The invention discloses an epoxidized soybean oil maleimide, and a preparation method and an application thereof. The preparation method comprises the following steps: adding epoxidized soybean oil, maleimidoalkyl acid, a polymerization inhibitor, a catalyst and a good solvent into a reactor, carrying out a reaction at 80-150 DEG C in a N2 atmosphere for 4-15 h, pouring the obtained reaction solution into ether having a volume 5 times that of the obtained reaction system, centrifuging the obtained mixture, collecting the obtained supernatant, and carrying out rotary evaporation to remove etherand the solvent in order to obtain the final product. The epoxidized soybean oil maleimide is different from other cross-linking agents, is a novel cross-linking agent with two reactive groups comprising an epoxy group and a maleimido group, can form a double-cure cross-linking network structure in the curing process, can significantly improve the mechanical properties and the toughness of materials, and broadens the application field of the epoxidized soybean oil.