Oxadiazole is heterocyclic compound having 2 nitrogen and 1 oxygen atom together to form a heterocyclic ring with five members. The current research was based on the design, synthesis and evaluation of anti-fungal activity of novel derivatives of 1,3,4-oxadiazole. The 1,3,4-oxadiazole derivatives based on physicochemical properties i.e., melting point, Rf value, FTIR, NMR and Mass spectroscopy and evaluated for the anti-fungal potential through well diffusion method against fungus strains i.e., Aspergillus niger, Penicillium notatum, Candida albicans and Rhizopus species. In results, the highest Rf values in C2, C3, C4, and C6 were found to be 0.73, 0.72, 0.73, and 0.71, respectively. The compound C4 exhibited the highest melting point, measuring between 184 and 188ºC. Highest anti-fungal activity was observed against the C. albicans as 7.59, 6.28, 7.11 and 7.38, in the C3, C4, C5 and C6, respectively. It can be said that 1,3,4-oxadiazole derivatives has more anti-fungal potential on C. albicans as compared to A. niger. It might be due to destruction of cell wall and/ nucleic acid of fungal strains. In conclusion, C3 and C6 were most prominent synthetic derivative among others. It also showed optimum physicochemical properties and anti-fungal potential when tested on the selected fungal strains. It showed a remarkable anti-fungal potential against different 4 fungal strains used.