During the larval development process of insects, juvenile hormone (JH) is essential for regulating various aspects of larval life, including growth, reproduction, and behavior, throughout their larval stage. The larval stage of Spodoptera frugiperda, when it consumes plant-derived metabolites, develops into pupae, but these pupae are unable to molt successfully. In this way, plant-derived metabolites contain or produce inhibitors of juvenile hormone, thereby disrupting the development of insect larvae and making them vulnerable to harm. Therefore, in this study, we established an in vitro screening system using yeast cells transformed with the Met-SRC juvenile hormone receptor of S. frugiperda. Through this system, we were able to identify juvenile hormone disruptors from plant-derived metabolites and confirm their developmental inhibitory effects on the larvae of S. frugiperda.