A series of new nicotinic acid hydrazones were synthesized by the condensation of compound (2) with different aromatic/heteroaromatic aldehydes in acidic condition. Compound (2) was obtained by the reaction of nicotinic acid hydrazide with 4-aminoacetophenone under reflux condition and characterized by the Infra-Red (IR), Proton Nuclear Magnetic Resonance (1H-NMR), Mass spectral and elemental analysis. All the title compounds were screened for antimicrobial activity (Five strains: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Candida albicans), in silico and molecular docking studies. The results revealed that compounds 3f, 3d and 3g exhibited 3.12 μg/ml, 6.25 μg/ml and 12.5 μg/ml as Minimum Inhibitory Concentration (MIC) towards E. coli, P. aeruginosa, C. albicans strains respectively, which is comparable to standard (3.12 and 6.25 μg/ml as MIC respectively) and also showed good docking score (around-6.50) than standard ciprofloxacin (-4.74) predicted by XP GLIDE module of Schrodinger suite against FAB protein. All the derivatives obeys Lipinski rule of five and also compound 3c have higher Pa value towards anti-tuberculosis predicted by Prediction of Activity Spectrum of Substances (PASS) online tool.