WO2005023218A1

WO2005023218A1 - Emulsion of perfluororganic combination with a function of gastransport

Info

Publication number: WO2005023218A1
Application number: PCT/KR2004/002235
Authority: WO
Inventors: Nikolay Kukushkin; Vladimir Kaptsov; Alla Sklifas; Eduard Pyagay
Original assignee: Krist Co., Ltd.
Priority date: 2003-09-04
Filing date: 2004-09-03
Publication date: 2005-03-17
Also published as: KR100862605B1; KR20060086356A; RU2240108C1

Abstract

Emulsion of perfluororganic combination with gas-transport properties is disclosed. The present emulsion comprises: a highly excreted PFC in amount 25~50 vol. %; a high-molecular perfluoreted additive in amount 2.5~20 vol. %; phospholipids in amount 2~10 mas. %; and a wide spectrum stabilizing additive. Especially, a ratio of the high-molecular perfluoreted additive to the highly excreted PFC varies from 1:2 to 1:10. Thus, the present has the following advantages: decreased average diameter of particles, higher dispersiveness, higher stability at storage, long storage, and simplification of requirement for storage, and so on.

Description

EMULSION OF PERFLUORORGANIC COMBINATION WITH A FUNCTION OF GASTRANSPORT

TECHNICAL FIELD

The proposed invention is concerned to the medicine and colloid chemistry and it can be used in a field of production of blood substitutes according to its gas-transporting function and chemically based on the emulsion of perfluorocarbons (PFCs). Our prospective invention can be used in medicine and pharmacy for production of the highly refined emulsions of the perfluorocarbons which are basic components in oxygen/CO -1ransporting blood substitutes. Moreover our system can be useful for cosmetics industry and scientific research practice for preparation of ointments and creams based on high refined emulsions of PFCs. BACKGROUDART

The emulsions of PFCs are compound colloid systems which consist of; dis persion medium based on water soluble surface active materials and dispersion phase based on PFCs. PFCs are major components of the given emulsion. They are chemic ally inert organic compounds and have high capacity to dissolve respiratory gases sue h as oxygen and carbon dioxide. PFCs are not soluble in water and therefore they m ust be emulsified during preparation of the blood substitute media. The PFCs' particle s are phagocytized by cells of the reticuloendothelial system and accumulate in differe nt organs. The half-excretion period is the quality performance of PFCs emulsions. Th e process of accumulation of PFCs in organism is a dose-dependent. In addition PFC s own properties (for example, the steams pressure, solubility in lipids, etc.) are also essential for this process. As a SAM (surface active materials) are used nontoxic high-molecular substa nces in particular proxanols (foreign analogues - pluronics and phospholipids). The am ount of proxanols in PFCs emulsion has to be possibly nrjuimalized and it is necessa ry only for dispersion and stabilization of PFCs. Influence of PFS emulsion on an organism, on the one hand, is mediated through physical and chemical influence of SAMs on cells of the organism. On the other hand, emulsion as the dispersion system has an influence to the whole organism. In particular, emulsion should not contain the big amount of large particles because of high risk of the capillary clotting and embolism. The toxicity, reactivity, average diameter of PFCs emulsion particles and stability at the storage are the important parameters of quality of PFCs emulsion. The PFCs emulsion toxicity is estimated by LD₅₀ parameter. The PFCs LD ₀ estimates the doze of emulsion injected to the animal (in m& per kg) at with survive 50 % of experimental animals. The PFCs emulsions should have long term storage and conditions of the storage should be simple. The storage parameters of the PFCs emulsion are estimated by change of average diameter of particles at the storage Vvithin 1~2 years. Dispersion and stability of emulsions PFCs depends on properties of the SAMs and from affinity of PFCs to SAMs. For example, while the phospholipids are used as SAMs, the temperature of dissolubility PFCs in hexane which serves as a lipophility measurement of the PFCs. Thus, below-mentioned parameters are detemiining at the characteristic emulsions PFCs

1) Toxicity is measured by LD₅₀ values 2) The half-excretion time period of PFCs from organism 3) Oxygen capacity of emulsion 4) Average diameter of emulsion particles 5) Change of average diameter of emulsion particles while autoclave sterilization 6) Change of average diameter of emulsion particles for 12 months at storage temperature from +4°C up to +30°C degrees 7) Dispensability of emulsion and change it changes during the storage. Based on described above parameters it is possible to determine the advantage of one PFCs emulsion from another. Nowadays are known PFCs emulsions of the first and second generations. The PFCs emulsions of the first generation were produced based on non-ionized SAMs or with addition of small amount of phospholipids and had the PFCs concentration no more than 20 vol.%. From these properties outgoes the limited oxygen capacity of PFCs: no more than 12 vol.%. The increasment of the PFCs concentration in emulsions of the first generation remains impossible due to sharp raise of viscosity. According to described characteristics only emulsions of the first generation with PFCs concentration no more than 10 vol.% were applied to the clinics. Perftoran (RUSSIA) is such an example (Patent RF #2070033, cl. A61 K 9/10,1994.). The emulsions of the second generation are based on phospholipid SAMs and have the PFCs concentration no more than 50 vol.%. Despite of the high PFCs concentration they have low viscosity. The oxygen capacity of these emulsions can achieve up to 26 vol.%. However, the low stability and comparatively larger average diameter of the particles are the disadvantage of these emulsions. Perftluordecalin (PFD) is the most widespread and easily accessible perfluroorganic component and can be used for medical and biological purposes. Widely known PFCs emulsion containing PFD in the amount of 10~20 vol. % and stabilized by proxanols (pluroniks) and phospholipids (Patent Great Britain GB #1361068, cl. A 61 K27/00, 1974; Clark L.C. & other, Ring versus straight chain perfluorocarbon emulsions for perϋision media, Microvascular Research, 1974, 8, p.320~340.). Those emulsions are concern to the first generation. Injected intravenously emulated PFD, as well as others PFCs, accumulate in a liver, spleen and bone marrow, thus it's not metabolizing in biochemical reactions. From an organism unchanged molecules of PFD are excreted with exhaled air within 7 day of half-life period. Relative harmlessness PFCs emulsions, containing PFD, were shown in numerous experiments, since RFfeyer" research (Federat. Proc, 1975, v. 34, p.1499-1505.). In mentioned above studies has been demonstrated the survivalence of rats after total blood replacement to the PFCs emulsion. However, it's well known fact, that the emulsion PFD stabilized by 3-4 % proxanol 268 solution is poorly stable in time and it's laminating in several days of storage at temperature +4^°C. The PFD emulsion, stabilized by combined

SAMs such as proxanol and phospholipids is more stable however, it has the same disadvantages. It has been established, that an addition to PFD any other perfluorocarbons increases the storage emulsion period. In PFCs emulsions, containing a mixture of two PFCs (so-called binary mixes PFCs) where one of components is fast excreted PFCs and another one is excreted from an organism rather slowly. Slowly excreted component improves stability of the emulsions at storage and reduces parameters of toxicity. Well-known PFCs emulsion of the first generation contains PFD in amount of 20 mas. % and also contains high molecular additive C16 HBPO (perfluoroperhyώrofluoroanthren) in amount of 1 mas. % stabilized by pluronic F68 in amount of 4 mas. % (Patent Great Britain GB #2171330, cl.B 01 F 17/42, 1986.; K.C. Lowe. Emulsified perfluorochemicals for oxygen-transport to tissues: effects on lymphoid system and immunological competence. Oxygen Transport to Tissue X. Plenum Press N.Y.-L., 1988, p.655-663.). The addition of the small amount of peφhorated polycyclic hydrocarbon-C16 HBPO which has higher temperature of boiling than the basic perfluorocaibon (PFD) slightly raises a period of storage of this emulsion (more than one month at temperature +4°C). However, this emulsion has the same disadvantages as are typical for PFD emulsions, such as: -low stability at storage -low oxygen capacity -it's reactive.

The known PFCs emulsion containing PFD and perfluor-n- methylcyclohexilpiperidin (PFMCP) in the ratio PFD/PFMCP=2/1 with concentration of the dispersive phase 10-20 vol. %, and stabilized by proxanol 268 in amount 4-8 mas. %, or proxanol 268 in amount 4-8 mas. % together with phospholipids of an egg yolk (lecitiiin) in amount 0.5 mas.% (Inventor's certificate USSR #1298976, cl. A 61 K 9/10, 1987.). The described emulsion has the average particles size at 0.06-0.126 microns and discretion of diameters of the particles from 0.05 up to 0.3 microns. The content of fluorine ions in this emulsion is not more than 2.3x10^"6 Mol/nii The present PFCs emulsion has toxicity LD₅₀=136 Hi g for mice. The known PFCs emulsion containing PFD and PFMCP in the ratio 2 tol with concentration of dispersion phase 10-20 vol. %, and stabilized by proxanol 268 in amount 4-8 mas. %( Patent RF #2070033, kl. A61 K 9/10,1994). The trading name of this emulsion is

"Perftoran". This emulsion after manufacturing has the average size of particles at 0.006-0.11 microns and discretion of diameter of particles from less than 0.1 up to 0.3 microns. The contents of the fluorine ions in this emulsion is 11 ~ 12 x 10^"6 M71 , the toxicity index LD₅o = 136-158 ml I kg for mice. Given emulsion have several disadvantages, such as: -low oxygen capacity -low stability at the storage (no more than 30 days at +4 ^°C) -for long storage the freezing (not less-18 ^°C) is necessary - It's reactive Nowadays the emulsions of the second generation are created in which new PFCs or already known PFCs are used. They are emulated only by phospholipids. The addition of phospholipids as emulators allows receiving emulsion with high content of the phluorocarbon phase at sufficient low viscosity and high oxygen capacity. These emulsions have lower reactivity. The second generation of PFCs emulsions containing perfluoroocthylbromide (PFOB) in amount from 20 to 125 mas. % and stabilized by phospholipids in amount of

5mas.% (Patent EPO #0231070, cl. A 61 K 9/10, 1987). A half-excretion period of PFOB lasts 4 days. The average size of the emulsion particles is 277 nm and 95 % of particles in size are less than 400 nanometers. This emulsion is stable during 18 months at temperature +4^°C.

After 15 months and 22 days the average diameter of particles increases up to 406 nanometers. The index of stability-Dt Do which is designating in how many times the average diameter of emulsion particles has increased after t months of storage, for this emulsion is equal to 1.4.

The preparation based on this emulsion is known under the trademark Oxygent (USA). It has oxygen capacity equal to 21 vol. %. The emulsion Oxygen has the following disadvantages: -relatively large average diameter of emulsion particles (more than 250 nm) -insufficiently high stability at the storage.

The most especially relative to declared PFCs emulsion is emulsion, described in the patent J ° 2088217 (Patent RF #2088217, cl. A 61 K 9/10, 31/02), and we took this emulsion as a prototype. This emulsion PFC is emulsion of the second generation. It contains PFD and PFOB in amount of 40-50 voV % and PFMCP in amount of 1-10 vol. % and stabilized by phospholipids in amount 2-6 vol. %. Average diameter of particles of this emulsion is equal to 232 nanometers. After 11 months of storage at +4°C the average diameter of particles of this emulsion increases in 23 %, it means that the index of stability is equal to 1.23. This emulsion has an oxygen capacity up to 22 vol. %. Disadvantages of this emulsion are following: -the large average diameter of particles -the high contents of large particles (14.9 % of particles have diameter more than 400 nanometers, look table 1 of the true application) -insufficiently high stability at storage.

DISCLOSUREOFTHEINVENTION

The purpose of present invention is the reduction of the average particles size, the increasement of dispersion of the emulsion, increasement of emulsion stability at storage, and decreasment of the requirements of the storage conditions. The defined purposes are achieved by addition to the emulsion of components such as: the special stabilizing additive of a wide spectrum, highly excreted perfluorocaibon in amount 25-50 vol. %, high-molecular perfluoreted additive in amount of 2.5-20 vol. % and phospholipids in amount of 2-10 mas. %. Thus the ratio of the contents of high-molecular perfluorated additive to highly excreted perfluorated component additives ranges from 1:2 till 1:10. As the stabilizing additive of a wide spectrum fat acids with CIO on C22 and/or their salts, or triglycerides are used. In the suggested emulsion PFD - highly excreted perfluorocarbon is the base of perfluorocarbone phase and it provides the transport of major amount of dissolved gases. The second perfluocarbone component PFMCP the high- molecular perfluorocarbone additive is applied by as a transporter of gases, and as the stabilizator of emulsions. Phospholipids of egg yolk or soy are used as emulators of perfluorocarbone. Special stabilizing additives are supplied for reduction of the average diameter of emulsion particles and also for the increasment of stability during the long storage. The emulsion includes antioxidants, for example, L-tocopherol in concentration range from 0.01 to 0.05 g/ . Except for that the emulsion can include salts as: sodium chloride (6 g/ 1 ), potassium chloride - 0.39 g/ i , magnesium chloride 0.19g/ i , sodium hydro carbonate - 0.65 g/ 1 , sodium hydro phosphate - 0.2 g/ 1 and glucose - 2 g/ 1 . Along of incomplete representation of characteristics of the Prototype (curves of distributions of particles in the diameters are not presented) there is no data of the storage at room temperature (20-24 ^°C) and at temperature of the bodies of experimental animals (approximately +40 ^°C). We also prepared control emulsion according to structure and way of preparation of the prototype. The control emulsion, as well as offered was prepared in the homogenizer at high pressure (pressure range 40-60 MPa) in aseptic conditions providing sterility and apyrogenicy of emulsion. Average diameter of emulsion particles and distribution of particles in the diameters were measured by photon-correlation spectrometer: "Autosizer II " (" Malvem " Great Britain). Prepared emulsions were sterilized by autoclave sterilization under the pressure of 1 arm (heating up to +120 ^°C) within 60 minutes, and further it was stored: (a) in a refrigerator at +4^°C, (b) in a room at +20-24 ^°C and (c) in a thermostat at +40 ^°C. Sharp toxicity (LD₅₀) of emulsions was determined on mice by a standard technique.

BRIEF DESCRIPTION OFTΗE DRAWINGS

Fig. 1 shows the examples of dependence curves of average particles diameter of various emulsions from a storage time of these emulsions in conditions +4^°C. Fig.2 shows the examples of the dependence curves of average particles diameter of the various emulsions from a storage time of those emulsions at room temperature conditions (+20-24 ^°C). Fig. 3 shows the examples of the dependence curves of average particles diameter of various emulsions from a storage time of these emulsions in thermostat (+40 ^°C). Fig. 4 shows the examples of distributional curves of particles of the freshly prepared emulsions. Fig.5 shows the distributional curves of the fresh prepared emulsions. Fig. 6 shows the distributional curves examples of the presented emulsion after 12 months preservation at room temperature +20-24 ^°C .

BEST MODES FOR CARRYING OUT THE INVENTION Now, preferred embodiment of the present invention will be described in detailwith reference to the annexed drawings. Fig. 1 shows the examples of dependence curves of average particles diameter of various emulsions from a storage time of these emulsions in conditions +4^°C. The horizontal axis X represents a storage time in days, the vertical axis is an average diameter of emulsions particles in nanometers. The curve 1 shows the dependence of average diameter of emulsion particles of Prototype, taken from the reference (Patent RF #2088217, cl. A 61 K 9/10, 31/02). The curve 2 represents the dependence of the average diameter of emulsion particles which are structurally completely similar to the Prototype emulsion and it was taken as a control emulsion. The control emulsion was made by mixture of the following ingredients: PFD in concentration of 36 vol.%, the concentration of PFMCP - 4 vol.%. In this emulsion, the ratio

PFMCP/PFD was 1/9. The control emulsion was stabilized by addition of phospholipids of egg yolk in amount of 5 mas. %. The curve 3 shows the dependence of average diameter of particles of the emulsion consisted of PFD - 36 vol. %., concentration PFMCP - 4 vol. %, the ratio of PFMCP/PFD was 1/9 and was stabilized by phospholipids of egg yolk in amount 5 mas. % and palmitic acids in the amount of 0.1 mas.. %. The curve 4 shows the dependence of average diameter of particles of the emulsion consisted of PFD - 36 vol.%., concentration PFMCP - 4 vol. %, the ratio of PFMCP/PFD was 1/9 and was stabilized by phospholipids of egg yolk in amount 5 mas. % and sodium palmitat in the amount of 0.1 mas.. %. From Fig. 1, follows that samples of suggested emulsion have smaller diameter and lower speed of integration in comparison with the Prototype and control emulsion. Next, Fig. 2 shows the examples of the dependence curves of average particles diameter of the various emulsions from a storage time of those emulsions at room temperature conditions (+20-24 TJ). The same designations as on Fig.1 are submitted. From Fig.2 become achieved, that samples of the performed emulsion have smaller diameter and lower integration speed in compare to the same parameters of control emulsion. Fig.3 shows the examples of the dependence curves of average particles diameter of various emulsions from a storage time of these emulsions in thermostat (+40 TJ). The same designations, as on Fig.l are submitted. From Fig. 3, follows that samples of suggested emulsion have smaller diameter and lower speed of integration in compare with the control emulsion. Fig. 4 shows the examples of distributional curves of particles of the freshly prepared emulsions. The curve 1 is corresponding to the emulsion which is st cturally completely similar to the Prototype (control emulsion): the amount of PFD - 36 vol. %., concentration PFMCP - 4 vol. %, PFMCP/PFD ratio = 1/9, stabilized by phospholipids of egg yolk in the amount of 5 mas.%. The curve 2 corresponds to the emulsion with the amount of incorporating PFD is - 32 vol. %., concentration PFMCP - 4 vol. %. PFMCP PFD ratio = 1/9, stabilized by addition of phospholipids of egg yolk in amount 5 mas.% and palmitic acids in amount of 0.1 mas. %. The horizontal axis X represents the average diameter of emulsions particles in nanometers, the vertical axis represents the number of particles corresponding to such diameter (%). Fig. 4 represents that the sample of the actually presented emulsion is ftinctionally distributed and contains lower amount of large particles in compare to the control emulsion. Fig. 5 shows the distributional curves of the fresh prepared emulsions. The curve 1: emulsion are completely analogous to the prototype emulsion (control emulsion) by consistence: the concentration of PFD-36 vol.%, the concentration of PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mas.%. The curve 2 graphically represents the emulsion which consists on PFD-36 vol.%, PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mass%, and contains an additionally stabilized by sodium palmitat-O.l mass%. The axial significance is the same as at Fig.4. Fig. 5 shows that the sample of the presented emulsion has narrow distribution and lower amount of large particles in compare to the control emulsion. Fig. 6 shows the distributional curves examples of the presented emulsion after 12 months preservation at room temperature +20-24 TJ. The curve 1 represents emulsion completely similar by its components to the prototype emulsion (control emulsion). The concentration of PFD-36 vol.%, the concentration of PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mas.%. The curve 2 graphically represents the emulsion which consists on PFD-36 vol.%, PFMCP-4 vol.%, PFMCP PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mass%, and contains an additionally stabilized by sodium palmitat-O.l mass%. The axial significance is similar to Fig. 4. Fig. 6 shows that the sample of the presented emulsion has narrow distribution and lower amount of large particles in compare to the control emulsion. Table 1 shows the results of comparative analysis of the prototype and prospective emulsion.

Table 1. Comparative analysis of the prototype and prospective emulsion

In Table 1, Indications are used as follows: D₀-the mean diameter of particles of the fresh prepared emulsion. D₁₂ is the mean diameter of the emulsion particles after 12 months preservation at +4 ^°C , +20-24 ^°C and +40 ^°C . D π/Do-stability index (the difference in the mean diameter of the emulsion particles after 12 months storage at +4^°C, +20-24 ^°C and +40 ^°C degrees respectively. The control emulsion is completely analogous to the prototype emulsion by consistence: the concentration of PFD-36 vol.%, the concentration of PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mas.%. The components of the presented emulsion with palmitic acids are: PFD - 36 vol. %, concentration PFMCP - 4 vol.%, the ratio of PFMCP/PFD is 1/9, stabilized by phospholipids of egg yolk in amount 5 mas.%, and contains palmitic acids in the amount of 0.1 mas.%. The components of the presented emulsion with sodium palmitat are: PFD-36 vol.%, PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk phospholipids in the amount of 5 mass%, and contains an additionally stabilized by sodium palmitat-0.1 mas.%. Table 1 shows the difference between mean diameters of the emulsion particles after 12 month storage at +4^°C. The mean diameters of emulsion particles of the prototype and control emulsion increased in 23% and 18% respectively. In contrast, the mean diameter of the particles in the samples of the prospective emulsion increased only in 4% and 8% respectively. Thus, the average enlargement speed of the emulsion particles in presented samples was 3-6 times lower than the average enlargement speed of the particles of the prototype and 2~5 times lower than the average enlargement speed of the particles of the control emulsion. The analogous results were received after storage the emulsion under temperature severity: +20 ^°C

- +40 ^°C. The storage at +40 ^°C such as the storage at +24 TJ decreased the average enlargement speed of the emulsion particles in 2 times in compare to the control emulsion.

The comparison of the present emulsion with the prototype emulsion under analogous conditions was impossible due to the absence of the corresponding data. The similar to the described above results were also received for the different samples of the presented emulsion which contained the different stabilizing ingredients (data not shown). Table 1 demonstrates that samples of the presented emulsion in compare to the control emulsion have lower values in the average particles diameter. The narrow functional distribution of the presented samples shows lower amount of the large particles. The number of particles which have more than 400nm in diameter is decreased in 2-4 times in the samples of the prospective emulsion compare to control. The analogous results have been received for other samples of the prospective emulsion. Thus, the application of the additives essentially increases the stability of the emulsion during preservation. Hereinafter, examples of preparation and components of the prospective perfluorocarbon emulsion according to the present invention are explained.

Example 1. In emulsion with the concentration of PFD-32 vol.%, and the concentration of PFMCP-8 vol.%, PFMCP PFD ratio-1/4, and stabilized by yolk phospholipids in the amount of 5 mas.%, we added stabilizing substance - palmitic acid in amount of 0.1 mas%. The emulsion was prepared by following recipe. After evaporation in the rotor-evaporator of the 10% lecithin solution, the salt hydration of the dry lecithin residue was carried in the presents of the palmitic acid during the hydration procedure. The L-tocopherol in the concentration 0.02 g 1 was also added during the hydration. Then the actual mixture was homogenized in the plimger homogenizer with drop-by-drop introduction of the PFCs, and 60 MPa (600kg/cm²) pressure during the time of homogenizer intake cycles (10-20 homogenizer cycles). As a result of homogenization we received the emulsion where the average particles diameter was equal to 198 nm. After sterilization of the actual emulsion in autoclave (120 TJ, latm) for 60 minutes the mean diameter of the particles of the emulsion was equal to 201nm. 12 month later after the storage at room temperature the mean diameter of the particles became 221nm.The stability index was 1.09 for the present emulsion and LD₅₀ for mice was

140itt2/kg. For the control emulsion LD₅₀ was 136ra-β/kg.

Example 2. In emulsion with the concentration of PFD-32 vol.%, and the concentration of

PFMCP-8 vol.%, PFMCP/PFD ratio - 1/4, stabilized by yolk lecithin in the amount of 5 mas.%, we added stabilizing substance- sodium palmitat in amount of 0.1 mas%. The emulsion was prepared as described in example 1. After preparation the mean diameter of the emulsion particles was 195 nm. After 12 month of storage the average diameter of the particles became equal to 219nm. The stability index of the emulsion in this case was 1.12.

LD₅₀ for mice was equal to 146 ml/kg. For the control emulsion LD₅₀ was 136 inM g.

Example 3. In emulsion with the concentration of PFD-32 vol.%, and the concentration of PFMCP-8 vol.%, PFMCP/PFD ratio is 1/4, stabilized by yolk lecithin in the amount of 5 mas.% we added stabilizing substance-trioletat glycerol in the amount of 0.1 mas.%. The emulsion was prepared as described in example 1. After preparation the average diameter of the emulsion particles was 210 nm. After 12 month of storage at the room temperature the average diameter of the particles increased up to 238nm. The stability index of this emulsion per yearwas 1.13. LD₅₀ for mice was equal to 136 ni-β/kg. For the control emulsion LD₅₀ was

136 ni-P/kg. Example 4. In emulsion with the concentration of PFD-20 vol.%, and the concentration of PFMCP-5 vol.%, PFMCP/PFD ratio is 1/4, stabilized by yolk lecithin in the amount of 2 mas.%, the stabilizing substance- palmitic acid was added in the amount of 0.05 mas.%. The emulsion was prepared as described in example 1. After preparation the average diameter of the emulsion particles was 221 nm. After 12 month of storage at the room temperature the mean diameter of the particles increased up to 265nm. The stability index of this emulsion per year was equal to 1.2. LD₅₀ for mice was equal to 158 ml/kg. For the control emulsion LD₅₀ was 136 inMcg.

Example 5. In emulsion with the concentration of PFD 0 vol.%, and the concentration of PFMCP-10 vol.%, PFMCP/PFD ratio is 1/4, stabilized by yolk lecithin in the amount of 10 mas.%, the stabilizing substance- sodium palmitat was added in the amount of 0.25 mas.%.

The emulsion was prepared as described in example 1. After preparation the average diameter of the emulsion particles was 190 nm. After 12 month of storage at the room temperature the average diameter of the particles increased to 205 nm. The stability index of this emulsion per year was equal to 1.08. LD₅₀ for mice was equal to 138 niM g. For the control emulsion LD₅o was 136 mMcg.

Example 6. In emulsion with the concentration of PFD-32 vol.%, and the concentration amount of PFMCP-4 vol.%, PFMCP/PFD ratio is 1/9, stabilized by yolk lecithin in the amount of 6 mas.%, the stabilizing substance- rrioletat glycerol was added in the amount of 0.2 mas.%. The emulsion was prepared as described in example 1. After preparation the average diameter of the emulsion particles was 213 nm. After 12 month of storage at the room temperature the average diameter of the particles increased to 245nm. The stability index of this emulsion per year was equal to 1.15. LD₅₀ for mice was equal to 150 iπM g. For the control emulsion LD o was 136 m-d/kg.

Example 7. In emulsion with the concentration of PFD-30 vol.%, and the concentration amount of PFMCP-10 vol.%, PFMCP/PFD ratio is 1/3, stabilized by yolk lecithin in the amount of 8 mas.%, we added stabilizing substance-sodium palmitat in the amount of 0.125 mas.%. The emulsion was prepared as described in example 1. After preparation the average diameter of the emulsion particles was equal to 185 nm. After 12 month of storage at the room temperature the average diameter of the particles increased up to 196 nm. The stability index of this emulsion per year was 1.06. LD₅o for mice was equal to 140 inMcg. For the control emulsion LD₅o was 136 ml/kg.

As has been shown on mentioned figures, tables and examples above, the performed perfluorocarbon emulsion with gas-transportation properties has several preferences as: -decreased average diameter of the emulsion particles -higher dispersion (narrow distribution) -higher stability at storage (higher index of stability) -low toxicity -can be sterilized in autoclave and the average diameter of particles after sterilization (+120 ^°C for 60 rnin) practically does not vary -does not demand the special equipment for long storage -is inexpensive in manufacturing as it consists from easily synthesized PFCs. While the invention has been shown and described with reference to certain preferred embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.

INDUSTRIAL APPLICABILITY The present invention can be used in medicine and pharmacy for production of the highly refined emulsions of the perfluorocarbons which are basic components in oxygen- transporting blood substitutes. Moreover our system can be useful for cosmetics industry and scientific research practice for preparation of ointments and creams based on high refined emulsions of PFCs.

Claims

WHAT IS CLAIMED IS:

1. An emulsion of perfluororganic combination containing perfluorocarbon (PFC) and phospholipids with gas-transport properties, the emulsion comprising: a highly excreted PFC in amount 25 - 50 vol.%; a high-molecular perfluoreted additive in amount 2.5 -20 vol.%; phospholipids in amount 2-10 mas.%; and, a wide spectrum stabilizing additive in amount 0.05 - 0.25 mas.%, taken from fat acids group with Cl 0-C22 and/or their salts of alkaline metals or triglicerids.

2. The emulsion of claim 1, wherein a ratio of the high-molecular perfluoreted additive to the highly excreted PFC varies from 1 :2 to 1 : 10.

3. The emulsion of claim 1 or 2, wherein the highly excreted PFC is PFD (perfluordecalin), and the high-molecular perfluoreted additive is perfluoro-n-methylcyclohexylpiperidine.