Hydrophobic anticancer drug 5-fluorouracil (5-FU) has been included in the carboxymethyl modified beta-cyclodextrin (CMCD), and the inclusion complex (5-FU/CMCD) was further intercalated into galleries of a zinc aluminum layered double hydroxide (ZnAl-LDH) by the ion-exchange method. Powder X-ray diffraction (XRD), Fourier transform infrared (FT-IR), and UV-vis spectroscopy indicate a successful intercalation of 5-FU/CMCD into the LDH gallery. The release behavior of 5-FU from drug/CMCD-LDH composite at different pH values was studied. It was found that 5-FU was released faster in pH 7.2 than in the acidic mediums (pH 4.8), and the released amount was higher. The introduction of CMCD into the LDH matrix could prolong the drug release time in comparison to that from LDH without CMCD, which can be attributed to the inclusion of 5-FU by the CMCD cavity. Studies on mathematical modeling of drug release show that the release of 5-FU from the drug/CMCD-LDH composite follows the Korsmeycr-Peppas equation very well at different pH values. The drug/CMCD inclusion complex intercalated LDH composite in this work provides a supramolecular formulation for controlled release behavior, which can be potentially applied for nonionic and hydrophobic drugs.