RU-2861489-C1 - METHOD FOR MANUFACTURING BLOOD-SUBSTITUTING PERFLUOROCARBON EMULSION

RU2861489C1RU 2861489 C1RU2861489 C1RU 2861489C1RU-2861489-C1

Abstract

FIELD: medicine; pharmaceuticals. SUBSTANCE: invention relates to the creation of artificial blood substitutes. A method for manufacturing a blood-substituting perfluorocarbon emulsion comprises preliminary preparation of three concentrated components – a perfluorocarbon component, a phospholipid component and a 10-fold phosphate-buffered saline, then adding water to the obtained phosphate-buffered saline, adding the phospholipid component and mixing, dispersing the resulting mixture in a high-pressure homogeniser until a homogeneous light liquid is formed, then adding dropwise the perfluorocarbon component until its volume fraction reaches 10-20 vol.%, and subjecting the resulting mixture to homogenisation in a high-pressure homogeniser at a pressure of 600-800 atm for 10-18 cycles of full circulation of the mixture in the apparatus. Wherein, a three-component mixture of perfluorodecalin, perfluorooctyl bromide and perfluoro-3-propylamine, taken in a volume ratio of 1:0.7:0.3, is used as the perfluorocarbon component, and the phospholipid component is a mixture of soy phospholipid and polyglyceryl-3-polyricinoleate in bidistilled water, wherein the soy phospholipid and polyglyceryl-3-polyricinoleate are contained in a ratio of 1:0.3. The soy phospholipid is introduced into the emulsion at the rate of 0.10 g per 1 ml of the perfluorocarbon component. EFFECT: obtaining a reproducible composition and dispersity of a blood-substituting perfluorocarbon emulsion with a particle size of 100-200 nm, having stability of colloidal properties during storage for at least 3 months at 4-8 °C. 1 cl, 2 tbl, 3 ex

Inventors

FAJZULLIN LEONID ZAKIEVICH
Aprosin Yurij Dmitrievich
Tereshina Elena Vladimirovna
Ilyasova Natalya Aleksandrovna
SUKHIKH GENNADIJ TIKHONOVICH

Dates

Publication Date: 20260505
Application Date: 20241220

Claims (1)

A method for producing a blood-substituting perfluorocarbon emulsion, characterized in that during its implementation an emulsion is obtained containing 10-20 vol.% of a perfluorocarbon component in a phosphate-buffered saline pH 7.2-7.4, wherein a three-component mixture of perfluorodecalin, perfluorooctyl bromide and perfluoro-3-propylamine taken in a volume ratio of 1:0.7:0.3 is used as the perfluorocarbon component, and a phospholipid component is used as the emulsifier, which is a mixture of soy phospholipid and polyglyceryl-3-polyricinoleate in bidistilled water, wherein soy phospholipid and polyglyceryl-3-polyricinoleate are contained in a ratio of 1:0.3, soy phospholipid is introduced into the emulsion at the rate of 0.10 g per 1 ml of perfluorocarbon component, and the method includes the preliminary preparation of three concentrated components - a perfluorocarbon component, a phospholipid component and a 10-fold phosphate-buffered saline, wherein water is first added to the resulting phosphate-buffered saline, the phospholipid component is introduced and mixed, then the resulting mixture is dispersed in a high-pressure homogenizer until a homogeneous light liquid is formed, after which the perfluorocarbon component is added dropwise until its volume fraction reaches 10-20 vol.%, and the resulting mixture is homogenized in a high-pressure homogenizer at a pressure of 600-800 atm for 10-18 cycles of a complete turnover of the mixture in the apparatus, wherein the perfluorocarbon component and the phospholipid component are pre-sterilized, the resulting perfluorocarbon emulsion is sterilized by autoclaving at a temperature of 125 °C for 8 minutes, and the gas space in the container with the said components and emulsion is replaced inert gas.

Description

The invention relates to the fields of medicine and pharmaceuticals. The subject of the invention is an emulsion comprising: a physiological saline buffer, a mixture of phospholipids with glycerol in liposomal form, and perfluorocarbon compounds (perfluorodecalin, perfluoropropylamine, perfluorooctyl bromide). The emulsion is prepared in two stages. In the first stage, a liposomal fraction is prepared in an aqueous-salt buffer, which is a mixture of plant phospholipid with polyglyceryl-3-polyricinoleate. In the second stage, the liposomal phospholipid fraction is poured into the reservoir of a high-pressure homogenizer, and a solution of perfluorocarbons in phosphate-buffered saline is added dropwise at 100-200 atm. Homogenization is then carried out at 600-800 atm for 10-18 cycles of complete mixture rotation in the apparatus at an extrusion frequency of 30-50 per minute. The technical result of the claimed method consists in the production of a blood-substituting perfluorocarbon emulsion, reproducible in composition and dispersion, with a particle size of 100-200 nm, which has stable colloidal properties when stored for at least 3 months at 4-8 °C. Introduction Growing environmental and social tensions worldwide, leading to emergencies and disasters, and the increasing number of local military conflicts both domestically and internationally, are contributing to the rise in severe injuries, accompanied by significant blood loss. Despite years of research aimed at finding a substitute for allogeneic blood, red blood cell transfusions have been and remain the standard for increasing oxygen delivery to patients with anemia. However, the challenges of procuring and storing sufficient quantities of donor blood remain unresolved. Therefore, the development of artificial blood substitutes that facilitate oxygen transfer is one of the most important areas of modern transfusion medicine. Currently, perfluorocarbons (PFCs) are considered the most promising artificial oxygen carriers due to their chemical inertness, harmlessness to living organisms, and ability to dissolve and transport large volumes of gases – O2 , CO2 , NO – to tissues and cells. Due to their chemical inertness and the high strength of the carbon-phosphorus bond, PFCs dissolve gases () not through chemical bonds, but by van der Waals forces. Therefore, gas transport is regulated by their portion pressure in the environment, in particular, in blood vessels. The most detailed description of the physicochemical properties and gas transport function of perfluorocarbons is presented in the review by Riess JG and Kuznetsova IN [1, 2]. The inherent hydrophobicity of PFCs prevents them from acting as gas carriers themselves, so they are emulsified by coating them with surfactants such as phospholipids, albumin, etc. An important factor in creating an emulsion is the choice of emulsifier, which determines its physical properties: particle size, gas permeability, and half-life. Of the many contenders for emulsifiers, animal and plant phospholipids are currently considered the best due to their physiological suitability for the body and their ability to form complexes with PFCs in the form of liposomal structures through which gases can freely pass both inward and outward. Research conducted by Kuznetsova I.N. showed that the dissolution of gases in PFC emulsions and their release into cells or tissues occurs according to the same principle as in free PFC complexes [2]. There are various methods for producing emulsions, including ultrasonic and mechanical methods. A disadvantage of ultrasonic treatment is that the resulting waves can break intermolecular bonds, particularly the C-F bond in fluorocarbons. This results in the formation of fluoride ions, which can be toxic to the body when introduced into the bloodstream. For producing thin-layer emulsions, the most effective method is to extrude a mixture of PFC and emulsifier through briefly opened fine orifices under high pressure. High-pressure homogenizers are used for this purpose. Using a high-pressure homogenizer to process the PFC/FL mixture allows for the production of emulsions ranging in size from 0.07 to 0.20 µm [3]. Patent RU 2745290 C2 [3], which is the prototype of the present invention, describes in detail the basic principles of producing PFC emulsions and their physicochemical properties. Perfluorocarbons (PFD, PFZPA, and PFOB) were used as components of the PFC emulsion, egg or soy phospholipid served as an emulsifier, and castor oil stabilized the emulsion. The emulsion was produced by extrusion using a high-pressure homogenizer at 600-800 atm. These same principles were largely used in the present invention. However, when reproducing the technology described in patent RU 2745290 C2 [3], the resulting emulsion was unstable and contained large particles larger than 1000 µm. Our studies showed that these structures are associates of FL with castor oil. Therefore, we made significant changes to