Site-specific colon drug delivery is immense in the treatment of colon-localized diseases. In this study, prodrugs of 5-aminosalicylic acid (5-ASA), the drug of choice in ulcerative colitis (UC) were synthesized by conjugating with chitosan (MW 30, 80 and 300 kDa) to achieve targeted delivery to the colon. FT-IR and UV-Vis analyses confirmed the successful synthesis and varying loading capacities of 5-ASA with a maximum of 11.32 ± 2.0 %. The obtained conjugates (7-9) exhibited gradual drug release properties, with up to 30 % of the drug content after 24 h, in the simulated colonic fluid containing rat gastrointestinal (GI) tract homogenates, demonstrating their colon-specific and slow-release properties. The conjugates were non-toxic to normal human colon epithelial cells at concentrations up to 5-10 μg/mL, suggesting a favorable safety profile. Additionally, conjugate (7) was tested for efficacy in a mouse model of ulcerative colitis. Necropsy results showed no significant structural changes or lesions in treated animals. Histopathological analysis revealed mild, multifocal lymphoplasmacytic infiltrates and scattered eosinophils in the lamina propria, indicating a low level of inflammation. Overall, the findings suggest that 5-ASA-azobenzyl-chitosan conjugates hold promise as a potential therapeutic option for ulcerative colitis and other localized colon disorders, with minimal side effects due to reduced systemic exposure.