The high incidence of antimicrobial-resistant (AMR) infections in recent decades has made the development of novel antimicrobial medications one of the medicinal chemists' top priorities. In this study, synthesis, in vitro antimicrobial activity, electrochemical studies, in silico pharmacokinetic ADME parameters, molecular docking, and molecular dynamic simulations were all performed on several 1,3,4-oxadiazole derivatives. The minimum inhibitory concentrations (MIC) of these compounds were evaluated against eleven species of gram-positive bacteria, gram-negative bacteria and fungal pathogens using azithromycin as the reference antibacterial agent, voriconazole and fluconazole as the reference antifungal agents. Both compounds 4d and 4f were found to be approximately as effective as azithromycin against E. faecalis and E. coli, respectively while three compounds showed antifungal activity against C. parapsilopsis, with MIC values that were identical to fluconazole for 4g and 4i, and to voriconazole for 4j. Also, the most potent derivatives-double stranded DNA (dsDNA) interactions were evaluated using an electrochemical technique. Compounds 4d, 4f and 4i were found to disrupt the structure of dsDNA however compound 4h wasn't able to bind to dsDNA. In addition, the structure-activity relationship (SAR) of compounds 4f and 4d were elucidated by molecular modeling studies.