The aim of this study was to prepare cationic liposomes with encapsulated azithromycin, perform physical characterization and determine the drug release profile. Cationic liposomes (DOPC/DODAB; DPPC/DODAB; DPPC/DC-Chol) with encapsulated azithromycin were prepared by the film hydration method. Homogenization of liposome dispersions was performed using an ultrasonic probe sonicator with a frequency of 40 Hz for 20 seconds. After sonication, the resulting liposomes were with a mean diameter of 300 nm and with much better size distribution (PI ranging from 0.360 to 0.471). The measured positive value of zeta potential in a range of +56.7 to +75.2 mV indicated the formation of physically stable liposomal dispersions. In vitro release studies were performed on the Franz diffusion cell during 24 hours and the amount of the released azithromycin was determined by HPLC method. For comparison, aqueous-ethanolic solution of azithromycin as the control and the anionic-SPC liposomes were prepared and tested under the same conditions. After 24 hours, all of the tested cationic liposomes released approximately the same amount (%) of azithromycin. From the previously obtained data for anionic liposomes, it is evident that they have released significantly less amount of azithromycin, but they have achieved a controlled and prolonged release, as opposed to cationic liposomes where such release is achieved only partially. It could be concluded that it is necessary to do additional in vitro release testing in order to optimize the conditions for achieving complete controlled and prolonged release of the drug from cationic liposomes.