As potential antimicrobial agents, 4-fluoroaniline and diethyl (ethoxymethylene)malonate were used to efficiently synthesize a new quinolone (3-9) series. The (HNMR)-H-1 and C-13 NMR, IR, mass spectrometry, and elemental analysis of all compounds were used to identify them. The final compounds 3-9 were in vitro screened for antimicrobial activity against gram negative-gram positive Escherichia coli, Yersinia pseudotuberculosis, Pseudomonas aeruginosa, Staphylococcus aureus, Enterococcus faecalis, Bacillus cereus, Mycobacterium smegmatis and fungal strains Candida albicans, Saccharomyces cerevisiae. The best antimicrobial activity was demonstrated by compounds 9a and 9b. Antimicrobial resistance and microbial infection could benefit from quinolone derivatives ' ability to inhibit the growth of microorganisms, according to this study.