Objectives: In the present study the electrospinning technique was used for production of poly (butylene adipate-co-terephthalate) nanofibers loaded with 5-fluorouracil (5-FIT) and curcumin in treatment of colorectal cancer. Methods: Studied processing and formulation variables included: voltages, polymer concentration, injection rate, distances between the needle tip and the collector plate, concentration of 5-FU, curcumin, and the polymer on the nanofiber features. The cytotoxicity of the nanofibers was studied on the HT29 colorectal cancer cells. Results: The results showed that changes in the drug release profiles were not significant but diameter of the nanofibers and drug entrapment percent were statistically affected by the polymer concentration and its injection rate. Cell culture results showed that the nanofibers loaded with the combination of the drugs had higher cytotoxicity against HT29 cells in comparison with the single-drug-loaded nanofibers and the drug solutions. Conclusion: Ecoflex nanofibers loaded with 5-FU and curcumin may be useful in topical application in colorectal cancers.