Spodoptera frugiperda has emerged as a major invasive pest worldwide. The utilization of chemical pesticides not only poses numerous ecological concerns but also fosters resistance in S. frugiperda. In this study, we designed and synthesized three novel thiothiazolidinone compounds (6a, 7b, and 7e) and incorporated innovative thiothiazolidinone structural elements into the piperine skeleton. Treatment with compounds 6a and 7e resulted in the blackening and agglomeration of oviduct eggs within the ovaries of certain female moths, impeding the release of normal eggs. The levels of vitellogenin and vitellogenin receptor, along with three trehalase inhibitors, exhibited a dynamic equilibrium state, leading to no discernible change in egg production but a notable increase in the generation of low-hatching-rate egg fragments. Compared with the injection of 2%DMSO, the eclosion rate of 6a injection was significantly decreased, as followed the spawning time and longevity were prolonged or significantly prolonged in the trehalase inhibitors of 6a, 7b, and 7e. We aimed to investigate the regulatory impacts of three new pepper thiothiazolidinone compounds on the reproduction of S. frugiperda, and to authenticate the efficacy of novel alginase inhibitors in inhibiting the reproduction of S. frugiperda. This research endeavors to aid in the identification of efficient and steadfast trehalase inhibitors, thereby expediting the research and development of potent biological pesticides.