In the present study, a series of substituted 1,3,4-thiadiazole derivatives 4(a-o), 5(a-m) and 6(a-j) were synthesized and characterized by IR, 1H NMR, 13C NMR and mass spectroscopic technique. The synthesized compounds were evaluated for their in vitro anti-mycobacterial activity against the Mycobacterium tuberculosis H37Rv and resistance MDR-TB strain. Among the compounds tested N-(5-(4-nitrophenyl)-1,3,4-thiadiazol-2-yl)furan-2-carboxamide (4h) showed significant inhibitory activity with MIC of 9.87 μM (H37Rv strain) and 9.87 μM (MDR-TB strain) compared to isoniazide [MIC of 3.64 μM (H37Rv) and >200 μM (MDR-TB strain)] and rifampin [MIC of 0.152 μM (H37Rv) and 128 μM (MDR-TB strain)]. In addition, these compounds have also been assessed for their cyto-toxicity to a mammalian Vero cell line using the MTT assay. The result shows that these compounds exhibit anti-tubercular activity at non-cytototoxic concentrations.