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Antifungal activities of MOFs (metal organic frameworks) have been demonstrated in studies, and improvement in efficiency of fungal inactivation is a critical issue in the application of MOFs. In this study, we employed 4,4′-azopyridine (AZPY) in the construction of MOF to improve its antifungal activity. Three-dimensional (3D) copper metal organic framework containing glutarate (Glu) and AZPY (<b>Cu(AZPY)-MOF</b>) was synthesized by a solvothermal reaction. Glutarates bridge Cu₂ dinuclear units to form two-dimensional (2D) layers, and these layers are connected by AZPY to form a 3D framework. When spores of two fungi, <i>Candida albicans</i> and <i>Aspergillus niger</i>, were treated with <b>Cu(AZPY)-MOF</b> for one day, number of CFU (colony forming unit) was continuously reduced over treated MOF concentrations, and maximum 2.3 and 2.5 log₁₀CFU reductions (approximately 99% reduction) were observed in <i>C. albicans</i> and <i>A. niger</i>, respectively. Small amounts of Cu^II ions and AZPY released from <b>Cu(AZPY)-MOF</b> were not critical for fungal inactivation. Our results indicate that the level of antifungal activity of <b>Cu(AZPY)-MOF</b> is greater than that of Cu-MOF without AZPY constructed in our previous study, and intercalation of AZPY is able to improve the antifungal activity of <b>Cu(AZPY)-MOF</b>.