Inflammatory bowel disease (IBD) is a chronic disease characterized by the inflammation of the gastrointestinal system (GIS). Electrospun nanofibers are commonly used in pharmaceutical technology due to many advantages such as high porosity, loading efficiency, large surface area, and stability. Budesonide is the first choice drug in the treatment of IBD and is used as a model drug. In this study, it was aimed to obtain controlled release nanofibers for colon targeting by using anionic, pH-sensitive Eudragit S100 (ES100) and cationic, timedependent Eudragit RL100 (ERL100) polymers. The conductivity, surface tension, and viscosity of the polymer solutions were analysed before electrospinning process. After production, the surface morphology, drug release kinetics, water contact angle, swelling index, and mucoadhesion studies of nanofibers were evaluated. Drug release studies were carried out at pH 1.2, 6.8, and 7.4. The formulation containing ES100:ERL100 (95:5) showed the most suitable drug release as a colon-specific drug delivery system with a minor release at pH 1.2, 6.8, and major release at pH 7.4. As a result, electrospun nanofibers obtained by mixing anionic and cationic polymethacrylate polymers may be a promising alternative as a drug delivery system in the treatment of IBD and other intestinal diseases.