RESUMO
Cpl-1, an endolysin derived from Cp-1 phage has been found to be effective in a number of in-vitro and in-vivo pneumococcal infection models. However its lower bioavailability under in-vivo conditions limits its applicability as therapeutic agent. In this study, Cpl-1 loaded chitosan nanoparticles were set up in order to develop a novel therapeutic delivery system to counter antibiotic resistant S. pneumoniae infections. Interactions of chitosan and Cpl-1 were studied by in-silico docking analysis. Chitosan nanoparticles and Cpl-1 loaded chitosan nanoparticles were prepared by using ionic gelation method and the process was optimized by varying chitosan:TPP ratio, pH, stirring time, stirring rate and Cpl-1 concentration. Chitosan nanoparticles and Cpl-1 loaded chitosan nanoparticles were characterized to ascertain successful formation of nanoparticles and entrapment of Cpl-1 into nanoparticles. Chitosan nanoparticles and Cpl-1 loaded nanoparticles were also evaluated for nanoparticle yield, entrapment efficiency, in-vitro release, stability, structural integrity of Cpl-1, in-vitro bioassay, swelling studies, in-vitro biodegradation and heamolysis studies. Mucoadhesion behavior of chitosan nanoparticles and Cpl-1 loaded nanoparticles was explored using mucous glycoprotein assay and ex-vivo mucoadhesion assay, both preparations exhibited their mucoadhesive nature. Cellular cytotoxicity and immune stimulation studies revealed biocompatible nature of nanoparticles. The results of this study confirm that chitosan nanoparticles are a promising biocompatible candidate for Cpl-1 delivery with a significant potential to increase bioavailability of enzyme that in turn can increase its in-vivo half life to treat S. pneumoniae infections.
Assuntos
Portadores de Fármacos/química , Composição de Medicamentos/métodos , Endopeptidases/administração & dosagem , Nanopartículas/química , Pneumonia Pneumocócica/tratamento farmacológico , Proteínas Virais/administração & dosagem , Células A549 , Administração Intranasal , Animais , Bacteriófagos/enzimologia , Disponibilidade Biológica , Quitosana/química , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/farmacocinética , Avaliação Pré-Clínica de Medicamentos , Liberação Controlada de Fármacos , Endopeptidases/química , Endopeptidases/isolamento & purificação , Endopeptidases/farmacocinética , Estudos de Viabilidade , Meia-Vida , Humanos , Masculino , Teste de Materiais , Camundongos , Simulação de Acoplamento Molecular , Pneumonia Pneumocócica/imunologia , Pneumonia Pneumocócica/microbiologia , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/farmacocinética , Streptococcus pneumoniae/isolamento & purificação , Streptococcus pneumoniae/virologia , Proteínas Virais/química , Proteínas Virais/isolamento & purificação , Proteínas Virais/farmacocinéticaRESUMO
Antimicrobial resistance in bacterial respiratory tract pathogens is a rapidly evolving and increasingly disconcerting problem. Major factors that have contributed to resistance are inappropriate prescribing of antibiotics for viral infections and the use of antibiotics with poor activity. The treatment of respiratory tract infections is significantly affected by resistance in organisms such as Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Resistance to beta-lactams, sulfonamides, and macrolides continues to rise. Evidence-based guidelines, founded on clinical and bacteriological outcomes, are imperative to treat patients effectively, to limit the spread of these pathogens, and to minimize further development of resistance. Pharmacokinetic and pharmacodynamic parameters have recently been shown to correlate with clinical outcome, and offer a more rational approach to predicting antimicrobial efficacy and determining clinically relevant susceptibility breakpoints.