC, also resistant to colistin) by irreversibly disrupting the cell membranes as shown by propidium iodide incorporation, using flow cytometry and live cell imaging. As proof of concept, in vivo toxicity studies showed that mice tolerated a systemic dose of up to 30 mg/kg peptide and were protected with a single 5 mg/kg intravenous (IV) dose against an otherwise lethal intraperitoneal injection of PA239. Efficacy was also demonstrated in an immune-compromised Klebsiella pneumoniae infection model using a daily dose of 4mg/kg E35 systemically for 2 days. This framework defines the determinants of efficacy of helical AMPs composed of only cationic and hydrophobic amino acids and provides a path for a potential departure from the restriction to topical use of AMPs toward systemic application. 
    more » « less
  2. null (Ed.)
    As we are on the cusp of the “post-antibiotic” era due to rapid spread of drug resistant bacteria, there is an urgent need for new antimicrobials that are not susceptible to bacterial resistance mechanisms. In this review, we will discuss the recent development of “polymer therapeutics” with antimicrobial activity. Learning from host-defence peptides, we propose the biomimetic design of synthetic polymers to target bacterial cell membranes, which act by compromising the membrane integrity. The discussion is extended to the future challenges and opportunities of antimicrobial polymers for clinical applications. 
    more » « less
  3. Abstract

    The emergence and spread of antimicrobial resistance highlights the urgent need for new antibiotics. Organoarsenicals have been used as antimicrobials since Paul Ehrlich’s salvarsan. Recently a soil bacterium was shown to produce the organoarsenical arsinothricin. We demonstrate that arsinothricin, a non-proteinogenic analog of glutamate that inhibits glutamine synthetase, is an effective broad-spectrum antibiotic against both Gram-positive and Gram-negative bacteria, suggesting that bacteria have evolved the ability to utilize the pervasive environmental toxic metalloid arsenic to produce a potent antimicrobial. With every new antibiotic, resistance inevitably arises. ThearsN1gene, widely distributed in bacterial arsenic resistance (ars) operons, selectively confers resistance to arsinothricin by acetylation of the α-amino group. Crystal structures of ArsN1N-acetyltransferase, with or without arsinothricin, shed light on the mechanism of its substrate selectivity. These findings have the potential for development of a new class of organoarsenical antimicrobials and ArsN1 inhibitors.

     
    more » « less
  4. The application of animal manures to cropland is an important nutrient recycling strategy in many parts of the world. Commonly, aggregated manure wastes contain chemical stressors including veterinary antimicrobials, heavy metals, and antimicrobial resistance genes (ARGs) that can stimulate the development and proliferation of antimicrobial resistance (AMR). While the presence of antimicrobials in manure is well-documented, the co-occurrence of other potentially impactful chemical stressors in swine manure remains underreported. This study quantifies and analyzes correlations between antimicrobials, metals, and certain ARGs present in manure samples from swine farms in Iowa, United States. Relationships between chemical stressors and different stages of swine production or feed composition are also investigated. Results revealed substantial levels of tetracyclines [up to 1,260 µg g −1 dry weight (d.w.) of manure for oxytetracycline] detected in all samples. Tiamulin, two ionophores (monensin and lasalocid), and one macrolide (tilmicosin) were detected at maximum class concentrations of 9.4, 0.547, and 0.472 µg g −1 d.w., respectively. The median relative abundances of ermB and tetM were 0.13 and 0.17 copies g −1 wet weight (w.w.) manure (normalized to 16S gene), respectively. Additionally, high levels of copper (Cu), iron (Fe), and zinc (Zn) were detected in all samples, with maximum concentrations of 887, 1,900, and 2,100 µg g −1 d.w., respectively. Notably, uranium (U) was detected in 11 samples, at concentrations up to 0.77 µg g −1 . A global analysis of AMR-stressor relationships using Spearman’s rank correlation indicates Cu, and Ba are the most positively and significantly correlated with cytotoxic anhydrotetracycline (ATC) and/or anhydrochlortetracycline (ACTC) concentrations in all tested facilities (Cu-ATC: ρ = 0.67, p = 0.0093; Cu-ACTC: ρ = 0.75, p = 0.0022; Ba-ATC: ρ = 0.84, p = 0.0002). Interestingly, ermB and tetM genes were strongly, positively correlated to each other ( ρ = 0.92, p < 0.0001), suggesting possible co-selection, despite the absence of correlation between ARGs and tetracycline concentrations. This study demonstrates the complexity of interactions between antimicrobials, metals, and ARGs in multiple manure storage pits prior to cropland application. 
    more » « less
  5. null (Ed.)
    Hydraphiles are synthetic amphiphiles that form pores in bilayer membranes. A study was undertaken to determine if the formation of pores could assist the penetration of antibiotics into bacteria. The disruption of ion homeostasis by the pore-formers leads to microbial toxicity. Co-administration of hydraphiles at concentration ≤ ½ MIC and antimicrobials to E. coli or P. aeruginosa showed potency enhancements of up to 30-fold. A possible mechanism is the enhancement of antibiotic influx owing to membrane disruption and/or altering the ion balance within the bacterial cells. 
    more » « less