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Controlled release, on the contrary traditional treatment, means the gradual release of a substance from a system in order to maintain its concentration in a medium over a period of time. In drug release systems, the controlled release of the drug to the diseased area has importance in terms of treatment and toxicity. Especially drugs with high toxic properties such as 5-Fluorouracil (5-FU) have an important place in controlled drug release technology. In order to complete the treatment successfully, it is important to develop controlled drug release systems to eliminate the possible side effects of anticancer drugs such as 5-FU and to increase the therapeutic effect. The aim of this study is to develop a unique biodegradable and biocompatible bionanocomposite film for the controlled release of 5-FU. For this purpose, bentonite loaded chitosan hydroxypropyl methylcellulose bionanocomposite films were synthesized using solution casting and solvent evaporation techniques. The chemical bond structure of the bionanocomposite films and the effect of bentonite addition on the amine and hydroxyl groups in the structure were determined by Fourier transform infrared. Morphological properties and compatibility of polymers and bentonite were characterized by using scanning electron microscopy. The degree of sorption of the bionanocomposite films was evaluated by swelling tests. As the bentonite content in the nanocomposite film increased from 5% to 25%, the sorption degree of film increased from 2.99% to 65.05%. In addition, the effects of bentonite addition of the synthesized bionanocomposite films and pH of medium on the 5-FU loading and encapsulation efficiency were investigated. Controlled release studies were carried out by using 5-FU loaded nanocomposite films. The obtained results showed that the usage of bentonite in nanocomposite drug carrier films were improved the release properties. After four days, a release value was obtained as 40.12% at pH=7.4 in the 5 wt. % bentonite loaded films. [ABSTRACT FROM AUTHOR]