Effect of Different Drug and Antibacterial Peptide Combinations Against 'M. Tuberculosis' Using In Vitro and Ex Vivo Techniques. Analysis in Exponential Phase and Latency

Duration

01/01/2017 - 30/03/2017

Funded by

Ministerio de Ciencia, Innovación y Universidades. Instituto de Salud Juan Carlos III. Unión Europea. Este proyecto está cofinanciado por el Fondo Europeo de Desarrollo Regional (FEDER). “Una manera de hacer Europa”

Objectives

1. Determine the synergistic activity of in vitro methods and the efficacy using a human macrophage model (THP-1 cell line) of the following combinations: Levofloxacin+clofazimine+PA-824; quinolone UB-8902 +clofazimine+PA-824; moxifloxacin+clofazimine+PA-8242.
2. Determine the MIC of a collection of 14 antimicrobial peptides against M tuberculosis.
3. Study the activity of the peptide and drug combinations of objectives 1 and 2 against clinical strains of M tuberculosis in latency.

Methods

We will use 10 multiresistant and 10 susceptible clinical strains of M. tuberculosis. The MIC of individual drugs will be determined using solid culture medium 7H11. The efficacy of the proposed combinations will be analyzed with the chequerboard method using solid culture medium and with the killing curves method, using liquid (7H9) and solid (7H11) culture medium.

Moreover, the combinations will be analyzed in a human macrophage model (THP-1 cell line). In this model the intracellular drug concentrations will be determined. On the other part, the MIC of a collection of 14 antimicrobial peptides will be analyzed, using liquid medium 7H9 and alamar blue reading.

Lastly, the effect of drugs alone and in combination and also the antimicrobial peptides, will be studied in a latency model based on nutrient starvation using PBS as culture medium.

Total Funding

69,575.00 euros

Project Code

PI13/01752