RESUMO
The synthetic antimicrobial peptide SET-M33 has strong activity against bacterial infections caused by Gram-negative bacteria. It is currently in preclinical development as a new drug to treat lung infections caused by Gram-negative bacteria. Here we report its strong anti-inflammatory activity in terms of reduced expression of a number of cytokines, enzymes, and signal transduction factors involved in inflammation triggered by LPS from Pseudomonas aeruginosa, Klebsiella pneumoniae, and Escherichia coli Sixteen cytokines and other major agents involved in inflammation were analyzed in macrophages and bronchial cells after stimulation with LPS and incubation with SET-M33. The bronchial cells were obtained from a cystic fibrosis patient. A number of these proteins showed up to 100% reduction in expression as measured by RT-PCR, Western blotting, or Luminex technology. LPS neutralization was also demonstrated in vivo by challenging bronchoalveolar lavage of SET-M33-treated mice with LPS, which led to a sharp reduction in TNF-α with respect to non-SET-M33-treated animals. We also describe a strong activity of SET-M33 in stimulating cell migration of keratinocytes in wound healing experiments in vitro, demonstrating a powerful immunomodulatory action generally characteristic of molecules taking part in innate immunity.
Assuntos
Anti-Inflamatórios/farmacologia , Brônquios/metabolismo , Fibrose Cística/metabolismo , Fatores Imunológicos/farmacologia , Lipopolissacarídeos/toxicidade , Animais , Fibrose Cística/genética , Fibrose Cística/patologia , Citocinas/metabolismo , Humanos , Camundongos , Células RAW 264.7RESUMO
A synthetic antimicrobial peptide was identified as a possible candidate for the development of a new antibacterial drug. The peptide, SET-M33L, showed a MIC90 below 1.5 µM and 3 µM for Pseudomonas aeruginosa and Klebsiella pneumoniae, respectively. In in vivo models of P. aeruginosa infections, the peptide and its pegylated form (SET-M33L-PEG) enabled a survival percentage of 60-80% in sepsis and lung infections when injected twice i.v. at 5 mg/Kg, and completely healed skin infections when administered topically. Plasma clearance showed different kinetics for SET-M33L and SET-M33L-PEG, the latter having greater persistence two hours after injection. Bio-distribution in organs did not show significant differences in uptake of the two peptides. Unlike colistin, SET-M33L did not select resistant mutants in bacterial cultures and also proved non genotoxic and to have much lower in vivo toxicity than antimicrobial peptides already used in clinical practice. The characterizations reported here are part of a preclinical development plan that should bring the molecule to clinical trial in the next few years.
Assuntos
Antibacterianos/uso terapêutico , Peptídeos Catiônicos Antimicrobianos/uso terapêutico , Klebsiella pneumoniae/fisiologia , Pneumonia/tratamento farmacológico , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/fisiologia , Sepse/tratamento farmacológico , Pele/efeitos dos fármacos , Animais , Peptídeos Catiônicos Antimicrobianos/química , Colistina/uso terapêutico , Modelos Animais de Doenças , Farmacorresistência Bacteriana , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Polietilenoglicóis/química , Pele/microbiologia , Pele/patologiaRESUMO
The increasing frequency of multidrug-resistant bacteria and a recent slowing in the development of new antimicrobial agents place mankind in a state of emergency with regard to the threat of new bacterial infections. Antibacterial peptides (AMPs) are considered an important class of molecules to develop against bacteria. AMPs have been known for many years but very few have yet been extensively used in clinical practice, mainly because of their general toxicity and manufacturing cost. Now, thanks to new technologies for screening and development, interest in these molecules has grown. Many new AMPs have been discovered and some are under evaluation for the development of new antibacterial therapeutics. Here we review the major AMPs currently used in clinical practice and others in the phase of preclinical and clinical development.