The drug delivery application of Porous metal–organic frameworks (MOFs) have been investigated due to their unique structures which are built of inorganic nodes and organic ligands. In present study, Uio-67-5-FU was successfully prepared by applied for delivery of 5-fluorouracil (5-FU). Using variety of analytical methods including FTIR, FESEM, EDS, and the prepared nanostructure was characterized. Results revealed the placement of the drug in zeolite is well done and also the in vitro loading and releasing studies, for 5-FU was evaluated. In addition, Based on the in vitro cytotoxicity results, Uio-67-5Fu was able to increase cytotoxicity compared to that of 5-Fu on HT-29 cancerous cells indicating the highlighted role of this drug delivery system. .