A new series of oxadiazole analogues was synthesized starting from 2aminopyridine. The compounds were characterized by infrared (IR), nuclear magnetic resonance (NMR), and mass spectral analyses followed by determination of their anticancer and antimicrobial activities. Three compounds were tested for in vitro anticancer activity against NCI-60 human cell lines of nine different panels including leukemia, non-small lung cancer, colon cancer, CNS cancer, melanoma, ovarian cancer, renal cancer, prostate cancer, and breast cancer according to the National Cancer Institute (NCI, USA) Protocol at 10 µM. The compounds N-{[5-(4chlorophenyl)-1,3,4-oxadiazol-2-yl]methyl}pyridin-2-amine (5c), N-{[5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl]methyl}pyridin-2-amine (5f), and N-{[5-(3,4-dimethoxyphenyl)-1,3,4-oxadiazol-2-yl]methyl}pyridin-2-amine (5g) showed anticancer with higher selectivity towards HOP-92 (Non-Small Cell Lung Cancer). N-{[5-(4-Fluorophenyl)-1,3,4-oxadiazol-2-yl]methyl}pyridin-2-amine (5b) showed maximum antibacterial activity with minimum inhibitory concentration (MIC) of 4-8 µg/mL, while N-{[5-(4-methoxyphenyl)-1,3,4-oxadiazol-2-yl]methyl}pyridin-2-amine (5f) showed maximum antifungal activity with MIC 4 µg/mL.