Abstract: Trehalose is a non-reducing disaccharide, which exists widely in nature and possesses important biological functions. It is not only an energy storage substance participating in energy metabolism, but also a structure substance participating in the formation of fungal cell walls and insect exoskeletons. Meanwhile, trehalose can regulate the pathogenicity of pathogens, and the growth and development of insects. Trehalose synthetase is mainly responsible for catalyzing the biosynthesis of trehalose, which includes two functional domains: trehalose-6-phosphate synthetase and trehalose-6-phosphate phosphatase. More and more recent studies showed that trehalose synthetase has a great potential as a novel target of pesticides. This review summaried the protein crystal structures, substrate binding modes, and chemical structures of reported inhibitors for trehalose-6-phosphate synthetase and trehalose-6-phosphate phosphatase. It is expected to provide guidance for the screening of inhibitors, the design and the modification of lead compounds, and the development of new pesticides targeting trehalose synthetase in the future.