116 research outputs found
Oxidative Polymerization of Dopamine: A High-Definition Multifunctional Coatings for Electrospun Nanofibers - An Overview
The invention that catecholamines undergo oxidative polymerization under alkaline conditions and form adhesive nanocoatings on wide variety of substrates has ushered their potential utility in engineering and biomedical applications. The oxidative polymerization of catecholamines can be triggered by light, chemical and physical methods, thus representing one of the widely explored surface coating methods. The overall objectives of this chapter are to compile the various methods of accomplishing surface coatings and compare the structural diversity of catecholamines. The progress achieved so far on polydopamine (pDA) coatings on electrospun polymers will be discussed. Finally, we will summarize the research efforts on catecholamine coatings for biomedical applications as well as their potential as a high definition coating method
Eggshell Matrix Protein Mimetics: Elucidation of Molecular Mechanism of Goose Eggshell Calcification using Designed Peptides
Model peptides were designed, synthesized and conducted a detailed structure-property study to unravel the molecular mechanism of goose eggshell calcification. The peptides were designed based on the primary structural features of the eggshell matrix proteins ansocalcin and OC-17. In vitro CaCO₃ crystal growth experiments in presence of these peptides showed calcite crystal aggregation as observed in the case of the parent protein ansocalcin. The structure of these peptides in solution was established using intrinsic tryptophan fluorescence studies and quasi-elastic light scattering experiments. The structural features are correlated with observed results of the in vitro crystallization studies.Singapore-MIT Alliance (SMA
Designer peptides to understand the mineralization of calcium salts
Recently, we reported the extraction, purification and amino acid sequence of ansocalcin, the major goose eggshell matrix protein. In vitro studies showed that ansocalcin induces spherical calcite crystal aggregates. We designed two peptides using the unique features of the sequence of ansocalcin and the role of these peptides in CaCO₃ crystallization was investigated. The peptides showed similar activities as compared to ansocalcin, but at a higher concentration. The full characterization of the peptides and a rational for the observed morphology for the calcite crystals are discussed in detail.Singapore-MIT Alliance (SMA
Deposition of Thin Films of Biocompatible Calcium Carbonate via Template-Driven Mineralization
Natural bone is a composite of collagen based hydrogel and inorganic dahilite crystals. The unusual combination of a hard inorganic material and an underlying elastic hydrogel network endows native bone with unique mechanical properties, such as low stiffness, resistance to tensile and compressive forces and high fracture toughness. Throughout the cavities of the bone, there are bone cells and myriads of soluble and extracellular matrix components that are constantly involved in the bone formation and remodeling process. Among the extra cellular component the acidic matrix proteins that are attached to the collagen scaffold play important templating and inhibitory roles during the mineralization process. It would be interesting to generate such functional scaffolds that mimic a template driven mineralization and which can assist cell adhesion, proliferation, migration and differentiation. Towards this direction, we have chosen one synthetic (Nylon 66 membrane) and one natural (eggshell membrane) scaffold and carried out a template driven mineralization of CaCO₃ as model systems. The surface modifications were carried out by the pre-adsorption of acidic polymers before the deposition of the CaCO₃. The deposition of the crystalline calcium carbonate on these modified templates were archived from a supersaturated solution of Ca(HCO₃)₂.Singapore-MIT Alliance (SMA
Strategies in Translating the Therapeutic Potentials of Host Defense Peptides
The golden era of antibiotics, heralded by the discovery of penicillin, has long been challenged by the emergence of antimicrobial resistance (AMR). Host defense peptides (HDPs), previously known as antimicrobial peptides, are emerging as a group of promising antimicrobial candidates for combatting AMR due to their rapid and unique antimicrobial action. Decades of research have advanced our understanding of the relationship between the physicochemical properties of HDPs and their underlying antimicrobial and non-antimicrobial functions, including immunomodulatory, anti-biofilm, and wound healing properties. However, the mission of translating novel HDP-derived molecules from bench to bedside has yet to be fully accomplished, primarily attributed to their intricate structure-activity relationship, toxicity, instability in host and microbial environment, lack of correlation between in vitro and in vivo efficacies, and dwindling interest from large pharmaceutical companies. Based on our previous experience and the expanding knowledge gleaned from the literature, this review aims to summarize the novel strategies that have been employed to enhance the antimicrobial efficacy, proteolytic stability, and cell selectivity, which are all crucial factors for bench-to-bedside translation of HDP-based treatment. Strategies such as residues substitution with natural and/or unnatural amino acids, hybridization, L-to-D heterochiral isomerization, C- and N-terminal modification, cyclization, incorporation with nanoparticles, and “smart design” using artificial intelligence technology, will be discussed. We also provide an overview of HDP-based treatment that are currently in the development pipeline
Host defense peptides at the ocular surface: roles in health and major diseases, and therapeutic potentials.
Sight is arguably the most important sense in human. Being constantly exposed to the environmental stress, irritants and pathogens, the ocular surface - a specialized functional and anatomical unit composed of tear film, conjunctival and corneal epithelium, lacrimal glands, meibomian glands, and nasolacrimal drainage apparatus - serves as a crucial front-line defense of the eye. Host defense peptides (HDPs), also known as antimicrobial peptides, are evolutionarily conserved molecular components of innate immunity that are found in all classes of life. Since the first discovery of lysozyme in 1922, a wide range of HDPs have been identified at the ocular surface. In addition to their antimicrobial activity, HDPs are increasingly recognized for their wide array of biological functions, including anti-biofilm, immunomodulation, wound healing, and anti-cancer properties. In this review, we provide an updated review on: (1) spectrum and expression of HDPs at the ocular surface; (2) participation of HDPs in ocular surface diseases/conditions such as infectious keratitis, conjunctivitis, dry eye disease, keratoconus, allergic eye disease, rosacea keratitis, and post-ocular surgery; (3) HDPs that are currently in the development pipeline for treatment of ocular diseases and infections; and (4) future potential of HDP-based clinical pharmacotherapy for ocular diseases
Teixobactin analogues reveal enduracididine to be non-essential for highly potent antibacterial activity and lipid II binding
Abstract. Teixobactin is a highly promising antibacterial depsipeptide consisting of four D-amino acids and a rare L-allo-enduracididine
amino acid. L-allo-enduracididine is reported to be important for the highly potent antibacterial activity of teixobactin. However, it is also a
key limiting factor in the development of potent teixobactin analogues due to several synthetic challenges such as it is not commercially
available, requires a multistep synthesis, long and repititive couplings (16-30 hours). Due to all these challenges, the total synthesis of
teixobactin is laborious and low yielding (3.3%). In this work, we have identified a unique design and developed a rapid synthesis (10 min
μwave assisted coupling per amino acid, 30 min cyclisation) of several highly potent analogues of teixobactin with yields of 10-24% by
replacing the L-allo-enduracididine with commercially available non-polar residues such as leucine and isoleucine. Most importantly, the
Leu10-teixobactin and Ile10-teixobactin analogues have shown highly potent antibacterial activity against a broader panel of MRSA and
Enterococcus faecalis (VRE). Time-kill kinetics data indicate that both these compounds are superior to vancomycin against MRSA (16
times more potent). Furthermore, these synthetic analogues displayed identical antibacterial activity to natural teixobactin (MIC 0.25 μg/ml)
against MRSA ATCC 33591 despite their simpler design and ease of synthesis. Detailed NMR analyses have provided us with further
insight into the 3D structures of these important analogues. We have confirmed lipid II binding and measured the binding affinities of
individual amino acid residues of Ala10-teixobactin towards geranyl pyrophosphate (a lipid II mimic) by NMR to understand the nature and
strength of binding interactions of the amino acid residues. An antagonization assay further confirms a lipid II mediated mode of action.
Contrary to current understanding, we have shown that a cationic amino acid at position 10 is not essential for target (lipid II) binding and
potent antibacterial activity of teixobactin. We thus provide strong evidence contrary to the many assumptions made about the mechanism of
action of this exciting new antibiotic. Introduction of a non-cationic residue at position 10 allows for tremendous diversification in terms of
the design and synthesis of highly potent teixobactin analogues and lays the foundations for the development of teixobactin analogues as
new drug-like molecules to target MRSA and Mycobacterium tuberculosis
Interaction of gelatin with polyenes modulates antifungal activity and biocompatibility of electrospun fiber mats
Evaluation of Host Defense Peptide (CaD23)-Antibiotic Interaction and Mechanism of Action: Insights from Experimental and Molecular Dynamics Simulations Studies
Background/aim: Host defense peptides (HDPs) have the potential to provide a novel solution to antimicrobial resistance (AMR) in view of their unique and broad-spectrum antimicrobial activities. We had recently developed a novel hybrid HDP based on LL-37 and human beta-defensin-2, named CaD23, which was shown to exhibit good in vivo antimicrobial efficacy against Staphylococcus aureus in a bacterial keratitis murine model. This study aimed to examine the potential CaD23-antibiotic synergism and the secondary structure and underlying mechanism of action of CaD23. Methods: Peptide-antibiotic interaction was evaluated against S. aureus, methicillin resistant S. aureus (MRSA), and Pseudomonas aeruginosa using established checkerboard and time-kill assays. Fractional inhibitory concentration index (FICI) was calculated and interpreted as synergistic (FIC1.0 and £4), or antagonistic (FIC>4). SYTOX green uptake assay was performed to determine the membrane-permeabilising action of CaD23. Molecular dynamics (MD) simulations were performed to evaluate the interaction of CaD23 with bacterial and mammalian mimetic membranes. Circular dichroism (CD) spectroscopy was also performed to examine the secondary structures of CaD23. Results: CaD23-amikacin and CaD23-levofloxacin combination treatment exhibited a strong additive effect against S. aureus SH1000 (FICI=0.60-0.69) and MRSA43300 (FICI=0.56- 0.60) but an indifferent effect against P. aeruginosa (FIC=1.03-1.15). CaD23 (at 25μg/ml; 2xMIC) completely killed S. aureus within 30 mins. When used at sub-MIC concentration (3.1μg/ml; 0.25xMIC), it was able to expedite the antimicrobial action of amikacin against S. aureus by 50%. The rapid antimicrobial action of CaD23 was attributed to the underlying membrane-permeabilising mechanism of action, evidenced by the SYTOX green uptake assay and MD simulations studies. MD simulations revealed that cationicity, alpha-helicity, amphiphilicity and hydrophobicity (related to the Trp residue at C-terminal) play important roles in the antimicrobial action of CaD23. The secondary structures of CaD23 observed in MD simulations were validated by CD spectroscopy. Conclusions: CaD23 is a novel alpha-helical, membrane-active synthetic HDP that can enhance and expedite the antimicrobial action of antibiotics against Gram-positive bacteria when used in combination. MD simulations serves as a powerful tool in revealing the peptide secondary structure, dissecting the mechanism of action, and guiding the design and optimisation of HDPs
Cysteines and Disulfide-Bridged Macrocyclic Mimics of Teixobactin Analogues and Their Antibacterial Activity Evaluation against Methicillin-Resistant Staphylococcus Aureus (MRSA)
Teixobactin is a highly potent cyclic depsipeptide which kills a broad range of multi-drug resistant, Gram-positive bacteria, such as Methicillin-resistant Staphylococcus aureus (MRSA) without detectable resistance. In this work, we describe the design and rapid synthesis of novel teixobactin analogues containing two cysteine moieties, and the corresponding disulfide-bridged cyclic analogues. These analogues differ from previously reported analogues, such as an Arg10-teixobactin, in terms of their macrocyclic ring size, and feature a disulfide bridge instead of an ester linkage. The new teixobactin analogues were screened against Methicillin-resistant Staphylococcus aureus and Methicillin-sensitive Staphylococcus aureus. Interestingly, one teixobactin analogue containing all l-amino acid building blocks showed antibacterial activity against MRSA for the first time. Our data indicates that macrocyclisation of teixobactin analogues with disulfide bridging is important for improved antibacterial activity. In our work, we have demonstrated the unprecedented use of a disulfide bridge in constructing the macrocyclic ring of teixobactin analogues
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