Development of new antimicrobial peptides and peptidomimetics and mechanism of resistance to peptide antibiotics
Nowadays in the world there is a very big problem associated with two factors related to each other. The first factor is the increase in the resistant of certain bacteria, especially the bacteria from the ESKPAE group. The second factor is the dramatically decrease of new antibiotics approved by the...
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| Tipo de recurso: | tesis doctoral |
| Estado: | Versión publicada |
| Fecha de publicación: | 2014 |
| País: | España |
| Institución: | CBUC, CESCA |
| Repositorio: | TDR. Tesis Doctorales en Red |
| OAI Identifier: | oai:www.tdx.cat:10803/285375 |
| Acceso en línea: | http://hdl.handle.net/10803/285375 |
| Access Level: | acceso abierto |
| Palabra clave: | Antibiòtics Antibióticos Antibiotics Pèptids Péptidos Peptides Resistència als medicaments Resistencia a los medicamentos Drug resistance Bacteris Bacterias Bacteria Ciències Experimentals i Matemàtiques 547 |
| Sumario: | Nowadays in the world there is a very big problem associated with two factors related to each other. The first factor is the increase in the resistant of certain bacteria, especially the bacteria from the ESKPAE group. The second factor is the dramatically decrease of new antibiotics approved by the FDA. These two problems show that there is an urgent need to find new antibiotics active against these resistant bacteria. In this thesis, we have tackled two different topics closely related in the race to find new antimicrobials. The first topic tackled was the knowledge of the mechanism of resistance of Gram-negative (A. nosocomialis) and Gram-positive (S. mitis) bacteria. The two antibiotics studied were peptides, colistin and daptomicin, these two peptides are resistant to A. nosocomialis and S. mitis, respectively. Both peptides had a similar mechanism of action related to the membrane of bacteria, therefore we are going to focus just in the modifications in the membrane of the strains resistant to the antibiotic peptides. In S. mitis it was observed, using proteomic techniques, that two proteins related with the membrane were observed. These two proteins has some homologue domains to several proteins involved in daptomycin resistant in S. aureus and Enterococci. In A. nosocomialis, the bacteria showed a high tolerance to colistin, and at 8 mg/L an inflexion point is observed. In this inflexion point, the MIC of colistin, against bacteria increase from <0.1 mg/L to 128 mg/L. These bacteria with high resistance to colistin showed no production of LPS due to the fact that mutations and a stop codon in lpxD gene were observed. This gene is involved in the synthetic pathway of the LPS. Apart from the understanding of the mechanism of action of peptide antibiotics, we have proposed several peptides and peptidomimetics against Acinetobacter species. We have used two different approaches. The first approach is the normal approach, testing several peptides or peptidomimetics against the desired bacteria. The first peptides tested were commercially available, and we found mastoparan that was active against both colistin-susceptible and colistin-resistant A. baumannii. This peptide was optimized specially in terms of stability in human serum. After several in vivo trials we did not observe any activity of the peptides tested, however we found a very strong bindoing with some proteins present in the human serum. Frog skin secretions peptides were also tested against colistin-susceptible and colistin-resistant Acinetobacter species, the results obtained were really interesting specially in two peptides. The last peptides tested were peptidomimetics. These peptidomimetics act as an antimicrobial peptide, with two different faces, one face with a cation charge and the other very amphipathic. These peptidomimetics are analogues from the original structure of cholic acid, the structure was modified in order to have antibacterial activity that was found in colistin-susceptible and colistin-resistant A. baumannii, K. pneumonia and P. aeruginosa. The second approach was completely different, in this case the idea was to block the virulence of bacteria caused by OmpA. This protein is involved in the adherence between bacteria and host cells, therefore several hexacylcic peptides were synthesized in order to inhibit the action of this protein. The results obtained were satisfactory, obtaining good activity in both in vitro and in vivo. |
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