Katsay O. Development of the technology to obtain the liposomal cytochrome C for the treatment of ophthalmic diseases

In the thesis presented the results of an experimental study on the development of the composition and the development of optimal technology for the production of liposomal cytochrome C. A method was proposed of producing a liposomal cytochrome C. The critical steps of this method: obtaining a lipid film; hydration of this film in a solution of cytochrome C until obtaining an emulsion of multilamellar vesicles (MLV); high-pressure homogenization of MLV emulsion to getting small unilamellar vesicle (SUV). For evelopment, we have chosen a phosphate salt buffer with pH 6.8, which has no toxic properties and widely uses in the manufacture of drugs. The influence of liposome composition was evaluated on the encapsulation efficiency and particle size during the creation of the liposomal form of cytochrome C. As a result of studies, the ratio of phosphatidylcholine: dipalmitoylphosphatidylglycerol (1.20: 1.0) was proposed, which provides the maximum encapsulation of cytochrome C (95.88%). The necessity to choose the form of lyophilization for solution preparation was shown with the example of stability evaluation of aqueous cytochrome C-containing liposomes and freeze-dried cytochrome C-containing liposomes. During the observation period, the aqueous solution shows the growth of degradation products (free fatty acids and lysophosphatidylcholine) by 202.5% and 409.7%, respectively, 16 compared with the reference values, as well as the decrease of the content of phosphatidylcholine and dipalmitoylphosphatidylcholine by 1.5 and 1.0%. At the same time, there were no significant changes in the composition of FD-Cyt-LS samples. The composition of freeze-dried cytochrome C-liposomes contained per 1 vial: 2.025 mg of cytochrome C, 18 mg of egg phosphatidylcholine, 15 mg of DPPG, 180 mg of lactose monohydrate, 2.34 mg of potassium dihydro phosphate, 24.4 disodium hydrophosphate. The optimum industrial technology of obtaining freeze-dried liposomal cytochrome C is substantiated and developed.