A new series of 2-(3-Arylureido) benzimidazole derivatives (5a-j) were synthesized via sequential oxidative cyclisation of o-phenelyne diamine 1and 4-nitrobenzaldehyde 2, reduction followed by the reaction of resulting amine 3with different arylisocynates 4. All the synthesized compounds were screened for their in-vitro proinflammatory cytokines TNF-α and IL-6 inhibition and antimicrobial activity (antibacterial and antifungal). The compounds 5c, 5e and 5g found to be potent TNF-α and IL-6 inhibitor as compared to the standard dexamethasone but at the MIC of 10 μM, while the compounds 5d found to be moderately active as compared to standard dexamethasone. The remaining compounds were found to have low, very low or no activity at all at the MIC of 10 μM. The antimicrobial activity data revealed that compounds 5b, 5f, and 5g found to be potent antibacterial and antifungal agents. Notably, the compounds 5f and 5g, exhibited 1.5-2.5 fold antibacterial and antifungal activity to that of control drugs Micanazole and fluconazole almost against all the gram positive and gram negative bacteria and fungi and thus found to be more potent even than the standard drugs.