A series of substituted N-(quinolin-4-yl)ethanediamine phenyl urea derivatives of biological interest were prepared by sequential quinoline synthesis, chlorination, and substitution reaction followed by reaction of resulting amine with different aryl isocyanates. All synthesized compounds (1–13) were screened for their pro-inflammatory cytokines (TNF-a and IL-6) and antimicrobial activity (antibacterial and antifungal). Biological activity evaluation study revealed that among all the compounds screened, compounds 4 and 6 were found to have promising anti-inflammatory activity (up to 78–71 % TNF-a and 96–90 % IL-6 inhibitory activity) at a higher concentration of 10 lM with reference to standard dexamethasone (72 % TNF-a and 86 % IL-6 inhibitory activities at 1 lM). Compounds 6, 8, 10, and 11 overall exhibited promising antimicrobial activity at MIC values ranging from 10 to 30 lg/mL against all the selected pathogenic bacteria and fungi.