WO2008082321A1 - Emulsion of perfluoroorganic compounds for medical use and a methods for the preparation and the use thereof - Google Patents

Abstract

The invention relates to biomedicine. The inventive emulsion of perfluoroorganic compounds, which is used for intravascular or local injection, comprises a rapidly eliminated perfluor carbon-perfluor decalin, a low eliminated perfluorinated tertiary amine of perfluor-N-(4-methyl-cyclohexyl)-piperidine and a physiologically acceptable salt-aqueous solution. The critical solution temperatures of all components of the perfluoroorganic compounds dissolved in hexane (TcrH) differ from each other at a value which is equal to or less than 2-4°C, thereby increasing the stability of the emulsions of perfluoroorganic compound. The emulsion ofperfluoroorganic compounds is stabilised by a mixture of polyoxyethylene-polyoxypropylene block copolymers and comprises only the entirely fluorinated impurities of the perfluoroorganic compounds, thereby reducing the toxicity thereof. The inventive emulsion producing method consists in mixing and dispersing the mixture of liquid perfluoroorganic compounds with the aqueous solution of a stabilising agent and in subsequently reducing it in a high pressure homogeniser in such a way that submicron emulsion particles are obtained. The inventive method for treating the blood system diseases consists in injecting the perfluoroorganic compound emulsion into the blood current or into a lymphatic channel.

Description

 An emulsion of perfluororganic compounds for medical use, a method for its preparation and a method for its use.

The claimed group of inventions relates to the field of biomedicine, transfusiology, pharmacology, biophysics, in particular to medicines based on an emulsion of perfluororganic compounds (PFOS), intended for the treatment of systemic and local disorders of the blood flow, hypoxic and ischemic conditions, improving the mass transfer of gases and metabolites between blood and tissues, maintaining the function of isolated organs and tissues, reducing the phenomena of inflammation, conducting infusion-transfusion therapy in shock and blood loss.

LIST OF CONVENTIONS, SYMBOLS, UNITS AND TERMS USED IN THIS APPLICATION

US Saturated affinity separation

CA surfactant

P-168, P-268, block copolymer of polyoxyethylene and polyoxypropylene

P1F-268 - Proxanol 16.8, Proxanol 268, Pluronic F68

POP Polyoxypropylene block of proxanol

POE Po Proxyanol Block

PFD Perfluorodecalin

PFDE Perfluorodighexyl ether

PFMCP Perfluoromethylcyclohexylpiperidine

PFOB Perfluorooctyl bromide

PFOS Organofluorine compounds

PFTA Perfluorinated Tertiary Amines PFTBA Perfluorotributylamine

PFTPA Perfluorotripropylamine PFU Perfluorocarbons PFE Perforated Esters

RES Reticuloendothelial system

CA Stabilizing Agent

T _kp g Critical dissolution temperature in hexane

Phospholipids ø average particle diameter

I _R reactogenicity index

PFOS used in biology and medicine are completely fluorinated organic compounds of various classes, including perfluorocarbons (PFCs) - compounds containing only fluorine and carbon atoms, perforated tertiary amines (PFTA), in which, along with fluorine and carbon atoms, there is a nitrogen atom, and also perforated esters, in the structure of which there are oxygen atoms. In recent years, bromine-containing PFOS have also been used. Liquid PFOS is transparent, colorless, odorless, about 2 times heavier than water. The high bond strength C-F (485.6 kJ / mol) ensures the chemical stability of PFOS and weak intermolecular interaction forces (1,2). The latter property provides an increased ability of PFOS to dissolve various gases.

PFOS practically does not dissolve in H ₂ O, but some PFOS are slightly soluble in lipids - lipophilic PFOS. All types of PFOS are not subject to metabolic transformations in humans and animals. The question of the stability and cleavage of a bromine atom and, therefore, the possibility of decay of a molecule left without bromine has not been discussed in the literature. Lipophilic PFOS, and among them the most lipophilic is perfluorooctyl bromide (PFOB), due to physical interaction with biological membranes and hydrophobic portions of protein molecules, can have a pronounced effect on the metabolism and many functions of cells, tissues and the body as a whole (3,4,5,6) . According to L. Clark (7), later confirmed by A.N. Sklifas et al. (8), the inclusion of other lipophilic PFOS in the emulsion along with lipophilic PFD can lead to the death of animals several months after the intravenous administration of such emulsions.

It is known that in order to eliminate this fatal effect, it is proposed to add a lipophobic organo perfluorine compound, for example, perftop-N- (4-meticyclohexyl) - piperidine in lipophilic organofluorine compounds (8, 9). However, this raises the problem of selecting an emulsifier, since lipophilic organo perfluorine compounds are stabilized by phospholipids, and lipophobic organo perfluorine compounds are stabilized by block copolymers of ethylene oxide and propylene oxide (Pluronic F-68 according to the Western classification; proxanol 268 according to the Russian nomenclature). In addition, the use of a mixture of substances that differ sharply in their physicochemical properties (lipophilic and lipophobic) significantly reduces the stability of the emulsion during storage (freezing, thawing, or long-term storage in an unfrozen state) and when the emulsion enters the bloodstream. Calculations and model experiments (10) give reason to believe that this is due to the clustering of an insufficiently homogeneous mixture of substances inside the particles of the emulsion and the constant turbulent motion of the fluorocarbon phase near the adsorption layer of the surface-active component, the stabilizing agent. Introduced intravenously in the form of emulsions PFOS quite quickly removed from the bloodstream. The bulk of PFOS is excreted through the lungs. Therefore, those PFOS that have a relatively high vapor pressure (they, as a rule, are more lipophilic) can cause emphysema up to rupture of individual alveoli and impaired gas exchange and blood flow in the lung tissue. Up to 30% of PFOS emulsions introduced into the body are captured by the cells of the reticuloendothelial system (RES) - mainly in the liver, spleen, bone marrow, lymph nodes, and are also partially dissolved in adipose tissue and membrane phospholipids of almost all cells. The PFOS accumulated in this way is retained in organs and tissues for a period of several days (PFOB, PFD) to several years (PFBA), depending on the physicochemical properties of PFOS and the dose of the emulsion introduced. The rate of PFOS removal from organs depends on a number of interconnected physico-chemical parameters: structure, molecular weight (MM) ₅ , boiling point, vapor pressure, and mainly on a parameter defined as the critical temperature for the dissolution of PFOS in hexane (T _cr g). T _{cr k} is the temperature at which equal volumes of the test compound and hexane are mixed with each other. _{Tcrg is} considered as a measure of the relative solubility of PFOS in lipids, which characterizes both their degree of dissolution in membrane lipids, the rate of their passage through membranes, and the degree of affinity for hydrophobic portions of protein molecules. The lower T, φg, the lower MM and the higher the vapor pressure, the better this PFOS is soluble in lipids, diffuses faster and is more easily and quickly excreted from the body. A good correlation was observed between the indicated parameters and the half-life (tsc) of PFOS from RES cells. Currently, the attention of researchers engaged in the development and study of PFOS emulsions is focused on a relatively small number of compounds (Table 1).

Table 1

The critical temperature of dissolution in hexane (T _cr g), vapor elasticity (P) and half-life from organs (tш) for various PFOS [1].

.

.

In the manufacture of emulsions for intravascular administration, most specialists prefer rapidly ascending lipophilic PFOS (2,10, 11,12,13,14), which have a relatively small T _cr value, including high lipophilic one. PFOB .. However, in the composition of “Fluosol DA”, produced in 70-80 tons by the company Greep Сrosss Corroratiop (Jarap), along with with quickly outputting PFD entered and relatively slowly displayed

PFTPA (12). The composition of Oxygent, developed by Alliapse Theareutic (USA), includes high lipophilic PFOB and PFDB (13D4). The commercial pharmacopoeial preparation Pperftopan, developed in the USSR and released into the pharmacy network by the company Pertofopan NPF, along with rapidly excreted PFD, contains PFMTSP, which lingers relatively long in RES cells (6).

It should be noted that the PFOS emulsions themselves, despite the chemical and metabolic inertness of their constituent components, also have pronounced biological activity, which is determined not only by the properties of the components themselves, but also largely depends on the phase boundary formed by the surface-active substance (CA) , the presence of CA or its dispersion in the aqueous phase, on the stability of the adsorption layer, as well as on the degree of dispersion of the emulsion, which can play the role of a huge sorption and exchange catalysis ically active surface. The emulsion introduced into the body actually introduces a huge hydrophobic phase into the bloodstream and into membranes, which can act as a medium for various reactions between compounds having increased solubility in the hydrophobic phase. In other words, the PFOS emulsion can stimulate microcatalysis phenomena occurring in the hydrophobic phase or at the phase boundary.

Since liquid PFOS does not dissolve in water and polar compounds that play a crucial role in the homeostasis of biological objects do not dissolve in them, so far PFOS can be introduced into the bloodstream only in the form of a finely dispersed emulsion. The ability of PFOS and PFOS emulsions to participate in gas exchange is determined by the physical dissolution of gases, and in accordance with the law. Henry the amount of dissolved gas is directly proportional to its partial _. pressure. The amount of gas dissolved in the PFOS emulsion depends on the volumetric content of the fluorocarbon phase and does not depend on the particle size. Thus, the amount of experimentally determined gas dissolved in PFOS emulsions is almost equal to the calculated values obtained by adding the gas content in each phase separately (the amount of dissolved oxygen in the aqueous phase plus the amount of dissolved gas in PFOS) at a given partial pressure of gas. Thus, the amount of any gas contained in the PFOS emulsion can be calculated in accordance with the physical laws of their solubility, based on the partial pressure of the gas and the volume ratio of the PFOC / H ₂ O fractions. Due to the submicron size of the PFOS emulsions, when introduced into the bloodstream, more than an order of magnitude increases the surface area involved in gas exchange. Together with an acceleration of the processes of oxygen saturation and release, which is almost ten times faster than the hemoglobin in erythrocytes, this contributes to a significant acceleration of oxygen and carbon dioxide mass transfer, even if the dose of an emulsion containing only 10 vol% PFOS introduced into the bloodstream does not exceed 1-5 % of the volume of circulating blood.

The therapeutic efficacy of PFOS emulsions, described in experimental and clinical studies performed with Fluosol-DA (12) and Perftoran (6.15), is limited by their possible side effects. The following factors limit the therapeutic use of PFOS emulsions as drugs. Firstly, back in 1971, Fujita (12) found that when the particle size of the emulsion is more than OD μm, their toxicity sharply increases, which is probably due to. thromboembolic complications. With an increase in the proportion of large particles (with an average diameter exceeding 0.4 μm) from 3% to 10%, the LD ₅₀ for PFOS emulsions decreases by more than 2 times. The significance of particle size for the tolerance of emulsions was shown by S.I. Vorobyov (16), who established that an increase in particle size, even if their fraction is only 1-2%, leads to reactogenicity, which manifests itself in the development of anaphylactoid reactions after the introduction of the first drops of emulsion into the bloodstream. Manifestations of anaphylactoid reactions can be different in severity - lungs, mediators and. severe: 6t of redness of the skin and mild allergic manifestations up to respiratory arrest.

According to the results of the study of fat emulsions and Fluosol DA, most researchers believe that reactogenicity is determined by the nature of the stabilizing agent - CA, used for dispersion PFOS and forming an adsorption layer around the particles. It is believed that the non-ionic CA block copolymer of polyoxyethylene (POE) and polyoxypropylene (POP) Pluronic F 68, formally analogous to proxanol 268, is responsible for the reactogenicity, and that its replacement with natural phospholipids can completely eliminate the problems of reactogenicity. However, it is known that fat emulsions stabilized by natural phospholipids are mostly reactogenic. Therefore, the reactogenicity of PFOS emulsions cannot be eliminated only through the use of phospholipids as an emulsifier and stabilizer of PFOS particles. According to the patent (17), the reactogenicity of PFOS emulsions, in addition to the degree of dispersion, is determined primarily by the surface properties of emulsified particles, i.e., by the state of the emulsifier layer stabilizing the particles. The leading parameters in this case are the bond strength of CA with the oil fluorocarbon "core" of the emulsion particles, the nature of the location of CA molecules on the surface, their packing density, the stability of adsorption properties with respect to proteins and other biologically active molecules in the bloodstream. To obtain the desired compliance, the said patent empirically suggests selecting the composition of the oil “core” composition of PFOS and the CA interacting with it. However, the empirical selection and established as a result of the combination of PFOS. and CA in any case leave uncertain a wide range of variations in the stability of PFOS emulsions due to the variability of chemical technologies for producing PFOS and CA, as well as seemingly insignificant deviations in the dispersion technology.

The main reasons for variations in CA composition are the very nature of polymer and phospholipid CAs. An important destabilizing factor is also the presence of constant turbulent flows inside the PFOS oil phase that arise when mixing dissimilar POPs included in the oil core: from a mixture of lipophilic and lipophobic PFOS that differ significantly in T _cr r.

In order to increase, stability and reduce the reactogenicity of PFOS emulsions according to the patent of the Russian Federation N ° 2206319 (18), which is selected in As a prototype, it was proposed to increase the uniformity of the “oil” core by using PFOS additives close to the main components of PFD and PFMCP in structure and slightly reducing the differences between PFD and PFMCP in terms of T _cr g- However, the range of differences in Tcr values between PFOS used in the composition according to the patent of the Russian Federation N ° 2206319 remains more than 10 ⁰ C - quite large. An experimental analysis performed using a thermal imaging highly sensitive matrix detector showed that with such a gap in terms of _Tcrg in the mixture of PFOS used in the emulsion according to RF patent N ° 2206319, very intense turbulent movements in the fluorocarbon phase are observed.

The objective of the claimed group of inventions related by a single inventive concept is to create a stable PFOS emulsion primarily by reducing the differences in T ^ g values inside the fluorocarbon self-assembly due to adsorption separation, in accordance with the type of PFOS used in the core of the emulsion particles and changing the composition of the stabilizing agent, by way. The solution of this problem is achieved using the original method of obtaining emulsion.

. The essence of the invention in terms of the emulsion of perfluororganic compounds (PFOS) for biological and medical purposes, is that it contains a mixture of perfluorocarbons (PFCs), the main component of which is perfluorodecalin (PFD) and a mixture of perfluorinated tertiary amines (PFTA), the main component of which is perfect-N- (4-methylcyclohexyl) -piperidine (PFMTSP), having lower than PFD, the rate of excretion from the body, a stabilizing agent and a physiologically acceptable water-salt solution with substrates of energy metabolism, all the components of PFOS have a critical temperature of dissolution in hexane (T _cr g) ₅ differing by no more than 2-4 ° C, and the stabilizing agent is a mixture of block copolymers from the group of block copolymers of polyoxyethylene-polyoxypropylene, with an average mass content polyoxypropylene 20%, while the PFD mixture contains only fully fluorinated PFU impurities obtained by the synthesis of PFD, and the PFMTSP mixture contains only fully fluorinated PFTA impurities, as well as at least one a component from the group: mixtures of perfluorotripropylamine (PFTPA) only with fully fluorinated impurities of PFTA, mixtures of perfluorotributylamine (PFTA) only with completely fluorinated impurities of PFTA, mixtures of perfluorodecyl ether (PFDE) only with fully fluorinated impurities of perfluorinated ethers (P).

In special cases, the implementation of the emulsion as perfluorocarbons contains a mixture of perfluorinated coproducts of perfluorodecalin, having an average half-life of at least 12 days and a range of T _cr in the range of 22-29 ° C, purified from hydrogen-containing and unsaturated impurities, as perfluorinated tertiary amines it It comprises a mixture of perfluorotripropylamine koproduktov having an average half-life of the organism for at least 60 days and _KP range T g in the range 41- 46 ° C, purified from hydrogen and unsaturated ca. This perforated or as tertiary amines it contains a mixture koproduktov pepftop-N-4- (metiltsiklogekcil) -pipepidina having an average half-life of the organism at least 80 days and _KP range T g in the range 32-40 ° C, purified from hydrogen and unsaturated impurities, as a slowly excreted PFOS, it contains a mixture of co-products of perfluorodecyl ether (PFDE) having an average half-life of at least 500 days and a range of T _cr in the range of 49-53 ° C, purified from hydrogen-containing and unsaturated impurities, as erftorirovannyh tertiary amines it may comprise a mixture of perfluorotributylamine koproduktov having an average half-life of the organism at least 900 days, and the range T g _KP within 56-60 ° C, purified from hydrogen and unsaturated impurities contained therein ratio rapidly eliminated slowly and are excreted PFOS ranges from 5: 1 to 1: 5; as a stabilizing agent, it contains a mixture of polyoxyethylene (POE) -polyoxypropylene (POP) copolymers selected by saturation affinity separation (NAS) with a molecular weight of 5-13 ys. Yes, as stabilizing agent it contains a mixture selected by US POE and POP POELOP copolymers with a ratio ranging from 78:22 to 82:18, the content of stabilizing agent therein is 1- 5% for viscosity 2-5 spoise, and a physiologically acceptable water-salt solution includes NaCl, KCl, MgCl ₂ , NaHCO ₃ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , D-glucose to ensure isosmosis in the final dosage form, while maintaining the ratio of Na ions and K at a level of 10: 1 - 30: 1 and a pH in the range of 6.9 - 8.0, the water-salt part of the finished final dosage form contains PO -150 mM NaCl, 4-5 mM KCl, 1-3 mM MgCl ₂ 6-20 mm NaHCO ₃ , 1-2 mm NaH ₂ PO ₄ 1-2 mm Na ₂ HPO ₄ and 10-15 mm D-glucose, and for use as an anti-ischemic protector, the water-salt portion of the drug contains 4-15 mM KCl, salts of tri-, di- and mon carboxylic acids, including 1-7 mm sodium citrate, 1-7 mm sodium isocitrate, 1-7 mm sodium succinate, 1-7 mm sodium ketoglutarate, 1-7 mm sodium pyruvate, 1-7 mm β-hydroxybutyrate sodium, 1-7 mm glutamate sodium, 1-7 mm sodium aspartate.

The invention in terms of a method for preparing an emulsion of PFOS (in accordance with the above set of characteristics of an emulsion) for biological and medical purposes, includes mixing and dispersing a mixture of liquid PFOS with an aqueous solution of a stabilizing agent, followed by grinding in a high pressure homogenizer to a submicron particle size of the emulsion, and:

- a mixture of PFOS having differences in T _{cr of} not more than 2-3 ° C, is poured into a container containing a solution of a mixture of CA in a ratio of 10-30 volume parts of PFOS to 90-70 volumes of a solution of a mixture of CA,

- the resulting inhomogeneous mixture was subjected to the procedure of US carried out at a temperature of 50-60 ° C _e comprising obtaining a coarse dispersion PFOS CA solution by stirring at a high speed mixer with a rotor speed of 1000-3000 rpm, the resulting coarse dispersion was extruded in a homogenizer under pressure of 100-300 kg / cm ² in an atmosphere that does not contain oxygen, and the coarse dispersion of PFOG is separated from the aqueous solution of CA, the precipitate of the coarse dispersion is mixed with a fresh portion of the CA solution from such a calculation that the volume fraction of PFOS was 10-30%,

- while the described operation US is repeated at least 2 times,

- after which the emulsion is passed repeatedly through the homogenizer at a pressure of 300-700 kg / cm ^2, - the resulting submicron emulsion is diluted with water-salt solution so that the final concentration of PFOS is A-12 volume% and the emulsion has an osmotic pressure in the range of 270-330 mOsm.

In particular cases of the method implementation, mixtures of liquid PFCs and PFTA or PFE are obtained in a ratio of 5: 1 to 1: 5, subjected to washing after mixing with pyrogen-free water-alcohol solutions and water, and mixtures of denser liquid PFOS are separated from the water-alcohol solution and water , filtered under pressure of an oxygen-free gas, poured and sealed into vials for filling and autoclaving, CA is prepared in the form of a 10-30% aqueous solution of copolymers of polyoxyethylene-polyoxypropylene - proxanol 168 and proscanol. 268 taken in accordance At a ratio of 10: 1 to 1:10, depending on the composition of the composition of the PFOS mixture, the CA solution is purified by passing through a carbon sorbent and a sterilizing filter under the pressure of an oxygen-free gas, the CA solution is heated for 8-24 hours and cooled to 16- 2O ⁰ C to completely restore the transparency of the solution, the aqueous solution of a mixture of polyoxyethylene-polyoxypropylene copolymers is heated at a temperature that is selected in the range of ± 2 ⁰ C from the cloud point established experimentally for the obtained mixture CA.

The essence of the invention in terms of a method for treating diseases of the circulatory system is that they introduce into the bloodstream or into the lymphatic duct quick-acting organofluorine compounds, characterized in that PFOS emulsion is introduced into the bloodstream as fast-moving organofluorine compounds (in accordance with the above set of emulsion characteristics).

In particular cases, the implementation of the method for the treatment of the phenomena of hypoxia, ischemia, edema, inflammation in shock, blood loss, thrombo-embolic lesions, functional and organic disorders of vascular patency, tissue damage is carried out by intravenous administration of PFOS emulsion in a dose of 0.5-30 ml per 1 kg of weight body, to suppress secondary alterations in inflammation and wound healing, as well as to accelerate the healing of traumatic, trophic and surgical wounds, are administered intravenously with PFOS emulsion in doses of 0.3-6 ml per kg body weight and / or topically in 0.5-50 ml portions subcutaneously, intramuscularly, the depth around the damaged area, or in the surrounding inflamed and injured tissues of the cavity or biological fluids, in particular intrapleural, intraperitoneal, intralumbal, into the common lymphatic duct at a dose of 1-100 ml, as well as as a cleansing or application when treating open wound surfaces, and to improve systemic and local blood flow, regulate systemic arterial pressure and regional perfusion pressure, facilitate and accelerate mass transfer through gases, substrates and metabolites, intravascular or local injection of PFOS emulsion into tissues is performed, including perfusion and non-perfusion protection of organs and tissues isolated or disconnected from the bloodstream.

Thus, the reduction of the main process of destruction of PFOS emulsions caused by molecular diffusion is achieved in the claimed composition as in other patents (12, 17, 18) by introducing into the fluorocarbon base a component with low vapor elasticity (for example, PFMCP or PFBA) and less soluble in water, having a higher boiling point and slowing down this process. But at the same time, unlike the prototype, in the present invention, the “window” of the gap between the physicochemical characteristics of the various PFOS included in the core of the emulsion droplets is eliminated as much as possible, and the adsorption layer of the emulsion particles is formed from the CA mixture by self-assembly - by the method of saturation adsorption separation (US).

The PFOS emulsion for biological and medical purposes contains a mixture of rapidly releasing perfluorocarbons (PFCs), with the main component of PFD, and a mixture of perfluorinated tertiary amines (PFTA) with the main component of pperftop-N- (4-methylcyclohexyl) -piperidine (PFMCP) with lower speeds from the body, a stabilizing agent and a physiologically acceptable water-salt solution with energy metabolism substrates, PFOS also contain PFD mixtures; all fully fluorinated PFC impurities obtained in the synthesis of PFD; in the PFMCP mixture, all completely fluorinated PFTA impurities, and / or additionally contains perfluorotripropylamine (PFPA) with all its fully fluorinated PFTA impurities, and / or additionally contains perfluorotributylamine ( PFTBA) with all the fluorine impurities inherent in it when receiving PFTA, and / or additionally contains perfluorodihexyl ether (PFDE) with all inherent in it when receiving completely fluorinated impurities perfluorinated ethers (PPE) and a stabilizing agent in a mixture bloksoporimerov, with possible formation of selected mixtures of PFOS homogenous structure inside the emulsion droplets, due to the fact that PFOS mixture components have small differences in the critical solution temperature in hexane (T _KP g) at intervals of not over 2-4 ° C.

The PFOS emulsion is characterized by the fact that a mixture of all perfluorinated perfluorodecalin co-products having an average half-life of at least 12 days and a Tgr range of 22-29 ° C, purified of hydrogen-containing and unsaturated impurities, is used as rapidly excreting PFCs.

The PFOS emulsion is characterized in that, as PFTA, it contains a mixture of all PFPA co-products having an average half-life of at least 60 days and a range of T _cr 41–46 ° C, purified from and containing hydrogen. unsaturated impurities.

The PFOS emulsion is characterized in that PFTA contains a mixture of PFMCP co-products having an average half-life of at least 80 days and a T ^ r range of 32-40 ° C, purified from hydrogen-containing and unsaturated impurities.

The PFOS emulsion is characterized by the fact that as a slowly excreted PFOS it contains a mixture of co-products of PFDE having an average half-life from the body of at least 500 days and a range of T ^ g 49- 53 ⁰ C, purified from hydrogen-containing and unsaturated impurities.

The PFOS emulsion is characterized in that as PFTA it contains a mixture of co-products of perfluorotributylamine (PFTBA) having an average the half-life of the body is not less than 900 days, and the range of T _to rg 56-

60 ⁰ C, purified from hydrogen-containing and unsaturated impurities.

The PFOS emulsion is characterized in that the ratio of rapidly ascending and slowly removing PFOS is 5: 1 or 1: 5. The PFOS emulsion is characterized in that as a stabilizing agent it contains a mixture of copolymers of polyoxyethylene (POE) - polyoxypropylene (POP) with a molecular weight of 5-13 thousand Da, formed (selected) by saturation affinity separation (NAS).

The PFOS emulsion is characterized in that it contains a mixture of POE and POP copolymers selected by the NAS method as a stabilizing agent with a POE: POP weight ratio in the range from 78:22 to 82:18.

The PFOS emulsion is characterized in that the content of the stabilizing agent in the aqueous phase is 1-5%, to ensure a viscosity of 2-5 cPoise. The PFOS emulsion is characterized in that a physiologically acceptable water-salt solution includes NaCl, KCl, MgCl ₂ , NaHCO ₃ , NaH ₂ PO ₄ , Na ₂ HPO ₄ and D-glucose to ensure isosmoticity in the final dosage form, while maintaining the ratio of Na ions and K at a level of 8: 1 - 30: 1 and a pH in the range of 6.9 - 8.0. The PFOS emulsion is characterized in that the water-salt portion of the finished final drug. the form contains 110-150 mM NaCl, 4-5 mM KCl, 1-3 MM MgCl ₂ 6-20 MMNaHCO ₃ , 1-2, MM NaH ₂ PO ₄ 1 -2 MMNa ₂ HPO ₄ and 10-15 mm D-glucose .

The PFOS emulsion is characterized in that when used as an anti-ischemic protector, the water-salt portion contains 4-15 mm KCl, 10-20 mm D-glucose and salts of tri-, di- and monocarboxylic acids, including 1-7 mm citrate sodium, 1-7 mm sodium isocitrate, 1-7 mm sodium succinate, 1-7 mm sodium sodium ketoglutarate, 1-7 mm sodium pyruvate, 1-7 mm sodium β-hydroxybutyrate, 1-7 mm sodium glutamate, 1- 7 mM sodium aspartate. A method of preparing an emulsion of PFOS. for biological and medical purposes, it is realized by mixing and dispersing a mixture of liquid PFOS with an aqueous solution of a stabilizing agent, followed by grinding in a high pressure homogenizer to a submicron particle size of the emulsion, and:

- a mixture of PFOS containing components differing in T _{cr of} not more than 4 ° C, is poured into a container containing a solution of a mixture of CA in a ratio of 10-30 volume parts of PFOS to 90-70 volumes of a solution of a mixture of CA,

- the resulting heterogeneous mixture is subjected to the NAS procedure, carried out at a temperature of 50-60 ° C, including obtaining a coarse dispersion of PFOS in a solution of a mixture of CA by mixing on a high-speed mixer at a rotor speed of 1000-3000 rpm; the resulting coarse dispersion is subjected to extrusion in a homogenizer under a pressure of 100-300 kg per square. cm, in an oxygen-free atmosphere, and a coarse dispersion of PFOS is separated from an aqueous solution of CA; the obtained precipitate of a coarse dispersion is mixed with a fresh portion of a CA solution from such a calculation that the volume fraction of PFOS is 10-30%,

- while the described operation US is repeated at least 2 times,

- after which the emulsion is passed repeatedly through a homogenizer at a pressure of 300-700 kg per square. cm, - the resulting submicron emulsion is diluted with a water-salt solution so that the final concentration of PFOS is 4-12 vol% and the emulsion has an osmotic pressure in the range of 270-330 mOsm per liter. .

The method of preparing the PFOS emulsion is implemented in such a way that mixtures of liquid PFCs and PFTA or PFE are obtained in a ratio of 5: 1 to 1: 5, subjected to washing after mixing, with pyrogen-free water-alcohol solutions and water, and mixtures of denser liquid PFOS and water are separated. -alcohol solution and water, filtered under pressure of a gas not containing oxygen, poured and. cork into bottles for rolling and autoclaving.

The method for preparing the PFOS emulsion is implemented in such a way that the CA mixture is prepared in the form of a 10-30% aqueous solution of polyoxyethylene-polyoxypropylene block copolymers - a mixture of proxanol 168 and proxanol 268, taken in a ratio of 10: 1 to 1:10 depending on the composition of the PFOS mixture, clean the CA solution by passing through a carbon sorbent and a sterilizing filter under the pressure of an oxygen-free gas, warm the CA solution for 8-24 hours and cooled to 16-20 ⁰ C to completely restore the transparency of the solution.

The method of preparing the PFOS emulsion is implemented in such a way that an aqueous solution of a mixture of block copolymers of polyoxyethylene-polyoxypropylene is heated at a temperature that is selected in the range of ± 20 ^° C from the cloud point established experimentally for the obtained mixture CA.

The method of using PFOS emulsion to reduce the effects of hypoxia, ischemia, edema, inflammation in shock, blood loss, thromboembolic lesions, functional and organic vascular obstruction, tissue damage is realized by intravenous administration in a dose of 0.5-30 ml per 1 kg of body weight.

The method of using PFOS emulsion to suppress secondary alteration in inflammation and wound healing, as well as to accelerate the healing of traumatic, trophic and surgical wounds, is implemented by intravenous administration in doses of 0.3-6 ml per kg body weight and / or topically in portions of 0.5-50 . ml subcutaneously, intramuscularly, in depth around the damaged area, or into the cavity or biological fluids surrounding the inflamed and injured tissues, in particular intrapleural, intraperitoneally, intralumbally, into the common lymphatic duct at a dose of 1-100 ml, as well as a cleaning or application fluid when processing open wound surfaces.

The method of applying the PFOS emulsion to improve systemic and local blood flow, regulate systemic blood pressure and regional perfusion pressure, facilitate and accelerate mass transfer through gases, substrates, and metabolites is realized by intravascular or local injection into tissues, as well as by perfusion and perfusion-free protection of those isolated or disconnected from blood flow of organs and tissues.

The reduction of the main process of destruction of PFOS emulsions caused by molecular diffusion is usually achieved by introducing fluorocarbon base component PFOS with low vapor pressure

(for example, PFMTSP or PFTA) and less soluble in water, having a higher boiling point and slowing down this process. However, this leads to the appearance of inhomogeneities in the fluorocarbon phase, which reduce the stability of the emulsions. In order to increase the stability of PFOS emulsions, the present invention uses such PFOS mixtures that significantly increase the uniformity of the “oil phase” and contribute to the formation of the adsorption layer from the CA mixture by saturation affinity separation, when physical self-assembly and enrichment of the adsorption layer of the emulsion particles are most suitable for this PFOS compositions by CA molecules contained in the emulsion of the general CA mixture repeatedly updated during the preparation of the emulsion.

Judging by the diagrams of T _cr for PFOS according to RF patent N ° 2206319, there is a gap of more than 10 ⁰ C between them, which indicates the presence of quite significant differences in the physicochemical properties of the components of the “oil” kernel, and therefore it is not physically homogeneous, which is manifested in the clustering of the fluorocarbon phase and in the constant turbulent movement of clusters inside this phase. In order to increase the uniformity of the fluorocarbon phase in our invention, PFOS compositions are used, which practically form an almost continuous series of T _cr values and thereby significantly increase the uniformity of the “oil” phase, and increase the stability of PFOS emulsions. At the same time, the rates of removal of PFOS from RES cells and lipids change somewhat: the half-life of PFD when introduced as part of the proposed mixtures increases by an average of 10-15%, that is, it is not 12-13 days (typical for PFD outside these mixtures), but 14-15 days, while, on the contrary, the initial excretion rate of PFTA and PFDE increases by 10-15%, and therefore the half-life decreases by 5-7%.

Improving the uniformity of the oil phase is achieved at the stage of selection of PFOS. It is known that obtaining individual PFOS on an industrial scale is the most expensive step in the process of converting organic compounds in PFOS. Studies conducted by V.A. Arkhipov and K.N. Makarov showed that the toxicity of PFOS for human cells depends only on impurities of hydrogen-containing and unsaturated compounds - that is, on the presence of incompletely fluorinated substances, even if only one hydrogen atom or one unsatisfactory connection. Examples of formulas of such compounds found in insufficiently purified PFOS impurities are given below in the form of a structural diagram of some toxic impurities of under-fluorinated PFOS molecules and their radicals.

The most important point from the point of view of safety is that when the impurities of completely fluorinated substances vary in the PFD, the toxicity of the drug does not change if there were no underfluorinated PFOS in it: the mixture remains completely safe for animal cells (table

2). Table 2.

The effect of purified PFOS and PFOS containing impurities of under-fluorinated fluorocarbon compounds in an amount of 1 ppm.

Similar studies were performed for other types of PFOS, in particular for PFTA and PFE, and the same conclusions were drawn. This led to the proposal not to isolate the individual compounds PFD, PFMTSP, PFTPA, PFTA, PFDE and their co-products (close homologs, isomers and impurities), but to obtain these PFOS surrounded by minor impurities synthesized with them, isomers and homologues close in T _cr , but at the same time it is tough to ensure and control the release of the resulting substances from under-fluorinated impurities so that the level of under-fluorinated impurities does not exceed 1 part per million (1 ppm). In such a situation, the production efficiency of PFOS for the yield of a useful product also increases sharply, and accordingly, the cost of the process for obtaining PFOS drops without compromising their biological safety. Moreover, close homologues, isomers, and impurities (let's call them co-products) form an expanded field around the main components in terms of T ^ r values, which allows one to form PFOS compositions more uniform in physicochemical properties with a given field width T _crg (Fig. 2). At the same time, the stability of emulsions increases (Table 3).

For example, Table 3 shows a comparison of the contribution of a decrease in ΔT _kp g to the stability of PFOS emulsions with the contribution of a decrease in vapor pressure of PFOS. In the case when a mixture of PFBA and PFD was used, the stability of the PFOS emulsion is achieved due to 10 times less vapor pressure of PFTA (as compared to PFD), since PFBA diffuses weakly and prevents the diffusion of PFDA (example 4, Table C), despite by a large value of ΔT _kr g • If in this situation, reduce the value of ΔT _kr g by adding other PFTA having impurities of the products (according to the claimed invention), despite the loss in vapor pressure (which is several times higher for PFMTSP and PFTPA, than PFTBA) there is an additional increasing the stability of PFOS emulsions (example 5, table.Z). In measure 5, despite the replacement of part of PFBA with PFTA with higher vapor elasticity, instead of the expected decrease in the stability of emulsions, there was a significant increase in the stability of the new composition due to a decrease in ΔT _cr g inside the PFOS core. Figure l shows diagrams of critical dissolution temperatures of various PFOS in hexane.

The use of such PFOS with extended boundaries _Tcrg led to an unexpected biological effect: the rate of elimination of individual components from RES cells changed. So, if an individual PFD had a T / 2 of 12-13 days, then in the presence of PFMTSP and PFTA, this value for PFD was extended to 14-15 days. On the contrary, if an individual PFMCP was withdrawn from E / 2 of the order of 90 days, then in the presence of PFD, the T / 2 value for PFMCP decreased to 80 days, which allows us to understand the previously obtained data for T / 2 variations for PFMCP, previously presented in the scientific literature without taking into account the fact that the data were obtained in the case of T / 2 equal to 90 days for an individual substance, and T / 2 of the order of 65 days (according to our data this is a sharply underestimated estimate) was obtained for the PFD / PFMTSP mixture with a component ratio of 2: 1.

Comparison of the dispersion and stability of PFOS emulsions obtained according to previous patents (example 1 according to RF patent Ne 2070033), according to the prototype (example 2 according to RF patent Ne 2206319) with PFOS emulsions obtained according to the claimed invention (groups 3, 5 - table 3). Stability was checked during storage, freezing-thawing and when mixed with 'solutions of dextran (polyglucin) with an average molecular weight of 60,000 Da. The results are shown in table 3.

Table 3

Pi shows the level of significance of differences when compared with the group corresponding to the index number at (n = 9-10).

Matching the type of stabilizing agent (CA) of the PFOS composition plays a key role in dispersing PFOS, maintaining the stability and biological activity of PFOS emulsions.

To obtain PFOS emulsions, mainly two types of CA are used: 1) non-ionic block copolymer of polyoxyethylene (POE) ¹ With polyoxypropylene (POP) in the form of Proxanol-268 or its analog Pluronic F-68, as well as phospholipids (PL) isolated from egg yolk or soy, etc. i For proxanol 268, a centrally located hydrophobic POP chain is connected at its ends to two POE chains. According to the accepted nomenclature of the names of block copolymers, the number 8 in the name of proxanol 268 corresponds to the fact that the average total mass of POE blocks in the molecule is 80%, and the number 26 means that the average MM of POP, which is on average 20% of the mass of the proxanol molecule, is equal to 2600 D. Thus, the average MM of the proxanol 268 block copolymer as a whole is 13000 D, with the ratio of PEO and POP blocks on average 80% and 20%, respectively. Recall that in the synthesis of a block copolymer, as in the synthesis of any polymer, far from identical in MM and in the ratio of the individual blocks of the molecule are obtained. Therefore, they are characterized by an average MM, the presence of a molecular weight distribution that varies even in one batch, and the average ratio of POP and POE blocks.

Proxanol interacts with the hydrophobic part, that is, through POP with PFOS, and with water, the hydrophilic part, with the POE block. The stabilizing effect of proxanol and Pluronic is due to the steric effect of the protective film formed by CA molecules around PFOS particles. Moreover, along with CA bound in the adsorption layer, a significant part up to 50% of CA molecules form various micellar structures free of PFOS in the aqueous phase. Between the CA ₅ molecules located in the adsorption layer of the emulsion particles and in the aqueous phase in the micelles, the dynamic equilibrium is established, which is necessary to stabilize the adsorption layer on the one hand, and on the other hand, which violates the packing density of CA molecules in the adsorption layer during long-term storage. Due to differences in the compositions of CA bound in the adsorption layer of PFOS particles and free micellar CA, substitution as a result of the exchange of molecules of the adsorption layer by molecules from the micelles of the aqueous phase is unfavorable to maintain the stability of the adsorption layer. In the presence of sharp differences, the achievement of an effective (maintaining a stationary level and type of CA in the adsorption layer of particles) dynamic equilibrium between CA molecules in the particles and in the aqueous phase is practically unattainable. Note that when used as an emulsion stabilizer

PFOS of natural PLs, there are also problems of heterogeneity of the substance used both in composition and in quality (presence of polar components, free fatty acids, lysoform).

When choosing and determining the quality of the block copolymers taken as CA, we proceeded from an assessment of two criteria: stability of emulsions

PFOS and the degree of damage to biological membranes. It turned out that proxanol 268 provides for the same composition selected by us

PFOS significantly greater stability of emulsions than proxanol 168, apparently due to the more pronounced detergent (amphiphilic) properties of proxanol 268: on average, an emulsion from a mixture of PFD with PFMCP with co-products of both components taken in a 2: 1 ratio, stabilized with proxanol 268 without freezing was stored longer than in the case of using proxanol 168: the average doubling period of the average particle size (at 0.07 μm in the initial emulsion) was 45 + 4 days with proxanol 268 and only 30 ± 3 days with proxanol 168 (p <0.02).

However, proxanol 168 does not damage biological membranes due to weak detergent properties, in particular, it does not cause a separation of reactions oxidative phosphorylation upon addition to isolated mitochondria, a drop in the transmembrane electrochemical potential of hydrogen ions on the inner membrane, and suppression of the phosphorylated respiration of liver mitochondria (Fig. 3). On the contrary, proxanol 268 causes a marked decrease in the parameters of oxidative phosphorylation when added to isolated mitochondria and a decrease in the rate of phosphorylating respiration is evidence of membrane damage. Damaging activity against biological membranes in Pluropis F68 is even more pronounced (Fig. 3), despite the fact that formally it should simply be an analogue of proxanol 268.

Figure 2 shows the effect of various additives (50 μl) to rat liver mitochondria on the preservation of phosphorylated respiration during succinate oxidation (5 mM). Additives: non-fluorinated PDF (N-PFD), purified from non-fluorinated PDF impurities with co-products (PDF) and 4% aqueous solutions of Pluropis F68 (Pl F68), proxanol 268 (Pr268) and proxanol 168 (Pr 168). The incubation medium was 250 mM sasarose, 3 mM KH ₂ PO ₄ (pH 7.4), 1 mM MgCl ₂ , 15 mM KCl, 200 μm ADP. The concentration of MX 3 mg protein per ml. The cell volume is 2 ml. The incubation temperature is 26 ° C.

Since in the synthesis of proxanols the ratio of POE and POP blocks is only 80:20 on average, and in industrial batches of raw materials this ratio can vary in one direction or another, we conducted a study in which we determined the effect of variations of this ratio on the stability of emulsions and damage to biological membranes . It turned out that an increase in the proportion of POP above 18% is absolutely necessary to obtain more stable emulsions; a decrease in the proportion of POP below 18% reduces the retention time of the emulsions with proxanol 268 by a third, and sposanol 168 by 2 times. An increase in the proportion of POP block above 22% is unacceptable due to increased damage to biological membranes, and then both proxanol approach in their damaging properties to Pluris F68 (Fig. 3). Thus, the range of permissible variations in the selected types of proxanol was determined. In order to combine a sufficiently high stability of emulsions with reduced damaging ability in relation to biological of the objects represented by membrane structures at most levels of the organization of the body, we used a mixture of proxanols 268 and 168. It was found that emulsions containing PFBA along with PFD and other PFOS can be stabilized without a significant loss in stability with an increase in the proportion of proxanol 168, and these emulsions have reduced damaging ability with respect to biological objects, such as isolated perfused organs: heart and kidney, compared with emulsions with a high proxanol content of 268. But these emu Lysia can hardly be introduced into the body, because PTTBA has too long a delay period in the cells of RES (T / 2 is 900 days). For PFOS emulsions containing more lipophilic than PFBA components, proxanol 268 is more acceptable as CA, however, as indicated, damage to biological membranes increases. Ultimately, we came to the conclusion that for each PFOS composition, it is necessary to select its own mixture of proxanols 168 and 268, which is very laborious, since it requires additional tests to select a mixture of proxanol each time. The way out of this difficulty is achieved: by using a self-assembly procedure based on the use of physico-chemical affinity between the PFOS composition taken and the proxanol mixture 168 and 268. In the process of repeated self-assembly, the adsorption layer of PFOS dispersion particles is sequentially saturated with the CA composition that has the highest affinity to the taken PFOS composition, it is only important to speed up the saturation process, which is distilled by raising the temperature in the mixer to 50-60 ° C (depending on the proxanol mixture taken and its rate perature turbidity) and the update procedure iyuvtornostyu aqueous phase CA, unreacted particles PFOS dispersion.

So, to improve the stability of the emulsion during storage in the claimed invention, a saturating affinity separation method (HAC) is used, which allows optimizing the composition of the adsorption layer of the particles of the emulsion in accordance with the composition of the PFOS - “oil” core of particles and reducing the differences between CA in the adsorption layer of particles emulsion and micellar CA aqueous phase. US is made by several consecutive sessions, including mechanical mixing taken to disperse the PFOS composition in a volume of 10-

30 parts with 90-70 parts of a 20-30% CA solution, representing a mixture of proxanol 168 (average MM of about 8000 D) and proxanol 268 (average MM of about 13000 D). An inhomogeneous mixture having two layers of liquids: the heavy lower layer of liquid PFOS and the upper layer of micellar solution CA _{5 are} mixed on a high-speed mixer at a rotor speed of 1-3 thousand rpm with immersion of a rotating blade, turbine or pestle of a magnetic stirrer in the dense phase of PFOS, from. subsequent extrusion of the coarse dispersion using a homogenizer under a pressure of 100-300 kg per square. cm when blowing into the homogenizer of a gas that does not contain oxygen. The resulting coarse dispersion of PFOS is precipitated by gravity or centrifugation. Then, on the separatory funnel and / or using a centrifugal separator, the actual PFOS dispersion is separated from the aqueous solution of micellar composition CA that does not bind to PFOS, which is subsequently discarded. The separated coarse dispersion of PFOS is mixed with a fresh portion of an aqueous solution of CA so that the volume fraction of PFOS is again 10-30%. The process of dispersing and separating the dispersion of PFOS from an aqueous solution of CA is repeated again 1-3 times. The resulting pre-emulsion is passed repeatedly through a homogenizer at a pressure of 300-700 kg per square. cm to obtain an average particle size of the emulsion in the range from 0.05 to 0.1 μm when the emulsion leaves the particle size on a plateau.

Thus, in the process of obtaining the emulsion, it is PFOS due to physicochemical affinity that the CA molecules that have the highest affinity for them are selected from the solution of the mixture of POE-POP copolymers. Replacing the spent CA solution with a fresh solution allows not only to optimize the composition of the adsorption layer, but also to minimize differences between the compositions of CA in the adsorption layer and in the micellar aqueous part of the emulsion. Due to this, additional stabilization of the adsorption layer and increase in the stability of the PFOS emulsion as a whole are achieved in comparison with the prototype (Tables 3 and 4).

The obtained PFOS submicron emulsion is immediately mixed with a water-salt solution, in which the presence of salt ions promotes the formation of a relative negative charge on the radicals of the block copolymer molecules that are turned into the aqueous phase, thereby providing additional stabilization of the adsorption layer, since this forms a kind of double electric layer of dipoles of water molecules at the surface of the particles of the PFOS emulsion. As a result, the diffusion of non-polar PFOS molecules through the charged adsorption layer is additionally slowed down and thereby the molecular distillation of the “oil” phase from small particles to larger particles (Ostwald ripening of emulsions) is reduced, as well as additional electrostatic forces that repel particles from each other (electrical “dissipation”) ), which prevents the adhesion of the particles of the emulsion.

The effect of the NAS procedure when using a mixture of proxanol 168 with proxanol 268 in various PFOS formulations for the time (day) of doubling the average particle size with an initial particle size of 0.06 + 0.01 μm is illustrated in Table 4.

Table 4

P is indicated when compared with the prototype. In the claimed invention, the water-salt solution contains 1.5-2 times higher concentrations of salts, compared with the prototype, so that after mixing the submicron emulsion, with water-salt solution, when the final concentration of PFOS is reduced to 4-12 volume%, the aqueous part of the emulsion was isosmotic to the extracellular fluid, in particular, blood plasma and lymph, and contained sodium, potassium, bicarbonate, chlorine, phosphate, and magnesium ions, as well as substrates necessary for maintaining energy and plastic metabolism. Salts of NaCl, KCl, MgCl ₂ , NaHCO ₃ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , D-glucose, carboxylic acids and amino acids are used in the dosage form. The water-salt solution ensures the maintenance of the osmotic pressure at the level of 270-330 mOsm in the final dosage form, the ratio of Na and K ions at the level from 10: 1 to 30: 1 and the pH in the range from 6.9 to 8.0. In this case, the water-salt part of the finished final dosage form contains PO -150 mM NaCl, 4-5 mM KCl, 1-3 mM MgCl ₂ 6-20 mM NaHCO ₃ , 1-2 mM NaH ₂ PO ₄ 1-2 mM Na ₂ HPO ₄ and 10-15 mM D-glucose and can be supplemented with substrates that simultaneously play an anti-ischemic protective role, in particular salts of tri-, di- and monocarboxylic acids, including 1-7 mm sodium citrate, 1-7 mm sodium isocitrate, 1-7 mm sodium succinate, 1-7 mm sodium β-hydroxybutyrate; as well as those capable of converting to succinate during hypoxia and ischemia, 1-7 mm sodium α-ketoglutarate, 1-7 mm sodium pyruvate, 1-7 mm sodium malate, 1-7 mm sodium glutamate, 1-7 mm sodium aspartate.

The procedure for obtaining PFOS emulsion after NAS is completed by homogenization under high pressure, which can lead to damage, destruction and peroxidation of the molecules of the stabilizing agent. Therefore, it is necessary to take measures of protection against "oxidative stress". This is achieved by the fact that the process of preparing the components and dispersing - NAS and homogenization is carried out in an oxygen-free environment.

The proposed method allows, by updating the CA solution, to select from a mixture of CA having a wide molecular weight distribution and variations in the ratio of polyoxyethylene and polyoxypropylene blocks, CA molecules that have the highest affinity for the PFOS composition used, regardless of variations in the composition of the mixture of PFOS and CA, which increases the stability, chemical uniformity and calibration of the resulting PFOS emulsions, and also reduces the likelihood of CA peroxides and thereby increases the safety of emulsions. The PFOS emulsion obtained by the proposed method is more stable during storage and when it enters the bloodstream, and also has a viscosity close to blood viscosity, which ensures shear stress in the parietal blood flow and normalizes the tone of blood vessels, which is necessary to improve mass transfer between intravascular fluid and tissues both for oxygen and carbon dioxide, and for substrates and metabolites. It is proposed to use the PFOS emulsion as an agent for intravascular and interstitial administration with the aim of treating systemic and local disturbances in blood flow, hypoxic and ischemic conditions, improving the mass transfer of gases and metabolites between blood and tissues, maintaining the function of isolated organs and tissues, reducing inflammation, and conducting infusion transfusion therapy for shock and blood loss, as well as for hydrophobic anti-ischemic modification of membranes and maintaining microcatalysis occurring in gy Drophobic phase and at the interface.

Just like the prototype, the resulting finished dosage form is stored in a frozen state to slow down molecular distillation, stabilize the adsorption layer and prevent particles from sticking together. After thawing, despite maintaining the particle size for 2–3 weeks, it is recommended that, in the case of intravascular administration, use a thawed preparation stored at 2–4 ° C for 2–3 days. With interstitial or intracavitary administration, a thawed preparation can be used for 2 to 3 weeks.

More specific examples regarding the composition of the emulsion and the implementation of the method of its manufacture, as well as the method of application are given below. Example 1

The emulsion consists of a mixture of PFD (fast-releasing PFOS) containing, along with PFD, all fully fluorinated impurities, which are accompany the main substance in the synthesis, and a mixture of slowly excreted

PFOS - PFTA or PFE, for example, PFMTSP (NsI total ratio of components 2 parts PFD: 1 part PFMTSP) or a mixture of PFMTSP and PFTPA (N-! 2 total ratio of components 2: 0. 5: 0.5), or a mixture of PFMTSP, PFTPA and PFDE (NaZ total ratio of components 1: 1: 0.5), or a mixture of PFMCP and PFDE (Na4 total ratio of components 1: 1: 1), or PFMCP and PFBA (Na5 total ratio of components 1: 1: 2) containing with the main PFTA and PFE all fully fluorinated impurities accompanying the main substance in the synthesis. After sterilization and pyrogenation, the prepared PFOS mixtures are mixed with CA, which is a mixture of copolymers of proxanol 268 and proxanol 168, taken in different ratios, in particular for the NaI composition in the ratio 5: 1, for N ° 2 - 5: 2, for N ° 3 - 2: 1, for N ° 4 - 1: 1, for N ° 5 - 1: 2. The dispersion is obtained by the NAS method with 4 repetitions of the NAS procedure and subsequent extrusion at a pressure of 400 kg per cm ² - 6 cycles, then 3 cycles at 700 kg per cm ² , and then 2 cycles at 500 kg per cm ² : Emulsions with an average particle size from 0.6 to 0.8 microns. Example 2 The PFOS emulsion described in Example 1 contains one part of a mixture of PFD (fast-flowing PFOS) and two parts of a mixture of 1 slowly excreted PFOS NaI, or Na2, or Na 3, or Na4 or Na 5 stabilized by a mixture of a mixture of copolymers of proxanol 268 and proxanol 168, taken in different ratios, in particular for the NaI composition in a ratio of 1: 1, for Na 2 - 1: 2, for Na 3 - 1: 4, for Na4 1: 6, for Na 1:10. The dispersion is produced by the NAS method. The dispersion is obtained by the NAS method with 2 repetitions of the NAS procedure, including extrusion under a pressure of 100 kg per cm ² and then 200 kg per cm ² , followed by extrusion of the NAS product at a pressure of 400 kg per cm ² g 3 cycles, then 2 cycles with 600 kg per cm ² , and then 2 cycles at 500 kg per cm ² . Emulsions with an average particle size of 0.50 to 0.75 microns are obtained. The LD50 value of such emulsions reaches 165-200 ml per kg of mouse body weight with intraperitoneal administration. Example 3 ^{• • •} • ^• - The emulsions described in Example 1 or 2 contain PFOS components having a half-life of 12 to 900 days from animal RES cells after intravenous administration at a dose of 30 ml per kg of body weight. Moreover, the ΔТкрГ range for the components of the PFOS core in emulsions according to examples 1 and 2 is for NeI less than 2 ° C in the range from 22 ° C to 41 ⁰ C and the half-life is about 85 days, for N ° 2 ΔТкрГ does not exceed 3 ° C in in the range from 22 ° C to 47 ° C and a half-life of about 74 days, for N ° 3 and N ° 4 ΔТкрГ does not exceed 4 ° C in the range from 22 ° C to 56 ° C and a half-life of about 450 days, for N ° 5, the maximum value of ΔТкрГ is 4 ° C in the range from 22 ° C to 60 ⁰ C and the half-life of about 850 days.

Example 4

The emulsion described in Example 1 contains fast-releasing PFOS and slowly releasing PFOS in a ratio of 5: 1 (sample A) or in a ratio of 1: 5 (sample B) and a stabilizing agent in which for sample A it has a POE to POP ratio of 78:22 and for Sample B - 82:18. The average particle size in emulsions is 0.08 μm, the duration of perfusion of an isolated rat heart according to Languedorf using 11 mM glucose as a substrate is without loss of the initial spontaneous frequency (at the level of 250 cuts per minute) for sample A (with a high content of POP block in CA) 3 hours , and for sample B (with a low POP content) - 9 hours. Conclusion: to maintain an isolated heart, an emulsion is preferable, in which a mixture of proxanols with a low POP block content of up to 18% is used as a stabilizing agent. Example 5

When diluting the PFOS emulsion according to examples of 1,2,3,4 aqueous-saline solution, the content of PFOS was 4, 10 or 12 volume% depending on the amount of added water-salt solution, while the content of the stabilizing agent in the aqueous phase was 1%, respectively. 4% and 5%, and the viscosity of the finished product based on the PFOS emulsion reached 1.5 cPoise, 2.6 cPoise and 5 cPoise, respectively. When replacing a rat with 30% of the volume of circulating (BCC) blood with these emulsions in non-narcotic Wistar rats (animal weight 290-320 g) with implanted telemetry sensors, the following dynamics of mean arterial pressure was observed (table 5)

Table 5

BP * indicates the average of 3 animals.

As can be seen from the data presented in the table, an increase in the volume content of PFOS and the viscosity of the emulsion contributes to a longer retention of blood pressure after blood substitution.

Example 6

The finished product based on the emulsion PFOS according to examples 1-5 contains 110 mm NaCl, 5 mm KCl, 2 mm MgCl ₂ 8 mm NaHCO ₃ , 2 mm NaH ₂ PO ₄

1 mm Na ₂ HPO ₄ and 20 mm D-glucose at a content of 12 volume% PFOS. ι With massive hemorrhage in 55% of the circulating blood volume in dogs with the NsI emulsion (5 animals) and N ° 2 (6 animals) according to Example 1, all dogs survive, despite the significant difference in the shift of buffer bases, up to a deficit of 12, and growth blood lactate / pyruvate ratio up to 20. Example 7

The finished product based on the emulsion PFOS according to examples 1-5 contains 120 mm NaCl, 5 mm KCl, 1 mm MgCl ₂ 6 mm NaHCO ₃ , 1.2 mm NaH ₂ PO ₄ 1 mm Na ₂ HPO ₄ and 10 mm D-glucose with a content of 4 volume% PFOS. Emulsions with a low content of PFOS were administered intravenously to rats and their effect was tested at a dose of 1-2 ml on regional blood flow. Shown that, despite the small doses of PFOS administered, such doses of the emulsion are sufficient to activate blood flow in the event of edema or trauma in animals (see example 16 for details). The increase in blood flow, determined by a laser dopperometer, averages from 30% to 90%, depending on the degree of the initial blood flow disturbance. Example 8

The finished product based on the PFOS emulsion according to examples 1-5 contains 120 mm NaCl, 5 mm KCl ₅ 3 mm MgCl ₂ 6 mm NaHCO ₃ , 1.2 mm NaH ₂ PO ₄ 1 mm Na ₂ HPO ₄ and 20 mm D-glucose 1 mM sodium citrate, 1 mM sodium isocitrate, 1 mM sodium succinate, 1 mM sodium α-ketoglutarate, 1 mM sodium pyruvate, 1 mM sodium β-hydroxybutyrate, 1 mM sodium glutamate, 1 mM sodium aspartate with 8 volume% PFOS. The emulsion obtained was used to preserve a Langendorff-perfused heart of a rabbit on a special perfusion bench: 3 hearts comparison group (emulsion according to the prototype), 3 hearts experiment (emulsion N ° 2 and JVa 3 according to example 1 with salt and substrate obtained according to this example), 3 heart - control with water-salt solution with a composition of salts and substrates, as in this example. The retention time of the initial heart rate without stimulation in the control group was 1.5-2 hours, in the comparison group (according to the prototype) - 4-5 hours; in the experimental group according to the claimed invention - 6-7 hours. Example 9

The finished product based on the PFOS emulsion according to examples 1-5 contains 150 mM NaCl, 15 mM KCl, 3 mM MgCl ₂ 12 mM NaHCO ₃ , 1.2 mM NaH ₂ PO ₄ 1 mM Na ₂ HPO ₄ and 20 mM D-glucose 7 mM sodium citrate, 7 mM sodium isocitrate, 7 mM sodium succinate, 7 mM sodium α-ketoglutarate, 7 mM sodium pyruvate, 7 mM sodium β-hydroxybutyrate, 7 mM sodium glutamate, 7 mM sodium aspartate with 4 volume% PFOS. Used for cardioplegia of an isolated rat heart at a temperature of 18-20 ⁰ C. It was shown that, in contrast to conventional water-salt and pharmaco-cold cardioplegia, when an hour and after cardiac arrest, electrical and contractile ability are restored in no more than 50% of cases and the contractile recovery value activity does not exceed 75-80% (10 animals), when using emulsions PFOS composition N ° 4 and N ° 5 from example 1 with the specified the substrate-salt composition restored electrical and contractility in more than 70% of cases and the recovery rate of contractile activity was not lower than 90% (10 animals, p <0.02) /

Example 10 The resulting emulsion JN ° 4 according to example 1 with the substrate-salt composition according to example 8 was used as a perfusion composition and according to example 9 was used as a cardioplegic composition on an isolated rabbit heart in the following mode: normothermic perfusion with the emulsion according to example 8 for 30 minutes at at a temperature of 37 ° C (perfusion preservation procedure), then changing the perfusion solution to an emulsion, cardioplegic composition with the substrate-salt composition according to Example 9 and perfusion at 20 ° C until cardiac arrest (cardioplegia procedure), then 1 hour, the heart was disconnected from the bloodstream and placed in a bath with an emulsion of a cardioplegic composition, after an hour the heart was again perfused with an emulsion with a substrate-salt composition in Example 8. In the comparison group, an emulsion of the prototype was used as a perfusion composition and a cardioplegic composition with a full substrate-salt composition according to the prototype, supplementing in the latter case, the concentration of potassium ions up to 15 mm. The initial diastolic and systolic pressure, the degree of restoration of pulse pressure and the level of ATP and creatine phosphate in the heart tissue were measured 30 minutes after reconstructive perfusion at 37 ° C. In the experiment and in the comparison group of 5 experiments. It turned out that in the comparison group (prototype), diastolic pressure was 20% higher (evidence of residual contracture), and systolic pressure 10% lower (evidence of less contractile recovery

, myocardial abilities), respectively, the value of pulse pressure is lower on average by 30 + 10% (p <0.05) than in the group with the claimed formulation of emulsions

PFOS. The total content of adenyl nucleotides fell in the experimental group (the claimed formulation) by 30 + 5%, and in the comparison group (pototype) by 40 + 6% (differences are not significant), while in the comparison group (prototype) the ratio of ATP / AMP = 4.2 ± 0.5, whereas for the group with the claimed composition the ratio of ATP / AMP = 6.5 + 0.7 (p <0.05), respectively, in the group with The claimed PFOS emulsion compositions have a higher creatine phosphate concentration in heart tissue by 25 ± 7% (p <0.02). Thus, the claimed composition improves 30% recovery of the contractile function of the heart after cardioplegic arrest and helps preserve the energy status of the tissue judging by the ratio of ATP / AMP and the level of creatine phosphate.

Example 11 (a method of obtaining an emulsion of PFOS and the finished product). In a sterilized thermostatically controlled reactor (1) with a volume of 20 l with nitrogen injection while filling the reactor and with an integrated stirrer, pour with light stirring (200-300 rpm) 11 l of an aqueous sterile pyrogen-free solution of surface-active substances (CA) containing 10% proxanol 168 and 20% proxanol 268 (total CA concentration of 30%) and 4 l of sterilized pyrogen-free mixture of PFOS purified from under-fluorinated impurities containing 2 l of perfluorodecalin (PFD) with its inherent set of perfluororganic impurities, 1.5 l of perftopop-N - (4th ticyclohexyl) -piperidine with its inherent set when preparing a set of organofluorine impurities and a set of 0.5 l of a mixture of perfluorinated tertiary amines containing up to 40% as main substances, perftop-l-propyl-3,4-dimethylpiperolidine, up to 16% perftop-1 -propyl-3-metipiperidine, up to 40% perfluorotripropylamine with their inherent when receiving a set of perfluororganic impurities. Next, the NAS procedure is carried out. The poured mixture of PFOS and a stabilizing agent is intensively mixed at a stirrer speed of about 1000 rpm, heated to a temperature of 50 ° C, stirring is continued for 10 minutes. Then the mixer is stopped and allowed to settle - settle on the bottom of the coarse dispersion of PFOS. The supernatant in a volume of 9 l was pumped out of the reactor and 9 l of fresh solution of the initial CA mixture was added. I turn on the mixer again at a speed of up to 2000 rpm and heat the contents of the reactor to 50 ° C, continue stirring for 10 minutes. Then the mixer is stopped again, allowed to settle to the bottom of the coarse dispersion of PFOS. A supernatant in a volume of 6 L was pumped out of the reactor and 6 L of fresh CA stock solution was added. The cycle of vigorous stirring is again repeated with heating for 10 min. and the resulting dispersion is sent to the receiving tank of the high pressure homogenizer (2), in which the extrusion is carried out, under pressure in the range of 100-200 kg per square meter. cm, maintaining the temperature in the extruder valve area not higher than 60 ° C. The emulsion obtained at the exit from the homogenizer is separated in a centrifugal separator (3) from a solution of CA ₅ not bound to PFOS particles. The supernatant is drained and the emulsion concentrate is again poured into thermostatic reactor 1 and then the volume of the liquid phase in the reactor is adjusted to 15 L with a fresh solution of the initial CA solution (the volume of the added solution is about 4 L). The resulting mixture of the pre-emulsion and the CA solution is intensively mixed at a stirrer speed of 3000 rpm at a temperature of 50 ° C and fed to the recirculation loop of the homogenizer, in which extrusion is carried out under a pressure of 400 rbOQ kg per square meter. see maintaining the temperature in the area of the valve of the extruder not higher than 60 ⁰ C. The emulsion is passed through the extruder at least 6 times under the control of light transmission until it reaches a plateau. After the light transmission reaches the plateau, the emulsion goes through another homogenization cycle under a pressure of 400 kg per sq. Cm and an extruder temperature not exceeding 50 ° C. The resulting submicron emulsion is discharged into a sterile thermostatically controlled reactor (collection tank 4) with a stirrer, at the same time 1 ml of the submicron emulsion is taken for operational control (5-7 min) using a laser nanosizer (type N5, Verlip Сootter company), and to determine the volume of the fluorocarbon phase using a hematocrit centrifuge. If the average particle size exceeds 100 nm, the emulsion from the reactor 4 is again subjected to 3-4 extrusion cycles until then, at kA, the average particle size in a freshly prepared submicron emulsion does not decrease to 60-80 nm. Then the submicron emulsion is poured into the collection tank 4 in a volume of about 14 liters. 14 l of submicron emulsion contains about 26.6 volume percent of the fluorocarbon phase. To the obtained submicron emulsion, 23.24 l of sterile pyrogen-free concentrate of the water-salt composition is added with gentle stirring (100 rpm) so that, after the final dosage form is obtained, it contains PO mM NaCl, 5 mM KCl, 1 mM MgCl ₂ in the aqueous phase of the preparation 6 mM NaHCO 3, 1.2 mM NaH ₂ PO ₄ 1 mM Na ₂ HPO ₄ and 10 mM D-glucose. The resulting drug it is poured using an automatic machine, it is poured under pressure through a filter with a pore size of 0.4 μm in a tangential flow into sterilized vials, sealed with rubber stoppers with caps for running in and left for 6-8 hours at a temperature of 2 ÷ 4 ⁰ C. After measuring the average particle size (64 nm ) and their distribution (see Fig. 2), PFOS content (10 vol%), pyrogenicity (total Δ <0.6 ° C in three rabbits for 3 hours) and osmosis (300 mOsm), the vials are marked and placed in the freezer chamber for storage at minus temperature -17 -20 ° C. The LD ₅₀ value for the PFOS emulsion obtained in this example was 165-170 ml per kg of mouse body weight. Under the same conditions of administration, the emulsion obtained by the prototype had an LD _5O value of 140-150 ml per kg of mouse body weight. After intravenous administration by rabbits of the PFOS emulsion obtained in this example at a dose of 20 ml per kg, including after 2–3 repeated administration with an interval of 2 weeks, all rabbits survived for 4 months of observation. Example 12

The method according to example 11, characterized in that the initial mixture of PFOS containing 2 parts of a mixture of PFD, 0.8 parts of a mixture of PFBA and 0.2 parts of a mixture of PFMTSP after washing is washed with a pyrogen-free aqueous-alcoholic solution (BCP): mix the mixture of PFOS and BCP in equal volumes shake for 5 minutes and then separate the BCP phase. The remaining PFOS mixture is washed with aggrogenic water: the mixture of PFOS and water is mixed in equal volumes, shaken for 5 minutes and then the aqueous phases are separated. Then repeat the washing with pyrogen-free water again. The washed PFOS mixture is filtered through a filter with a pore size of 0.4 μm under pressure of nitrogen not containing oxygen and poured into glass vials. PFOS is poured into each bottle not more than 2/3 of the total volume of the bottle. The bottles are sealed with rubber stoppers for running-in and sterilized by autoclaving at 120 ° C. A stabilizing agent for a given composition of PFOS mixture is prepared by mixing 1 part of proxanol 268 and 1 part of proxanol 268 in the form of a 20% solution c _. pyrogen-free water. The resulting CA solution was passed through an activated granular carbon column and then through a cellulose acetate filter with a pore size of 1-2 μm by oxygen pressure not containing oxygen. Using a turbidimeter determine the cloud point of the resulting solution of a stabilizing agent when heated from room temperature at a rate of 1 degree per minute. The recorded cloud point, in this case, was 70 ° C. This temperature is used to heat the proxanol solution in the reactor, where the temperature retention accuracy is ± 2 ° C. The solution is heated for 8 hours, then slowly cooled to 18 ° C for 6 hours and check the degree of transparency on the nephelometer. A fully cooled solution should have the same light scattering as the initial solution before heating. Then carry out the NAS procedure, as described in example 10, despite the fact that the obtained submicron emulsion is divided into two portions. The first portion is diluted with a water-salt composition obtained according to example 8 in such a way that the PFOS content is 10 volume% (preparation for perfusion preservation of organs). The second portion of the emulsion is diluted with water-salt solution according to example 9 in such a way that the concentration of PFOS is 4% by volume (the drug for cardioplegia is the temporary preservation of the heart’s blood stream disconnected from blood flow).

The resulting preparation based on PFOS emulsion is intended for the treatment of systemic and local blood flow disorders, hypoxic and ischemic conditions, improvement of mass transfer of gases and metabolites between blood and tissues, maintenance of the function of isolated organs and tissues, reduction of inflammation, and infusion-transfusion therapy in shock and blood loss .

PFOS emulsion is used to reduce the phenomenon of hypoxia, ischemia, edema, inflammation in shock, blood loss, thrombotic-embolic lesions, functional and organic disorders of vascular patency, tissue damage by intravenous administration in a dose of 0.5-30 ml per 1 kg of body weight.

PFOS emulsions are used to suppress secondary alteration in inflammation and wound healing, as well as to accelerate the healing of traumatic, trophic and surgical wounds by intravenous administration in doses of 0.3 - b ml per kg body weight and / or topically in 0.5-50 ml portions subcutaneously. , intramuscularly, in depth around the damaged area, or in the surrounding inflamed and injured tissues of the cavity or biological fluids, in particular, intrapleural, intraperitoneal, intralumbal, into the common lymphatic duct at a dose of 1-100 ml, as well as a cleaning or application fluid when treating open wound surfaces. PFOS emulsion is used to improve systemic and local blood flow, to regulate systemic arterial pressure and regional perfusion pressure, to facilitate and accelerate mass transfer through gases, substrates and metabolites by intravascular or local injection into tissues, as well as during perfusion and non-perfusion protection of organs isolated or disconnected from the bloodstream and tissues.

Example 13 (method of using an emulsion for perfusion preservation of the kidneys)

PFOS emulsions composition of Na 4 and Na 5, according to example 1 (the production method according to examples 11 and 12, respectively) and the emulsion obtained according to the prototype, were used as perfusion formulations to maintain isolated dog kidneys in recirculation mode with normothermic perfusion with change of perfusate every 10 hours. The experiment was performed on 6 dogs in each group, a total of 18 dogs, weighing from 10 to 22 kg. As substrates in emulsion N ° 4, 10 mM D-glucose, 1 mM sodium citrate, 1 mM sodium isocitrate, 1 mM sodium succinate, 1 mM sodium sodium ketoglutarate, 1 mm sodium sodium pyruvate, 1 mm sodium β-hydroxybutyrate, 7 were used. mm glutamate sodium, 7 mm sodium aspartate. The time of preservation of the kidney before the rise in perfusion pressure (before the development of tissue edema) was 38 ± 3 hours (emulsion JN4) and 40 ± 3 hours (emulsion N ° 5), whereas when using the emulsion PFOS according to the prototype it was 34 + 2 hours. During perfusion, the emulsion was changed every 9 hours. The functioning of the preserved kidneys was tested in each case by replanting the perfused kidney at the moment of perfusion termination (30 minutes after the start of the exponential increase in the perfused pressure) to the recipient dog (under inhalation anesthesia, with premedication of promedol and sodium gamma hydroxybutyrate) to the femoral artery and femur . The restoration of blood flow, the elasticity of the kidney and the restoration of diuresis were recorded. In the experience with the prototype restoration of blood flow and diuresis was observed in 4 cases out of 6, with an emulsion N ° 4 in 5 dogs out of 6, with an emulsion N ° 5 in all 6 dogs, the transplant began to give urine immediately after restoration of blood flow.

Example 14. The use of the emulsion PFOS according to example 12 to suppress inflammation by intravenous administration in doses or locally in portions intramuscularly, in depth around the damaged area. The experiment was performed on 48 Wistar male rats anesthetized with thiopental and oxybutyrate, which, using a dental drill, applied a calibrated wound to the soft and bone tissues of the thigh. Then the bone wound was filled with bone crumb, and the soft muscle tissue, fascia and skin were sutured with tightening sutures. In the control group, animals were injected with saline in the tail vein at a dose of 1.5 ml per kg of body weight (group 1), or intramuscularly 0.3 ml above and below the wound (group 2); in groups 3 and 4, PFOS emulsion was administered, respectively in group 3 - intravenously, in group 4 intramuscularly. In all groups of animals, 4 animals were slaughtered on days 3, 5 and 7, at the indicated times in each group. Thigh tissues were fixed in formalin and then in Canadian balsam, after which soft tissues and bone tissue were subjected to pathomorphological examination. In the control groups (administration of saline solution intravenously and locally) on day 3, pronounced leukocyte infiltration phenomena were observed, leukocyte infiltrates penetrated the soft layers with thick layers, including those surrounding the initially undamaged tissues, the lesion site increased due to secondary alteration with a large number of osteoclasts along the periphery bone wounds; leukocyte infiltration decreased significantly on the 5th day, the first signs of connective tissue regeneration appeared and the number of osteoclasts decreased, the first osteoblastic elements appeared; on the 7th day, signs of lymphoid and epithelioid infiltration persisted, the number of osteoblasts and elements of the procollagenic framework of the connective tissue increased. In group 3 with intravenous administration of the emulsion already on the 3rd day the influence of leukocyte infiltration, ubiquitous lymphoid infiltration is weakly expressed, there are no phenomena of secondary alteration; 5 days in soft and bone tissue signs epithelioid infiltration and pronounced signs of the beginning of the regeneration of connective tissue elements; to ^"7 day amid widespread regeneration of connective elements appear separate differentiable cells with structures characteristic of the original tissue. In group 4, with intramuscular emulsion PFOS pathological pattern closer to that in group 3, despite the fact that the emulsion was administered not into the systemic circulation Thus, the administration of PFOS emulsion has a pronounced anti-inflammatory effect, reduces leukocyte infiltration and practically eliminates the phenomena of secondary alteration In case of inflammation and wound healing due to which there is no increase in the damage zone and regenerative healing processes begin earlier, these anti-inflammatory and wound healing processes are more pronounced with intravenous administration of PFOS emulsion, but these phenomena are unexpectedly bright in nature with local administration of PFOS emulsion.

Example 15. The treatment of fat and air embolism in rabbits. The PFOS emulsion obtained in Example 11 was used to treat fatty and airborne embolism in rabbits, which are especially sensitive to impaired brain vessels. Rabbits weighing 3-4 kg were injected through the ear vein with 10 ml of a coarsely dispersed emulsion of corn oil in water with an average particle size of 100-200 microns and visible air bubbles. After 3-5 minutes, the animals rolled their eyes, they fell on their side, one rabbit showed a disturbance in the respiratory rhythm according to Chain-Stokes, and one animal developed apnea - breathing stopped. 20 minutes after the rabbit lost consciousness, or 3-4 minutes after the appearance of Cheyne-Stokes breathing, or immediately after stopping breathing using a catheter previously fixed in the ear vein, rabbits were injected with PFOS emulsion at a dose of 4 ml per kg of body weight. The therapeutic effect in the form of removing animals from a coma and a state of clinical death (as a result of respiratory arrest) developed almost immediately “on the needle”: rhythmic breathing was restored, rabbits regained consciousness, opened their eyes, pupil reflexes were restored. In the control experiment with the introduction of physical. no solution of the therapeutic effect was observed: rabbits with Chain-Stokes breathing and apnea died.

Example 16. Regulation of regional blood flow and improvement of blood flow in the affected area. In rats after trauma to the thigh according to Example 13, tissue edema and impaired local blood flow on the injured hip were observed on days 1-3. Emulsion PFOS or physical. the solution was administered intravenously at a dose of 2 ml per kg of body weight or 0.2 ml locally at 4 points around the wound, retreating 5-8 mm. After 30 minutes, the systemic blood flow in the region of large vessels proximal to the wound and local blood flow directly above the sutured wound, as well as in the opposite, not damaged limb, were measured using an ultrasonic dopplerometer. Intravenous administration of the PFOS emulsion. The experiment was performed on 40 male Wistar rats weighing 250-300 g. The rats were administered as controls, which were injected with 0.9% sodium chloride solution intravenously (group 3, n = 10) and intramuscularly (group 4, n = 10). In all rats, the state of blood flow in the initial state was additionally evaluated prior to drug administration. Dopplerographic research was carried out on an ultrasonic diagnostic apparatus "Biomedical AU-5" (Italy) using a linear sensor with a frequency of 10-13 MHz according to the standard technique. Doppler mapping was performed based on the analysis of the direction and energy of the flow in the femoral arteries and in the tissues of the thigh before and after the introduction of the emulsion PFOS and physical. solution.

With the intravenous administration of PFOS emulsion, the maximum systolic blood flow velocity in the femoral artery exceeded the original more than 3 times (p <0.05), the minimum diastolic linear blood flow velocity along the vessel increased 2.5 times (p <0.05), the average speed - 4.5 times (p <0.05). At the same time, the resistance index decreased by 26.0% (p <0.05), which indicates a decrease in total peripheral vascular resistance. The pulse index significantly exceeded the background value by 2.02 times. When mapping, there was a significant increase in arterial and venous blood flow compared to the original in both the femoral artery and in the affected tissues of the thigh, while on the unaffected limb there was a clear tendency to decrease blood flow after intravenous administration of the PFOS emulsion. After intramuscular injection of PFOS emulsion increased blood flow velocity, but differences from control (saline solution) were not significant. When mapping, a significant increase in arterial and venous blood flow was visually observed compared with the initial in the femoral artery and in the tissues of the affected hip of rats. Energy Doppler mapping showed an increase in red blood cell mass both in the area of tissue infiltration with PFOS emulsion and with intravenous administration.

After intravenous administration physical. solution, the maximum systolic blood flow velocity in the femoral artery was 139.9%, the minimum diastolic linear blood flow velocity along the vessel was 161.1%, the average speed was 191.8% of the initial value (p <0.05). However, these changes in blood flow velocities were observed with a practically constant index of resistance, which indicates the preservation of spastic vascular tone in the microcirculation system. The pulse index characterizing the state of blood supply in the wound area exceeded the background value by 26.8% (p> 0.05). Differences in blood flow rates and indices of resistance and pulse with intramuscular injection of physical. solution in comparison with the initial background is not detected. When mapping, a slight increase in arterial blood flow was visually observed compared with the original in the femoral artery and in the affected tissues of the thigh. Energy Doppler mapping showed a slight increase in the mass of red blood cells in the area of tissue infiltration nat. solution and after its intravenous administration.

Thus, intravenous administration of PFOS emulsion in small doses significantly improves systemic and local blood flow in the wound area, while even a slight decrease in local blood flow occurred on the unaffected limb, which is in good agreement with the hypothesis of O. Rafikova et al. (2004) on the role of the phenomena of hydrophobic NO-dependent microcatalysis, manifested in improving blood flow, and the possibility of decreasing blood flow during NO sequencing with PFOS emulsion, which can cause a decrease in blood flow (as happened in our experiment on the healthy side). In this regard, the discovery turned out to be very significant significant effect on local blood flow after local administration of emulsion

PFOS.

Example 17

Intracavitary administration of PFOS emulsion to reduce inflammation. Male rats weighing 250-280 g under thiopenatol anesthesia underwent laparotomy and subsequent suturing of the abdominal wound with a clean but not sterilized instrument - without observing aseptic rules (5 animals control, 10 animals experiment, 5 for each emulsion PFOS). As a result, animals of the control group, which were intraperitoneally injected with 3 ml of physical. solution 24 and 48 hours after surgery, after 72 hours, pronounced infiltrative changes in the abdominal cavity and effusion in the form of neutrophilic activated white blood cells were observed. At the same time, the isolated leukocytes had a 3-5-fold increased ability to generate reactive oxygen species detected using a chemoluminometer in the presence of luminol after the addition of formyl myristyl acetate. When introduced into the abdominal cavity after 24 and 48 hours in 3 ml of PFOS emulsion obtained according to the claimed invention (samples N ° 2 and N ° 3 in example 1), the inflammation in the abdominal cavity was sharply reduced in animals, the effusion was so small that it was difficult to obtain a suspension of leukocytes, and the obtained leukocytes increased the production of reactive oxygen species after the addition of formyl myristyl acetate only 1.5-2 times. Pathomorphological analysis of organs in no case: neither in control nor in experience revealed the appearance of septic foci. Thus, intracavitary administration of an emulsion contributed to a decrease in the inflammatory process in the abdominal cavity without the use of antibiotics, suppressed leukocyte hyperactivity, without causing any septic complications that could be expected in connection with the suppression of PFOS emulsion of leukocytes penetrating into the abdominal cavity. Subsequent healing proceeded in the control with the formation of a rather pronounced hypertrophic scar, while in animals that were injected with PFOS emulsion twice intraperitoneally, the suture was soft and even without scarring. As a result, a stable PFOS emulsion was created, an effective method for its preparation and an effective method for its use.

Information sources:

1 K.N. Makarov et al., Zh. All-Union Chemical Society. DI. Mendeleev, 1985, t.Z., JУ ° 4, p. 387-394.

2 J.G. Riess // Scheme. Rev., 2001, v. Yul, Xa 9, p. 2797-2914.

3 E. Maevsky, Biological activity of perfluorinated carbons and their emulsions. In Sat _. Perfluorinated Carbon in Biology and Medicine, ed. By F.F. Beloyartsev. 1980, Pushchino, p. 121-125

4 Samples V.V. et al. Biophysical mechanisms of toxicity of fluorocarbon emulsions // Biophysics, 1994., vol. 39, no. 4, p. 732-737.

5 O. Rafikova at al., Sirsilatiop. 2004; vl 10: p. 3573-3580. 6 Mayevsku E.I. et al., IP "Substitutions" Ed. b RM Wipslow Amstördam, L, NY T ₅ Acad. Press Elsevier, 2006, Charter 26, p. 288-297.

7 Clark L.CJr, et al., Ip. Mat. Res. Soc. Sum. Roc. 1989, V.l 10, P.129-134

8 A.N. Sklifas et al. Study of the toxicity mechanism of perfluorodecalin emulsions for rabbits. // In Sat. “Productive active media for medicine and biology (new aspects of research).)) 1993,

Pushchino, from 129-135

9 Kaptsov B.B .; Samples V. In .; Kukushkin N.I. and etc.; RF patent JNb 2088217, 1997.08.27.

10 Ivanitsky G.R., Vorobyov SI., Deev A.A. “Life” of perfluorocarbon emulsion. // Sat “Physiological activity of fluorine-containing compounds (experiment and clinic).)) 1995, Pushchino, p. 5, -32;

11 Ivanitsky G.P. // Biophysics on the threshold of the new millennium: perfluorocarbon media and gas transport substitutes. // Russian Biomedical Technologies - 2000, vol. 1, November, p. 9-54]

12 Love KC IP "Substitutions" Ed. b RM Wipslow Amstördam, L, NY T, Acad. Press Elsevier, 2006, Charter 25, p. 276-287. Keirert R.E. IP Adv. Exp Honey. Biol. Ohhugep transport to Tissue XXV. Ed

T. Harrison and J. ICluver. Academis / Plepum Publishers, 2003, v. 530, Charter 29, p. 207-213. Keirert R.E. ip "Vloud Substitutes" Ed. bu R.M. Wiпslów Amstördam, L, N. Y. T, Acad. Press Elsever, 2006, Charter 28, p. 312-324. Collection "Perfluorocarbon compounds in medicine and biology), ed. G.R.Ivanitsky, E.B. Zhibyrta E.I. Maevsky, Pushchino, 2004, 267 p. Vorobiev SI., Kutyshenko SI., Sklifas A H. et al. Competentactivating effect of prefluorocarbon emulsions: a review. // Biocompatibility, 1995, t.Z, N ° l-2, p.51-62. Kuznetsova I.H., Mayevsky E.I. RF patent 2259819, publication date 2005.09.10. Mayevsky E.I., Ivanitsky G.P., Makarov K.N. et al. RF Patent N ° 2206319, publication date 2003.06.20, prototype.

Claims

Claim 1. An emulsion of organofluorocarbon compounds (PFOS) for biological and medical purposes, containing a mixture of perfluorocarbons (PFCs), the main component of which is perfluorodecalin (PFD) and a mixture of perfluorinated tertiary amines (PFTA), the main component of which is Perftop-N- (4-methylcyclohexyl ) -piperidine (PFMTSP), having lower than PFD, the rate of excretion from the body, a stabilizing agent and a physiologically acceptable water-salt solution with energy exchange substrates, characterized in that all the components of PFOS are known eniya critical solution temperature in hexane (T g _KP), which differ by no more than 2-4 ° C, and the stabilizing agent is a mixture of block copolymers from the group of block copolymers of polyoxyethylene-polyoxypropylene having an average polyoxypropylene containing 20 mass%., while the PFD mixture contains only completely fluorinated PFC impurities obtained by the synthesis of PFD, and the PFMCP mixture contains only fully fluorinated PFTA impurities, as well as at least one component from the group: mixtures of perfluorotripropropylamine (PFPA) with only lnostyu fluorinated impurities PPTA mixture of perfluorotributylamine (PFTBA) with only fully fluorinated impurities PPTA mixture perftordetsilovogo ester (PFDE) only with fully fluorinated impurities perfluorinated ethers (PPE). 2. The PFOS emulsion according to claim 1, characterized in that, as perfluorocarbons, it contains a mixture of perfluorinated coproducts of perfluoro decalin having an average half-life of at least 12 days and a range of T _cr in the range of 22-29 ⁰ C, purified from hydrogen-containing and unsaturated impurities. 3. The PFOS emulsion according to claim 1, characterized in that, as perfluorinated tertiary amines, it contains a mixture of all perfluorotripropylamine co-products having an average elimination half-life of at least 60 days and a range of T _cr in the range 41-46 ⁰ C, purified from hydrogen-containing and unsaturated impurities. 4. The PFOS emulsion according to claim 1, characterized in that as perforated tertiary amines it contains a mixture of coproducts of perftop-N-4- (methylcyclohexyl) piperidine having an average half-life of at least 80 days and a range of T _{cr of} within 32-40 ° C, purified from hydrogen-containing and unsaturated impurities. 5. The PFOS emulsion according to claim 1, characterized in that it contains a mixture of perfluorodecyl ether co-products (PFDE) having a mean half-life of at least 500 days and a range of T _cr from 49-53 ° C as slow-eliminating PFOS. purified from hydrogen-containing and unsaturated impurities. 6. The PFOS emulsion according to claim 1, characterized in that, as perfluorinated tertiary amines, it contains a mixture of perfluorotributylamine co-products having an average elimination half-life of at least 900 days and a range of T _cr in the range of 56 ° C, purified from hydrogen-containing and unsaturated impurities. 7. The PFOS emulsion according to claim 1, characterized in that the ratio of rapidly ascending and slowly removing PFOS contained therein is from 5: 1 to 1: 5. 8. The PFOS emulsion according to claim 1, characterized in that it contains a mixture of copolymers of polyoxyethylene (POE) -polyoxypropylene (POP) with a molecular weight of 5-13 thousand Da selected as a stabilizing agent. 9. The emulsion PFOS according to any one of paragraphs. 1.8, characterized in that as a stabilizing agent it contains a mixture of POE and POP copolymers selected by the NAS method with a POELOP ratio in the range from 78:22 to 82:18. 10. The emulsion of PFOS according to any one of paragraphs. 1.8, characterized in that the content of the stabilizing agent in it is 1-5%, to provide a viscosity of 2-5 spoises. 11. The emulsion of PFOS according to any one of paragraphs. 1-8, characterized in that a physiologically acceptable aqueous salt solution includes NaCl, KCl, MgCl ₂ , NaHCO ₃ , NaH ₂ PO ₄ , Na ₂ HPO ₄ , D-glucose to ensure isosmosis in the final dosage form, while maintaining the ratio of Na and K ions at a level of 10: 1 - 30: 1 and a pH in the range of 6.9 - 8.0. 12. The PFOS emulsion according to claim 1, characterized in that the water-salt portion of the finished final dosage form contains 110-150 mM NaCl ₅ 4-5 mM KCl, 1-3 mM MgCl ₂ 6-20 mM NaHCO ₃ , 1-2 mM NaH ₂ PO ₄ 1-2 mM Na ₂ HPO ₄ and 10-15 mM D-glucose. 13. The emulsion of PFOS according to any one of paragraphs. 1.12, characterized in that, for use as an anti-ischemic protector, the water-salt portion of the preparation contains 4-15 mm KCl, salts of tri-, di- and monocarboxylic acids, including 1-7 mm sodium citrate, 1- 7 mM sodium isocitrate, 1-7 mm sodium succinate, 1-7 mm sodium sodium ketoglutarate, 1-7 mm sodium pyruvate, 1-7 mm sodium β-hydroxybutyrate, 1-7 mm sodium glutamate, 1-7 mm sodium aspartate . 14. A method of preparing an emulsion of PFOS according to claim 1 for biological and medical purposes, comprising mixing and dispersing a mixture of liquid PFOS with an aqueous solution of a stabilizing agent, followed by grinding in a high pressure homogenizer to a submicron particle size of the emulsion, characterized in that - a mixture of PFOS having differences in T _{cr of} not more than 2-3 ° C, is poured into a container containing a solution of a mixture of CA in a ratio of 10-30 volume parts of PFOS to 90-70 volumes of a solution of a mixture of CA, - the obtained heterogeneous mixture is subjected to the NAS procedure, carried out at a temperature of 50-60 ° C, including obtaining a coarse dispersion of PFOS in a CA solution by stirring on a high-speed mixer at a rotor speed of 1000-3000 rpm, the obtained coarse dispersion is extruded in a g-genogenesis under pressure of 100-300 kg / cm ² in an atmosphere containing no oxygen, and the coarse dispersion was separated from the aqueous solution PFOS CA, the resulting precipitate coarse dispersion is mixed with a fresh portion of solution CA from this calculation to Ob mnaya PFOS fraction was 10-30%, - while the described operation US is repeated at least 2 times, - after which the emulsion is passed repeatedly through the homogenizer at a pressure of 300-700 kg / cm ^2, - the resulting submicron emulsion is diluted with water-salt solution so that the final concentration of PFOS is 4-12 vol% and the emulsion has an osmotic pressure in the range of 270-330 mOsm. 15. A method of preparing an emulsion of PFOS according to claim 14, characterized in that mixtures of liquid PFCs and PFTA or PFE are obtained in a ratio of 5: 1 to 1: 5, after washing, they are washed with pyrogen-free water-alcohol solutions and water, the mixtures are separated by more dense liquid PFOS from a water-alcohol solution and water, filtered under pressure of a gas that does not contain oxygen, poured and sealed into bottles for rolling and autoclaving. 16. The method of preparation of the PFOS emulsion according to claim 14, characterized in that the CA mixture is prepared in the form of a 10-30% aqueous solution of copolymers of polyoxyethylene-polyoxypropylene - proxanol 168 and proxanol 268, taken in a ratio of 10: 1 to 1:10, depending from the composition of the composition of the PFOS mixture, the CA solution is purified by passing through a carbon sorbent and a sterilizing filter under oxygen-free gas pressure, the CA solution is heated for 8-24 hours and cooled to 16-2O ⁰ C to completely restore the transparency of the solution. 17. The method of preparing the PFOS emulsion according to claim 14, characterized in that the aqueous solution of the mixture of polyoxyethylene-polyoxypropylene copolymers is heated at a temperature that is selected in the range of ± 20 ^° C from the cloud point established experimentally for the obtained CA mixture. 18. A method of treating diseases of the circulatory system, in which fast-releasing organofluorine compounds are introduced into the bloodstream or lymphatic duct, characterized in that the emulsion PFOS according to claim 1 is introduced into the bloodstream as fast-releasing organofluorine compounds. 19. The method according to p. 18, characterized in that for the treatment of the phenomena of hypoxia, ischemia, edema, inflammation in shock, blood loss, thrombo-embolic lesions, functional and organic disorders vascular patency tissue damage is carried out by intravenous administration of PFOS emulsion in a dose of 0.5-30 ml per 1 kg of body weight. 20. The method according to p. 18, characterized in that for the suppression of secondary alteration in inflammation and wound healing, as well as to accelerate the healing of traumatic, trophic and surgical wounds, intravenous administration of PFOS emulsion is carried out in doses of 0.3-6 ml per kg body weight and / or topically, in portions of 0.5-50 ml subcutaneously, intramuscularly, in depth around the damaged area, or into the surrounding inflamed and injured tissues of the cavity or biological fluids, in particular intrapleural, intraperitoneal, intralumbal, into the general lymphatic otok at a dose of 1-100 ml, as well as the cleaning liquid or the processing applicative open wound surfaces. 21. The method according to p. 18, characterized in that in order to improve systemic and local blood flow, regulate systemic blood pressure and regional perfusion pressure, facilitate and accelerate mass transfer through gases, substrates and metabolites, intravascular or local injection of PFOS emulsion into tissues is carried out, including including, with perfusion and non-perfusion protection of organs and tissues isolated or disconnected from the bloodstream.