[go: up one dir, main page]

WO2008082321A1 - Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation - Google Patents

Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation Download PDF

Info

Publication number
WO2008082321A1
WO2008082321A1 PCT/RU2006/000714 RU2006000714W WO2008082321A1 WO 2008082321 A1 WO2008082321 A1 WO 2008082321A1 RU 2006000714 W RU2006000714 W RU 2006000714W WO 2008082321 A1 WO2008082321 A1 WO 2008082321A1
Authority
WO
WIPO (PCT)
Prior art keywords
pfos
emulsion
mixture
solution
sodium
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/RU2006/000714
Other languages
English (en)
Russian (ru)
Other versions
WO2008082321A8 (fr
Inventor
Evgeny Ilich Maevsky
Genrikh Romanovich Ivanitskiy
Kirill Nikolaevich Makarov
Lev Lvovich Gervits
Viktor Vasilievich Moroz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Individual
Original Assignee
Individual
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to PCT/RU2006/000714 priority Critical patent/WO2008082321A1/fr
Publication of WO2008082321A1 publication Critical patent/WO2008082321A1/fr
Publication of WO2008082321A8 publication Critical patent/WO2008082321A8/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/131—Amines acyclic
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/02—Halogenated hydrocarbons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/075—Ethers or acetals
    • A61K31/08—Ethers or acetals acyclic, e.g. paraformaldehyde
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4453—Non condensed piperidines, e.g. piperocaine only substituted in position 1, e.g. propipocaine, diperodon
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A61K9/0026—Blood substitute; Oxygen transporting formulations; Plasma extender
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/08—Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/10—Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

Definitions

  • 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.
  • PFOS perfluororganic compounds
  • 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.
  • PFCs perfluorocarbons
  • PFTA perforated tertiary amines
  • 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.
  • PFOB
  • 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.
  • RES reticuloendothelial system
  • 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.
  • tsc half-life
  • T cr g The critical temperature of dissolution in hexane (T cr g), vapor elasticity (P) and half-life from organs (t ⁇ ) for various PFOS [1].
  • 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).
  • 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.
  • the PFOS emulsion can stimulate microcatalysis phenomena occurring in the hydrophobic phase or at the phase boundary.
  • PFOS 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.
  • 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.
  • 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.
  • 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.
  • the reactogenicity of PFOS emulsions cannot be eliminated only through the use of phospholipids as an emulsifier and stabilizer of PFOS particles.
  • 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.
  • the said patent empirically suggests selecting the composition of the oil “core” composition of PFOS and the CA interacting with it.
  • 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.
  • CA composition 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.
  • 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.
  • PFOS perfluororganic compounds
  • PFCs perfluorocarbons
  • PFTA perfluorinated tertiary amines
  • PFMTSP perfluorinated tertiary amines
  • T cr g critical temperature of dissolution in hexane
  • 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
  • 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 ko istov 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 ko etcov 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 ko apparatusov 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
  • 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
  • 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%,
  • 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.
  • 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.
  • 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).
  • 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
  • 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.
  • a component with low vapor elasticity for example, PFMCP or PFBA
  • 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 perfluo
  • 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-
  • PFTBA perfluorotributylamine
  • 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).
  • POE polyoxyethylene
  • POP polyoxypropylene
  • 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 ⁇ -hydroxybutyrate, 1-7 mm sodium glutamate, 1- 7 mM sodium aspartate.
  • a method of preparing an emulsion of PFOS 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
  • 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%,
  • 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.
  • 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 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.
  • 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%.
  • Table 3 shows a comparison of the contribution of a decrease in ⁇ T kp g to the stability of PFOS emulsions with the contribution of a decrease in vapor pressure of PFOS.
  • 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 ⁇ T kr g • If in this situation, reduce the value of ⁇ T 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).
  • CA stabilizing agent
  • 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.
  • POP polyoxypropylene
  • PL phospholipids isolated from egg yolk or soy, etc.
  • proxanol 268 a centrally located hydrophobic POP chain is connected at its ends to two POE chains.
  • 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.
  • 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.
  • 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.
  • a significant part up to 50% of CA molecules form various micellar structures free of PFOS in the aqueous phase.
  • proxanol 268 provides for the same composition selected by us
  • 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).
  • 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 ⁇ l) 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 ⁇ m ADP.
  • the cell volume is 2 ml.
  • the incubation temperature is 26 ° C.
  • 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).
  • proxanol 268 is more acceptable as CA, however, as indicated, damage to biological membranes increases.
  • proxanol 268 is more acceptable as CA, however, as indicated, damage to biological membranes increases.
  • 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.
  • 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.
  • HAC saturating affinity separation method
  • 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 ⁇ m when the emulsion leaves the particle size on a plateau.
  • 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.
  • 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.
  • 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 ⁇ -hydroxybutyrate; as well as those capable of converting to succinate during hypoxia and ischemia, 1-7 mm sodium ⁇ -ketoglutarate, 1-7 mm sodium pyruvate, 1-7 mm sodium malate, 1-7 mm sodium glutamate, 1-7 mm sodium aspartate.
  • substrates that simultaneously play an anti-ischemic protective role in particular salts of tri
  • 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.
  • 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.
  • 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.
  • Example 1 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.
  • 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.
  • the ⁇ influence beau ⁇ 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 ⁇
  • the half-life is about 85 days, for N ° 2 ⁇
  • N ⁇
  • N ⁇ 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 ⁇ intention beau ⁇ does not exceed 4 ° C in the range from 22 ° C to 56 ° C and a half-life of about 450 days, for N °
  • 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 ⁇ m, 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.
  • 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.
  • 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.
  • BP * indicates the average of 3 animals.
  • 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
  • Example 7 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 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.
  • 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.
  • 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 ⁇ -ketoglutarate, 1 mM sodium pyruvate, 1 mM sodium ⁇ -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.
  • 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 ⁇ -ketoglutarate, 7 mM sodium pyruvate, 7 mM sodium ⁇ -hydroxybutyrate, 7 mM sodium glutamate, 7 mM sodium aspartate with 4 volume% PFOS.
  • 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.
  • 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 PFOS.
  • 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).
  • 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 terti
  • 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 ⁇ ootter company), and to determine the volume of the fluorocarbon phase using a hematocrit centrifuge.
  • 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.
  • the resulting drug is poured using an automatic machine, it is poured under pressure through a filter with a pore size of 0.4 ⁇ m 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.
  • PFOS content 10 vol%
  • pyrogenicity total ⁇ ⁇ 0.6 ° C in three rabbits for 3 hours
  • osmosis 300 mOsm
  • 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.
  • BCP pyrogen-free aqueous-alcoholic solution
  • the washed PFOS mixture is filtered through a filter with a pore size of 0.4 ⁇ m 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 ⁇ m by oxygen pressure not containing oxygen.
  • 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.
  • 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.
  • 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 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.
  • 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.
  • mice 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.
  • groups 3 and 4 PFOS emulsion was administered, respectively in group 3 - intravenously, in group 4 intramuscularly.
  • 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.
  • 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.
  • Example 16 Regulation of regional blood flow and improvement of blood flow in the affected area.
  • tissue edema and impaired local blood flow on the injured hip were observed on days 1-3.
  • 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).
  • 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.
  • 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.
  • 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.
  • mapping 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.
  • 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.
  • 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.

Landscapes

  • Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Hematology (AREA)
  • Dermatology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Heart & Thoracic Surgery (AREA)
  • Cardiology (AREA)
  • Vascular Medicine (AREA)
  • Diabetes (AREA)
  • Urology & Nephrology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Hydrogenated Pyridines (AREA)
  • Medicinal Preparation (AREA)

Abstract

L'invention concerne le domaine biomédical. L'émulsion de composés organiques perfluorés, utilisée pour l'injection intravasculaire ou locale, est constituée d'un perfluorocarbone éliminable rapidement (décaline perfluorée) et d'une amine tertiaire perfluorée éliminable lentement perfluor-N-(4-méthyl-cyclohexyl)-pipéridine ainsi que d'une solution saline aqueuse physiologiquement acceptable. Les températures critiques de dissolution dans l'hexane (T[crit]H) de tous les composants des composés organiques perfluorés différent entre elles de 2 à 4°C au maximum, ce qui augmente la stabilité des émulsions de composés organiques perfluorés. L'émulsion de composés organiques perfluorés est stabilisée par un mélange de copolymères bloc polyoxyéthylène-polyoxypropylène et ne comprend que des impuretés entièrement fluorées de composés organiques perfluorés, ce qui réduit leur toxicité. Le procédé de fabrication de l'émulsion comprend le mélangeage et la dispersion d'un mélange de composés organiques perfluorés liquides avec la solution aqueuse d'un agent de stabilisation puis la réduction des particules d'émulsion dans un homogénéisateur à une taille submicronique. Le procédé de traitement de maladies du système sanguine consiste en l'injection de l'émulsion dans la circulation sanguine ou dans un canal lymphatique de l'émulsion de composés organiques perfluorés.
PCT/RU2006/000714 2006-12-28 2006-12-28 Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation Ceased WO2008082321A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/RU2006/000714 WO2008082321A1 (fr) 2006-12-28 2006-12-28 Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/RU2006/000714 WO2008082321A1 (fr) 2006-12-28 2006-12-28 Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation

Publications (2)

Publication Number Publication Date
WO2008082321A1 true WO2008082321A1 (fr) 2008-07-10
WO2008082321A8 WO2008082321A8 (fr) 2008-12-11

Family

ID=39588849

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU2006/000714 Ceased WO2008082321A1 (fr) 2006-12-28 2006-12-28 Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation

Country Status (1)

Country Link
WO (1) WO2008082321A1 (fr)

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4866096A (en) * 1987-03-20 1989-09-12 Air Products And Chemicals, Inc. Stable fluorochemical aqueous emulsions
US4943595A (en) * 1981-09-08 1990-07-24 The Green Cross Corporation Perfluorochemicals and process for preparing the same
RU2199311C2 (ru) * 2001-04-26 2003-02-27 Воробьев Сергей Иванович Состав перфторуглеродного кровезаменителя на основе эмульсии перфторорганических соединений для медико-биологических целей
RU2206319C2 (ru) * 2000-07-20 2003-06-20 Открытое акционерное общество Научно-производственная фирма "Перфторан" Эмульсия перфторорганических соединений для медицинских целей, способ ее приготовления и способы лечения и профилактики заболеваний с ее использованием
RU2259819C1 (ru) * 2004-03-01 2005-09-10 Кузнецова Ирина Николаевна Эмульсия перфторорганических соединений медицинского назначения и способ её получения

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4943595A (en) * 1981-09-08 1990-07-24 The Green Cross Corporation Perfluorochemicals and process for preparing the same
US4866096A (en) * 1987-03-20 1989-09-12 Air Products And Chemicals, Inc. Stable fluorochemical aqueous emulsions
RU2206319C2 (ru) * 2000-07-20 2003-06-20 Открытое акционерное общество Научно-производственная фирма "Перфторан" Эмульсия перфторорганических соединений для медицинских целей, способ ее приготовления и способы лечения и профилактики заболеваний с ее использованием
RU2199311C2 (ru) * 2001-04-26 2003-02-27 Воробьев Сергей Иванович Состав перфторуглеродного кровезаменителя на основе эмульсии перфторорганических соединений для медико-биологических целей
RU2259819C1 (ru) * 2004-03-01 2005-09-10 Кузнецова Ирина Николаевна Эмульсия перфторорганических соединений медицинского назначения и способ её получения

Also Published As

Publication number Publication date
WO2008082321A8 (fr) 2008-12-11

Similar Documents

Publication Publication Date Title
EP0627913B1 (fr) Augmentation de l'apport d'oxygene associee a l'hemodilution
AU629832B2 (en) Improved emulsions of highly fluorinated organic compounds
US4423077A (en) Perfluorochemical emulsion artificial blood
KR101623993B1 (ko) 소생액
Krafft et al. The design and engineering of oxygen-delivering fluorocarbon emulsions
JPH11507378A (ja) 酸素飽和状態（ｏｘｇｅｎａｔｉｏｎ ｓｔａｔｕｓ）をモニターすることによって簡便化された血液希釈法
JPH02198562A (ja) 組織酸素低下および虚血性障害治療のための酸素添加合成栄養物の脈管外循環装置
US5061484A (en) Perfluorochemical emulsion with stabilized vesicles
US4497829A (en) Process for preparing perfluorochemical emulsion artificial blood
DK152256B (da) Fremgangsmaade til fremstilling af en stabil bloderstatning
KR20090023620A (ko) 혈액 대용물 및 여타 치료적 용도를 위한 최적화 플루오로카본 에멀션
Geycr Perfluorochemicals as oxygen transport vehicles
RU2259819C1 (ru) Эмульсия перфторорганических соединений медицинского назначения и способ её получения
RU2393849C2 (ru) Эмульсия перфторорганических соединений медицинского назначения, способ ее приготовления и способ ее применения
RU2070033C1 (ru) Способ получения перфторуглеродных эмульсий для медицинских целей
US6562872B1 (en) Emulsion of perfluoroorganic compounds for medical purposes, a process for the preparation thereof and methods for treating and preventing diseases with the use thereof
CN1056746C (zh) 含有效成分的冻干乳液
JPS58502204A (ja) 合成全血およびその製造方法
WO2008082321A1 (fr) Émulsions de composés organiques perfluorés médicinaux, procédé de préparation et procédé d'utilisation
RU2199311C2 (ru) Состав перфторуглеродного кровезаменителя на основе эмульсии перфторорганических соединений для медико-биологических целей
EP0417104A1 (fr) Emulsion perfluorochimique contenue dans des vesicules stabilises
Kuznetsova About Mechanism of Gas Transport Function and Side Reactions of Perfluorocarbon Nano-dispersions
RU2775474C1 (ru) Газотранспортный перфторуглеродный гемокорректор и способ его получения
WO2020209752A1 (fr) Émulsion de composés de perfluorocarbone et procédé de production
CA2315877C (fr) Facilitation de l'oxygenation associee a une hemodilution

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 06851104

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 06851104

Country of ref document: EP

Kind code of ref document: A1