In the search to find new therapeutically solutions for the tuberculosis treatment, we designed and synthesized several thiourea derivatives. We evaluated the effect on various strains of Mycobacterium tuberculosis and the acute toxicity on mice. We also performed an in silico molecular docking study in order to analyse possible interactions with certain bacterial molecular targets. All tested compounds showed a good tuberculostatic effect, no acute toxicity in mice and optimal predicted protein-ligand complex conformations, being promising lead molecules for the development of future thiourea derivatives.