Ten novel 1,3,4-oxadiazole analogs (A-J) were prepared and tested in terms of their antibacterial and antioxidant properties. The obtained compounds were identified by the determination of melting points, thin-layer chromatography, and the confirmation of molecular formulas, whereas the identification of representative compounds proceeded by adding the UV- Visible spectroscopy, Fourier-transform infrared spectroscopy (FT-IR), and nuclear magnetic resonance (¹H- and ¹C-NMR) methods. The spectral analyses proved the successful construction of the oxadiazole nucleus and the existence of the typical functional groups. The synthesized derivatives were evaluated with respect to the Gram-negative bacteria (Escherichia coli and Klebsiella pneumoniae) and Gram-positive bacterium (Staphylococcus aureus and Micrococcus luteus) regarding their antibacterial activity based on the agar diffusion test with 100 µg/mL, 500 µg/mL, and 1000 µg/mL concentrations of the derivatives. In a concentration-dependent fashion, several compounds showed a significant level of antibacterial activity. Out of them, methyl-substituted (B) and chloro-substituted (D) counterparts exhibited similar inhibition zones as the standard drug ciprofloxacin. The DPPH free radical scavenging assay was used to determine the antioxidant activity. These findings revealed moderate-strong antioxidant potential of most compounds, with some halogenated derivatives having lower IC50 values compared to ascorbic acid. Possible structure-activity relationship analysis showed that electron-withdrawing and electron-donating substituents have a significant impact on biological activity. All in all, the results demonstrate substituted 1,3,4-oxadiazole derivatives as promising candidates in the further development as antimicrobial and antioxidant agents.