Alice McCloskey

Ultrashort Cationic Naphthalene-Derived Self-Assembled Peptides as Antimicrobial Nanomaterials

Self-assembling dipeptides conjugated to naphthalene show considerable promise as nanomaterial structures, biomaterials, and drug delivery devices. Biomaterial infections are responsible for high rates of patient mortality and morbidity. The presence of biofilm bacteria, which thrive on implant surfaces, are a huge burden on healthcare budgets, as they are highly resistant to current therapeutic strategies. Ultrashort cationic self-assembled peptides represent a highly innovative and cost-effective strategy to form antibacterial nanomaterials. Lysine conjugated variants display the greatest potency with 2% w/v NapFFKK hydrogels significantly reducing the viable Staphylococcus epidermidis biofilm by 94%. Reducing the size of the R-group methylene chain on cationic moieties resulted in reduction of antibiofilm activity. The primary amine of the protruding R-group tail may not be as readily available to interact with negatively charged bacterial membranes. Cryo-SEM, FTIR, CD spectroscopy, and oscillatory rheology provided evidence of supramolecular hydrogel formation at physiological pH (pH 7.4). Cytotoxicity assays against murine fibroblast (NCTC 929) cell lines confirmed the gels possessed reduced cytotoxicity relative to bacterial cells, with limited hemolysis upon exposure to equine erythrocytes. The results presented in this paper highlight the significant potential of ultrashort cationic naphthalene peptides as future biomaterials.

Evolution of antimicrobial peptides to self-assembled peptides for biomaterial applications.

Biomaterial-related infections are a persistent burden on patient health, recovery, mortality and healthcare budgets. Self-assembled antimicrobial peptides have evolved from the area of antimicrobial peptides. Peptides serve as important weapons in nature, and increasingly medicine, for combating microbial infection and biofilms. Self-assembled peptides harness a “bottom-up” approach, whereby the primary peptide sequence may be modified with natural and unnatural amino acids to produce an inherently antimicrobial hydrogel. Gelation may be tailored to occur in the presence of physiological and infective indicators (e.g. pH, enzymes) and therefore allow local, targeted antimicrobial therapy at the site of infection. Peptides demonstrate inherent biocompatibility, antimicrobial activity, biodegradability and numerous functional groups. They are therefore prime candidates for the production of polymeric molecules that have the potential to be conjugated to biomaterials with precision. Non-native chemistries and functional groups are easily incorporated into the peptide backbone allowing peptide hydrogels to be tailored to specific functional requirements. This article reviews an area of increasing interest, namely self-assembled peptides and their potential therapeutic applications as innovative hydrogels and biomaterials in the prevention of biofilm-related infection.

Peptide Therapeutics and the Pharmaceutical Industry: Barriers Encountered Translating from the Laboratory to Patients

Peptides are receiving increasing interest as clinical therapeutics. These highly tunable molecules can be tailored to achieve desirable biocompatibility and biodegradability with simultaneously selective and potent therapeutic effects. Despite challenges regarding up-scaling and licensing of peptide products, their vast clinical potential is reflected in the 60 plus peptide-based therapeutics already on the market, and the further 500 derivatives currently in developmental stages. Peptides are proving effective for a multitude of disease states including: type 2 diabetes (controlled using the licensed glucagon-like peptide-1 receptor liraglutide); irritable bowel syndrome managed with linaclotide (currently at approval stages); acromegaly (treated with octapeptide somatostatin analogues lanreotide and octreotide); selective or broad spectrum microbicidal agents such as the Gram-positive selective PTP-7 and antifungal heliomicin; anticancer agents including goserelin used as either adjuvant or monotherapy for prostate and breast cancer, and the first marketed peptide derived vaccine against prostate cancer, sipuleucel-T. Research is also focusing on improving the biostability of peptides. This is achieved through a number of mechanisms ranging from replacement of naturally occurring L-amino acid enantiomers with D-amino acid forms, lipidation, peptidomimetics, N-methylation, cyclization and exploitation of carrier systems. The development of self-assembling peptides are paving the way for sustained release peptide formulations and already two such licensed examples exist, lanreotide and octreotide. The versatility and tunability of peptide-based products is resulting in increased translation of peptide therapies, however significant challenges remain with regard to their wider implementation. This review highlights some of the notable peptide therapeutics discovered to date and the difficulties encountered by the pharmaceutical industry in translating these molecules to the clinical setting for patient benefit, providing some possible solutions to the most challenging barriers.

Potential strategies for the eradication of multidrug-resistant Gram-negative bacterial infections

Antimicrobial resistance is one of the leading threats to society. The increasing burden of multidrug-resistant Gram-negative infection is particularly concerning as such bacteria are demonstrating resistance to nearly all currently licensed therapies. Various strategies have been hypothesized to treat multidrug-resistant Gram-negative infections including: targeting the Gram-negative outer membrane; neutralization of lipopolysaccharide; inhibition of bacterial efflux pumps and prevention of protein folding. Silver and silver nanoparticles, fusogenic liposomes and nanotubes are potential strategies for extending the activity of licensed, Gram-positive selective, antibiotics to Gram-negatives. This may serve as a strategy to fill the current void in pharmaceutical development in the short term. This review outlines the most promising strategies that could be implemented to solve the threat of multidrug-resistant Gram-negative infections.

Anti-biofilm activity of ultrashort cinnamic acid peptide derivatives against medical device-related pathogens.

The threat of antimicrobial resistance has placed increasing emphasis on the development of innovative approaches to eradicate multidrug‐resistant pathogens. Biofilm‐forming microorganisms, for example, Staphylococcus epidermidis and Staphylococcus aureus, are responsible for increased incidence of biomaterial infection, extended hospital stays and patient morbidity and mortality. This paper highlights the potential of ultrashort tetra‐peptide conjugated to hydrophobic cinnamic acid derivatives. These peptidomimetic molecules demonstrate selective and highly potent activity against resistant biofilm forms of Gram‐positive medical device‐related pathogens. 3‐(4‐Hydroxyphenyl)propionic)‐Orn‐Orn‐Trp‐Trp‐NH2 displays particular promise with minimum biofilm eradication concentration (MBEC) values of 125 µg/ml against methicillin sensitive (ATCC 29213) and resistant (ATCC 43300) S. aureus and activity shown against biofilm forms of Escherichia coli (MBEC: 1000 µg/ml). Kill kinetics confirms complete eradication of established 24‐h biofilms at MBEC with 6‐h exposure. Reduced cell cytotoxicity, relative to Gram‐positive pathogens, was proven via tissue culture (HaCaT) and haemolysis assays (equine erythrocytes).

Self-assembling ultrashort NSAID-peptide nanosponges: multifunctional antimicrobial and anti-inflammatory materials

Peptide-based materials are receiving significant attention for use within biomedicine due to their high chemical and functional versatility enabling tailoring of their structure to replicate the properties of host tissue and the extracellular matrix. This paper studies the design, synthesis and characterization of NSAID-peptide conjugates. Attachment of NSAIDs to a diphenylalanine–dilysine (FFKK-OH) peptide sequence generates supramolecular hydrogel forming molecules with antimicrobial and anti-inflammatory properties. NSAID-peptides demonstrate broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria implicated in a variety of antimicrobial resistant nosocomial infections including Staphylococcus aureus and Pseudomonas aeruginosa. Naproxen-peptides show particular promise, forming biocompatible nanofibrous viscoelastic hydrogel nanosponges composed of β-sheet secondary structures at low concentrations (0.4% w/v). Conjugation of the peptide motif FFKK-OH to naproxen increases selectivity for COX-2 enzyme, implicated in chronic wound scar-tissue formation. Our findings suggest that ultrashort NSAID-peptides have potential use as multifunctional materials for a range of biomedical applications. This includes as topical agents for treatment of chronic wounds, where a profile of persistent inflammation, pain and the presence of infection has been proven to be detrimental to successful wound repair. This work may also serve as a template for the design of future medical device coatings with tailored antimicrobial and anti-inflammatory properties.

Ultrashort self-assembling Fmoc-peptide gelators for anti-infective biomaterial applications

Biomaterial‐related infections have a significant impact on society and are a major contributor to the growing threat of antimicrobial resistance. Current licensed antibiotic classes struggle to breakdown or penetrate the exopolysaccharide biofilm barrier, resulting in sub‐therapeutic concentrations of antibiotic at the surface of the biomaterial, treatment failure and increased spread of resistant isolates. This paper focuses for the first time on the ability of ultrashort Fmoc‐peptide gelators to eradicate established bacterial biofilms implicated in a variety of medical device infections (Gram‐positive: Staphylococcus aureus, Staphylococcus epidermidis and Gram‐negative Escherichia coli, Pseudomonas aeruginosa). The effect of increasing the cationicity of FmocFF via addition of di‐lysine and di‐orntithine was also studied with regard to antibacterial activity. Our studies demonstrated that Fmoc‐peptides (FmocFF, FmocFFKK, FmocFFFKK, FmocFFOO) formed surfactant‐like soft gels at concentrations of 1% w/v and above using a method of glucono‐δ‐Lactone pH induction. The majority of Fmoc‐peptides (0.5–2% w/v) demonstrated selective action against established (grown for 24 h) biofilms of Gram‐positive and Gram‐negative pathogens. These results are likely to increase the clinical translation of short‐peptide gelator platforms within the area of anti‐infective biomaterials including as wound dressings and coatings for prostheses, catheters, heart valves and surgical tubes. In the long term, this will lead to wider treatment choices for clinicians and patients involved in the management of medical device infections and reduce the burden of antimicrobial resistance. Copyright