Ceragenins Research Articles

In Vitro Activities of the Cationic Steroid Antibiotics CSA-13, CSA-131, CSA-138, CSA-142, and CSA-192 Against Carbapenem-resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosa is an important opportunistic pathogen that is difficult to treat because of the antibiotic resistance that has developed in recent years. Increasing carbapenem resistance has led to a rise in hospital infections caused by this bacterium. As a result, researchers have begun to search for new molecules. Ceragenins are the general name for membrane-acting cationic steroid antimicrobial molecules that have activity similar to that of antimicrobial peptides. In this study, we investigated the in vitro activities of the cationic steroid antibiotics (CSAs) CSA-13, CSA-131, CSA-138, CSA-142, CSA-192, and colistin on carbapenem-resistant Pseudomonas aeruginosa (CRPA). Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined by broth dilution method. The MIC50 (μg/mL) values of CSA-13, CSA-131, CSA-138, CSA-142, CSA-192, colistin, and meropenem were 8, 4, 8, 16, 32, 1, and 16, respectively. The MBC values were equal to or twice the MIC values. CSA-131 and CSA-138 appear to be good candidates for CRPA treatment. However, the lack of stability, efficacy, and pharmacokinetic properties of CSA requires further research in the future in vivo and in vitro.

Ceragenins exhibiting promising antimicrobial activity against various multidrug resistant Gram negative bacteria

Ceragenins are novel promising agents for the treatment of infections caused by multi-drug resistant microorganisms. Since colistin resistance has become a worldwide problem, the need for new treatment agents has been increasing steadily. Therefore, this study aimed to investigate in vitro antimicrobial activities of ceragenins (Cationic Steroid Antibiotics) (CSA-8, CSA-13, CSA-142 and CSA-192) against multidrug resistant Gram negative isolates from Turkey. Experiments were performed by using broth microdilution method against Klebsiella pneumoniae, Morganella morganii, Pseudomonas aeruginosa and Stenotrophomonas maltophilia isolates. All microorganisms except for three isolates were identified as multidrug resistant. Among tested ceragenins, CSA-13 showed the best results (MIC: 8-64 µg/ml). Nevertheless, the antimicrobial activity of CSA-8 was not significant. In conclusion, ceragenins appear to be a good candidate as antimicrobial therapy in the presence of multidrug (including colistin) resistant microorganisms.

In vitro Activities of Cationic Steroid Antibiotics, CSA-13, CSA-131, CSA-138, CSA-142 and CSA-192 against carbapenem-resistant Pseudomonas aeruginosa

In vitro Activities of Cationic Steroid Antibiotics, CSA-13, CSA-131, CSA-138, CSA-142 and CSA-192 against carbapenem-resistant Pseudomonas aeruginosa

Antibacterial activity and safety of commercial veterinary cationic steroid antibiotics and neutral superoxidized water.

Antibiotic resistance of bacteria common to the ocular surface is an evolving problem. Thus, novel treatment options with new modes of action are required. We investigated the antibacterial activity and safety of three commercially available topical veterinary ophthalmic products (cationic steroid antibiotics, products A and B, and a neutral superoxidized water, product C) to determine their potential use as antimicrobial alternatives. The minimum inhibitory concentrations (MIC) of the three products were determined against 17 antibiotic resistant bacterial clinical isolates from the ocular surface. Using a standard cytotoxicity assay, the products at varying concentrations were evaluated with a corneal fibroblast cell line and a macrophage-like cell line to determine their potential toxic effect in vitro. The commercial ophthalmic solutions, ofloxacin 0.3%, tobramycin 0.3% and gentamicin 0.3% were used as positive controls for the MIC and tobramycin 0.3% was used as positive control for the cytotoxicity assays. For the MIC, Product C showed no inhibition of growth for any organisms, while Products A and B showed inhibition of growth similar to slightly less than the positive controls. For the cytotoxicity assays, Product C exhibited minimal toxicity while Products A and B exhibited toxicity similar to the controls. In conclusion, Product C had no antibacterial activity in these assays, while Products A and B had antibacterial profiles similar to slightly less than common topical ophthalmic antibiotics and cytotoxicity profiles similar to common topical ophthalmic antibiotics. To our knowledge, this is the first report on the antibacterial activity and safety of the cationic steroid antibiotics and superoxidized water.

A Novel Class of Antimicrobial Agents: Ceragenins (Cationic Steroid Antibiotics)

A Novel Class of Antimicrobial Agents: Ceragenins (Cationic Steroid Antibiotics)

Study of the effect of antimicrobial peptide mimic, CSA‐13, on an established biofilm formed by Pseudomonas aeruginosa

The formation of a Pseudomonas aeruginosa biofilm, a complex structure enclosing bacterial cells in an extracellular polymeric matrix, is responsible for persistent infections in cystic fibrosis patients leading to a high rate of morbidity and mortality. The protective environment created by the tridimensional structure reduces the susceptibility of the bacteria to conventional antibiotherapy. Cationic steroid antibiotics (CSA)-13, a nonpeptide mimic of antimicrobial peptides with antibacterial activity on planktonic cultures, was evaluated for its ability to interact with sessile cells. Using confocal laser scanning microscopy, we demonstrated that the drug damaged bacteria within an established biofilm showing that penetration did not limit the activity of this antimicrobial agent against a biofilm. When biofilms were grown during exposure to shear forces and to a continuous medium flow allowing the development of robust structures with a complex architecture, CSA-13 reached the bacteria entrapped in the biofilm within 30 min. The permeabilizing effect of CSA-13 could be associated with the death of the bacteria. In static conditions, the compound did not perturb the architecture of the biofilm. This study confirms the potential of CSA-13 as a new strategy to combat persistent infections involving biofilms formed by P. aeruginosa.

Effect of a low concentration of a cationic steroid antibiotic (CSA-13) on the formation of a biofilm by Pseudomonas aeruginosa

Cationic steroids like CSA-13 have been designed by analogy with antimicrobial cationic peptides and have bactericidal properties. The purpose of this work was to evaluate the effect of a low concentration (1 mg l(-1)) of CSA-13 on the formation of a biofilm by eight strains of Pseudomonas aeruginosa (four mucoid and four nonmucoid strains) on an inert surface. The biofilm formation was measured with the Crystal Violet method. CSA-13 inhibited the formation of a biofilm by three strains. The zeta potential varied among the strains. The inhibition by the cationic steroid analogue affected the populations of bacteria with the lowest zeta potential. P. aeruginosa bound a fluorescent, more hydrophobic analogue of CSA-13 but there was no correlation between this binding and the inhibition by CSA-13 of biofilm formation. The interaction of CSA-13 with bacteria did not modify their ability to produce rhamnolipids. A low concentration of CSA-13 inhibits the formation of a biofilm by P. aeruginosa through electrostatic interactions and without affecting the production of rhamnolipids. A low, nontoxic concentration of CSA-13 might be beneficial for the prevention of biofilm formation.

In VitroAmoebicidal Activity of a Ceragenin, Cationic Steroid Antibiotic-13, AgainstAcanthamoeba castellaniiand Its Cytotoxic Potential

Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Acanthamoeba keratitis is difficult to treat, without total efficacy in some patients because of cysts that are less susceptible than trophozoites to the usual treatments. Contact lens wearers are most at risk and account for some 95% of cases. Cationic steroid antibiotic (CSA)-13 is a small molecule aminosterol that has been shown to mimic the activity of endogenous antimicrobial peptides and has bactericidal activity based on membrane disruption. We investigated here the in vitro effectiveness of CSA-13 with a concentration of 100, 75, 50, and 25 mg/mL on proliferation of Acanthamoeba castellanii trophozoites and cysts and cytotoxic potential. CSA-13 was evaluated for its amoebicidal activity using an inverted light microscope at 1, 2, 4, 8, 12, and 24 h. For the determination of cytotoxicity of the CSA-13 on L929 cells, agar diffusion tests were performed. CSA-13 inhibited trophozoite growth in dose- and time-dependent ways. At 1 h, no viable trophozoites were observed in the presence of CSA-13 solution in a concentration 100 mg/mL in phosphate-buffered saline. Results of cytotoxicity experiments demonstrated that CSA-13 solution had mild toxicity at 100 mg/mL concentration on cells, whereas it had no toxicity at 75 mg/mL concentration. The findings of this experiment as in vitro ameboebicidal activity for Acanthamoeba suggest that CSA-13 has a potential to be used as a new agent in lens solutions to prevent Acanthamoeba growth and infections.

Interaction between tobramycin and CSA-13 on clinical isolates of Pseudomonas aeruginosa in a model of young and mature biofilms

The bactericidal activity of a cholic acid antimicrobial derivative, CSA-13, was tested against eight strains of Pseudomonas aeruginosa (both reference and clinical strains) and compared with the response to tobramycin. In planktonic cultures, the minimal inhibitory and minimal bactericidal concentrations of CSA-13 and tobramycin were in the 1-25 mg/L range except for one mucoid clinical strain which was much less sensitive to tobramycin (minimal bactericidal concentration, 65-125 mg/L). In young (24 h) biofilms, the sensitivity to CSA-13 was reduced (half-maximal concentration CSA-13 averaged 88 mg/L) and varied among the eight strains. The sensitivity to tobramycin was also very variable among the strains and some were fully resistant to the aminoglycoside. The combination of tobramycin with CSA-13 was synergistic in five strains. Only one strain showed antagonism between the two drugs at low concentrations of CSA-13. One reference and five clinical strains were tested in mature (12 days) biofilms. The effect of CSA-13 was delayed, some strains requiring 9 days exposure to the drug to observe a bactericidal effect. All the strains were tolerant to tobramycin but the addition of CSA-13 with tobramycin was synergistic in three strains. CSA-13 permeabilized the outer membrane of the bacteria (half-maximal concentration, 4.4 mg/L). At concentrations higher than 20 mg/L, it also permeabilized the plasma membrane of human umbilical vein endothelial cells. In conclusion, CSA-13 has bactericidal activity against P. aeruginosa even in mature biofilms and cationic steroid antibiotics can thus be considered as potential candidates for the treatment of chronic pulmonary infections of patients with cystic fibrosis. Considering its interaction with the plasma membrane of eukaryotic cells, less toxic derivatives of CSA-13 should be developed.

Anti-Trypanosomatid Activity of Ceragenins

Cationic steroid antibiotics (CSAs), or ceragenins, are amphiphilic compounds consisting of a cholic acid backbone that is attached to several cationic amines. In this study, we tested the hypothesis that CSAs possess antiparasitic activities with minimal to no effects on mammalian cells, and thus could be used as potential therapeutic agents against pathogenic trypanosomatids. To investigate this notion, we synthesized CSAs and determined their trypanocidal and leishmanicidal activities in vitro. The 3 ceragenins assayed, i.e., CSA-8, CSA-13, and CSA-54, showed several degrees of parasiticidal activity. CSA-13 was the most effective compound against Leishmania major promastigotes and Trypanosoma cruzi trypomastigotes, at LD(50) 4.9 and 9 microM, respectively. The trypanocidal activities of these ceragenins were also assessed by infectivity experiments. We found CSA-8 was more effective on T. cruzi intracellular amastigotes when the infected host cells were treated for 24 hr (LD(50), 6.7 microM). Macrophages and LLC-MK(2) (treated for 72 hr) showed relative low susceptibility to these compounds. Our results suggest that ceragenins are indeed promising chemotherapeutic agents against trypanosomatids, but they require further investigation.

Preparation, quality control and biological evaluation of 99mTc-labelled cationic steroid antibiotic (CSA-13)

Ceragenins (CSAs) are a new class of antimicrobial agents that display broad spectrum antibacterial activity and may find use in the treatment of various infections. A member of this class CSA-13 was labelled with 99mTc using SnCl2·H2O as a reducing agent. The labelling efficiency depended on the ligand/reductant ratio, pH and volume of the reaction mixture. The radiochemical purity and stability of 99mTc-CSA was determined by thin layer chromatography. Biodistribution studies of 99mTc-CSA were performed in Sprague Dawley rats. Liver and spleen uptake was quite high. A significantly higher accumulation of 99mTc-CSA was seen at sites of S. aureus infected rats. The results were compared with 99mTc-Ciprofloxacin.

Enhancement of the efficacy of erythromycin in multiple antibiotic-resistant gram-negative bacterial pathogens

To improve the efficacy of erythromycin, a hydrophobic antibiotic, against multiple antibiotic-resistant gram-negative bacterial pathogens by enhancing their outer membrane permeability. Fifty-one nonrepeat gram-negative bacterial pathogens of various genera, resistant to multiple antibiotics, including erythromycin, were selected by disc agar diffusion tests. The amphiphilic cationic steroid antibiotic, Ceragenin CSA-13, a potent permeabilizer of bacterial outer membranes, reduced the minimum inhibitory concentration of erythromycin in 92% of the bacterial pathogens selected for the test, when supplemented with erythromycin. A synergistic effect of Ceragenin CSA-13 and erythromycin in combination was also observed. Spectrofluorimetric study confirmed that Ceragenin CSA-13 acts by depolarizing the bacterial outer membrane. The toxicity of Ceragenin CSA-13 was evaluated to be insignificant by measuring 'median lethal dose' (LD(50)) on mouse model. Ceragenin CSA-13 may be useful as an agent to make erythromycin effective against infections caused by multiple antibiotic resistant gram-negative bacteria. The outcome of the study suggests erythromycin-Ceragenin combination as a new approach to overcome the problem associated with the rapid emergence of multi-drug-resistant pathogens. The insignificant toxicity of Ceragenin CSA-13, as found, supports the possibility of the application of this compound for human therapeutics.

Minimum requirements of hydrophobic and hydrophilic features in cationic peptide antibiotics (CPAs): pharmacophore generation and validation with cationic steroid antibiotics (CSAs)

Cationic peptide antibiotics (CPAs) are known to possess amphiphilic structure, by virtue of which they display lytic activity against bacterial cell membranes. Naturally occurring antimicrobial peptides contain a large number of amino acid residues, which limits their clinical applicability. Recent studies indicate that it is possible to decrease the chain-length of these peptides without loss of activity, and suggest that a minimum of two positive ionizable (hydrophilic) and two bulky groups (hydrophobic) are required for antimicrobial activity. By employing the HipHop module of the software package CATALYST, we have translated these experimental findings into 3-D pharmacophore models by finding common features among active peptides. Positively ionizable (PI) and hydrophobic (HYD) features are the important characteristics of compounds used for pharmacophore model development. Based on the highest score and the presence of amphiphilic structure, two separate hypothesis, Ec-2 and Sa-6 for Escherichia coli and Staphylococcus aureus, respectively, were selected for mapping analysis of active and inactive peptides against these organisms. The resulting models not only provided information on the minimum requirement of PI and HYD features but also indicated the importance of their relative arrangement in space. The minimum requirement for PI features was two in both cases but the number of HYD features required in the case of E. coli was four while for S. aureus it was found to be three. These hypotheses were able to differentiate between active and inactive CPAs against both organisms and were able to explain the experimental results. The hypotheses were further validated using cationic steroid antibiotics (CSAs), a different class of facial amphiphiles with same mechanism of antimicrobial action as that of CPAs. The results showed that CSAs also require similar minimum features to be active against both E. coli and S. aureus. These studies also indicate that the minimum feature requirements may be conserved for different strains of the same organism. Figure shows the mapping of an active cationic peptide antibiotic (CPA) mapped to the most acceptable hypothesis Sa6 against S. aureus.

Antibacterial Peptides - A Bright Future or a False Hope

The increasing number of antibiotic resistant bacterial strains presents an emerging world health problem that demands continued effort to develop new antibacterial compounds. Endogenous antibacterial peptides (ABPs) that are constitutively and/or inducibly produced in tissues exposed to external surroundings represent new candidates for devel- opment of such compounds. Most ABPs target bacterial membranes initially by electrostatic interactions between posi- tively charged amino acids and negatively charged molecules present on bacterial walls, followed by compromise of the permeability barrier of bacterial membranes through the formation of pores, leading to rapid cell death and efficient bacte- rial elimination. Other mechanisms, such as the inhibition of bacterial protein and DNA synthesis or receptor-mediated stimulation of host defense mechanisms are also ascribed to these molecules. The activities of ABPs correlate positively with a gradient of hydrophobicity along the peptide backbone, net positive charge at neutral pH, and secondary structure. More than 850 sequences with antibacterial activity have been described, and this number continues to grow with the ad- dition of newly discovered, as well as synthetic, peptides. Many strategies, including increasing net positive charge, in- creasing net hydrophobicity, conjugation of peptides with lipophilic acids, incorporation of carbonate bonds, and synthesis of their hybrids and truncated sequences that omit hemolytic regions, have been proposed to increase efficiency of syn- thetic ABPs. Additionally peptide-mimicking molecules such as cationic steroid antibiotics (CSAs) may be useful alterna- tives to natural ABPs. Current challenges for practical application of ABPs are the high cost of synthesis/isolation, inacti- vation by blood plasma, confinement by anionic polyelectrolytes, and possible unknown toxicity.

Cationic steroid antibiotics demonstrate DNA delivery properties

Recently, cationic steroids have been developed that display broad-spectrum antibacterial activity. These compounds, characterized by the presence of several amino groups, present a facially amphiphilic morphology. Formulations containing such steroids were tested for their ability to facilitate the uptake of a reporter plasmid into various cell lines. The results show that, when associated with the naturally occurring zwitterionic lipid dioleoyl-phosphatidylethanolamine (DOPE), cationic steroid antibiotics allow for transfection levels comparable to those obtained with DOTAP. The activity of the amphiphilic mixture was nearly unaffected by bafilomycin A1 and chloroquine treatment, suggesting a mechanism that is independent of the acidification process associated with endocytosis. Collectively, our results show that DNA delivery agents possessing strong antibacterial properties can be obtained by conjugating amino groups to a steroid nucleus.

Expanding the spectrum of activity of mupirocin to include gram-negative bacteria using cationic steroid antibiotics

Expanding the spectrum of activity of mupirocin to include gram-negative bacteria using cationic steroid antibiotics

Cationic steroid antibiotics (CSA) exhibit cytotoxic activity against vaccinia virus (VV)

Cationic steroid antibiotics (CSA) exhibit cytotoxic activity against vaccinia virus (VV)

Origins of Cell Selectivity of Cationic Steroid Antibiotics

A key factor in the potential clinical utility of membrane-active antibiotics is their cell selectivity (i.e., prokaryote over eukaryote). Cationic steroid antibiotics were developed to mimic the lipid A binding character of polymyxin B and are shown to bind lipid A derivatives with affinity greater than that of polymyxin B. The outer membranes of Gram-negative bacteria are comprised primarily of lipid A, and a fluorophore-appended cationic steroid antibiotic displays very high selectivity for Gram-negative bacterial membranes over Gram-positive bacteria and eukaryotic cell membranes. This cell selectivity of cationic steroid antibiotics may be due, in part, to the affinity of these compounds for lipid A.

Synthesis and characterization of peptide-cationic steroid antibiotic conjugates.

[reaction: see text] New cationic steroid antibiotics have been prepared by conjugating tripeptides to a triamino analogue of cholic acid. These compounds were synthesized on a solid phase in an indexed library that was screened for antibacterial activity against Gram-negative and Gram-positive bacteria. Selected compounds were synthesized on a larger scale, and minimum inhibition concentrations were measured. These results correlated very well with the screening of the indexed library of antibiotics. The most active antibiotics demonstrate a marked improvement over a related and well characterized cationic steroid antibiotic.

ChemInform Abstract: Design, Synthesis and Characterization of Cationic Peptide and Steroid Antibiotics

AbstractChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.