MICROCAPSULES CONTAINING LIVE MICROORGANISMS AND USE THEREOF

Abstract

A microcapsule for a topical treatment for improving skin and mucus membranes condition includes a matrix material in a solid state, in which a live probiotic microorganism is encapsulated, wherein the live microorganism is released from the matrix upon contact with a skin surface and/or mucous membranes of the human or warm-blooded animal body, wherein the matrix material has a melting or softening temperature selected from the range of 20-43 C., and wherein the live probiotic microorganism is present in the range of 0.001 to 80 wt. % from the weight of the matrix material.

Claims

1. A microcapsule for a topical treatment for improving skin and mucus membranes condition comprising: a matrix material in a solid state in which a live probiotic microorganism is encapsulated, wherein the live microorganism is released from the matrix upon contact with a skin surface and/or mucous membranes of the human or warm-blooded animal body, wherein the matrix material has a melting or softening at a temperature selected from the range of 20-43 C.-, and wherein the live probiotic microorganism is present in the range of 0.001 to 80 wt. % from the weight of the matrix material.

2. The microcapsule according to claim 1, wherein the water content in said microcapsule does not exceed 10 wt. %, preferably not more than 5 wt. %, or most preferably does not exceed 1 wt. %.

3. The microcapsule according to claim 1, wherein the matrix material is selected from the group comprising: solid lipids: solid animal and solid vegetable oils and fats, fully hydrogenated or partially hydrogenated vegetable and animal oils and fats, saturated fatty acids, partially hydrogenated or fully hydrogenated fatty acids, fatty acid esters, saturated, partially hydrogenated or fully hydrogenated monoglycerides, diglycerides and triglycerides, phospholipids, lecithins, partially hydrogenated or completely hydrogenated phospholipids and lecithins, lysolecithins and lysophosphatidylcholine; solid waxes: animal waxes, plant waxes, mineral waxes, synthetic waxes, wax esters, saturated and unsaturated fatty alcohols, fatty alcohol ethers/esters; solid saturated and unsaturated hydrocarbons (paraffins); solid silicones and silicone ethers/esters; polyol ethers/esters: glycerol ethers/esters, sorbitan, sorbitan stearate, glyceryl ricinoleate; polyglycerols and their ethers/esters, hydrophobic gelling agents: silicon dioxide, polyethylenes, and their mixtures.

4. The microcapsule according to claim 1, wherein the melting or softening point of the matrix is selected from the range of 26.5-35.0 C.

5. The microcapsule according to claim 1, wherein the microcapsule has a size of 100-7000 m and is prepared by mechanical grinding of a cooled suspension of the probiotic microorganism in the matrix material.

6. The microcapsule according to claim 1, wherein the microcapsule have has a size of 50-3000 m and is prepared by cooling of droplets of a suspension of the probiotic microorganism in the matrix material.

7. The microcapsule according to claim 1, wherein the microcapsule has a size of 100-2000 m and are is prepared by spraying the molten matrix material into the fluidized bed of the lyophilisate of the probiotic microorganism.

8. The microcapsule according to claim 1, wherein the microcapsule has a size of 20-1000 m and are is prepared by cooling of emulsified suspension of the probiotic microorganism in the matrix material.

9. The microcapsule according to claim 1, wherein the microcapsule have has a size of 20-1000 m and are prepared by cooling of the sprayed suspension of the lyophilisate of the probiotic microorganism in the matrix material.

10. The Microcapsule according to claim 1, wherein the microcapsule has a size of 250-5000 m and are is prepared by hot extrusion of the suspension of the probiotic microorganism in the matrix material.

11. The microcapsule according to claim 1, wherein the microcapsule additionally contains at least one shell and/or coating that melt or disintegrate when the microcapsule is applied to the skin or mucous membranes.

12. The microcapsule according to claim 11, wherein the shell is obtained by spraying a molten shell material on the microcapsule.

13. The microcapsule according to claim 12, wherein the melting point of the shell material is selected from a range of 28-72 C., preferably 35.5-54 C.

14. The microcapsule according to claim 13, wherein the shell material has the same melting point as the matrix material.

15. The microcapsule according to claim 14, wherein the amount of the shell material does not exceed 50% of the total weight of the microcapsule.

16. The microcapsule according to claim 13, wherein the shell material has a melting point exceeding the melting point of the matrix material.

17. The microcapsule according to claim 16, wherein the amount of the shell material does not exceed 20% of the total weight of the microcapsule.

18. The microcapsule according to claim 11, wherein the shell is obtained by spraying a solution or suspension of the shell material on the microcapsule.

19. The microcapsule according to claim 18, wherein the shell material is selected from the group comprising the same substances as the matrix material.

20. The microcapsule according to claim 18, wherein the shell material is selected from the group comprising cellulose ethers: hydroxymethylpropylcellulose (HPMC) and its derivatives, hydroxypropylcellulose, methylcellulose, ethylcellulose, carboxymethylcellulose (CMC), cellulose acetate phthalate, methacrylic acid and its derivatives (Eudragit), polyvinylpyrrolidone and its derivatives, polysaccharides and their derivatives: sodium alginate, gum arabic, gellan gum, starch, modified starch, guar gum, pectin, amidated pectin, carrageenan, chitosan, mesquite gum, agar gum, psyllium gum, tamarind gum, xanthan, locust bean gum; protein: wheat protein, soy protein, sodium caseinate, gelatin, zein, shellac, hyaluronic acid and its derivatives, any synthetic and natural water-soluble polymers, and mixtures thereof.

21. The microcapsule according to claim 20, wherein the amount of the shell material does not exceed 20% of the total weight of the microcapsule.

22. The microcapsule of claim 11, wherein the coating is obtained by powdering the microcapsule with a micronized coating material.

23. The microcapsule according to claim 22, wherein the micronized coating material is selected from the group of substances comprising the same substances as the matrix material or the shell material; or from the groups of substances comprising inorganic salts, metal oxides, talc, salts and esters of saturated and unsaturated fatty acids.

24. The microcapsule according to claim 1, wherein the microcapsule additionally contain at least one coloring agent and/or at least one substance suitable for use in pharmaceutical, food and cosmetic products.

25. The microcapsule according to claim 1, wherein the probiotic microorganism is selected from the group comprising Saccharomyces cerevisiae (including Saccharomyces boulardii), Bacillus subtilis, Bacillus coagulans, Bacillus amyloliquefaciens, Bifidobacterium bifidum, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium pseudocatenulatum, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus curvatus, Lactobacillus delbruckii subsp. Lactis, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus (GG), Lactobacillus sake, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus kefyr, Lactococcus lactis, Streptococcus thermophilus, Staphylococcus carnosus, Staphylococcus xylosus, Staphylococcus epidermidis, Streptococcus salivarius, Escherichia coli, Propionibacterium (including Propionibacterium freudenreichii), or members belonging to the types of Actinobacteria, Bacteriodetes, Cyanobacteria, Firmicutes, and Proteobacteria (including their genetically modified strains), as well as their combinations.

26. Use of the microcapsule according to claim 1 as an external pharmaceutical, cosmetic, cosmeceutical product for treatment of skin or mucous membrane of the subject in need thereof.

27. A composition containing the microcapsule according to claim 1, suitable for use as external pharmaceutical, cosmetic, cosmeceutical product for treatment of skin or mucous membrane of the subject in need thereof.

28. The composition according to claim 27, wherein composition contains the microcapsule in a dosage of 0.01 to 80 wt. % of the total weight of the composition.

29. The composition according to claim 27, wherein the composition has the form of a cream, lotion, gel, paste, solution, suspension, ointment, emulsion, or powder.

Description

SUMMARY OF THE DRAWINGS

[0077] FIG. 1 shows the view of the microcapsules prepared according to example 1.

[0078] FIG. 2 shows the view of the microcapsules prepared according to example 2.

[0079] FIG. 3 shows the view of the microcapsules prepared according to example 3.

[0080] FIG. 4 shows the view of the microcapsules prepared according to example 4.

[0081] FIG. 5 shows the view of the microcapsules prepared according to example 5.

[0082] FIG. 6 shows the view of the microcapsules prepared according to example 6.

[0083] FIG. 7 shows the view of the microcapsules with the shell prepared according to example 7.

[0084] FIG. 8 shows the view of the microcapsules with the shell prepared according to example 8.

[0085] FIG. 9 shows the view of the microcapsule with the shell prepared according to example 9.

[0086] FIG. 10 shows the view of the microcapsule with the shell prepared according to example 10.

[0087] FIG. 11 shows the view of the microcapsules with the shell prepared according to example 11.

[0088] The spirit of the invention is illustrated by the examples below.

EXAMPLE 1. UNCOATED MICROCAPSULES PREPARED BY THE PROCESS OF MECHANICAL GRINDING

[0089] 50 g of cocoa butter refined (Cargill, USA) with a melting point of 34 C. were melted in a water bath at a temperature of 40 C., 50 g of a mixture of grinded lyophilisate of Saccharomyces cerevisiae (95 wt. %) and sorbitan stearate (5 wt. %) (Angel Yeast Co. Ltd, China) were added and stirred until a homogeneous suspension is formed.

[0090] The resulting mixture cooled to room temperature, grinded on a knife mill to a particle size of 100-7000 m and sieved through a cascade of sieves, taking off fractions with particle size of 100-250 m, 500-1000 m, 1000-2000 m, 2000-3000 m, 3000-4000 m, 4000-5000 m, 5000-6000 m, 6000-7000 m.

[0091] The obtained fractions were processed on a spheronizer (marumerizer) to provide the microcapsules with a spherical shape.

[0092] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in the amount of 47.5 wt. % were prepared with a moisture content of 8.70.2%.

[0093] FIG. 1 shows the view of the 500-1000 m fraction. The view shows that microcapsules have the form of agglomerated particles of the lyophilisate of microorganisms.

[0094] Consumer properties of the prepared microcapsules are shown in Table 1.

[0095] In alternative embodiments, the homogeneous suspension was cooled below room temperature before grinding.

[0096] EXAMPLE 2. UNCOATED MICROCAPSULES PREPARED BY COOLING OF DROPLETS

[0097] 5 g of white beeswax (Koster Keunen), 2 g of micronized silicon dioxide of the AEROSIL 200 Pharma trademark (Evonik), 13 g of silicone Cosmetic Grade Fluid of the Dow Corning 556 trademark and 60 g of silicone Cosmetic Wax of the Dow Corning 2503 trademark were melted in a water bath at a temperature of 40 C. (softening point of the mixture was 24.7 C.), 20 g of grinded lyophilisate of Bacillus amyloliquefaciens (OOO NPF Research Center, Russia) were added and stirred until a homogeneous suspension is formed. The resulting mixture in microdrops (using a micropipette) was added to a liquid cooling agent (liquid nitrogen), the formed microcapsules were separated from the cooling agent, dried and sieved through a sieve, taking off fractions with a particle size of 100-250 m, 500-1000 m, 1000-2000 m, 2000-3000 m.

[0098] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 20 wt. % were prepared with a moisture content in the microcapsule of 2.40.1%.

[0099] FIG. 2 shows the view of the 500-1000 m fraction. The view shows that microcapsules have the form of spherical microparticles containing inclusions of the lyophilisate of microorganisms.

[0100] Consumer properties of the prepared microcapsules are shown in Table 1.

[0101] Alternative embodiment may use as a liquid cooling agent: cooled water, salt solutions, organic solvent or its mixture with water, CO2.

[0102] In alternative embodiments, a pipette, syringe, aspirator, electrospray generator, automatic device for droplets generation, encapsulator or 3D-printer were used for droplets generating.

EXAMPLE 3. UNCOATED MICROCAPSULES PREPARED BY PROCESS OF AGGLOMERATION IN A FLUIDIZED BED

[0103] 80 g of lyophilisate of Bifidobacterium bifidum, Bifidobacterium longum (OOO Bialgam, Russia) were placed in the working chamber of Mini-Glatt fluidized bed unit (Glatt) and a fluidized bed was formed from the resulting powder. 20 g of a mixture of triglycerides of the Suppocire trademark (Gattefosse) with a melting point of 33.7 C. was melted in a water bath at a temperature of 40 C., 1 mg of -carotene dye was added, and the fluidized bed was sprayed, for gluing of particles of the probiotic bacteria lyophilisate and forming microcapsules in the form of agglomerates. Then the prepared microcapsules were sieved through a sieve, taking off fractions with a particle size of 100-250 m, 500-1000 m, 1000-2000 m.

[0104] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 80 wt. % were prepared with a moisture content in the microcapsule of 5.70.3%.

[0105] FIG. 3 shows the view of the 500-1000 m fraction. The view shows that microcapsules have the form of agglomerated particles of the lyophilisate of microorganisms.

[0106] Consumer properties of the prepared microcapsules are shown in Table 1.

EXAMPLE 4. THE MICROCAPSULES PREPARED BY THE PROCESS OF COOLING AN EMULSION

[0107] 90 g of a mixture of hydrogenated palm oil (Oleochemicals) with a melting point of 32.9 C. melted in a water bath at a temperature of 45 C., 9.9 g of soybean lecithin and 0.1 g of the ground lyophilisate of Streptococcus thermophilus (FSUE Experimental Biofabrika, Russia) were added and stirred until a homogeneous suspension is formed. The resulting mixture (homogeneous suspension) was added to a beaker with water (1000 ml) warmed to 40 C., and the aqueous and oil phases intensively stirred until a homogeneous emulsion is formed, lowering the temperature of the resulting emulsion to 4 C. during stirring. Then the stirring was stopped, the resulting solids were separated from the aqueous phase and dried. The resulting powder was sieved through a sieve, taking off fractions with a particle size of 20-125 m, 125-250 m, 250-500 m, 500-1000 m.

[0108] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 0.1 wt. % were prepared with a moisture content of 3.10.1%.

[0109] FIG. 4 shows the view of microcapsules prior to fractionation (size of 20-1000 m). The view shows that microcapsules have the form of microspheres containing inclusions of the lyophilisate of microorganisms.

[0110] Consumer properties of the prepared microcapsules are shown in Table 1.

[0111] In alternative embodiment, the preparation and subsequent cooling of the emulsion (oil in water) were carried out under controlled conditions using microfluidic grids.

EXAMPLE 5. MICROCAPSULES PREPARED BY THE PROCESS OF SPRAY COOLING

[0112] 90 g of a mixture of mono-, di-, and triglycerides of the Witepsol W35 trademark (Oleochemicals) with a melting point of 34.4 C. was melted in a water bath at a temperature of 40 C., 10 g of crushed lyophilisate of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum (OOO Bialgam, Russia) was added, and stirred until a homogeneous suspension is formed. The resulting mixture was sprayed in a stream of cold air, then the resulting powder was divided in a microparticle classifier, taking off fractions with a particle size of 20-125 m, 125-250 m, 250-500 m, 500-1000 m.

[0113] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in the amount of 10 wt. %, were prepared with a moisture content of 1.70.1%.

[0114] FIG. 5 shows the view of the 500-1000 m fraction. The view shows that microcapsules have the form of regular-shaped microspheres containing inclusions of the lyophilisate of microorganisms.

[0115] Consumer properties of the prepared microcapsules are shown in Table 1.

[0116] In alternative embodiments, spraying was performed in a stream of cold nitrogen, gaseous carbon dioxide or inert gas.

[0117] In alternative embodiments, spraying was carried out into a liquid cooling agent, which may be chilled water, salt solutions, organic solvent or its mixture with water, liquid nitrogen, or carbon dioxide.

[0118] In alternative embodiments, airless spray nozzles, two-, three-, four-, or five-phase nozzles, ultrasonic nozzles, or a rotating disk can be used to spray the suspension.

EXAMPLE 6. MICROCAPSULES PREPARED BY PROCESS OF HOT EXTRUSION

[0119] 80 g of a mixture of paraffin cosmetic of the Depilflax trademark with sorbitan stearate (Fine Organics) in a ratio of 19:1 (softening temperature of the mixture is 42 C.) was melted in a water bath at a temperature of 65 C., 20 g of grinded lyophilisate of Escherichia coli (FSUE SPA Microgen, Russia) were added and stirred until a homogeneous suspension is formed. The resulting mixture was cooled to 42-45 C. and pushed through a sieve with a diameter of holes 500 m. The resulting intermediate was cooled to room temperature and processed on a spheronizer (marumerizer) to provide the particles with spherical shape.

[0120] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 20 wt. % were prepared with a moisture content of 0.9%.

[0121] FIG. 6 shows the view of the microcapsules. The view shows that microcapsules have the form of oval-spherical microparticles containing inclusions of the lyophilisate of microorganisms.

[0122] Consumer properties of the prepared microcapsules are shown in Table 1.

[0123] In alternative embodiments, the process was carried out on an automatic or semi-automatic devicean extruder.

[0124] In alternative embodiments, the intermediate was cooled below room temperature.

EXAMPLE 7. MICROCAPSULES WITH A LIPID SHELL

[0125] 82 g of microcapsules prepared as described in example 1 were placed in the working chamber of Mini-Glatt unit (Glatt) and a fluidized bed was formed.

[0126] 18 g of solid fat of the Witepsol E85 trademark (Oleochemicals) with a melting point of 43.1 C. was melted in a water bath at 60 C., 1 mg of (3-carotene dye was added and sprayed into the fluidized bed of the microcapsules for coating.

[0127] FIG. 7 shows the view of microcapsules (size of 500-1000 m). The view shows that microcapsules have the form of microspheres containing a core and a shell.

[0128] Consumer properties of the prepared microcapsules are shown in Table 1.

[0129] The microcapsules with the shell constituting about 18% of the total weight of the capsule were prepared.

[0130] In alternative embodiments, the coating can be applied in a drum coater.

[0131] In alternative embodiments, microcapsules prepared according to examples 2-6 can be taken.

EXAMPLE 8. MICROCAPSULES WITH A WAX SHELL

[0132] 91 g of microcapsules prepared as described in example 4 were placed in the working chamber of Mini-Glatt unit (Glatt) and a fluidized bed was formed.

[0133] 9 g of synthetic wax of the Ceralene 692C trademark (Euroceras) with a melting point of 59 C. was melted in a water bath at a temperature of 80 C., 1 mg of -carotene dye was added and sprayed into the fluidized bed of the microcapsules for coating.

[0134] FIG. 8 shows the view of microcapsules (size of 500-1000 m). The view shows that microcapsules have the form of microspheres coated with the dense shell.

[0135] Consumer properties of the prepared microcapsules are shown in Table 1.

[0136] The microcapsules with the shell constituting about 9% of the total weight of the capsule were prepared.

[0137] In alternative embodiments, microcapsules prepared according to examples 1-3 and 5-7 can be taken.

EXAMPLE 9. MICROCAPSULES WITH AN ETHYLCELLULOSE SHELL

[0138] 98 g of microcapsules prepared as described in example 5 (fraction 500-1000 m), were placed in the working chamber of Mini-Glatt unit (Glatt) and a fluidized bed was formed.

[0139] 2 g of cellulose ethyl ether of the Ethocel trademark (DOW) were dissolved in 20 ml of 96% ethanol at a temperature of 60 C. with stirring, 1 mg of eosin dye was added, and the resulting solution was sprayed into the fluidized bed of microcapsules for coating.

[0140] The microcapsules with the shell constituting about 2% of the total weight of the capsule were obtained.

[0141] FIG. 9 shows the view of microcapsules (size of 500-1000 m). The view shows that microcapsules have the form of microspheres coated with the dense shell.

[0142] Consumer properties of the prepared microcapsules are shown in Table 1.

[0143] In alternative embodiments, supercritical fluid (carbon dioxide) can be used as a solvent for the shell material.

[0144] In alternative embodiments, microcapsules prepared according to examples 1-4 and 6-8 can be taken.

EXAMPLE 10. MICROCAPSULES WITH HYDROXYPROPYLMETHYLCELLULOSE (HPMC) SHELL

[0145] 95 g of microcapsules prepared as described in example 6 were placed in the working chamber of Mini-Glatt unit (Glatt) and a fluidized bed was formed.

[0146] 5 g HPMC of the Vivapharm E6 trademark (JRS Pharm) were dissolved in 50 ml of water, 1 mg of indigo carmine dye was added, and the resulting solution was sprayed into the fluidized bed of microcapsules for coating.

[0147] The microcapsules with the shell constituting about 5% of the total weight of the capsule were prepared.

[0148] FIG. 10 shows the view of the microcapsule (size of 900 m). The view shows that microcapsules have the form of microspheres coated with the shell.

[0149] Consumer properties of the prepared microcapsules are shown in Table 1.

[0150] In alternative embodiments, microcapsules prepared according to examples 1 and 3-9 can be taken.

EXAMPLE 11. MICROCAPSULES WITH A SHELL OBTAINED BY THE PROCESS OF POWDERING

[0151] 95 g of microcapsules prepared as described in example 3 (500-1000 m fraction) and then 5 g of powdering agent (a mixture of micronized cosmetic paraffin of the Depilflax trademark (particle size of 20-40 m) and micronized magnesium stearate (particle size of 20-30 m)) were placed in the working chamber of Mini-Glatt unit (Glatt) and a fluidized bed was formed by allowing to stand for 10 minutes at a temperature of 27 C.

[0152] FIG. 11 shows the view of microcapsules (size of 500-1000 m). The view shows that microcapsules have the form of spherical agglomerates.

[0153] Consumer properties of the prepared microcapsules are shown in Table 1.

[0154] In alternative embodiments, the temperature of the air forming the fluidized bed can be increased to values equal to the melting point of the matrix or shell materials, wherein the temperature increase can be both short-term and permanent.

[0155] In alternative embodiments, microcapsules prepared according to examples 1, 2 and 4-10 can be taken.

EXAMPLE 12. POLYMERIC MICROCAPSULES WITH LACTOBACILLUS

[0156] 10 g of lyophilisate of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum (OOO Bialgam, Russia) were dispersed in 20 g of sterile aqueous solution of sodium chloride (0.85 wt. %) and 20 g of sterile aqueous solution of sodium alginate (5 wt. %) were added to this solution. The resulting solution was sprayed into 500 ml of sterile aqueous solution of calcium gluconate (3 wt. %) with the Buchi B-390 encapsulator (nozzle size 500 m). The prepared microcapsules were separated from the solution, washed with 500 ml of sterile aqueous solution of sodium chloride (0.85 wt. %), frozen at a temperature of 82 C. and lyophilized in TFD-5503 unit (Ilshin).

[0157] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in the amount of 60 wt. % were prepared with a moisture content of 5.9%.

[0158] Consumer properties of the prepared microcapsules are shown in Table 1.

EXAMPLE 13. POLYMERIC MICROCAPSULES WITH BIFIDOBACTERIUM

[0159] The preparation of the microcapsules was carried out as described in example 12, except that lyophilisate of Bifidobacterium bifidum, Bifidobacterium longum (OOO Bialgam, Russia) was used.

[0160] Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 60 wt. % were prepared with a moisture content of 4.4%.

[0161] Views of microcapsules were obtained using an optical microscope Mikmed-5 (JSC LOMO). The moisture content was determined on the MA-100 instrument (Sartorius AG).

[0162] Evaluation of consumer properties of microcapsules was carried out as follows: 40 g of the microcapsules prepared in examples 1-13 were placed in plastic jars for cosmetics with a screw cap and placed into storage in the climate chamber KFB 115 (Binder) at a temperature of 201.0 C. and no light. After 90 days, the jars were opened, and appearance of the microcapsules and the presence or absence of an unpleasant odor were recorded. 200 mg of microcapsules were applied with a spatula as a thin layer on the inner surface of the wrist of the left hand of healthy volunteers (6 persons) with a normal body temperature (36.6-36.7 C.) and kept for 60 seconds (without rubbing on the skin), with observing the changes in the shape and physical state of microcapsules and recording the time of complete deformation from the moment of application to the skin. Then the experiment with rubbing was carried out: 200 mg of microcapsules were applied to the inner surface of the wrist of the left hand of healthy volunteers (6 persons) with a normal body temperature (36.6-36.7 C.) and rubbed on the skin with the palm of the right hand. At the same time, the time of melting of microcapsules and the presence or absence of an abrasive effect (sense of scratching caused by the presence of solid microparticles when rubbing on the skin) were recorded.

[0163] Consumer properties of the prepared microcapsules are shown in Table 1.

TABLE-US-00001 TABLE 1 Consumer properties of microcapsules prepared according to examples 1-13. Disintegrating when applied to the skin without with Storage Unpleasant Abrasive rubbing, rubbing, No. Sample period Appearance odor effect sec sec 1 Microcapsules 90 days Brown no no 23 6 4 1 prepared agglomerates according to Example 1 (500-1000 m fraction) 2 according to 90 days Cream- no no 13 4 1 Example 2 coloured (500-1000 m microspheres fraction) 3 according to 90 days Pink-brown no no 38 10 4 1 Example 3 agglomerates (500-1000 m fraction) 4 according to 90 days Cream- no no 22 2 2 1 Example 4 coloured (500-1000 m microspheres fraction) 5 according to 90 days Yellow no no 19 5 3 2 Example 5 microspheres (500-1000 m fraction) 6 according to 90 days Cream- no no no 5 2 Example 6 coloured (diameter microspheres 500 m) 7 according to 90 days Yellow no no no 7 3 Example 7 microspheres (diameter 500-1000 m) 8 according to 90 days Yellow no no no 12 4 Example 8 microspheres (diameter 500-1000 m) 9 according to 90 days Yellow no no no 9 3 Example 9 microspheres (diameter 500-1000 m) 10 according to 90 days Deep blue no no no 10 4 Example 10 microspheres (diameter 500-1000 m) 11 according to 90 days Beige- no no 37 12 4 2 Example 11 coloured (diameter microspheres 500-1000 m) 12 according to 90 days Grey-beige no yes no no Example 12 microspheres (diameter 500 m) 13 according to 90 days Grey-beige no yes no no Example 13 microspheres (diameter 500 m)

[0164] Table 1 shows that the polymer microcapsules prepared according to examples 12 and 13 do not soften and have an abrasive effect when applied to the skin, which makes their use as an external dosage form and cosmetic product unacceptable. The microcapsules prepared according to examples 1-11 have satisfactory consumer properties (no unpleasant odor and no abrasive effect) and are capable to disintegrate when applied to the skin, which indicates their effectiveness as a means of delivering microorganisms for external and cosmetic use. In addition, Table 1 shows that, depending on the nature of the matrix and shell material, microcapsules provided herein (examples 1-11) have different disintegration times when applied to the skin, which makes it possible to use them variably to solve various biopharmaceutical problems related to delivery of live microorganisms to the skin and mucous membranes.

EXAMPLE 14. COMPARISON OF CONSUMER PROPERTIES AND STABILITY OF THE MICROCAPSULES PROVIDED IN THE PRESENT INVENTION, POLYMERIC MICROCAPSULES AND NON-ENCAPSULATED LYOPHILISATE OF MICROORGANISMS FORMULATED AS A GEL

[0165] Consumer properties were evaluated by applying the microcapsules provided according to the present invention prepared as described in example 5 (fraction with size of 500-1000 m), polymer microcapsules prepared as described in example 12, and non-encapsulated lyophilisate of microorganisms Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum (manufactured by OOO Bialgam, Russia) formulated in an amount of 4 wt. % as a water-containing gel based on polyethylene oxide (this gel can be used as a vehicle for external pharmaceuticals). Then the organoleptic properties and actual number of viable cells of probiotic microorganisms in the gel samples were evaluated during storage at 18 C.

[0166] The number of microorganisms in the samples was determined according to GOST R 56139. The comparison results are shown in Table 2.

TABLE-US-00002 TABLE 2 Comparison of consumer properties and stability of microcapsules provided in the present invention, polymeric microcapsules and non-encapsulated lyophilisate of microorganisms Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum formulated as a gel. Number of viable Storage Unpleasant Abrasive microorganisms, No. Sample period Appearance odor effect U/g 1 Microcapsules 0 days Colorless no no 8.8 10.sup.6 according to the transparent gel present invention containing prepared suspended according to spherical example 5 microinclusions formulated as gel 5 days The same no no 8.4 10.sup.6 10 days The same no no 8.3 10.sup.6 30 days The same no no 7.5 10.sup.6 60 days The same no no 7.4 10.sup.6 90 days The same no no 7.1 10.sup.6 182 days The same no no 6.8 10.sup.6 364 days The same no no 6.1 10.sup.6 2 Polymer 0 days Pinkish no yes 2.4 10.sup.6 microcapsules transparent gel prepared containing according to suspended example 12 spherical formulated as gel microinclusions 5 days Gel turbidity is weak 8.5 10.sup.5 observed, colour is light brown 10 days Disintegration Yes 3.3 10.sup.4 of microparticles is observed, gel is turbid and non-transparent, the colour is brown 30 days The same Yes 3 Non-capsulated 0 days Non- no no 1.4 10.sup.7 lyophilis ate of transparent microorganisms brown gel with formulated as gel the signs of layering 5 days The same Yes no 8.4 10.sup.4 10 days The same Yes 1.3 10.sup.3 30 days The same Yes

EXAMPLE 15. COMPARISON OF CONSUMER PROPERTIES AND STABILITY OF THE MICROCAPSULES PROVIDED IN THE PRESENT INVENTION, POLYMERIC MICROCAPSULES, AND NON-ENCAPSULATED LYOPHILISATE OF MICROORGANISMS FORMULATED AS A CREAM

[0167] Consumer properties were evaluated by formulating the microcapsules provided in the present invention prepared as described in example 11, polymer microcapsules, prepared as described in example 13, and non-encapsulated lyophilisate of microorganisms Bifidobacterium bifidum, Bifidobacterium longum (manufactured by OOO Bialgam, Russia) as a cosmetic cream (the composition of the cream: purified water, glycerol monostearate, cetyl alcohol, stearyl alcohol, diethylene glycol stearate, PEG-400 stearate, dipropylene glycol, polysorbate 20, PEG-40, hydrogenated castor oil, hydroxypropyl guar, magnesium silicate) in an amount of 4 wt. %. Then the organoleptic properties and actual number of viable cells of probiotic microorganisms in cream samples were evaluated over time during storage at 18 C.

[0168] The number of microorganisms in the samples was determined according to GOST R 56139. The comparison results are shown in Table 3.

TABLE-US-00003 TABLE 3 Comparison of consumer properties and stability of the microcapsules provided in the present invention, polymeric microcapsules, and non-encapsulated lyophilisate of microorganisms Bifidobacterium bifidum, Bifidobacterium longum formulated as a cream. Abrasive effect when applied Number of viable Storage Unpleasant to the microorganisms, No. Sample period Appearance odor skin U/g 1 Microcapsules 0 days White cream no no 9.5 10.sup.9 according to the containing present invention suspended prepared spherical according to microinclusions example 11 5 days The same no no 8.6 10.sup.9 formulated as 10 days The same no no 8.2 10.sup.9 cream 30 days The same no no 7.5 10.sup.9 60 days The same no no 7.4 10.sup.9 90 days The same no no 7.2 10.sup.9 182 days The same no no 6.8 10.sup.9 364 days The same no no 6.3 10.sup.9 2 Polymer 0 days Light beige no yes 4.3 10.sup.8 microcapsules cream prepared containing according to suspended example 13 spherical formulated as microinclusions cream 5 days Light brown Yes 4.4 10.sup.7 cream 10 days The same Yes 8.1 10.sup.4 30 days The same Yes 3 Non-capsulated 0 days Light brown no no 7.0 10.sup.9 lyophilis ate of cream, without microorganisms inclusions formulated as 5 days Browningi s Weak 3.5 10.sup.5 cream observed 10 days The same Yes 8.3 10.sup.3 30 days The same Yes

[0169] Tables 2 and 3 show that non-encapsulated microorganisms formulated as a water-containing gel and cosmetic cream quickly lose their viability (low stability) and cause a change in consumer properties of the dosage form for external use (gel) and cosmetic composition (cream)discoloration and appearance of an unpleasant odor. Use of polymer microcapsules does not allow to increase the stability of microorganisms and prevent the change in consumer properties of the product. Use of microcapsules provided in the present invention makes it possible to obtain a stable product for external use with satisfactory organoleptic properties, which allows to use microorganisms as an active component in the composition of external pharmaceutical and cosmetic products.

EXAMPLE 16. MICROCAPSULES PREPARED BY THE PROCESS OF SPRAY COOLING

[0170] 80 g of saturated fatty acids Cosmobase A, with a melting temperature of 29.2 C. (OOO CosmoLab, Russia), were melted in a water bath under the temperature of 40 C. 10 g of beeswax (OOO Pasechnic Russia) and 10 g of grinded lyophilisate of Staphylococcus Epidermidis (OOO CosmoLab, Russia) were added and stirred until a homogeneous suspension is formed. The resulting mixture was sprayed to a liquid cooling agent (liquid nitrogen). The formed microcapsules in liquid nitrogen were placed in a refrigerant at a temperature of 3 C. for 20 hours. The resulting powder was sieved through a cascade of sieves, taking off fractions with particle size of 250-500 m and 500-1000 m. Uncoated microcapsules containing lyophilisate of live encapsulated microorganisms in an amount of 10 wt. % were prepared with a moisture content in the microcapsule of 4.90.6%.

[0171] Consumer properties of microcapsules provided in the Example 16 are set forth in Table 4.

TABLE-US-00004 TABLE 4 Abrasive effect Disintegrating when when applied to the skin applied without with Storage Unpleasant to the rubbing, rubbing, No Sample period Appearance odor skin sec sec 1 Microcapsules 90 days Beige-colored no no 26 4 5 2 prepared microspheres according to example 16 (250-500 m fraction)

[0172] Microcapsules containing live microorganisms and their use disclosed in the present invention are intended for use in medicine and cosmetology as a pharmaceutical and cosmetic product for external use for the normalization of microflora and the functional state of the skin and mucous membranes during aging, damage by environmental factors (UV rays, wind, low temperatures, injuries) and pathological processes (microbial infections, inflammatory and allergic diseases, metabolic disorders). At the time of use, when applied to the skin, the microcapsules melt under the action of body temperature, releasing live microorganisms, where the latter have a probiotic effect. In addition, the matrix material, when applied to the skin, has an independent protective, moisturizing and nourishing effect. Use of the microcapsules provided herein makes its possible to isolate microorganisms from water and other active substances contained in the pharmaceutical or cosmetic composition, to prevent the processes of growth of microorganisms inside the package during storage of cosmetic product, thereby eliminating such consumer disadvantages as low stability, short shelf life, unpleasant odor.