LD50 (mouse, i.p.) = 191mg/kg

The cellular viability for a macrophage cell line J774 was determined by Mosmans´s MTT microcultured tetrazolium assay. We evaluated non-infected or infected macrophages with Mycobaterium bovis Bacillus Calmette-Guerin (BCG) in the presence and absence of test compounds (1,12-Dodecanediamine). The cells were plated in flat bottom 96 well plates (2.5 10^6 cells/well/100 mkL) cultured for 24 h in a controlled atmosphere (CO2 5% at 37 C), and non-adherent cells were washed by gentle flushing with RPMI 1640 supplemented with fetal bovine serum (10%) and gentamicin (25 lg/mL). Adherent cells were infected or not with BCG (2.5 10^6 UFC/well/100 mkL) cultured in the presence of medium alone, tween 20 (3%) (live and dead controls, respectively) or different concentrations of compounds (1.0, 10.0 and 100 mkg/mL) in a triplicate assay. After 48 h, stock MTT solution (5 mg/mL of saline; 20 mL/well) was added to the culture and 4 h later, the plate was centrifugate for 2 min at 2800 rpm, supernatant was discharged and Dimethyl sulfoxide (DMSO) (100 mkL/well) was added for formazan crystals solubilization and the absorbance was read at 540 nm in a plate reader.