Objectives: 5-Flourouracil loaded to MWCNTs as carbon nanotubes possess pronounced imminent in cancer therapy. The rationale is to conjugate the drug with MWCNTs and to prepare novel carrier delivery system for cancer to enhance the release of drug from F-MWCNTs-5Fu composite and to recognize conceivable effects and release of anticancer drug. Methods: Multi walled carbon nanotubes empower covalent and Non covalent functionalization. Multi-functionalized MWCNTs loaded with anticancer drug. Formulations are considered according to DOE. The characterization studies of FMWCNTs-5Fu composite were completed by FT-IR, SEM, TGA, and TEM. The analysis entrust the increase in particle size, and zeta potential. Drug loading efficiency and In-vitro release of drug was statistical analyzed. Results: Our study consists of 6 factors with 3 levels, for designing the experiments L18 orthogonal array needs to be selected based on number of degree of freedom (DOF). Total number of DOF = (No of levels – 1)× No of main factors =(6-1)*1+(3-1) =11 neglecting the interaction factors, So total number of (DOF) + 1 is the minimum number of experiments. SEM and TEM images explicate that, the drug loaded to pristine MWCNTs and established by particle size analyzer. The drug entrapment efficiency outcome indicates more than 60% of drug entrapment was achieved. Conclusion: Functionalization process help to boost the release of drug in controlled manner and upsurges the dispersion of the formulations. Prepared Functionalized MWCNTs-5Fu approaches with heightened development in medical use. FT-IR spectrum exemplifies the drug attached to MWCNTs, Functionalized MWCNTs-5Fu formulations releases the drug with good percentage and the obtained results were analyzed statistically.