The aim of the current work was to formulate as well as evaluate colon targeted matrix tablet using Azathioprine. Excipients include Eudragit S100, Microcrystalline cellulose, Lactose, Talc, Magnesium stearate. Quantities of the excipients were chosen by carrying out FT-IR method. Preformulation studies were performed so as to study the nature as well as compatibility of Active Pharmaceutical Ingredient with that of the excipients by physical observations along with FT-IR studies. The tablets were processed by direct compression method making use of the selected excipient quantities. Pre-compression parameters like tapped density, bulk density, Hauser’s ratio, compressibility index, and compressibility index. The formulated Azathioprine matrix tablets were coated using pan coating method with enteric polymer Eudragit S 100. The final tablets were evaluated for weight variation, thickness, hardness, and drug content, friability and disintegration time and in-vitro dissolution studies. Cumulative percentage drug release found to be 64.24 % (lowest) for F1 to 97.66 (highest) for F9 up to 8 hr. as the sustained release is main aim, formulation F1 was selected a optimized formulation. Formulation F1 containing enteric coated matrix tablet of Azathioprine would be a promising formulation that will aid in achieving the purpose of treating inflammatory bowel diseases (ulcerative colitis) without any gastric irritation or ulcers, which is useful for patients having prehistory of ulcerative colitis.