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Urease may rapidly hydrolyze urea, causing a significant amount of ammonia to be released and decreasing the efficacy of urea fertilizers. However, urease inhibitors (UIs) can inhibit the interaction between urease and urea so that ammonium nitrogen can be absorbed by crops for a long time. In this paper, we developed and fabricated two Cu-based coordination polymers (Cu-CPs) to function as UIs by combining two aromatic carboxylic acids and a bis-pyridine-bis-amide ligand, which demonstrate outstanding urease inhibitory efficacy. Cu-CP-1 exhibits a pronounced inhibitory effect on the inhibitory activity of jack bean urease while the semi-inhibitory concentration (IC 50) value was 0.74 ± 0.01 μM, whereas the IC 50 value of Cu-CP-2 was 4.77 ± 0.51 μM. Furthermore, inhibitory activity against urease, kinetic, molecular docking, and comparisons of the title Cu-CPs and other complex-based materials have been comprehensively investigated. More importantly, the oxygen atoms of μ 2 -OH and –COO play important roles in urease inhibition activity. The structure–function relationship were further discussed which indicates that Cu-CPs can be used as a kind of UIs. By using different O-donor second auxiliary ligands, we have designed and synthesized two Cu-based CPs to operate as urease inhibitors which show excellent urease inhibitory activity. [Display omitted] • The development of UIs has been a matter of great concern to researchers. • Two Cu-CPs have been synthesized by using diverse O-donor ligands. • The Cu-CPs are served as UIs and show excellent urease inhibitory activity. • Inhibitory activity, kinetic, molecular docking of the Cu-CPs have been studied. [ABSTRACT FROM AUTHOR]