Synthesis and biological evaluation of a series of aryl triazoles as firefly luciferase inhibitors

As the most studied bioluminescent system, firefly luciferase is widely applied in many aspects, such as developing small molecule probes, bioluminescent imaging, high-throughput screening, dual luciferase reporters, etc. Considering that a false positive phenomenon often emerges while researchers conduct high-throughput screening based on firefly luciferase, and that the triazole core is a privileged scaffold in drug design and development, we herein report a series of triazoles with potent inhibitory activity in vitro and in vivo, comparable to that of the well-known inhibitor resveratrol. More interesting, a kinetics study disclosed that these triazoles exhibited a brand new inhibition mode, mixed noncompetitive for the substrate aminoluciferin and noncompetitive for ATP. Henceforth, these compounds can notify researchers for possible false positives. Moreover, they will shed light on luciferase structure-function mechanistic exploration and help expand its application in various areas.