FIELD: medicine.
SUBSTANCE: invention refers to medicine, more specifically to creation of new liposomal forms of combined antibacterial preparations for treating extremely dangerous infectious diseases. Invention represents method of producing liposomes with encapsulated doxycycline hydrochloride and surface localised monoclonal antibodies, including creation of emulsion of lipids and antibacterial preparation and subsequent production of liposomes, characterised by that for increasing number of encapsulated antibiotic and additional immobilisation on surface of liposomes monoclonal antibodies, batch in 63.3 mg mixture of lipids phosphatidylcholine, cholesterol in ratio of 7:3 is dissolved in 4.3 ml of chloroform and mixed with 100 mg of antibiotic dissolved in 0.01 M of phosphate buffer with pH 7.2 in volume of 1.5 ml, treated in rotary mixer with power of 500 W at 15,000 rpm for 3 minutes until formation of emulsion, completely removed organic solvent in rotary evaporator, produced gel added with 5 ml of 0.01 M of phosphate buffer with of pH 7.2 and thoroughly stirred in flask in rotary evaporator, separating non-included antibiotics is carried out by centrifugation at 20000 rpm for 1 h in centrifuge, percentage of doxycycline immobilisation is determined by serial dilutions, at that, it amounted to 70 %, dimensions of liposomal preparation in electron microscopy study achieved from 600 to 800 nm, degree of oxidation of lipids membrane liposomes is 0.8 ± 0.02 units, that is followed by immobilising of immunoglobulins at outer membrane of liposomes, and obtained preparation is used for treating acute experimental melioidosis in laboratory animals. Production of emulsion in rotary mixer allows excluding toxic effect of titanium ions, reduced processing time and improving encapsulation of doxycycline hydrochloride for up to 70 %. Change in mode of immunoglobulin introduction in reaction mixture increases percentage of immobilisation on outer membrane of liposomes from 17 % to 30.9 %.
EFFECT: produced liposomes provide 80% survival rate of laboratory animals with acute experimental melioidosis.
3 cl, 1 tbl, 2 ex
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Authors
Dates
2016-04-27—Published
2015-03-24—Filed