In silico studies were conducted on 6-chloro-1H-imidazo[4,5-b]pyridine derivatives to select the best possible drug candidates based on drug properties and bioactivity score of the compounds. 6-chloro-1H-imidazo[4,5-b]pyridine derivatives were subjected to predict bioactivity prediction and drug likeness score on the basis of Lipinski’s rule through molinspiration cheminformatics software. Isoniazid was used as reference standard for comparing the molecular properties and bioactivity score. The results of new 6-chloro-1H-imidazo[4,5-b]pyridine derivatives were compared with Isoniazid to check the prospective of the optimized compounds. The best possible drug candidates were reported after comprehensive analysis on predicted cLogP, solubility, molecular weight, Topological Molecular Polar Surface Area (TPSA), drug- likeness, drug score properties and bioactivity score for different targets like G-protein-coupled Receptors (GPCR), ion channel, kinase, nuclear receptor, protease and enzymes. All the derivatives are found to be biologically important molecules with desirable molecular properties for drug likeness and possess good enzyme inhibitor activity. Thus these derivatives are considered to the best drug candidates for inhibition of mycobacterium tuberculosis synthetase. All the derivatives have not found to violate Lipinsiki’s rule and shows significant bioactivity score when comparing to isoniazid. Thus these derivatives have emerged as a promising anti-tuberculosis lead molecule.