Combination therapeutic composition of cabs mitochondria and graphene for psoriasis

Abstract

The present invention relates to a combination therapeutic composition of CABS mitochondria and graphene for psoriasis. It has been confirmed that the combined administration of mitochondria and graphene, of the present invention, reduced skin thickness, dead skin cells and erythema in a psoriasis animal model. In addition, it has been confirmed that inflammatory immune cell subtypes and tissue-resident memory cells, which are psoriasis pathogenesis immune cells, are modulated. Furthermore, it has been confirmed that the thickness of the epidermal layer is reduced.

Description

Combination therapeutic composition of CABS MITOCHONDRIA and graphene for psoriasis

[0001] The present invention relates to a therapeutic composition for psoriasis in combination with CABS mitochondria and graphene.

[0002] Psoriasis is a very common inflammatory skin disease affecting approximately 100 million people worldwide, with about 1–2% of the current population. It is an autoimmune disease. When psoriasis spreads significantly from localized areas, it can cover almost the entire body. Although the exact cause of psoriasis is not yet fully understood, genetic factors and the Koebner phenomenon—where localized psoriasis develops upon skin damage—are known to play a role. Furthermore, various environmental factors are suggested to exacerbate the condition. These include stress, the discontinuation of invasive corticosteroids, and other environmental factors. While psoriasis is not life-threatening, it causes psychological distress, loss of self-confidence, and depression, and requires long-term treatment.

[0003] In psoriasis, unlike a normal immune system, T cells become activated and secrete cytokines even in the absence of any antigens on the skin. These cytokines act on normal skin cells, which normally regenerate and mature over a cycle of approximately 28 days, causing them to regenerate and mature very rapidly over a cycle of 3 to 4 days. Skin cells that have regenerated and matured at such a rapid rate migrate to the skin surface, where dead cells accumulate in layers to form grayish-white scales. Additionally, blood vessels dilate and blood flow increases, causing the skin to turn red. Regarding T cell activation in psoriasis, it has been reported that Th-1 cells trigger the condition, and that the use of IL-10, which inhibits Th-1 cell activity, alleviates psoriasis symptoms. Furthermore, cytokines such as IL-1β, IL-6, IL-8, GM-CSF, and TNF-α induce psoriasis triggering, and increased concentrations of VEGT and Substances P have also been reported to be associated with psoriasis.

[0004] Common treatments for psoriasis include topical medications, phototherapy, and oral medications. Topical treatments include steroids, coal tar, anthralin, vitamin D3 and its analogues, retinoids, and sun exposure; however, these treatments carry side effects such as skin thinning, stretch marks, burns, irritation, and photosensitivity. Phototherapy involves the administration of psoralen in combination with ultraviolet B or ultraviolet A rays, but it has the disadvantages of accelerated skin aging and an increased risk of skin cancer. Oral medications, applied to the most severe cases of psoriasis, include methotrexate, oral retinoids, and cyclosporine. However, cyclosporine requires careful monitoring as it can cause kidney damage and blood pressure issues. While methotrexate has the advantage of being effective for both psoriasis and psoriatic arthritis, it must be used with caution as cumulative use can lead to liver cirrhosis or fibrosis. Additionally, the use of oral retinoids must be carefully controlled due to the potential for serious complications during childbirth in women. As such, conventional drugs used to treat psoriasis have side effects or drawbacks. Furthermore, drugs such as methotrexate and cyclosporine, which inhibit T-cell activity, are highly toxic. Consequently, biologics with fewer side effects than these drugs have been developed, including alefacept (Amevive), itanercept (Enbrel), infliximab (Remicade), and ipalizumab (Raptiva). Therefore, there is an urgent need for a psoriasis treatment that is not only effective but also highly safe, free from issues of side effects or toxicity.

[0005] Accordingly, the inventors confirmed that the combined administration of CABS mitochondria and graphene improves psoriasis, thereby completing the present invention.

[0006] The object of the present invention is to provide a composition for the prevention or treatment of skin diseases comprising mitochondria and graphene as active ingredients.

[0007] Another objective of the present invention is to provide a cosmetic composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0008] Another objective of the present invention is to provide a patch composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0009] Another objective of the present invention is to provide a method for preventing or treating a skin disease, comprising the step of administering a composition for preventing or treating the skin disease to an individual.

[0010] To achieve the above objective, the present invention provides a composition for the prevention or treatment of skin diseases comprising mitochondria and graphene as active ingredients.

[0011] In addition, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0012] In addition, the present invention provides a patch composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0013] In addition, the present invention provides a method for preventing or treating a skin disease, comprising the step of administering a composition for preventing or treating the skin disease to an individual.

[0014] It was confirmed that the combined administration of mitochondria and graphene according to the present invention reduces skin thickness, keratinization, and erythema in an animal model of psoriasis. Furthermore, it was confirmed that it regulates inflammatory immune cell subtypes and tissue-resident memory cells, which are immune cells responsible for the pathogenesis of psoriasis. Additionally, since it reduces the thickness of the epidermal layer, it can be usefully applied in related industries.

[0015] Figures 1 to 3 are figures confirming the characteristics of the nano-graphene oxide of the present invention (Figure 1: TEM analysis results, Figure 2: confirmation of average particle distribution, Figure 3: FTIR analysis results).

[0016] Figure 4 illustrates the experimental process for confirming the combined effect of mitochondria and graphene of the present invention.

[0017] Figure 5 is a figure showing the visual confirmation of the psoriasis improvement effect following the combined administration of mitochondria and graphene of the present invention.

[0018] Figure 6 is a figure quantifying keratin, erythema, and skin thickness in a psoriasis animal model following the combined administration of mitochondria and graphene of the present invention.

[0019] Figure 7 is a figure showing the frequency of Tc17 cells in a psoriasis animal model following the combined administration of mitochondria and graphene of the present invention by flow cytometry.

[0020] Figures 8 and 9 show the frequency of tissue-resident memory cells in a psoriasis animal model following the combined administration of mitochondria and graphene of the present invention by flow cytometry (A: CD8 TRM quantification, B: CD8 TRM 17 quantification).

[0021] Figure 10 is a figure showing the skin tissue of a psoriasis animal model after the combined administration of mitochondria and graphene of the present invention, confirmed by H&E staining.

[0022] Figure 11 is a figure quantifying the thickness of the epidermal layer of a psoriasis animal model following the combined administration of mitochondria and graphene of the present invention.

[0023] Embodiments of the present invention will be described in detail below with reference to the attached drawings. In the following description, detailed descriptions of technologies well known to those skilled in the art may be omitted. Furthermore, in describing the present invention, detailed descriptions of related known functions or configurations may be omitted if it is determined that such descriptions would unnecessarily obscure the essence of the present invention. Additionally, the terminology used in this specification is used to appropriately express preferred embodiments of the present invention, and may vary depending on the intent of the user or operator, or the conventions of the field to which the present invention belongs.

[0024] Therefore, the definitions of these terms should be based on the content throughout this specification. Throughout the specification, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0025] The present invention provides a composition for the prevention or treatment of skin diseases comprising mitochondria and graphene as active ingredients.

[0026] The term "graphene" in the present invention is a compound word formed by combining "graphite," meaning graphite, and the suffix "-ene," which in chemistry denotes a molecule having a carbon double bond. The graphene has a hexagonal honeycomb structure in which carbon atoms are connected infinitely. That is, it is a thin film structure in the form of a two-dimensional planar layer with a thickness of 0.35 nm, consisting of a single layer of carbon atoms. Although graphene has a simple structure, it exhibits excellent physicochemical properties in various dimensions, such as high electrical and thermal conductivity, high mechanical strength and hardness, unique optical properties, and chemical stability. The intracellular absorption of graphene is relatively limited due to its thin film form. Therefore, in terms of cell safety, it has advantages over carbon nanotubes, which are absorbed into cells through various pathways such as phagocytosis and membrane surface adsorption.

[0027] The term "prevention" as used in this invention refers to any act of suppressing the symptoms of a specific disease or delaying its progression through the administration of the composition of this invention.

[0028] The term "treatment" as used in this invention refers to any act of improving or beneficially altering the symptoms of a specific disease through the administration of the composition of this invention.

[0029] The pharmaceutical composition of the present invention may additionally include an adjuvant in addition to the active ingredient. Any adjuvant known in the art may be used without limitation, but, for example, Freund's complete or incomplete adjuvant may be further included to increase the effect.

[0030] The pharmaceutical composition according to the present invention may be prepared in a form in which an active ingredient is incorporated into a pharmaceutically acceptable carrier. Here, the pharmaceutically acceptable carrier includes carriers, excipients, and diluents commonly used in the pharmaceutical field. Pharmaceutically acceptable carriers that can be used in the pharmaceutical composition of the present invention are not limited to these, but may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

[0031] The pharmaceutical composition of the present invention may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external formulations, suppositories, or sterile injectable solutions, each according to conventional methods.

[0032] When formulating, the product may be prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Solid dosage forms for oral administration include tablets, pills, powders, granules, and capsules, and such solid dosage forms may be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc., with the active ingredient. In addition, lubricants such as magnesium stearate and talc may also be used in addition to simple excipients. Liquid dosage forms for oral administration include suspensions, liquid formulations, emulsions, and syrups, and may contain various excipients, such as wetting agents, sweeteners, flavoring agents, and preservatives, in addition to commonly used diluents such as water and liquid paraffin. Preparations for parenteral administration include sterile aqueous solutions, water-insoluble solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Water-insoluble solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Bases for suppositories may include Witepsol, Tween 61, cocoa paste, laurin paste, glycerogelatin, etc.

[0033] The pharmaceutical composition according to the present invention may be administered to an individual by various routes. Any mode of administration may be anticipated, for example, by oral, intravenous, intramuscular, subcutaneous, or intraperitoneal injection.

[0034] The dosage of the pharmaceutical composition according to the present invention is selected by taking into consideration the age, weight, gender, physical condition, etc. of the individual. It is obvious that the concentration of the active ingredient included in the pharmaceutical composition can be selected in various ways depending on the subject, and preferably, it is included in the pharmaceutical composition at a concentration of 0.01 to 5,000 μg / mL. If the concentration is less than 0.01 μg / mL, pharmaceutical activity may not appear, and if it exceeds 5,000 μg / mL, it may exhibit toxicity to the human body.

[0035] According to one embodiment of the present invention, the graphene may be nano-graphene oxide, and the nano-graphene oxide may have a particle size of 15 to 38 nm.

[0036] Furthermore, "graphene oxide" refers to a form in which oxygen is attached to graphene as it oxidizes and intervenes between carbon bonds. For example, when strong acids such as sulfuric acid are added to graphite and reacted, the graphene oxidizes and is replaced by oxygen, which is called "graphene oxide." Since oxidizing graphene increases its elasticity and it does not lose its electrical conductivity even when stretched or folded, it is a material attracting attention in the field of nanomaterials. Additionally, graphene oxide exhibits a relatively wide particle size range, ranging from tens of micrometers to as small as tens of nanometers, and graphene oxide with a size of tens of nanometers is named "nano-graphene oxide."

[0037] According to one embodiment of the present invention, the mitochondria may be mitochondria isolated from adipose tissue-derived mesenchymal stem cells (AM) or Catholic bone marrow stem cells (CABS).

[0038] According to one embodiment of the present invention, the composition may reduce the frequency of pathogenic immune cells.

[0039] According to one embodiment of the present invention, the immune cell may be a Tc17 or a tissue-resident memory cell (TRM), and the tissue-resident memory cell may be a CD8-positive TRM or a CD8-positive TRM 17.

[0040] According to one embodiment of the present invention, the skin disease may be psoriasis.

[0041] The "psoriasis" of the present invention is a type of skin disease characterized by red, millet-like rashes on the skin and layers of silvery-white, dandruff-like skin scales accumulating on the rashes, with the rashes gradually growing in size. Although the cause of psoriasis is not clearly known, it is known to be a chronic inflammatory disease caused by abnormalities in the body's immune system.

[0042]

[0043] In addition, the present invention provides a cosmetic composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0044] The term "improvement" as used in the present invention refers to any action that at least reduces parameters related to the condition being treated, such as the degree of symptoms.

[0045] A cosmetic composition containing the active ingredient of the present invention can be used in various ways for the prevention or improvement of skin diseases. Products to which the present composition can be added include, for example, cosmetics such as various creams, lotions, skin toners, essences, etc., as well as shampoos, rinses, cleansers, facial cleansers, soaps, treatments, packs, beauty solutions, etc.

[0046] The cosmetic composition of the present invention comprises a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extracts.

[0047] As for water-soluble vitamins, any that can be incorporated into cosmetics may be used, but preferably, examples include vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine hydrochloride, vitamin B12, pantothenic acid, nicotinic acid, nicotinamide, folic acid, vitamin C, vitamin H, etc., and their salts (thiamine hydrochloride, sodium ascorbate, etc.) or derivatives (sodium ascorbate-2-phosphate, magnesium ascorbate-2-phosphate, etc.) are also included in the water-soluble vitamins that can be used in the present invention. Water-soluble vitamins can be obtained by conventional methods such as microbial conversion methods, purification methods from microbial cultures, enzymatic methods, or chemical synthesis methods.

[0048] As for the useful vitamin, any that can be incorporated into cosmetics may be used, but preferably, examples include vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol), and their derivatives (ascorbin palmitate, ascorbin stearate, ascorbin dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherol nicotinate, vitamin E, DL-pantotenyl alcohol, D-pantotenyl alcohol, pantotenyl ethyl ether, etc.) are also included in the useful vitamin used in the present invention. The useful vitamin can be obtained by conventional methods such as microbial conversion, purification from microbial cultures, enzymes, or chemical synthesis.

[0049] As for the high molecular weight peptide, any material that can be incorporated into cosmetics may be used, but preferably, collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin, etc. The high molecular weight peptide can be obtained by purification through conventional methods such as purification from a microbial culture medium, enzymatic methods, or chemical synthesis methods, or it can be used by purifying it from natural materials such as the dermis of pigs or cattle, or silk fibers of silkworms.

[0050] As for the high molecular weight polysaccharide, any that can be incorporated into cosmetics may be used, but preferably, hydroxyethylcellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or its salt (sodium salt, etc.) may be used. For example, chondroitin sulfate or its salt, etc., can be used after purifying it from mammals or fish.

[0051] As for sphingolipids, any that can be incorporated into cosmetics may be used, but preferably, ceramides, phytosphingosines, sphingoglycolipids, etc. Sphingolipids can be obtained by purifying them by conventional methods or by chemical synthesis from mammals, fish, shellfish, yeast, or plants.

[0052] Any seaweed extract that can be incorporated into cosmetics may be used, but preferably, brown algae extract, red algae extract, green algae extract, etc., may be used. Additionally, calagenan, arginic acid, sodium alginate, potassium alginate, etc., purified from these seaweed extracts are also included in the seaweed extract used in the present invention. The seaweed extract can be obtained by purifying seaweed by conventional methods.

[0053] In addition to the essential ingredients mentioned above, the cosmetic composition of the present invention may also include other ingredients that are conventionally incorporated into cosmetic compositions as needed. Other ingredients that may be added include oil components, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohols, colorants, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, etc. Examples of oil components include ester-based oils, hydrocarbon-based oils, silicone-based oils, fluorinated-based oils, animal oils, plant oils, etc.

[0054] As ester-based oils, tri-2-ethylhexanoate glyceryl, 2-ethylhexanoate cetyl, isopropyl myristate, butyl myristate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, butyl stearate, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl myristate, isostearyl myristate, isostearyl palmitate, octyldodecyl myristate, isocetyl isostearate, diethyl sebacate, diisopropyl adipice, isoalkyl neopentanoate, tri(capryl, capric acid)glyceryl, tri-2-ethylhexanoate trimethylolpropane, triisostearate trimethylolpropane, tetra-2-ethylhexanoate pentaelislitol, cetyl caprylate, decyl lauricate, Hexyl laurate, Decyl myristate, Myristyl myristate, Cetyl myristate, Stearyl stearate, Decyl oleate, Cetyl lisinooleate, Isostearyl laurate, Isotridecyl myristate, Isocetyl palmitate, Octyl stearate, Isocetyl stearate, Isodecyl oleate, Octyldodecyl oleate, Octyldodecyl linoleate, Isopropyl isostearate, Cetostearyl 2-ethylhexanoate, Stearyl 2-ethylhexanoate, Hexyl isostearate, Ethylene glycol dioctanoate, Ethylene glycol dioleate, Propylene glycol dicaprine, Di(Capryl, Capric acid)propylene glycol, Propylene glycol dicaprylate, Neopentyl glycol dicaprine, Neopentyl glycol dioctanoate, Glyceryl tricaprylate, Glyceryl triundecylate, Glyceryl triisopalmitate, Glyceryl triisostearate, Octyldodecyl neopentanoate, Isostearyl octanoate, Octyl isononanoate, Hexyldecyl neodecanoate, Octyldodecyl neodecanoate, Isocetyl isostearate, Isostearyl isostearate, Octyldecyl isostearate, Polyglycerin oleic acid ester, Polyglycerin isostearic acid ester, Triisocetyl citrate, Triisoalkyl citrate, Triisooctyl citrate, Lauryl lactate, Myristyl lactate, Cetyl lactate, Octyldecyl lactate, Triethyl citrate, Acetyltriethyl citrate, Acetyltributyl citrate, Trioctyl citrate, Diisostearyl malate, Hydroxystearic acid 2-ethylhexyl, di2-ethylhexyl succinate, diisobutyl adipicate, diisopropyl sebacicate,Examples include ester systems such as dioctyl sebacate, cholesteryl stearate, cholesteryl isostearate, cholesteryl hydroxystearate, cholesteryl oleate, dihydrocholesteryl oleate, pitsteryl isostearate, pitsteryl oleate, isocetyl 12-stealloylhydroxystearate, stearyl 12-stealloylhydroxystearate, and isostearyl 12-stealloylhydroxystearate.

[0055] Examples of hydrocarbon oils include squalene, liquid paraffin, alpha-olefin oligomer, isoparaffin, ceresin, paraffin, liquid isoparaffin, polybutene, microcrystalline wax, and Vaseline.

[0056] Examples of silicone-based oils include polymethylsilicon, methylphenylsilicon, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane and methylcetyloxysiloxane copolymer, dimethylsiloxane and methylstealoxysiloxane copolymer, alkyl-modified silicone oil, amino-modified silicone oil, etc.

[0057] Examples of fluorinated oils include perfluoropolyethers.

[0058] Examples of animal or plant oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, safflower oil, soybean oil, corn oil, rapeseed oil, apricot kernel oil, palm kernel oil, palm oil, castor oil, sunflower oil, grapeseed oil, cottonseed oil, coconut oil, cucurbit oil, wheat germ oil, rice germ oil, shea butter, laurel oil, macadamia nut oil, meadowsweet oil, egg yolk oil, beef tallow, horse oil, mink oil, orange raffia oil, jojoba oil, candelilla wax, carnaba wax, liquid lanolin, hydrogenated castor oil, etc.

[0059] Examples of moisturizers include water-soluble low-molecular-weight moisturizers, oil-soluble molecular moisturizers, water-soluble polymers, and oil-soluble polymers.

[0060] Examples of water-soluble low molecular weight moisturizers include serine, glutamine, sorbitol, mannitol, pyrrolidone-sodium carboxylate, glycerin, propylene glycol, 1,3-butylene glycol, ethylene glycol, polyethylene glycol B (degree of polymerization n = 2 or more), polypropylene glycol (degree of polymerization n = 2 or more), polyglycerin B (degree of polymerization n = 2 or more), lactic acid, lactate, etc.

[0061] Examples of fat-soluble low-molecular-weight moisturizers include cholesterol and cholesterol esters.

[0062] Examples of water-soluble polymers include carboxyvinyl polymer, polyaspartate, tragacanth, xanthan gum, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, carboxymethylcellulose, water-soluble chitin, chitosan, dextrin, etc.

[0063] Examples of fat-soluble polymers include polyvinylpyrrolidone and eicocene copolymers, polyvinylpyrrolidone and hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, and polymeric silicones. Examples of emollients include long-chain acylglutamic acid cholesteryl esters, hydroxystearate cholesteryl, 12-hydroxystearate, stearic acid, rosin acid, and lanolin fatty acid cholesteryl esters.

[0064] Examples of surfactants include nonionic surfactants, anionic surfactants, cationic surfactants, and amphoteric surfactants.

[0065] Examples of nonionic surfactants include self-emulsifying monostearate glycerin, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE glycerin fatty acid ester, POE alkyl ether, POE fatty acid ester, POE hydrogenated castor oil, POE castor oil, POE and POP (polyoxyethylene and polyoxypropylene) copolymer, POE and POP alkyl ether, polyether-modified silicone, alkanolamide lauric acid, alkylamine oxide, hydrogenated soybean phospholipid, etc.

[0066] Examples of anionic surfactants include fatty acid soaps, alpha-acylsulfonates, alkylsulfonates, alkylallylsulfonates, alkylnaphthalenesulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphates, alkylamide phosphates, alkylloylalkyl taurine salts, N-acyl amino acids, POE alkyl ether carboxylates, alkyl sulfosuccinates, sodium alkyl sulfoacetate, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, etc.

[0067] Examples of cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyltrimethylammonium bromide, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, behenyltrimethylammonium bromide, benzalkonium chloride, diethylaminoethylamide stearate, dimethylaminopropylamide stearate, and quaternary ammonium salts of lanolin derivatives. Examples of amphoteric surfactants include carboxybetaine type, amidebetaine type, sulfobetaine type, hydroxysulfobetaine type, amidesulfobetaine type, phosphobetaine type, aminocarboxylate type, imidazoline derivative type, amideamine type, etc.

[0068] Organic and inorganic pigments include inorganic pigments such as silica, anhydrous silica, magnesium silicate, talc, sericite, mica, kaolin, bengala, clay, bentonite, titanium-coated mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide, calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine, chromium oxide, chromium hydroxide, calamine, and complexes thereof; Examples include polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenolic resin, fluoropolymer, silicone resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene and styrene copolymer, silk powder, cellulose, CI pigment yellow, CI pigment orange, and other organic pigments, as well as composite pigments of inorganic pigments and organic pigments.

[0069] As organic powders, metal soaps such as calcium stearate; metal alkyl phosphate salts such as sodium zinc cetylphosphate, zinc laurylphosphate, and calcium laurylphosphate; polyvalent metal salts of acyl amino acids such as calcium lauroyl-beta-alanine, zinc lauroyl-beta-alanine, and calcium lauroylglycine; polyvalent metal salts of amide sulfonic acids such as calcium lauroyl-taurine and calcium palmitoyl-taurine; N-acyl basic amino acids such as N-epsilon-lauroyl-L-lysine, N-epsilon-palmitoyllysine, N-alpha-palitoylolnitine, N-alpha-lauroylarginine, and N-alpha-hydrogenated beef tallow fatty acid acylarginine; and N-acyl polypeptides such as N-lauroylglycylglycine. Alpha-amino fatty acids such as alpha-aminocaprilic acid and alpha-aminolauric acid; polyethylene, polypropylene, nylon, polymethyl methacrylate, polystyrene, divinylbenzene and styrene copolymers, tetrafluoroethylene, etc. are examples.

[0070] As ultraviolet absorbers, para-aminobenzoic acid, ethyl para-aminobenzoate, amyl para-aminobenzoate, octyl para-aminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylate, benzyl cinnamonate, paramethoxycinnamonate-2-ethoxyethyl paramethoxycinnamonate, octyl paramethoxycinnamonate, mono-2-ethylhexaneglyceryl diparamethoxycinnamonate, isopropyl paramethoxycinnamonate, a mixture of diisopropyl and diisopropyl cinnamonate esters, urocanic acid, ethyl urocanic acid, hydroxymethoxybenzophenone, hydroxymethoxybenzophenonesulfonic acid and its salts, dihydroxymethoxybenzophenone, sodium dihydroxymethoxybenzophenonedisulfonate, dihydroxybenzophenone, tetrahydroxybenzophenone, Examples include 4-tert-butyl-4'-methoxydibenzoylmethane, 2,4,6-trianilino-p-(carbo-2'-ethylhexyl-1'-oxy)-1,3,5-triazine, 2-(2-hydroxy-5-methylphenyl)benzotriazole, etc.

[0071] Examples of disinfectants include hinokitiol, triclosan, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcinol, isopropylmethylphenol, azulene, salicylic acid, zincphyllithion, benzalkonium chloride, photosensitive element 301, mononitroguaiacol sodium, undecylenic acid, etc.

[0072] Examples of antioxidants include butylhydroxyanisole, propyl gallic acid, and elisorbic acid.

[0073] Examples of pH adjusters include citric acid, sodium citrate, malic acid, sodium malate, fumaric acid, sodium fumarate, succinic acid, sodium succinate, sodium hydroxide, sodium monohydrogen phosphate, etc.

[0074] Examples of alcohols include higher alcohols such as cetyl alcohol.

[0075] In addition, the ingredients that may be added are not limited to those mentioned above, and any of the above ingredients may be incorporated within a range that does not impair the purpose and effects of the present invention.

[0076] The cosmetic composition of the present invention may take the form of a solution, an emulsion, a viscous mixture, etc.

[0077] The ingredients included in the cosmetic composition of the present invention may include ingredients commonly used in cosmetic compositions as active ingredients, and include, for example, conventional auxiliary agents and carriers such as stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0078] The cosmetic composition for preventing or improving skin diseases according to the present invention can be prepared in any formulation commonly manufactured in the art, such as a lotion, cream, lotion, pack, foundation, lotion, beauty serum, hair cosmetic, etc.

[0079] Specifically, the cosmetic composition of the present invention includes formulations of skin lotion, skin softener, skin toner, milk lotion, astringent, lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, foundation, essence, nourishing essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, hair lotion, hair tonic, hair essence, hair shampoo, hair rinse, hair treatment, body lotion, and body cleanser.

[0080] In the case where the formulation of the present invention is a paste, cream, or gel, animal fibers, plant fibers, wax, paraffin, starch, tracanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as carrier components.

[0081] In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0082] In the case where the formulation of the present invention is a solution or emulsion, a solvent, a solvating agent, or an emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0083] In the case where the formulation of the present invention is a suspension, liquid diluents such as water, ethanol, or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used as carrier components.

[0084] In the case where the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolin derivative, or ethoxylated glycerol fatty acid ester, etc. may be used as a carrier component.

[0085]

[0086] In addition, the present invention provides a patch composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

[0087]

[0088] In addition, the present invention provides a method for preventing or treating a skin disease, comprising the step of administering a composition for preventing or treating the skin disease to an individual.

[0089] The composition for the prevention or treatment of skin diseases of the present invention may be administered to mammals, such as rats, mice, livestock, and humans, by various routes. Any mode of administration may be anticipated, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intradural, or intracerebroventricular injection.

[0090] The term "individual" as used in this invention refers to a subject requiring a method for the prevention, control, or treatment of a disease, and may be used without limitation and includes humans, dogs, monkeys, cats, rodents, e.g., mice, genetically modified mice, etc. More specifically, it refers to mammals such as humans or non-human primates, mice, rats, dogs, cats, horses, and cattle.

[0091] The present invention will be explained in more detail through the following examples. However, the following examples are intended only to illustrate the content of the present invention and do not limit the present invention.

[0092]

[0093] <Preparation Example 1> Preparation and Characterization of Nano-graphene Oxide

[0094] <1-1> Preparation of Nano-Graphene Oxide

[0095] To confirm the combined effect of CABS mitochondria and graphene of the present invention, nano-sized graphene oxide (NGO) was prepared. Specifically, graphite (Qingdao Kropfmuehl, China) was used as the carbon structure, and graphene oxide was synthesized using the Couette-Tayler flow method, in which a vortex is generated in a container containing a graphite mixture to allow sulfuric acid to penetrate the graphite layer and promote a chemical reaction. The sulfuric acid (H2SO4), sodium nitrate (NaNO3), and potassium permanganate (KMnO4) listed below were all purchased from Samjeon Sunyak. Graphite was mixed with sulfuric acid (H2SO4) in a ratio of 1:40 to 1:70, and sodium nitrate (NaNO3), which lowers the viscosity of the mixed solution to facilitate stirring, was mixed into the mixture in a ratio of 1:0.2 to 1:0.7 relative to graphite. At this time, sulfuric acid (H2SO4) is inserted between the interlayer structures of graphite, expanding the interlayer spacing.

[0096] The above mixture is mixed at 150 rpm to 500 rpm for 1 to 3 hours. Then, the above mixture and potassium permanganate (KMnO4) are mixed in a ratio of 1:5 to 1:15, and a rotational force of 1,200 rpm to 5,000 rpm is applied for 1 to 72 hours to carry out interlayer expansion and separation of graphite.

[0097] At this stage, it is important to properly manage the reaction time and rotational force by controlling the oxidation reaction to control the interlayer spacing and to produce single-layer graphene oxide. The graphene oxide produced by the above process is nanoscaled by applying ultrasound ranging from a minimum of 10 W to 900 W. The nanoscaled conditions vary depending on the size of the graphene oxide being nanoscaled.

[0098]

[0099] To enhance the frictional force of the obtained graphene oxide, DI water or PBS solvent is added in an amount of 50 to 100 times the solid content of the graphene oxide. A certain amount of graphene oxide mixed in the solvent is introduced into a nanofabrication device where physical methods are applied to reduce the particle size of the graphene oxide. Furthermore, the ratio of each material applied during the production of graphene oxide can act as a ratio that promotes the nanofabrication of the graphene oxide. In this experiment, nanofabrication of graphene oxide can be separated using physical methods in the nanofabrication device. Additionally, it can be separated by size through a centrifugation process. At this time, centrifugation can be performed at a rotation speed of 8,000 to 20,000 rpm depending on the size.

[0100]

[0101] <1-2> Characterization of Nano-Graphene Oxide

[0102] The characteristics of the nano-graphene oxide prepared in Preparation Example 1-1 above were confirmed. Specifically, the morphology of the nano-graphene oxide was confirmed using a transmission electron microscope (TEM). To this end, nano-graphene oxide was loaded onto a 400-mesh carbon-coated copper grid and analyzed using a Cs-corrected HRTEM (JEM-ARM200F, Cold FEG, JEOL Ltd, Japan). In addition, particle size analysis (CPS Disc Centrifuge; CPS Instruments, USA) was used to confirm the average particle size. At this time, nano-graphene oxide was separated by size using centrifugal precipitation in a liquid medium, and the precipitate was stabilized by a slight density gradient within the liquid. Finally, to confirm the chemical bonding characteristics, the Fourier transform infrared spectroscopy (FTIR) spectrum was analyzed using a Platinum FT-IR Spectrometer (ALPHA II, Bruker Corp., USA) to analyze the FT-IR peaks CO (1045 cm-1), C=O (1630 cm-1, 1729 cm-1), CH (2859 cm-1), and the OH peak generated during the nano-processing of nano-graphene oxide.

[0103]

[0104] As a result, as shown in Figures 1 to 3, the morphology of the nano-graphene oxide of the present invention was confirmed, and it was confirmed that the average particle size is 15.9 nm. In addition, it was confirmed that a carbohydrate peak is exhibited in FTIR.

[0105]

[0106] <Preparation Example 2> Preparation of CABS mitochondria and AM mitochondria

[0107] <2-1> Preparation of CABS Mitochondria

[0108] Mitochondria were isolated from the bone marrow-derived stem cells of the present invention to produce Catholic Bone Marrow Stem Cells (CABS) and CABS mitochondria. The 'CABS-mito' of the present invention refers to mitochondria isolated and purified from Catholic Bone Marrow Stem Cells; it was named by combining 'mitochondria' with CABS, the English abbreviation for the cell name (Catholic Bone Marrow Stem Cell). Specifically, cultured Bone Marrow-derived Mesenchymal Stem Cells were recovered, suspended in a mitochondrial isolation buffer, and physically lysed using a homogenizer. Subsequently, differential centrifugation was performed, and the supernatant was obtained after removing nuclei and cellular debris through low-speed centrifugation. Subsequently, only the mitochondrial pellet was selectively precipitated through high-speed centrifugation and obtained, and CABS mitochondria were isolated.

[0109]

[0110] <2-2> Preparation of AM Mitochondria

[0111] Adipose tissue-derived mesenchymal stem cells (AM) of the present invention produced AM mitochondria. The term 'AM-mito' used herein refers to mitochondria isolated and purified from adipose tissue-derived mesenchymal stem cells, and was named by taking the English initials of adipose tissue and mesenchymal, which are the origins of the cells. Specifically, the collected adipose tissue was minced and enzymatically dissociated by collagenase treatment, and then centrifuged to obtain the stromal vascular fraction (SVF). Adipose-derived mesenchymal stem cells (Adipose Derived-MSCs) exhibiting adherence to culture vessels were selectively proliferated from the SVF.

[0112] The secured cells were swollen in a hypotonic buffer and physically lysed, then the nuclear and cytoplasmic debris were sequentially removed through a continuous centrifugation process, and the mitochondrial fraction was finally isolated and purified from the supernatant.

[0113]

[0114] <Example 1> Confirmation of effect on improving psoriasis activity

[0115] To determine whether the combined use of CABS or AM mitochondria and graphene according to the present invention improves psoriasis, an animal model of psoriasis was constructed. Specifically, the dorsal hair of SKG mice, known to readily form an autoimmune environment, was removed. One day later, imiquimod, a psoriasis-inducing substance, was applied to the dorsal skin of the mice at a dose of 62.5 mg daily for six days. Additionally, on the third day after the application of imiquimod, 10 μg / ml of CABS mitochondria or AM mitochondria and 15.9 nm grade graphene nanooxide were diluted in saline to a concentration of 6.7 mpk and injected intraperitoneally. Scaling scores, redness scores, and thickness scores were measured from the time of psoriasis induction until the end of the experiment, and the mice were humanely sacrificed at the end of the experiment. The specific process of constructing the animal model is shown in Figure 4. As control groups, a mitochondrial administration group administered only CABS mitochondria or AM mitochondria and a graphene-alone administration group (Graphene 6.7 mpk) and a negative control group with psoriasis induced by imiquimod were used.

[0116]

[0117] As a result, as shown in Figures 5 and 6, compared to the negative control group, it was visually confirmed that the pathological activity of psoriasis was reduced in the mitochondrial treatment group treated with CABS or AM mitochondria and the graphene-alone treatment group, and that the keratin index, erythema index, and skin thickness index were reduced. In addition, in the group administered with the combination of mitochondria and graphene, it was confirmed that the keratin index, erythema index, and skin thickness index were significantly reduced compared to the mitochondrial treatment group and the graphene-alone treatment group, confirming that the combined administration of graphene enhances the psoriasis-improving effect of CABS or AM mitochondria.

[0118]

[0119] <Example 2> Confirmation of Immune Cell Subtype Regulation

[0120] To determine whether the combined use of CABS or AM mitochondria and graphene of the present invention improves psoriasis, the regulatory effects on immune cell subtypes and tissue-resident memory cells (TRMs) were examined. Specifically, spleen tissue was obtained from each group of mice sacrificed in Example 1, and single cells isolated from the spleen were stimulated with PMA lonobycin for 2 hours and then stimulated with Golgi stop for 2 hours, after which flow cytometry staining was performed. Subsequently, IL-17-positive CD8 T cells (Tc17), which are the immune cell subtypes causing psoriasis, and CD8-positive TRM (CD8 TRM) and CD8-positive TRM 17 (CD8 TRM 17) cells, which are TRM subtypes, were analyzed using flow cytometry.

[0121]

[0122] As a result, as shown in Figure 7, compared to the negative control group, the frequency of Tc17 cells was significantly reduced in the group treated with mitochondria, and within the mitochondria group, it was confirmed that the frequency of Tc17 cells was significantly reduced in the group treated with CABS mitochondria. In addition, in the group administered with mitochondria and graphene, it was confirmed that the frequency of Tc17 cells was significantly reduced compared to treatment with mitochondria alone.

[0123]

[0124] In addition, when examining the frequency of tissue-resident memory cells, it was confirmed that the frequency of CD8 TRM and CD TRM 17 cells was significantly reduced in the group treated with mitochondria compared to the negative control group. Furthermore, it was confirmed that the group treated with CABS mitochondria showed a superior regulatory effect on tissue-resident memory cells compared to AM mitochondria. Additionally, in the group administered with mitochondria in combination with graphene, it was confirmed that the frequency of tissue-resident memory cells was significantly reduced compared to the group administered with mitochondria alone (Figs. 8 and 9), confirming that the combination of mitochondria and graphene effectively regulates the pathogenic immune cell subtypes of psoriasis.

[0125]

[0126] <Example 3> Histological Analysis

[0127] Histological analysis was performed to determine whether the combined use of CABS or AM mitochondria and graphene of the present invention improves psoriasis. Specifically, skin tissue was obtained from each group of mice sacrificed in Example 1, sectioned, and then stained with Hematoxylin & Eosin (H&E) to measure the epidermal thickness.

[0128]

[0129] As a result, as shown in Figures 10 and 11, it was confirmed that the epidermal layer thickness was significantly reduced in the group treated with mitochondria compared to the negative control group. In addition, it was confirmed that the epidermal layer thickness was significantly reduced in the group treated with mitochondria alone compared to the group treated with mitochondria alone, thereby confirming that the combined administration of mitochondria and graphene effectively controls psoriasis.

[0130]

[0131] Therefore, it was confirmed that the combined administration of mitochondria and graphene of the present invention reduced skin thickness, keratinization, and erythema in an animal model of psoriasis. Furthermore, it was confirmed that it regulated inflammatory immune cell subtypes and tissue-resident memory cells, which are pathogenic immune cells of psoriasis. It was also confirmed that it reduced the thickness of the epidermal layer.

Claims

1. A composition for the prevention or treatment of skin diseases comprising mitochondria and graphene as active ingredients.

2. In Paragraph 1, The above graphene is a composition in which the graphene is nano-sized graphene oxide.

3. In Paragraph 2, A composition in which the above-mentioned nano-graphene oxide has a particle size of 15 to 38 nm.

4. In Paragraph 1, A composition in which the mitochondria are mitochondria isolated from adipose tissue-derived mesenchymal stem cells (AM) or Catholic bone marrow stem cells (CABS).

5. In Paragraph 1, The above composition is a composition that reduces keratin, erythema, and skin thickness.

6. In Paragraph 5, A composition in which reducing the skin thickness above is reducing the thickness of the epidermal layer.

7. In Paragraph 1, The above composition is a composition that reduces the frequency of pathogenic immune cells.

8. In Paragraph 7, A composition in which the immune cells are Tc17 or tissue-resident memory cells (TRM).

9. In Paragraph 8, A composition in which the tissue-resident memory cells are CD8-positive TRM or CD8-positive TRM 17.

10. In Paragraph 1, A composition in which the above skin disease is psoriasis.

11. A cosmetic composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

12. A patch composition for the prevention or improvement of skin diseases comprising mitochondria and graphene as active ingredients.

13. A method for preventing or treating a skin disease comprising the step of administering the composition of claim 1 to an individual.

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