WO2016122140A1 - Pharmaceutical composition for preventing or treating skin diseases, containing cell culture medium - Google Patents

Pharmaceutical composition for preventing or treating skin diseases, containing cell culture medium Download PDF

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WO2016122140A1
WO2016122140A1 PCT/KR2016/000276 KR2016000276W WO2016122140A1 WO 2016122140 A1 WO2016122140 A1 WO 2016122140A1 KR 2016000276 W KR2016000276 W KR 2016000276W WO 2016122140 A1 WO2016122140 A1 WO 2016122140A1
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skin
pharmaceutical composition
component
hydrochloride
cell culture
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PCT/KR2016/000276
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French (fr)
Korean (ko)
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김찬화
김영준
김현정
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주식회사 바이오코즈글로벌코리아
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Priority to KR1020167002821A priority Critical patent/KR20160102959A/en
Publication of WO2016122140A1 publication Critical patent/WO2016122140A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
    • A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4415—Pyridoxine, i.e. Vitamin B6
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A61K31/51—Thiamines, e.g. vitamin B1
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/14—Alkali metal chlorides; Alkaline earth metal chlorides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin

Definitions

  • the present invention relates to a composition for preventing or treating skin diseases, including a cell culture medium, or a composition for improving skin conditions, and a method for preventing or treating skin diseases using the composition, or a method for improving skin conditions.
  • allergic diseases are increasing in various age groups due to changes in dietary and residential life and exposure to chemical or biological harmful substances caused by environmental pollution. This is sensitive to the external environment harmless to the human body, it appears as a respiratory disease such as asthma or rhinitis or skin diseases such as contact dermatitis, atopic dermatitis.
  • the skin causes an allergic reaction by direct contact with external allergens by air, food, etc.
  • This allergic skin disease is caused by continuous exposure to external antigens such as food, bacteria, ticks, and climate factors.
  • Inflammatory diseases are caused by allergic reactions to various environmental factors.
  • Allergy here is a type of hypersensitivity reaction caused by imbalance, which is a response to a specific antigen called allergn.
  • the allergic reaction begins with histamine decarboxylated by L-histidine dicarboxylase, which is stored in granule form in mast cells or basophils, in which they bind to allergens, whereby the cells are degranulated, resulting in histamine and ⁇ -hexaxa. Secrete hexoxaminidase.
  • Atopic dermatitis one of allergic diseases, is a growing trend worldwide and is a chronic and recurrent skin disease that begins mainly in infancy and childhood.
  • the cause of atopic dermatitis is not yet clearly identified, but environmental factors, genetic predisposition, immunochemical reactions, and skin barrier abnormalities are considered as the main causes of the disease, and the symptoms of pruritus, dry skin, eczema, etc. It appears to be various.
  • the distribution and response patterns of skin lesions also vary according to the age of the patient.
  • drugs commonly used for the treatment of atopic dermatitis generally include topical steroids, local immunomodulators, systemic steroids, systemic immunosuppressants, and antihistamines, but there are risks of various side effects caused by the administration or administration of these drugs.
  • topical steroids local immunomodulators, systemic steroids, systemic immunosuppressants, and antihistamines
  • antihistamines drugs commonly used for the treatment of atopic dermatitis
  • drugs commonly used for the treatment of atopic dermatitis generally include topical steroids, local immunomodulators, systemic steroids, systemic immunosuppressants, and antihistamines, but there are risks of various side effects caused by the administration or administration of these drugs.
  • an excellent active ingredient that is safe and exhibits skin-improving effects such as dry skin and abnormal skin protective film.
  • DMEM / F12 Dulbecco's Modified Eagle Medium / F12
  • DMEM / F12 is a medium used for culturing cells, and is composed of various kinds of amino acids, vitamins, inorganic salts, and other substances, so that cells are stable for growth and differentiation.
  • the medium or a composition including some components of the medium can be used for treating skin diseases or improving skin condition.
  • Another object of the present invention is to provide a composition for improving skin condition comprising a cell culture medium or some components thereof, and a method for improving skin condition using the composition.
  • the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases, including cell culture medium.
  • the present invention also provides a method for preventing or treating skin diseases comprising the step of applying the pharmaceutical composition of the present invention to the skin of a subject in need thereof.
  • the present invention also provides a use of the pharmaceutical composition according to the invention for use in the manufacture of a medicament for the prevention or treatment of skin diseases.
  • the present invention provides a cosmetic composition for improving skin conditions comprising a cell culture medium.
  • the present invention also provides a method for improving the skin condition using the cosmetic composition of the present invention.
  • the present invention also provides a use of the cosmetic composition according to the invention for use in the preparation of a cosmetic for improving the skin condition.
  • the pharmaceutical composition comprising the cell culture medium of the present invention inhibits the degranulation of the skin and the expression of inflammatory response-related substances, promotes the expression of skin barrier-related substances, exhibits an effect on the recovery of damaged skin, preventing the skin disease Or it can be usefully used to treat or improve skin condition.
  • Example 1 is a graph showing the concentration change of ⁇ -hexosaminidase after treating the composition of Example 1 for each concentration.
  • Example 2 is a graph showing a change in the concentration of histamine after treating the composition of Example 1 for each concentration.
  • Figure 3 shows the change in expression of IL-6 at the mRNA level after the treatment of the composition of Example 1 by concentration.
  • Figure 4 shows the change in expression of serine palmitoyltransferase (SPT) at the mRNA level after treating the composition of Example 1 by concentration.
  • SPT serine palmitoyltransferase
  • Figure 5 shows the change in the expression level of the filaggrin (filaggrin) after the treatment of the composition of Example 1 by concentration.
  • Example 6 is a graph confirming the proliferation rate of cells damaged by sodium lauryl sulfate (SLS) after treating the composition of Example 2 and the extract of Comparative Example 1 by concentration.
  • SLS sodium lauryl sulfate
  • Example 7 is a graph confirming the proliferation rate of cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
  • Figure 8 shows the recovery of artificial skin damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
  • Example 9 is a graph confirming the expression level of IL-1 ⁇ in cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
  • Example 10 is a graph confirming the expression level of TNF- ⁇ of cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
  • the present invention provides a pharmaceutical composition for preventing or treating skin diseases, including cell culture medium.
  • the term "cell culture medium” or “culture medium” contains a component required by the cell for cell growth and survival in vitro , or contains a component that aids cell growth and survival.
  • the component may be a vitamin, an essential or non-essential amino acid, and a trace element.
  • the medium may be a medium used for culturing cells, preferably eukaryotic cells, more preferably mammalian cells.
  • cell culture media examples include, but are not limited to, DMEM / F12, DMEM, MEM ⁇ , and the like.
  • Cell culture medium according to the present invention is a serum-free medium (serum-free).
  • Serum-free medium refers to a culture medium that does not contain serum (eg, animal-derived serum) derived from animals, including humans.
  • Animal-derived serum provides universal nutrients for the growth of cells to be cultured, but contains unidentified trace components, making it difficult to analyze, establishing reproducible testing and production processes, and ensuring stability to the human body. There is no problem.
  • the cell culture medium according to the present invention is a serum-free medium to ensure the stability to the human body, it is possible to establish a reproducible test and production process.
  • the cell culture medium is composed of amino acid components, vitamin components, inorganic salt components and other components,
  • the amino acid component is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L-lysine hydrochloride, L-methionine, At least one amino acid selected from the group consisting of L-proline, L-serine, L-threonine and L-valine, or a combination thereof,
  • said vitamin component is at least one amino acid selected from the group consisting of i-inositol, thiamine hydrochloride, niacinamide and pyridoxine hydrochloride or combinations thereof,
  • the inorganic salt component is sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), potassium chloride (KCl), calcium chloride (CaCl 2 ) (anhydrous) and sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O) At least one amino acid selected from the group consisting of or a combination thereof,
  • the other component is preferably D-glucose (dextrose) or sodium pyruvate.
  • the cell culture medium is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L- Lysine hydrochloride, L-methionine, L-proline, L-serine, L-threonine, L-valine, i-inositol, thiamine hydrochloride, niacinamide, pyridoxine hydrochloride, sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), Potassium chloride (KCl), calcium chloride (CaCl 2 ) (anhydrous), sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O), D-glucose (dextrose) and sodium pyruvate .
  • Cell culture medium of the present invention may be included in 0.5 to 100% by weight, preferably 1.0 to 10% by weight based on the total weight of the pharmaceutical composition. Preferably from 5.0 to 8% by weight. If the composition is included in less than 0.5% by weight may have a problem that the effect is insignificant.
  • the amino acid component is used for the growth of cells as a raw material for protein synthesis, which is 0.1 to 0.2% by weight glycerin, 0.01 to 0.1% by weight L-alanine, 0.5 to 3% by weight based on the total weight of the composition L-arginine hydrochloride, 0.1-0.2 wt% L-cysteine hydrochloride-monohydrate, 2-3 wt% L-glutamine, 0.1-0.5 wt% L-histidine hydrochloride-monohydrate, 0.4-1.0 Wt-% L-lysine hydrochloride, 0.1-0.5 wt-% L-methionine, 0.1-0.5 wt-% L-proline, 0.1-0.3 wt-% L-serine, 0.3-0.6 wt-% L-threonine and 0.3 To 0.6% by weight of L-valine.
  • amino acid component and content included in the composition according to the present invention is 0.12 to 0.14 wt% glycerin, 0.02 to 0.04 wt% L-alanine, 1.01 to 1.06 wt% L-arginine hydrochloride, 0.11 to 0.15 wt% L-cysteine hydrochloride-monohydrate, 2.3-2.8 wt% L-glutamine, 0.20-0.25 wt% L-histidine hydrochloride-monohydrate, 0.5-0.8 wt% L-lysine hydrochloride, 0.1-0.3 wt% L-methionine, 0.1-0.3 wt% L-proline, 0.16-0.2 wt% L-serine, 0.35-0.40 wt% L-threonine and 0.35-0.40 wt% L-valinyl Can be.
  • amino acid component in the content within the above range can help the growth and maintenance of cells and improve the formulation stability.
  • the vitamin component serves to maintain the activity of the cells, which is 0.01 to 0.1% by weight of i-inositol, 0.01 to 0.1% by weight of thiamin hydrochloride, 0.005 to 0.03% by weight of niacinamide and 0.01 to 0.1% by weight of pyridoxine hydrochloride.
  • One embodiment of the vitamin component and content included in the composition according to the present invention is 0.08 to 0.09 wt% i-inositol, 0.01 to 0.02 wt% thiamine hydrochloride, 0.01 to 0.02 wt% niacinamide and 0.01 to 0.02 Wt% pyridoxine hydrochloride.
  • a vitamin component in a content within the above range can help maintain cell activity.
  • the inorganic salt component serves to regulate the expression of cell function, which is 40 to 55% by weight sodium chloride (NaCl), 10 to 20% by weight sodium hydrogencarbonate (NaHCO 3 ), 1 to 1 based on the total weight of the composition 3 weight percent potassium chloride (KCl), 0.5 to 1.0 weight percent calcium chloride (CaCl 2 ) (anhydrous) and 0.2 to 0.7 weight percent sodium hydrogen phosphate monohydrate (NaH 2 PO 4 —H 2 O). .
  • One embodiment of the inorganic salt component and content included in the composition according to the present invention is 45 to 50% by weight of sodium chloride (NaCl), 15 to 18% by weight of sodium bicarbonate (NaHCO 3 ), 2.0 to 2.5% by weight of Potassium chloride (KCl), 0.8-0.9 wt% calcium chloride (CaCl 2 ) (anhydrous) and 0.4-0.5 wt% sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O).
  • sodium chloride NaCl
  • NaHCO 3 sodium bicarbonate
  • KCl Potassium chloride
  • CaCl 2 calcium chloride
  • NaH 2 PO 4 -H 2 O sodium hydrogen phosphate monohydrate
  • the other components include a carbon source serving as an energy source or a component for maintaining the pH of the medium, which is 15 to 30% by weight of D-glucose (dextrose) or 0.3 to 0.5 based on the total weight of the composition. Weight percent sodium pyruvate.
  • composition according to the present invention may be 20 to 25% by weight of D-glucose (dextrose) or 0.35 to 0.4% by weight of sodium pyruvate.
  • Dextrose D-glucose
  • sodium pyruvate Other components included in the content within the above range, for example, the carbon source may generate energy to help cell growth, improve formulation stability, and the like.
  • the skin disease may be selected from the group consisting of atopic dermatitis, allergy, skin eczema, acne, psoriasis, itching and combinations thereof.
  • the pharmaceutical composition of the present invention may further include a component selected from the group consisting of an amino acid component, a vitamin component, an inorganic salt component, other components, and combinations thereof.
  • the amino acid component may be any amino acid component as long as it is an amino acid component used in a general animal cell culture medium.
  • an amino acid component used in a general animal cell culture medium.
  • the vitamin component may be any vitamin component as long as it is a vitamin component used in a general animal cell culture medium.
  • the vitamin component may be selected from the group consisting of biotin, calcium D-pantothenate, folic acid, riboflavin, vitamin B12, and combinations thereof. Can be.
  • the inorganic salt component may be any inorganic salt component as long as it is an inorganic salt component used in a general animal cell culture medium.
  • copper sulfate pentahydrate (CuSO 4 -5H 2 O) ferric sulfate heptahydrate (FeSO 4- ) 7H 2 O
  • magnesium chloride anhydrous
  • magnesium sulfate MgSO 4
  • disodium hydrogen phosphate Na 2 HPO 4
  • ZnSO 4 -7H 2 O zinc sulfate heptahydrate
  • the other component may be any component as long as it is a component used in a general animal cell culture medium, and may be selected from the group consisting of, for example, hypoxanthine Na, linolenic acid, lipoic acid, putrescine 2HCl, thymidine, and combinations thereof. Can be.
  • the pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier is commonly used in the manufacture of a drug, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but is not limited thereto.
  • composition of the present invention may further include a pharmaceutically acceptable additive selected from the group consisting of lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and combinations thereof.
  • a pharmaceutically acceptable additive selected from the group consisting of lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and combinations thereof.
  • the carrier may comprise from about 1% to about 99.99% by weight, preferably from about 90% to about 99.99% by weight, based on the total weight of the pharmaceutical composition of the present invention, wherein the pharmaceutically acceptable additive is about 0.1 Weight percent to about 20 weight percent.
  • compositions of the present invention may be administered parenterally, and may preferably be administered directly to the skin in a topical manner.
  • Formulations of the pharmaceutical compositions of the present invention may be in the form of external skin preparations such as transdermal injections, ointments, solutions, creams, payments, sprays, patches, and the like.
  • Suitable dosages of the pharmaceutical compositions of the present invention are determined in view of various related factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and reaction sensitivity.
  • the dosage should not be understood as limiting the scope of the invention in any aspect.
  • the present invention provides a method for preventing or treating a skin disease comprising applying the pharmaceutical composition to the skin of a subject in need thereof.
  • the present invention also provides the use of the pharmaceutical composition for use in the manufacture of a medicament for the prevention or treatment of skin diseases.
  • the subject to which the pharmaceutical composition may be applied may be a mammal, and specifically, a human.
  • the present invention provides a cosmetic composition for improving skin condition comprising a cell culture medium.
  • the cosmetic composition of the present invention has the same active ingredient as the above-mentioned pharmaceutical composition.
  • the cosmetic composition according to the present invention is selected from the group consisting of inhibiting the occurrence of wrinkles, inhibiting skin aging, improving skin elasticity, skin regeneration, wound or wound healing, corneal regeneration, skin irritation and combinations thereof, skin cells It can be used to protect the skin from a decrease or loss of function or to improve the skin condition, or to prevent or improve skin diseases.
  • the skin disease may be selected from the group consisting of atopic dermatitis, allergy, skin eczema, acne, psoriasis, itching and combinations thereof.
  • the cosmetic composition of the present invention can be applied directly to the skin for the purpose of improving the skin condition.
  • the cosmetic composition may be formulated into a cosmetic formulation commonly prepared in the art.
  • the cosmetic compositions are formulated, for example, in solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like.
  • the present invention is not limited thereto. More specifically, it may be formulated in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.
  • the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and mixtures thereof It may include a carrier component selected from the group consisting of.
  • the formulation of the cosmetic composition of the present invention is a powder or a spray
  • it may include a carrier component selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and mixtures thereof, in particular spray
  • a carrier component selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and mixtures thereof, in particular spray
  • chlorofluorohydrocarbon it may further include propane / butane or dimethyl ether.
  • the formulation of the cosmetic composition of the present invention may comprise a carrier component selected from the group consisting of solvents, solvating agents, emulsifying agents and mixtures thereof which are solutions or emulsions.
  • a carrier component selected from the group consisting of solvents, solvating agents, emulsifying agents and mixtures thereof which are solutions or emulsions.
  • examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycols, sorbitan fatty acid esters, mixtures thereof, and the like.
  • solvents selected from the group consisting of solvents, solvating agents, emulsifying agents and mixtures thereof which are solutions or emulsions.
  • examples thereof include water, ethanol, isopropanol, ethyl
  • the formulation of the cosmetic composition of the present invention is a suspension
  • a liquid diluent such as water, ethanol or propylene glycol
  • suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystal Carrier cellulose, aluminum metahydroxy, bentonite, agar, tragacanth and mixtures thereof.
  • the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide Carrier sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and mixtures thereof.
  • the "carrier component" in the cosmetic composition of the present invention is a compound or composition already known and used that may be included in cosmetic preparations, and is a component that is not toxic, unstable or irritant as long as the human body is suitable for contact with the skin.
  • the cosmetic composition of the present invention may further include an adjuvant selected from the group consisting of antioxidants, stabilizers, solubilizers, humectants, pigments, perfumes, sunscreens, colorants, surfactants, and combinations thereof.
  • the adjuvant is not limited to use as long as the adjuvant commonly used in the preparation of the cosmetic composition.
  • the present invention provides a method for improving the skin condition using the cosmetic composition.
  • the present invention also provides the use of said cosmetic composition for use in the preparation of a cosmetic for improving skin condition.
  • the method for improving the skin condition may include applying to the skin of the subject in need thereof.
  • the subject may be a mammal, specifically a human.
  • Applying to the skin may include applying or spraying the cosmetic composition according to the invention directly on the skin according to its form.
  • the application amount and the number of times of use of the cosmetic composition may be appropriately set according to the age, sex, use, degree of symptoms of the user, for example, the appropriate amount of the cosmetic composition at a frequency of 1 to 6 times a day. It can be applied to the skin.
  • the composition was prepared using ultrapure purified water as the solvent, and the concentration of serum-free DMEM / F12 medium (Gibco, USA) having the composition of Table 1 was 2%, 10%, 50%, or 100%, respectively.
  • each of the components shown in Table 2 below were dissolved in ultrapure water as a solvent to confirm that all the components were dissolved, and then filtered through a 0.22 ⁇ m filter. The filtrate was then added diluted with ultrapure purified water such that the final concentration in the cell culture medium was 0.5%, 1%, 2%, 5% or 8%.
  • Each component described below is a material listed in the cosmetic raw materials published by the Korea Food and Drug Administration, all were purchased from Sigma-Aldrich, USA.
  • March extract for use as a control was used to receive a 40% March extract from Cosmax Co., Ltd., a cosmetics company.
  • 40% of the extract of Machiwyeon treatment was diluted with ultrapure purified water so that the final concentration is 0.5%, 1%, 2%, 5% or 8%.
  • the concentration of ⁇ -hexosaminidase which is an indicator of degranulation initiated by an allergic reaction, was measured by the following method.
  • bas basophilic leukemia cells (Korean Cell Line Bank) in culture in a culture dish were treated with trypsin 0.5% (w / v) and separated from the culture dish.
  • 2 ⁇ 10 5 isolated basophilic mast cells were mixed with anti-DNP IgE (Sigma-Aldrich, USA) at a concentration of 0.5 mg / ml, and then placed in each well of a 96 well plate. After culturing in an incubator at 37 ° C. and 5% CO 2 for one day, the cells were completely adhered to the bottom of the 96 well plate, and then the culture medium was removed.
  • the Tyrode solution NaCl: 137 mmol / l, KCl: 2.7 mmol / l, CaCl 2 : 1.8 mmol / l, MgCl 2 : 1 mmol / l, glucose: 5.6 mmol / l, HEPES: 20 mmol / l, BSA: 0.1%, titrated to pH 7.3 with NaOH, sterilized with a filter at 4 ° C. Storage).
  • the washed cells were treated with DMEM / F12 medium of Example 1 by concentration, and reacted for 3 hours by adding 1 ⁇ M DNP-HSA solution (Sigma-Aldrich, USA). At this time, a solvent containing no DMEM / F12 medium was used as a negative control.
  • the histamine synthesis inhibitory effect was evaluated the histamine synthesis inhibitory effect.
  • the reaction product was obtained in the same manner as in Experimental Example 1, and the cell culture solution of the reaction solution was collected and measured using an ELISA assay kit for histamine (Oxford biomedical research, USA) according to the manufacturer's guidelines.
  • Example 1 In order to confirm the effect of the composition prepared in Example 1 on the inflammatory response or skin barrier caused by atopy, the inflammatory cytokine IL-6 (interleukin-6), the skin barrier component filaggrin, and the damaged skin barrier recovery The expression of serine palmitoyltransferase (SPT), an important factor in, was confirmed at the mRNA level.
  • SPT serine palmitoyltransferase
  • mRNA was extracted with trizol from cells obtained by treatment with 0.05% trypsin. Synthesis of cDNA from the extracted mRNA was performed according to the manufacturer's guidelines using ReverTra Ace® qPCR Master Mix (TOYOBO, Japan), wherein the primers used (Cosmogenetech, Korea) are shown in Table 3 below.
  • the expression of IL-6 which is an inflammation-associated substance, was suppressed depending on the concentration of DMEM / F12 medium, whereas the skin barrier-related substance, filaggrin and serine palmitoyltransferase
  • the DMEM / F12 medium has an effect of inhibiting the inflammatory response and improving the skin condition.
  • Example 2 In order to confirm how the composition prepared in Example 2 affects the proliferation of skin cells, recovery rate from damage by sodium lauryl sulfate (SLS) or UV was confirmed.
  • SLS sodium lauryl sulfate
  • human dermal fibroblast cells (Korean Cell Line Bank) being cultured in a culture dish were treated with trypsin 0.5% (w / v) and separated from the culture dish.
  • the separated fibroblasts 4 ⁇ 10 3 were put in each well of a 96 well plate.
  • a control a non-treated control, which was not treated with anything, and the gusset extract treatment group prepared in Comparative Example 1 were used.
  • CCK solution (Dojindo, Japan) was added to each well, followed by reaction for 2 hours, and absorbance at 450 nm using a model 680 microplate reader. Measured.
  • artificial skin was purchased by Tego Science Co., Ltd. for the preparation of the epidermal part.
  • 6 artificial skins were removed from the agar and transferred to a 6-well plate, followed by adding 3 ml of the preservation medium (maintenance medium, TA-MM001, Tego science, Korea) provided at 37 ° C. and 5% CO 2 conditions for one day. Stabilized in an incubator.
  • Five of the stabilized artificial skins were transferred to a new 6-well plate with 1 ml of PBS, exposed to 1 J of UVA (ultraviolet-A, radiation spectrum 315-400 nm) and washed twice with PBS.
  • Six artificial skins, including controls not treated with UVA, were placed in a new 6 well plate.
  • the cultured artificial skin was removed from the well, placed in a cassette, and then fixed in a 5% formaldehyde solution for one day, and stained with H & E to observe and photograph skin tissue.
  • the group treated with UVA only damaged the artificial skin compared to the group not treated, and when treated with the gut extract of Comparative Example 1 there was no change in the damaged artificial skin, but
  • the mixed composition of Example 2 was treated, it was confirmed that artificial skin damaged by UVA was restored to normal, so that the mixed composition of Example 2 or the DMEM / F12 medium containing the composition as a component reduced skin sensitivity. It can be seen that it can be used as.
  • composition prepared in Example 2 affects the expression of inflammatory cytokines IL-1 ⁇ (interleukin-1 ⁇ ) and TNF- ⁇ (tumor necrosis factor- ⁇ ) in artificial skin exposed to UVA.
  • IL-1 ⁇ interleukin-1 ⁇
  • TNF- ⁇ tumor necrosis factor- ⁇
  • Example 5 the artificial skin was treated with the composition of Example 2 and the extract of Portico of Comparative Example 1 and cultured for 3 days.
  • the measurement results are shown in FIGS. 9 and 10.
  • the group treated with UVA only increased the amount of IL-1 ⁇ and TNF- ⁇ compared to the group not treated, compared to the case treated with the gusset extract of Comparative Example 1
  • the mixed composition of Example 2 was treated to further inhibit the amount of IL-1 ⁇ and TNF- ⁇
  • the mixed composition of Example 2 of the present invention or the composition as a component It can be seen that DMEM / F12 medium can be used for the treatment of skin diseases such as atopic dermatitis.

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Abstract

The present invention relates to: a composition for preventing or treating skin diseases or for improving the condition of the skin, containing a cell culture medium; and a method for preventing or treating skin diseases or for improving the condition of the skin by using the composition. The composition according to the present invention inhibits the degranulation of the skin and the expression of substances related to inflammatory reactions, promotes the expression of substances related to the skin barrier, and exhibits an effect of facilitating damaged skin recovery, and thus the composition containing a cell culture medium can be useful in preventing or treating skin diseases or in improving the condition of the skin.

Description

세포 배양배지를 포함하는 피부 질환 예방 또는 치료용 약학 조성물Pharmaceutical composition for preventing or treating skin diseases, including cell culture medium
본 발명은 세포 배양배지를 포함하는 피부 질환 예방 또는 치료용, 또는 피부 상태 개선용 조성물과 상기 조성물을 이용한 피부 질환 예방 또는 치료방법, 또는 피부 상태 개선 방법에 관한 것이다.The present invention relates to a composition for preventing or treating skin diseases, including a cell culture medium, or a composition for improving skin conditions, and a method for preventing or treating skin diseases using the composition, or a method for improving skin conditions.
산업화, 도시화의 심화로 식생활과 주거생활의 변화, 환경오염에 의한 화학적 또는 생물학적 유해물질에 대한 노출로, 다양한 연령층에서 알레르기 질환이 크게 증가하고 있다. 이는 인체가 무해한 외부 환경에 대해서도 민감한 반응을 보이며, 천식이나 비염과 같은 호흡기 질환 또는 접촉성 피부염, 아토피성 피부염 등의 피부질환 등으로 나타나게 된다.Due to industrialization and urbanization, allergic diseases are increasing in various age groups due to changes in dietary and residential life and exposure to chemical or biological harmful substances caused by environmental pollution. This is sensitive to the external environment harmless to the human body, it appears as a respiratory disease such as asthma or rhinitis or skin diseases such as contact dermatitis, atopic dermatitis.
특히 피부는 외부 알레르기 유발 물질이 공기, 음식 등에 의해 직접적으로 접촉되어 과민한 면역 반응을 일으키게 되는데, 이와 같은 알레르기성 피부 질환은 음식물, 세균, 진드기, 기후환경인자 등 외부 항원에 지속적으로 노출되어 발병하는 염증 질환으로 개인의 유전적인 특성 때문에 다양한 환경 인자들에 대한 알레르기 반응이 개개인마다 다르게 나타난다.In particular, the skin causes an allergic reaction by direct contact with external allergens by air, food, etc. This allergic skin disease is caused by continuous exposure to external antigens such as food, bacteria, ticks, and climate factors. Inflammatory diseases are caused by allergic reactions to various environmental factors.
여기서 알레르기란, 면역 불균형에 의한 과민반응의 일종으로 알러젠(allergn)이라는 특정한 항원에 대하여 나타나는 반응이다. 알레르기 반응은, 히스티딘이 L-히스티딘 디카복시레이즈에 의해 디카복실레이션되어 형성된 히스타민이 비만세포나 호염구에 과립형태로 저장되는데 이들이 알러젠과 결합하여 시작되고, 이때 세포가 탈과립되어 히스타민과 β-헥소사미니데이즈(hexoxaminidase)를 분비한다.Allergy here is a type of hypersensitivity reaction caused by imbalance, which is a response to a specific antigen called allergn. The allergic reaction begins with histamine decarboxylated by L-histidine dicarboxylase, which is stored in granule form in mast cells or basophils, in which they bind to allergens, whereby the cells are degranulated, resulting in histamine and β-hexaxa. Secrete hexoxaminidase.
알레르기성 질환의 하나인 아토피성 피부염은 세계적으로 증가하는 추세로서, 주로 유아기 및 소아기에 시작되는 만성적이고 재발성의 피부질환이다. 아토피성 피부염의 발병 원인은 아직 확실하게 규명되지 않은 상태이나, 환경적 요인, 유전적 소인, 면역화학적 반응 및 피부보호막 이상 등이 주요 발병원인으로 여겨지고 있으며, 이의 증상도 소양증, 피부 건조증, 습진 등으로 다양하게 나타나고 있다. 또한 환자의 연령에 따라 피부 병변의 분포 및 반응 양상도 다르게 나타나고 있다.Atopic dermatitis, one of allergic diseases, is a growing trend worldwide and is a chronic and recurrent skin disease that begins mainly in infancy and childhood. The cause of atopic dermatitis is not yet clearly identified, but environmental factors, genetic predisposition, immunochemical reactions, and skin barrier abnormalities are considered as the main causes of the disease, and the symptoms of pruritus, dry skin, eczema, etc. It appears to be various. In addition, the distribution and response patterns of skin lesions also vary according to the age of the patient.
현재 아토피성 피부염의 치료를 위해서 많이 사용되고 있는 약물은 통상 국소 스테로이드, 국소 면역 조절제, 전신 스테로이드제, 전신면역억제제, 항히스타민제 등이 있으나, 이들 약물의 투여 또는 복용에 의해 각종 부작용의 발생 위험이 있다는 점에서 안전하고, 피부 건조증, 피부 보호막 이상 등의 피부 개선 효과를 나타내는 우수한 활성 성분의 개발이 요구되고 있다.Currently, drugs commonly used for the treatment of atopic dermatitis generally include topical steroids, local immunomodulators, systemic steroids, systemic immunosuppressants, and antihistamines, but there are risks of various side effects caused by the administration or administration of these drugs. In view of the above, there is a demand for development of an excellent active ingredient that is safe and exhibits skin-improving effects such as dry skin and abnormal skin protective film.
한편, DMEM/F12(Dulbecco's Modified Eagle Medium/F12)는 일반적으로 세포의 배양에 사용되는 배지로, 다양한 종류의 아미노산, 비타민, 무기염 및 기타 물질로 구성되어 있어 세포가 성장 및 분화에 있어 안정적인 환경을 조성하나, 상기 배지 또는 상기 배지의 일부 구성성분을 포함하는 조성물이 피부질환 치료 또는 피부상태 개선을 위해 사용될 수 있음은 알려진 바가 없다.On the other hand, DMEM / F12 (Dulbecco's Modified Eagle Medium / F12) is a medium used for culturing cells, and is composed of various kinds of amino acids, vitamins, inorganic salts, and other substances, so that cells are stable for growth and differentiation. However, it is not known that the medium or a composition including some components of the medium can be used for treating skin diseases or improving skin condition.
본 발명의 목적은 세포 배양배지 또는 이의 일부 구성성분을 포함하는 피부질환 치료용 약학 조성물, 및 상기 조성물을 이용하여 피부 질환을 예방 또는 치료하는 방법을 제공하는 것이다.It is an object of the present invention to provide a pharmaceutical composition for treating skin diseases comprising a cell culture medium or some components thereof, and a method for preventing or treating skin diseases using the composition.
본 발명의 다른 목적은 세포 배양배지 또는 이의 일부 구성성분을 포함하는 피부 상태 개선용 조성물, 및 상기 조성물을 이용하여 피부 상태를 개선하는 방법을 제공하는 것이다.Another object of the present invention is to provide a composition for improving skin condition comprising a cell culture medium or some components thereof, and a method for improving skin condition using the composition.
상기 목적을 달성하기 위하여, 본 발명은 세포 배양배지를 포함하는 피부 질환의 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of skin diseases, including cell culture medium.
상기 목적을 달성하기 위하여, 또한 본 발명은 본 발명의 약학 조성물을 이를 필요로 하는 대상의 피부에 도포하는 단계를 포함하는 피부 질환의 예방 또는 치료 방법을 제공한다.In order to achieve the above object, the present invention also provides a method for preventing or treating skin diseases comprising the step of applying the pharmaceutical composition of the present invention to the skin of a subject in need thereof.
상기 목적을 달성하기 위하여, 또한 본 발명은 피부 질환의 예방 또는 치료를 위한 약제의 제조에 사용하기 위한 본 발명에 따른 약학 조성물의 용도를 제공한다.In order to achieve the above object, the present invention also provides a use of the pharmaceutical composition according to the invention for use in the manufacture of a medicament for the prevention or treatment of skin diseases.
상기 다른 목적을 달성하기 위하여, 본 발명은 세포 배양배지를 포함하는 피부 상태 개선용 화장료 조성물을 제공한다.In order to achieve the above another object, the present invention provides a cosmetic composition for improving skin conditions comprising a cell culture medium.
상기 다른 목적을 달성하기 위하여, 또한 본 발명은 본 발명의 화장료 조성물을 이용하여 피부 상태를 개선하는 방법을 제공한다.In order to achieve the above another object, the present invention also provides a method for improving the skin condition using the cosmetic composition of the present invention.
상기 다른 목적을 달성하기 위하여, 또한 본 발명은 피부 상태를 개선하기 위한 화장료의 제조에 사용하기 위한 본 발명에 따른 화장료 조성물의 용도를 제공한다.In order to achieve the above another object, the present invention also provides a use of the cosmetic composition according to the invention for use in the preparation of a cosmetic for improving the skin condition.
본 발명의 세포 배양배지를 포함하는 약학 조성물은 피부의 탈과립화 및 염증반응 관련 물질의 발현을 억제하고, 피부장벽 관련 물질의 발현을 촉진하며, 손상된 피부의 회복에 효과를 나타내어, 피부질환의 예방 또는 치료나, 피부상태를 개선하는데 유용하게 사용될 수 있다.The pharmaceutical composition comprising the cell culture medium of the present invention inhibits the degranulation of the skin and the expression of inflammatory response-related substances, promotes the expression of skin barrier-related substances, exhibits an effect on the recovery of damaged skin, preventing the skin disease Or it can be usefully used to treat or improve skin condition.
본 발명의 상기 및 다른 목적과 특징들은 첨부된 도면과 함께, 하기 본 발명의 설명으로부터 명확해질 것이다:The above and other objects and features of the present invention will become apparent from the following description of the invention, taken in conjunction with the accompanying drawings:
도 1은 실시예 1의 조성물을 농도별로 처리한 후, β-헥소사미니데이즈의 농도 변화를 나타내는 그래프이다.1 is a graph showing the concentration change of β-hexosaminidase after treating the composition of Example 1 for each concentration.
도 2는 실시예 1의 조성물을 농도별로 처리한 후, 히스타민의 농도 변화를 나타내는 그래프이다.2 is a graph showing a change in the concentration of histamine after treating the composition of Example 1 for each concentration.
도 3은 실시예 1의 조성물을 농도별로 처리한 후, IL-6의 발현 변화를 mRNA 수준에서 확인한 것이다.Figure 3 shows the change in expression of IL-6 at the mRNA level after the treatment of the composition of Example 1 by concentration.
도 4는 실시예 1의 조성물을 농도별로 처리한 후, 세린 팔미토일트렌스퍼레이즈(SPT)의 발현 변화를 mRNA 수준에서 확인한 것이다.Figure 4 shows the change in expression of serine palmitoyltransferase (SPT) at the mRNA level after treating the composition of Example 1 by concentration.
도 5는 실시예 1의 조성물을 농도별로 처리한 후, 필라그린(filaggrin)의 발현 변화를 mRNA 수준에서 확인한 것이다.Figure 5 shows the change in the expression level of the filaggrin (filaggrin) after the treatment of the composition of Example 1 by concentration.
도 6은 실시예 2의 조성물 및 비교예 1의 추출물을 농도별로 처리한 후, 라우릴황산나트륨(sodium lauryl sulfate, SLS)으로 손상된 세포의 증식률을 확인한 그래프이다.6 is a graph confirming the proliferation rate of cells damaged by sodium lauryl sulfate (SLS) after treating the composition of Example 2 and the extract of Comparative Example 1 by concentration.
도 7은 실시예 2의 조성물 및 비교예 1의 추출물을 처리한 후, UV에 손상된 세포의 증식률을 확인한 그래프이다.7 is a graph confirming the proliferation rate of cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
도 8은 실시예 2의 조성물 및 비교예 1의 추출물을 처리한 후, UV에 손상된 인공피부의 회복을 나타낸 것이다.Figure 8 shows the recovery of artificial skin damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
도 9는 실시예 2의 조성물 및 비교예 1의 추출물을 처리한 후, UV에 손상된 세포의 IL-1α의 발현량을 확인한 그래프이다.9 is a graph confirming the expression level of IL-1α in cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
도 10은 실시예 2의 조성물 및 비교예 1의 추출물을 처리한 후, UV에 손상된 세포의 TNF-α의 발현량을 확인한 그래프이다.10 is a graph confirming the expression level of TNF-α of cells damaged by UV after treating the composition of Example 2 and the extract of Comparative Example 1.
이하, 본 발명을 보다 상세히 설명한다.Hereinafter, the present invention will be described in more detail.
본 발명은 세포 배양배지를 포함하는 피부 질환의 예방 또는 치료용 약학 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating skin diseases, including cell culture medium.
본 발명에 사용한 용어 “세포 배양배지”또는 “배양배지”는 시험관 내에서(in vitro) 세포 성장 및 생존을 위해 세포가 필요로 하는 성분을 함유하거나, 세포 성장 및 생존을 돕는 성분을 함유한다. 구체적으로, 상기 성분은 비타민, 필수 또는 비필수 아미노산, 및 미량 원소일 수 있다. 상기 배지는 세포, 바람직하게는 진핵세포, 더 바람직하게는 포유동물 세포의 배양에 사용되는 배지일 수 있다.As used herein, the term "cell culture medium" or "culture medium" contains a component required by the cell for cell growth and survival in vitro , or contains a component that aids cell growth and survival. In particular, the component may be a vitamin, an essential or non-essential amino acid, and a trace element. The medium may be a medium used for culturing cells, preferably eukaryotic cells, more preferably mammalian cells.
세포 배양배지의 예로는 DMEM/F12, DMEM, MEMα 등이 알려져 있으나, 이에 한정되지 않는다.Examples of cell culture media include, but are not limited to, DMEM / F12, DMEM, MEMα, and the like.
본 발명에 따른 세포 배양배지는 무혈청 배지(serum-free)이다. 무혈청 배지는 인간을 포함한 동물로부터 유래된 혈청(예를 들어, 동물 유래 혈청)을 함유하지 않는 배양 배지를 의미한다. 동물 유래 혈청은 배양하고자 하는 세포의 성장을 위한 범용적인 영양분을 제공하나, 미확인된 미량 성분을 포함하므로 분석이 어렵고, 재현성 있는 시험 및 생산공정을 확립하는데 어려움이 있으며, 인체에 대한 안정성을 보장할 수 없는 문제점이 있다. 그러나 본 발명에 따른 세포 배양배지는 무혈청 배지로 인체에 대한 안정성이 보장되고, 재현성 있는 시험 및 생산공정의 확립이 가능하다.Cell culture medium according to the present invention is a serum-free medium (serum-free). Serum-free medium refers to a culture medium that does not contain serum (eg, animal-derived serum) derived from animals, including humans. Animal-derived serum provides universal nutrients for the growth of cells to be cultured, but contains unidentified trace components, making it difficult to analyze, establishing reproducible testing and production processes, and ensuring stability to the human body. There is no problem. However, the cell culture medium according to the present invention is a serum-free medium to ensure the stability to the human body, it is possible to establish a reproducible test and production process.
상기 세포 배양배지는 아미노산 성분, 비타민 성분, 무기염 성분 및 기타 성분으로 구성되며,The cell culture medium is composed of amino acid components, vitamin components, inorganic salt components and other components,
a) 상기 아미노산 성분은 글리세린, L-알라닌, L-아르기닌 하이드로클로라이드, L-시스테인 하이드로클로라이드-모노하이드레이트, L-글루타민, L-히스티딘 하이드로클로라이드-모노하이드레이트, L-리신 하이드로클로라이드, L-메티오닌, L-프롤린, L-세린, L-트레오닌 및 L-발린으로 구성된 군으로부터 선택되는 적어도 하나의 아미노산 또는 이의 조합이고,a) the amino acid component is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L-lysine hydrochloride, L-methionine, At least one amino acid selected from the group consisting of L-proline, L-serine, L-threonine and L-valine, or a combination thereof,
b) 상기 비타민 성분은 i-이노시톨, 티아민 하이드로클로라이드, 나이아신아미드 및 피리독신 하이드로클로라이드로 구성된 군으로부터 선택되는 적어도 하나의 아미노산 또는 이의 조합이며,b) said vitamin component is at least one amino acid selected from the group consisting of i-inositol, thiamine hydrochloride, niacinamide and pyridoxine hydrochloride or combinations thereof,
c) 상기 무기염 성분은 염화나트륨(NaCl), 탄산수소나트륨(NaHCO3), 염화칼륨(KCl), 염화칼슘(CaCl2)(무수) 및 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O)로 구성된 군으로부터 선택되는 적어도 하나의 아미노산 또는 이의 조합이고,c) The inorganic salt component is sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), potassium chloride (KCl), calcium chloride (CaCl 2 ) (anhydrous) and sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O) At least one amino acid selected from the group consisting of or a combination thereof,
d) 기타 성분은 D-글루코즈(덱스트로즈) 또는 소듐 피루베이트인 것이 바람직하다.d) The other component is preferably D-glucose (dextrose) or sodium pyruvate.
본 발명의 일구체예에 의하면, 상기 세포 배양배지는 글리세린, L-알라닌, L-아르기닌 하이드로클로라이드, L-시스테인 하이드로클로라이드-모노하이드레이트, L-글루타민, L-히스티딘 하이드로클로라이드-모노하이드레이트, L-리신 하이드로클로라이드, L-메티오닌, L-프롤린, L-세린, L-트레오닌, L-발린, i-이노시톨, 티아민 하이드로클로라이드, 나이아신아미드, 피리독신 하이드로클로라이드, 염화나트륨(NaCl), 탄산수소나트륨(NaHCO3), 염화칼륨(KCl), 염화칼슘(CaCl2)(무수), 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O), D-글루코즈(덱스트로즈) 및 소듐 피루베이트으로 구성된 것이 더욱 바람직하다.According to one embodiment of the present invention, the cell culture medium is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L- Lysine hydrochloride, L-methionine, L-proline, L-serine, L-threonine, L-valine, i-inositol, thiamine hydrochloride, niacinamide, pyridoxine hydrochloride, sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), Potassium chloride (KCl), calcium chloride (CaCl 2 ) (anhydrous), sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O), D-glucose (dextrose) and sodium pyruvate .
본 발명의 세포 배양배지는 상기 약학 조성물의 총 중량을 기준으로 0.5 내지 100 중량%로 포함될 수 있고, 바람직하게는 1.0 내지 10 중량%로 포함할 수 있다. 바람직하게는 5.0 내지 8 중량%로 포함할 수 있다. 상기 조성물을 0.5 중량% 미만으로 포함하는 경우 그 효과가 미비한 문제점이 발생할 수 있다.Cell culture medium of the present invention may be included in 0.5 to 100% by weight, preferably 1.0 to 10% by weight based on the total weight of the pharmaceutical composition. Preferably from 5.0 to 8% by weight. If the composition is included in less than 0.5% by weight may have a problem that the effect is insignificant.
상기 조성물에 있어서, 아미노산 성분은 단백질 합성의 원료로 세포의 성장에 이용되며, 이는 조성물 총 중량을 기준으로 0.1 내지 0.2 중량%의 글리세린, 0.01 내지 0.1 중량%의 L-알라닌, 0.5 내지 3 중량%의 L-아르기닌 하이드로클로라이드, 0.1 내지 0.2 중량%의 L-시스테인 하이드로클로라이드-모노하이드레이트, 2 내지 3 중량%의 L-글루타민, 0.1 내지 0.5 중량%의 L-히스티딘 하이드로클로라이드-모노하이드레이트, 0.4 내지 1.0 중량%의 L-리신 하이드로클로라이드, 0.1 내지 0.5 중량%의 L-메티오닌, 0.1 내지 0.5 중량%의 L-프롤린, 0.1 내지 0.3 중량%의 L-세린, 0.3 내지 0.6 중량%의 L-트레오닌 및 0.3 내지 0.6 중량%의 L-발린을 포함할 수 있다.In the composition, the amino acid component is used for the growth of cells as a raw material for protein synthesis, which is 0.1 to 0.2% by weight glycerin, 0.01 to 0.1% by weight L-alanine, 0.5 to 3% by weight based on the total weight of the composition L-arginine hydrochloride, 0.1-0.2 wt% L-cysteine hydrochloride-monohydrate, 2-3 wt% L-glutamine, 0.1-0.5 wt% L-histidine hydrochloride-monohydrate, 0.4-1.0 Wt-% L-lysine hydrochloride, 0.1-0.5 wt-% L-methionine, 0.1-0.5 wt-% L-proline, 0.1-0.3 wt-% L-serine, 0.3-0.6 wt-% L-threonine and 0.3 To 0.6% by weight of L-valine.
본 발명에 따른 조성물에 포함되는 아미노산 성분 및 함량에 대한 일구체예는 0.12 내지 0.14 중량%의 글리세린, 0.02 내지 0.04 중량%의 L-알라닌, 1.01 내지 1.06 중량%의 L-아르기닌 하이드로클로라이드, 0.11 내지 0.15 중량%의 L-시스테인 하이드로클로라이드-모노하이드레이트, 2.3 내지 2.8 중량%의 L-글루타민, 0.20 내지 0.25 중량%의 L-히스티딘 하이드로클로라이드-모노하이드레이트, 0.5 내지 0.8 중량%의 L-리신 하이드로클로라이드, 0.1 내지 0.3 중량%의 L-메티오닌, 0.1 내지 0.3 중량%의 L-프롤린, 0.16 내지 0.2 중량%의 L-세린, 0.35 내지 0.40 중량%의 L-트레오닌 및 0.35 내지 0.40 중량%의 L-발린일 수 있다. 상기 범위 내의 함량으로 아미노산 성분을 포함함으로써 세포의 성장 및 유지에 도움을 주고 제형 안정성을 향상시킬 수 있다.One embodiment of the amino acid component and content included in the composition according to the present invention is 0.12 to 0.14 wt% glycerin, 0.02 to 0.04 wt% L-alanine, 1.01 to 1.06 wt% L-arginine hydrochloride, 0.11 to 0.15 wt% L-cysteine hydrochloride-monohydrate, 2.3-2.8 wt% L-glutamine, 0.20-0.25 wt% L-histidine hydrochloride-monohydrate, 0.5-0.8 wt% L-lysine hydrochloride, 0.1-0.3 wt% L-methionine, 0.1-0.3 wt% L-proline, 0.16-0.2 wt% L-serine, 0.35-0.40 wt% L-threonine and 0.35-0.40 wt% L-valinyl Can be. By including the amino acid component in the content within the above range can help the growth and maintenance of cells and improve the formulation stability.
상기 비타민 성분은 세포의 활성을 유지하는 역할을 하며, 이는 조성물 총 중량을 기준으로 0.01 내지 0.1 중량%의 i-이노시톨, 0.01 내지 0.1 중량%의 티아민 하이드로클로라이드, 0.005 내지 0.03 중량%의 나이아신아미드 및 0.01 내지 0.1 중량%의 피리독신 하이드로클로라이드를 포함할 수 있다.The vitamin component serves to maintain the activity of the cells, which is 0.01 to 0.1% by weight of i-inositol, 0.01 to 0.1% by weight of thiamin hydrochloride, 0.005 to 0.03% by weight of niacinamide and 0.01 to 0.1% by weight of pyridoxine hydrochloride.
본 발명에 따른 조성물에 포함되는 비타민 성분 및 함량에 대한 일구체예는 0.08 내지 0.09 중량%의 i-이노시톨, 0.01 내지 0.02 중량%의 티아민 하이드로클로라이드, 0.01 내지 0.02 중량%의 나이아신아미드 및 0.01 내지 0.02 중량%의 피리독신 하이드로클로라이드일 수 있다. 상기 범위 내의 함량으로 비타민 성분을 포함함으로써 세포 활성 유지에 도움을 줄 수 있다.One embodiment of the vitamin component and content included in the composition according to the present invention is 0.08 to 0.09 wt% i-inositol, 0.01 to 0.02 wt% thiamine hydrochloride, 0.01 to 0.02 wt% niacinamide and 0.01 to 0.02 Wt% pyridoxine hydrochloride. By including a vitamin component in a content within the above range can help maintain cell activity.
상기 무기염 성분은 세포 기능의 발현을 조절하는 역할을 하며, 이는 조성물 총 중량을 기준으로 40 내지 55 중량%의 염화나트륨(NaCl), 10 내지 20 중량%의 탄산수소나트륨(NaHCO3), 1 내지 3 중량%의 염화칼륨(KCl), 0.5 내지 1.0 중량%의 염화칼슘(CaCl2)(무수) 및 0.2 내지 0.7 중량%의 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O)를 포함할 수 있다.The inorganic salt component serves to regulate the expression of cell function, which is 40 to 55% by weight sodium chloride (NaCl), 10 to 20% by weight sodium hydrogencarbonate (NaHCO 3 ), 1 to 1 based on the total weight of the composition 3 weight percent potassium chloride (KCl), 0.5 to 1.0 weight percent calcium chloride (CaCl 2 ) (anhydrous) and 0.2 to 0.7 weight percent sodium hydrogen phosphate monohydrate (NaH 2 PO 4 —H 2 O). .
본 발명에 따른 조성물에 포함되는 무기염 성분 및 함량에 대한 일구체예는 45 내지 50 중량%의 염화나트륨(NaCl), 15 내지 18 중량%의 탄산수소나트륨(NaHCO3), 2.0 내지 2.5 중량%의 염화칼륨(KCl), 0.8 내지 0.9 중량%의 염화칼슘(CaCl2)(무수) 및 0.4 내지 0.5 중량%의 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O)일 수 있다. 상기 범위 내의 함량으로 무기염 성분을 포함함으로써 세포 기능 조절에 도움이 되고 의약품이나 화장품으로 제형화시키는 데 어려움이 없다.One embodiment of the inorganic salt component and content included in the composition according to the present invention is 45 to 50% by weight of sodium chloride (NaCl), 15 to 18% by weight of sodium bicarbonate (NaHCO 3 ), 2.0 to 2.5% by weight of Potassium chloride (KCl), 0.8-0.9 wt% calcium chloride (CaCl 2 ) (anhydrous) and 0.4-0.5 wt% sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O). By including an inorganic salt component in the content within the above range is helpful in regulating cell function and is difficult to formulate into pharmaceuticals or cosmetics.
상기 기타 성분은 에너지원으로 역할을 하는 탄소원 또는 배지의 pH를 유지하는 성분 등을 포함하는 것으로서, 이는 조성물 총 중량을 기준으로 15 내지 30 중량%의 D-글루코즈(덱스트로즈) 또는 0.3 내지 0.5 중량%의 소듐 피루베이트를 포함할 수 있다.The other components include a carbon source serving as an energy source or a component for maintaining the pH of the medium, which is 15 to 30% by weight of D-glucose (dextrose) or 0.3 to 0.5 based on the total weight of the composition. Weight percent sodium pyruvate.
본 발명에 따른 조성물에 포함되는 기타 성분 및 함량에 대한 일구체예는 20 내지 25 중량%의 D-글루코즈(덱스트로즈) 또는 0.35 내지 0.4 중량%의 소듐 피루베이트일 수 있다. 상기 범위 내의 함량으로 포함되는 기타 성분은, 예를 들어 탄소원은 에너지를 생성하여 세포 성장에 도움을 주고 제형 안정성을 향상시키는 등의 도움을 줄 수 있다.One embodiment for the other components and contents included in the composition according to the present invention may be 20 to 25% by weight of D-glucose (dextrose) or 0.35 to 0.4% by weight of sodium pyruvate. Other components included in the content within the above range, for example, the carbon source may generate energy to help cell growth, improve formulation stability, and the like.
상기 피부 질환은 아토피성 피부염, 알레르기, 피부 습진, 여드름, 건선, 가려움증 및 이의 조합으로 이루어진 군에서 선택되는 것일 수 있다.The skin disease may be selected from the group consisting of atopic dermatitis, allergy, skin eczema, acne, psoriasis, itching and combinations thereof.
본 발명의 약학 조성물은 아미노산 성분, 비타민 성분, 무기염 성분, 기타 성분 및 이의 조합으로 이루어진 군에서 선택되는 성분을 더 포함할 수 있다.The pharmaceutical composition of the present invention may further include a component selected from the group consisting of an amino acid component, a vitamin component, an inorganic salt component, other components, and combinations thereof.
상기 아미노산 성분은 일반적인 동물 세포 배양 배지에 사용되는 아미노산 성분이면 어떤 아미노산 성분이라도 사용할 수 있으며, 예를 들어, L-아스파라긴-모노하이드레이트, L-아스파르트산, L-시스틴 2HCl, L-글루탐산, L-이소류신, L-류신, L-페닐알라닌, L-트립토판, L-티로신 디소듐염 디하이드레이트 및 이의 조합으로 이루어진 군에서 선택될 수 있다.The amino acid component may be any amino acid component as long as it is an amino acid component used in a general animal cell culture medium. For example, L-asparagine-monohydrate, L-aspartic acid, L-cystine 2HCl, L-glutamic acid, L- Isoleucine, L-leucine, L-phenylalanine, L-tryptophan, L-tyrosine disodium salt dihydrate, and combinations thereof.
상기 비타민 성분은 일반적인 동물 세포 배양 배지에 사용되는 비타민 성분이면 어떤 비타민 성분이라도 사용할 수 있으며, 예를 들어, 바이오틴, D-판토텐산칼슘, 엽산, 리보플라빈, 비타민 B12, 및 이의 조합으로 이루어진 군에서 선택될 수 있다.The vitamin component may be any vitamin component as long as it is a vitamin component used in a general animal cell culture medium. For example, the vitamin component may be selected from the group consisting of biotin, calcium D-pantothenate, folic acid, riboflavin, vitamin B12, and combinations thereof. Can be.
상기 무기염 성분은 일반적인 동물 세포 배양 배지에 사용되는 무기염 성분이면 어떤 무기염 성분이라도 사용할 수 있으며, 예를 들어, 황산동 펜타하이드레이트(CuSO4-5H2O), 황산제이철 헵타하이드레이트(FeSO4-7H2O), 염화마그네슘(무수), 황산마그네슘(MgSO4)(무수), 인산수소이나트륨(Na2HPO4) 무수, 황산아연 헵타하이드레이트(ZnSO4-7H2O) 및 이의 조합으로 이루어진 군에서 선택될 수 있다The inorganic salt component may be any inorganic salt component as long as it is an inorganic salt component used in a general animal cell culture medium. For example, copper sulfate pentahydrate (CuSO 4 -5H 2 O), ferric sulfate heptahydrate (FeSO 4- ) 7H 2 O), magnesium chloride (anhydrous), magnesium sulfate (MgSO 4 ) (anhydrous), disodium hydrogen phosphate (Na 2 HPO 4 ) anhydrous, zinc sulfate heptahydrate (ZnSO 4 -7H 2 O) and combinations thereof Can be chosen from
상기 기타 성분은 일반적인 동물 세포 배양 배지에 사용되는 성분이면 어떤 성분이라도 사용할 수 있으며, 예를 들어, 히포크산틴 Na, 리놀렌산, 리포산, 푸트레신 2HCl, 티미딘 및 이의 조합으로 이루어진 군에서 선택될 수 있다.The other component may be any component as long as it is a component used in a general animal cell culture medium, and may be selected from the group consisting of, for example, hypoxanthine Na, linolenic acid, lipoic acid, putrescine 2HCl, thymidine, and combinations thereof. Can be.
본 발명의 약학 조성물은 약학적으로 허용가능한 담체를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier.
상기 약학적으로 허용되는 담체는 약품 제조 시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 활석, 스테아르산 마그네슘, 미네랄 오일 등을 포함할 수 있으나, 이에 한정되는 것은 아니다. The pharmaceutically acceptable carrier is commonly used in the manufacture of a drug, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose , Polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, and the like, but is not limited thereto.
또한, 본 발명의 약학 조성물은 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제, 및 이의 조합으로 이루어진 군에서 선택되는 약학적으로 허용가능한 첨가제를 추가로 포함할 수 있다.In addition, the pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive selected from the group consisting of lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, and combinations thereof.
상기 담체는 본 발명의 약학 조성물 총 중량을 기준으로 약 1 중량% 내지 약 99.99 중량%, 바람직하게는 약 90 중량% 내지 약 99.99 중량%로 포함될 수 있으며, 상기 약학적으로 허용가능한 첨가제는 약 0.1 중량% 내지 약 20 중량%로 포함될 수 있다. The carrier may comprise from about 1% to about 99.99% by weight, preferably from about 90% to about 99.99% by weight, based on the total weight of the pharmaceutical composition of the present invention, wherein the pharmaceutically acceptable additive is about 0.1 Weight percent to about 20 weight percent.
본 발명의 약학 조성물은 비경구로 투여될 수 있고, 바람직하게는 국소 투여 방식으로 피부에 직접 투여될 수 있다.The pharmaceutical compositions of the present invention may be administered parenterally, and may preferably be administered directly to the skin in a topical manner.
본 발명의 약학 조성물의 제형은 경피 투여 주사제, 연고제, 액제, 크림제, 결제, 스프레이제, 패취제 등과 같은 피부 외용제 형태일 수 있다.Formulations of the pharmaceutical compositions of the present invention may be in the form of external skin preparations such as transdermal injections, ointments, solutions, creams, payments, sprays, patches, and the like.
본 발명의 약학 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 여러 관련 인자에 비추어 결정되는 것이므로 상기 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 이해되어서는 아니 된다.Suitable dosages of the pharmaceutical compositions of the present invention are determined in view of various related factors such as formulation method, mode of administration, patient's age, weight, sex, morbidity, food, time of administration, route of administration, rate of excretion and reaction sensitivity. The dosage should not be understood as limiting the scope of the invention in any aspect.
따라서, 본 발명은 상기 약학 조성물을 이를 필요로 하는 대상의 피부에 도포하는 단계를 포함하는 피부 질환의 예방 또는 치료 방법을 제공한다.Accordingly, the present invention provides a method for preventing or treating a skin disease comprising applying the pharmaceutical composition to the skin of a subject in need thereof.
또한, 본 발명은 피부 질환의 예방 또는 치료를 위한 약제의 제조에 사용하기 위한 상기 약학 조성물의 용도를 제공한다.The present invention also provides the use of the pharmaceutical composition for use in the manufacture of a medicament for the prevention or treatment of skin diseases.
상기 약학 조성물이 도포될 수 있는 대상은 포유동물일 수 있고, 구체적으로는 인간일 수 있다.The subject to which the pharmaceutical composition may be applied may be a mammal, and specifically, a human.
본 발명은 세포 배양배지를 포함하는 피부 상태 개선용 화장료 조성물을 제공한다.The present invention provides a cosmetic composition for improving skin condition comprising a cell culture medium.
본 발명의 화장료 조성물은 앞서 전술한 바와 같은 약학 조성물과 유효성분이 동일하다.The cosmetic composition of the present invention has the same active ingredient as the above-mentioned pharmaceutical composition.
본 발명에 따른 화장료 조성물은 주름의 발생 억제, 피부 노화 억제, 피부 탄력 개선, 피부 재생, 상처 또는 창상 회복(wound healing), 각막 재생, 피부자극완화 및 이의 조합으로 이루어진 군에서 선택되는, 피부 세포의 기능 저하 또는 손실로부터 피부를 보호 또는 피부 상태를 개선하거나, 또는 피부 질환을 예방 또는 개선하는데 사용될 수 있다. 상기 피부 질환은 아토피성 피부염, 알레르기, 피부 습진, 여드름, 건선, 가려움증 및 이의 조합으로 이루어진 군에서 선택되는 것일 수 있다.The cosmetic composition according to the present invention is selected from the group consisting of inhibiting the occurrence of wrinkles, inhibiting skin aging, improving skin elasticity, skin regeneration, wound or wound healing, corneal regeneration, skin irritation and combinations thereof, skin cells It can be used to protect the skin from a decrease or loss of function or to improve the skin condition, or to prevent or improve skin diseases. The skin disease may be selected from the group consisting of atopic dermatitis, allergy, skin eczema, acne, psoriasis, itching and combinations thereof.
본 발명의 화장료 조성물은 피부 상태의 개선을 목적으로 피부에 직접 적용될 수 있다.The cosmetic composition of the present invention can be applied directly to the skin for the purpose of improving the skin condition.
상기 화장료 조성물은 당업계에서 통상적으로 제조되는 화장료 제형으로 제제화될 수 있다. 상기 화장료 조성물은 예를 들어, 용액, 현탁액, 유탁액, 페이스트, 겔, 크림, 로션, 파우더, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션 및 스프레이 등으로 제형화될 수 있으나, 본 발명이 이에 한정되는 것은 아니다. 보다 상세하게는, 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징폼, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형으로 제제화될 수 있다.The cosmetic composition may be formulated into a cosmetic formulation commonly prepared in the art. The cosmetic compositions are formulated, for example, in solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays, and the like. However, the present invention is not limited thereto. More specifically, it may be formulated in the form of a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.
본 발명의 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라가칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화아연 및 이의 혼합물로 이루어진 군에서 선택되는 담체 성분을 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide and mixtures thereof It may include a carrier component selected from the group consisting of.
본 발명의 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 락토스, 탈크, 실리카, 알루미늄 하이드록시드, 칼슘 실리케이트, 폴리아미드 파우더 및 이의 혼합물로 이루어진 군에서 선택되는 담체 성분을 포함할 수 있으며, 특히 스프레이인 경우에는 추가적으로 클로로플루오로하이드로카본, 프로판/부탄 또는 디메틸 에테르 등을 더 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a powder or a spray, it may include a carrier component selected from the group consisting of lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder and mixtures thereof, in particular spray In the case of chlorofluorohydrocarbon, it may further include propane / butane or dimethyl ether.
본 발명의 화장료 조성물의 제형이 용액 또는 유탁액인 용매, 용매화제, 유탁화제 및 이의 혼합물로 이루어진 군에서 선택되는 담체 성분을 포함할 수 있다. 이의 예로는, 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄 지방산 에스테르, 및 이의 혼합물 등을 들 수 있다.The formulation of the cosmetic composition of the present invention may comprise a carrier component selected from the group consisting of solvents, solvating agents, emulsifying agents and mixtures thereof which are solutions or emulsions. Examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic esters, polyethylene glycols, sorbitan fatty acid esters, mixtures thereof, and the like. Can be mentioned.
본 발명의 화장료 조성물의 제형이 현탁액인 경우에는 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미세결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가, 트라가칸트 및 이의 혼합물로 이루어진 군에서 선택되는 담체 성분을 포함할 수 있다.When the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystal Carrier cellulose, aluminum metahydroxy, bentonite, agar, tragacanth and mixtures thereof.
본 발명의 화장료 조성물의 제형이 계면-활성제 함유 클렌징인 경우에는 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성유, 라놀린 유도체, 에톡실화 글리세롤 지방산 에스테르 및 이의 혼합물로 이루어진 군에서 선택되는 담체 성분을 포함할 수 있다. When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing, an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, an isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide Carrier sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, ethoxylated glycerol fatty acid esters, and mixtures thereof.
본 발명의 화장료 조성물에서 "담체 성분"이란 화장품 제제에 포함될 수 있는 이미 공지되어 사용되고 있는 화합물 또는 조성물로서, 피부에 접촉 시 인체가 적응가능한 이상의 독성, 불안정성 또는 자극성이 없는 성분이다.The "carrier component" in the cosmetic composition of the present invention is a compound or composition already known and used that may be included in cosmetic preparations, and is a component that is not toxic, unstable or irritant as long as the human body is suitable for contact with the skin.
본 발명의 화장료 조성물은 담체 성분 이외에도, 항산화제, 안정화제, 용해화제, 보습제, 안료, 향료, 자외선 차단제, 발색제, 계면 활성제 및 이의 조합으로 이루어진 군에서 선택되는 보조제를 추가로 포함할 수 있다. 상기 보조제는 화장료 조성물의 제조에 통상적으로 사용되는 보조제라면 사용에 제한이 없다.In addition to the carrier component, the cosmetic composition of the present invention may further include an adjuvant selected from the group consisting of antioxidants, stabilizers, solubilizers, humectants, pigments, perfumes, sunscreens, colorants, surfactants, and combinations thereof. The adjuvant is not limited to use as long as the adjuvant commonly used in the preparation of the cosmetic composition.
따라서, 본 발명은 상기 화장료 조성물을 이용하여 피부 상태를 개선하는 방법을 제공한다.Therefore, the present invention provides a method for improving the skin condition using the cosmetic composition.
또한, 본 발명은 피부 상태를 개선시키기 위한 화장료의 제조에 사용하기 위한 상기 화장료 조성물의 용도를 제공한다.The present invention also provides the use of said cosmetic composition for use in the preparation of a cosmetic for improving skin condition.
상기 피부 상태를 개선하는 방법은 이를 필요로 하는 대상의 피부에 도포하는 단계를 포함할 수 있다. 상기 대상은 포유동물일 수 있고, 구체적으로는 인간일 수 있다.The method for improving the skin condition may include applying to the skin of the subject in need thereof. The subject may be a mammal, specifically a human.
피부에 도포하는 단계는 본 발명에 따른 화장료 조성물을 그 형태에 따라 피부에 직접 도포하거나, 분무하는 것을 포함할 수 있다. 이때, 상기 화장료 조성물의 도포량 및 하루 사용 횟수는, 사용자의 연령, 성별, 용도, 증상의 정도 등에 따라 적절하게 설정될 수 있고, 예를 들면, 상기 화장료 조성물의 적당량을 하루 1 내지 6회의 빈도로 피부에 도포할 수 있다.Applying to the skin may include applying or spraying the cosmetic composition according to the invention directly on the skin according to its form. At this time, the application amount and the number of times of use of the cosmetic composition may be appropriately set according to the age, sex, use, degree of symptoms of the user, for example, the appropriate amount of the cosmetic composition at a frequency of 1 to 6 times a day. It can be applied to the skin.
이하, 본 발명을 하기 실시예에 의거하여 좀 더 상세하게 설명하고자 한다. 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명의 범위가 이들만으로 제한되는 것은 아니다.Hereinafter, the present invention will be described in more detail based on the following examples. The following examples are only for illustrating the present invention, but the scope of the present invention is not limited thereto.
실시예Example 1 One
용매로서 초순수정제수를 사용하여 하기 표 1의 조성을 갖는 무혈청 DMEM/F12 배지(Gibco, USA)의 농도를 각각 2%, 10%, 50% 또는 100%로 하여 조성물을 제조하였다.The composition was prepared using ultrapure purified water as the solvent, and the concentration of serum-free DMEM / F12 medium (Gibco, USA) having the composition of Table 1 was 2%, 10%, 50%, or 100%, respectively.
무혈청 DMEM/F12의 조성Composition of serum free DMEM / F12
성분ingredient 농도(g/ℓ)Concentration (g / ℓ)
글리세린glycerin 0.018750.01875
L-알라닌L-alanine 0.004450.00445
L-아르기닌 하이드로클로라이드L-arginine hydrochloride 0.14750.1475
L-아스파라긴-모노하이드레이트L-asparagine-monohydrate 0.00750.0075
L-아스파르트산L-aspartic acid 0.006650.00665
L-시스테인 하이드로클로라이드-모노하이드레이트L-Cysteine Hydrochloride-Monohydrate 0.017560.01756
L-시스틴 2HClL-cystine 2HCl 0.031290.03129
L-글루탐산L-glutamic acid 0.007350.00735
L-글루타민L-Glutamine 0.3650.365
L-히스티딘 하이드로클로라이드-모노하이드레이트L-Histidine Hydrochloride-Monohydrate 0.031480.03148
L-이소류신L-isoleucine 0.054470.05447
L-류신L-leucine 0.059050.05905
L-리신 하이드로클로라이드L-Lysine Hydrochloride 0.091250.09125
L-메티오닌L-methionine 0.017240.01724
L-페닐알라닌L-phenylalanine 0.035480.03548
L-프롤린L-proline 0.017250.01725
L-세린L-serine 0.026250.02625
L-트레오닌L-threonine 0.053450.05345
L-트립토판L-Tryptophan 0.009020.00902
L-티로신 디소듐염 디하이드레이트L-tyrosine disodium salt dihydrate 0.055790.05579
L-발린L-valine 0.052850.05285
바이오틴Biotin 0.0000030.000003
D-판토텐산칼슘D-Calcium Pantothenate 0.002240.00224
엽산Folic acid 0.002650.00265
나이아신아미드Niacinamide 0.002020.00202
피리독신 하이드로클로라이드Pyridoxine Hydrochloride 0.002130.00213
리보플라빈Riboflavin 0.002190.00219
티아민 하이드로클로라이드Thiamine Hydrochloride 0.002170.00217
비타민 B12Vitamin B12 0.000680.00068
i-이노시톨i-inositol 0.01260.0126
염화칼슘(CaCl2)(무수)Calcium chloride (CaCl 2 ) (anhydrous) 0.11660.1166
황산동 펜타하이드레이트(CuSO4-5H2O)Copper sulfate pentahydrate (CuSO 4 -5H 2 O) 0.0000010.000001
황산제이철 헵타하이드레이트(FeSO4-7H2O)Ferric Sulfate Heptahydrate (FeSO 4 -7H 2 O) 0.0004170.000417
염화마그네슘(무수)Magnesium chloride (anhydrous) 0.028640.02864
황산마그네슘(MgSO4)(무수)Magnesium Sulfate (MgSO 4 ) (anhydrous) 0.048840.04884
염화칼륨(KCl)Potassium Chloride (KCl) 0.31180.3118
탄산수소나트륨(NaHCO3)Sodium bicarbonate (NaHCO 3 ) 2.4382.438
염화나트륨(NaCl)Sodium Chloride (NaCl) 6.99556.9955
인산수소이나트륨(Na2HPO4) 무수Disodium hydrogen phosphate (Na 2 HPO 4 ) anhydrous 0.071020.07102
인산수소나트륨 모노하이드레이트 (NaH2PO4-H2O)Sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O) 0.06250.0625
황산아연 헵타하이드레이트(ZnSO4-7H2O)Zinc Sulfate Heptahydrate (ZnSO 4 -7H 2 O) 0.0004320.000432
D-글루코즈(덱스트로즈)D-glucose (dextrose) 3.1513.151
히포크산틴 NaHippoxanthin Na 0.002390.00239
리놀렌산Linolenic acid 0.0000420.000042
리포산Lipoic acid 0.0001050.000105
푸트레신 2HClPutrescine 2HCl 0.0810.081
소듐 피루베이트Sodium pyruvate 0.0550.055
티미딘Thymidine 0.0003650.000365
실시예Example 2 2
하기 표 2에 기재된 각 성분을 용매로서 초순수정제수에 충분히 용해시켜 모든 성분들이 녹은 것을 확인한 뒤, 0.22 ㎛ 여과기로 여과하였다. 그 후, 상기 여과물을 세포 배양 배지 내의 최종 농도가 0.5%, 1%, 2%, 5% 또는 8%가 되도록 초순수정제수로 희석하여 첨가하였다. 하기 기재된 각 구성성분은 대한민국 식약처에서 고시한 화장품 원료집에 등재되어 있는 물질로서, 모두 시그마-알드리치(Sigma-Aldrich, USA)에서 구입하였다.Each of the components shown in Table 2 below were dissolved in ultrapure water as a solvent to confirm that all the components were dissolved, and then filtered through a 0.22 μm filter. The filtrate was then added diluted with ultrapure purified water such that the final concentration in the cell culture medium was 0.5%, 1%, 2%, 5% or 8%. Each component described below is a material listed in the cosmetic raw materials published by the Korea Food and Drug Administration, all were purchased from Sigma-Aldrich, USA.
혼합 조성물의 조성Composition of the Mixed Composition
성분ingredient 농도 (g/ℓ)Concentration (g / ℓ)
글리세린glycerin 0.018750.01875
L-알라닌L-alanine 0.004450.00445
L-아르기닌 하이드로클로라이드L-arginine hydrochloride 0.14750.1475
L-시스테인 하이드로클로라이드-모노하이드레이트L-Cysteine Hydrochloride-Monohydrate 0.017560.01756
L-글루타민L-Glutamine 0.3650.365
L-히스티딘 하이드로클로라이드-모노하이드레이트L-Histidine Hydrochloride-Monohydrate 0.031480.03148
L-리신 하이드로클로라이드L-Lysine Hydrochloride 0.091250.09125
L-메티오닌L-methionine 0.017240.01724
L-프롤린L-proline 0.017250.01725
L-세린L-serine 0.026250.02625
L-트레오닌L-threonine 0.053450.05345
L-발린L-valine 0.052850.05285
i-이노시톨i-inositol 0.01260.0126
티아민 하이드로클로라이드Thiamine Hydrochloride 0.002170.00217
나이아신아미드Niacinamide 0.002020.00202
피리독신 하이드로클로라이드Pyridoxine Hydrochloride 0.002130.00213
염화나트륨(NaCl)Sodium Chloride (NaCl) 6.99556.9955
탄산수소나트륨(NaHCO3)Sodium bicarbonate (NaHCO 3 ) 2.4382.438
염화칼륨(KCl)Potassium Chloride (KCl) 0.31180.3118
염화칼슘(CaCl2)(무수)Calcium chloride (CaCl 2 ) (anhydrous) 0.11660.1166
인산수소나트륨 모노하이드레이트 (NaH2PO4-H2O)Sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O) 0.06250.0625
D-글루코즈(덱스트로즈)D-glucose (dextrose) 3.1513.151
소듐 피루베이트Sodium pyruvate 0.0550.055
비교예Comparative example 1 One
대조군으로 사용하기 위한 마치현 추출물은 화장품 전문업체인 코스맥스㈜에서 40% 마치현 추출물을 제공받아 사용하였다. 상기 40%의 마치현 추출물을 처리할 때 초순수정제수로 희석하여 최종 농도가 0.5%, 1%, 2%, 5% 또는 8%가 되도록하였다.March extract for use as a control was used to receive a 40% March extract from Cosmax Co., Ltd., a cosmetics company. When the 40% of the extract of Machiwyeon treatment was diluted with ultrapure purified water so that the final concentration is 0.5%, 1%, 2%, 5% or 8%.
실험예Experimental Example 1:  One: 탈과립화Degranulation 정도 비교 Degree comparison
상기 실시예 1에서 제조된 조성물의 아토피성 피부염 치료 효과를 확인하기 위하여 알레르기 반응에 의해 시작되는 탈과립화의 지표인 β-헥소사미니데이즈의 농도를 다음과 같은 방법으로 측정하였다.In order to confirm the atopic dermatitis therapeutic effect of the composition prepared in Example 1, the concentration of β-hexosaminidase, which is an indicator of degranulation initiated by an allergic reaction, was measured by the following method.
먼저, 배양 접시에서 배양 중인 호염기성 비만세포(rat basophilic leukemia cell, 한국세포주은행)를 트립신 0.5%(w/v)로 처리하여 배양 접시로부터 분리시켰다. 상기 분리된 호염기성 비만세포 2×105개를 0.5 mg/㎖ 농도의 항-DNP IgE(Sigma-Aldrich, USA)와 함께 섞어준 후, 96 웰 플레이트의 각 웰에 넣어 주었다. 하루 동안 37℃ 및 5% CO2 조건의 인큐베이터에서 배양한 후 96 웰 플레이트 바닥에 세포가 완전히 붙으면 기존의 배양액을 제거한 후, 타이로드 용액(NaCl: 137 mmol/ℓ, KCl: 2.7 mmol/ℓ, CaCl2: 1.8 mmol/ℓ, MgCl2: 1 mmol/ℓ, 글루코즈: 5.6 mmol/ℓ, HEPES: 20 mmol/ℓ, BSA: 0.1%, NaOH로 pH 7.3으로 적정 후, 필터로 멸균하여 4℃에서 보관)을 사용하여 세척하였다. 상기 세척된 세포에 실시예 1의 DMEM/F12 배지를 농도별로 각각 처리하고, 1 μM DNP-HSA 용액(Sigma-Aldrich, USA)을 첨가하여 3시간 동안 반응시켰다. 이때, 음성 대조군으로는 DMEM/F12 배지가 포함되지 않은 용매를 사용하였다.First, bas basophilic leukemia cells (Korean Cell Line Bank) in culture in a culture dish were treated with trypsin 0.5% (w / v) and separated from the culture dish. 2 × 10 5 isolated basophilic mast cells were mixed with anti-DNP IgE (Sigma-Aldrich, USA) at a concentration of 0.5 mg / ml, and then placed in each well of a 96 well plate. After culturing in an incubator at 37 ° C. and 5% CO 2 for one day, the cells were completely adhered to the bottom of the 96 well plate, and then the culture medium was removed. Then, the Tyrode solution (NaCl: 137 mmol / l, KCl: 2.7 mmol / l, CaCl 2 : 1.8 mmol / l, MgCl 2 : 1 mmol / l, glucose: 5.6 mmol / l, HEPES: 20 mmol / l, BSA: 0.1%, titrated to pH 7.3 with NaOH, sterilized with a filter at 4 ° C. Storage). The washed cells were treated with DMEM / F12 medium of Example 1 by concentration, and reacted for 3 hours by adding 1 μM DNP-HSA solution (Sigma-Aldrich, USA). At this time, a solvent containing no DMEM / F12 medium was used as a negative control.
이어, 상기 반응물의 세포배양액 50 ㎕를 취하여 1 mM p-니트로페닐-N-아세틸-β-d-글루코사미니데가 포함된 0.1 M 시트르산나트륨(pH 4.5) 50 ㎕와 혼합한 뒤 이를 37℃에서 1시간 동안 반응시켰다. 상기 반응을 종료시키기 위해 0.1 M Na2CO3 및 0.1 M NaHCO3(pH 10)가 함유된 카보네이트 완충용액 200 ㎕를 처리한 뒤, 모델 680 마이크로플레이트 리더기(Bio-rad, USA)를 이용하여 405 nm의 파장에서 흡광도를 측정하였다.Subsequently, 50 µl of the cell culture solution of the reaction solution was taken and mixed with 50 µl of 0.1 M sodium citrate (pH 4.5) containing 1 mM p-nitrophenyl-N-acetyl-β-d-glucosaminide and then at 37 ° C. The reaction was carried out for 1 hour. To terminate the reaction, 200 μl of carbonate buffer containing 0.1 M Na 2 CO 3 and 0.1 M NaHCO 3 (pH 10) was treated, followed by 405 using a model 680 microplate reader (Bio-rad, USA). Absorbance was measured at a wavelength of nm.
음성 대조군의 흡광도 값을 기준으로 하여 농도별로 DMEM/F12를 처리한 군의 흡광도 값을 비교한 결과를 도 1에 나타내었다.The results of comparing the absorbance values of the groups treated with DMEM / F12 for each concentration based on the absorbance values of the negative control group are shown in FIG. 1.
도 1에서 보는 바와 같이, 실시예 1의 DMEM/F12 배지를 100%의 농도로 처리하였을 때 β-헥소사미니데이즈의 농도가 약 30% 감소하였으며, 2%, 10% 또는 50%의 농도로 처리하였을 때에도 유의적으로 β-헥소사미니데이즈의 농도가 감소되어 DMEM/F12 배지가 탈과립화를 억제하는 것을 확인할 수 있었다.As shown in FIG. 1, when the DMEM / F12 medium of Example 1 was treated at a concentration of 100%, the concentration of β-hexosaminidase was reduced by about 30%, and at a concentration of 2%, 10%, or 50%. When treated, the concentration of β-hexosaminidase was significantly decreased, and it was confirmed that DMEM / F12 medium inhibited degranulation.
실험예Experimental Example 2: 히스타민 합성 억제 효과 평가 2: Evaluation of Histamine Synthesis Inhibitory Effect
상기 실시예 1에서 제조된 조성물의 아토피성 피부염 치료 효과를 확인하기 위하여 히스타민 합성 억제 효과를 평가하였다. 먼저, 상기 실험예 1과 동일한 방법으로 반응물을 수득한 뒤, 상기 반응물의 세포 배양액을 모아 제조사의 가이드라인에 따라, 히스타민에 대한 ELISA 분석 키트(Oxford biomedical research, USA)를 사용하여 측정하였다.In order to confirm the atopic dermatitis therapeutic effect of the composition prepared in Example 1 was evaluated the histamine synthesis inhibitory effect. First, the reaction product was obtained in the same manner as in Experimental Example 1, and the cell culture solution of the reaction solution was collected and measured using an ELISA assay kit for histamine (Oxford biomedical research, USA) according to the manufacturer's guidelines.
음성 대조군의 히스타민 농도를 기준으로 하여 농도별로 DMEM/F12를 처리한 군의 히스타민 농도를 비교한 결과를 도 2에 나타내었다.The results of comparing the histamine concentration of the group treated with DMEM / F12 for each concentration based on the histamine concentration of the negative control group are shown in FIG. 2.
도 2에서 보는 바와 같이, 실시예 1의 DMEM/F12 배지를 100%의 농도로 처리하였을 때 히스타민의 농도가 약 30% 감소하였으며, 2%, 10% 또는 50%의 농도로 처리하였을 때에도 유의적으로 히스타민의 농도가 감소되어 DMEM/F12 배지가 히스타민의 합성을 억제하는 것을 확인할 수 있었다.As shown in FIG. 2, when the DMEM / F12 medium of Example 1 was treated at a concentration of 100%, the concentration of histamine was reduced by about 30%, and significant even when treated at a concentration of 2%, 10%, or 50%. As a result, the concentration of histamine was decreased, and it was confirmed that the DMEM / F12 medium inhibited the synthesis of histamine.
실험예Experimental Example 3: 염증반응 및  3: inflammatory response and 피부장벽Skin barrier 관련 물질의 발현 확인 Confirmation of expression of related substances
상기 실시예 1에서 제조된 조성물이 아토피에 의한 염증반응 또는 피부장벽에 미치는 영향을 확인하기 위하여 염증성 사이토카인인 IL-6(interleukin-6), 피부장벽 구성요소인 필라그린, 및 손상된 피부장벽 회복에 중요한 요소인 세린 팔미토일트렌스퍼레이즈(SPT)의 발현을 mRNA 수준에서 확인하였다.In order to confirm the effect of the composition prepared in Example 1 on the inflammatory response or skin barrier caused by atopy, the inflammatory cytokine IL-6 (interleukin-6), the skin barrier component filaggrin, and the damaged skin barrier recovery The expression of serine palmitoyltransferase (SPT), an important factor in, was confirmed at the mRNA level.
상기 실험예 1과 동일한 방법으로 반응물을 수득한 뒤, 0.05% 트립신을 처리하여 획득한 세포에서 트리졸(trizol)로 mRNA를 추출하였다. 상기 추출된 mRNA로부터 cDNA의 합성은 ReverTra Ace® qPCR Master Mix(TOYOBO, Japan)를 사용하여 제조사의 가이드라인에 따라 수행하였고, 이때 사용된 프라이머(Cosmogenetech, Korea)는 하기 표 3에 나타낸 바와 같다.After the reaction was obtained in the same manner as in Experimental Example 1, mRNA was extracted with trizol from cells obtained by treatment with 0.05% trypsin. Synthesis of cDNA from the extracted mRNA was performed according to the manufacturer's guidelines using ReverTra Ace® qPCR Master Mix (TOYOBO, Japan), wherein the primers used (Cosmogenetech, Korea) are shown in Table 3 below.
염증반응 및 피부장벽 관련 물질 발현 확인에 사용된 프라이머Primer used to identify inflammatory response and expression of substances related to skin barrier
이름name 서열(5'→3')Sequence (5 '→ 3') 서열번호SEQ ID NO:
IL-6 정방향IL-6 forward AAA TGC CAG CCT GCT GAC GAA CAAA TGC CAG CCT GCT GAC GAA C 서열번호 1SEQ ID NO: 1
IL-6 역방향IL-6 reverse AAC AAC AAT CTG AGG TGC CCA TAAC AAC AAT CTG AGG TGC CCA T 서열번호 2SEQ ID NO: 2
필라그린 정방향Pillar Green Forward TCT GTT CAG GAG CAG TCA AGGTCT GTT CAG GAG CAG TCA AGG 서열번호 3SEQ ID NO: 3
필라그린 역방향Reverse Pillargreen GCT TCA TGG TGA TGC GAC CAGCT TCA TGG TGA TGC GAC CA 서열번호 4SEQ ID NO: 4
SPT 정방향SPT forward GAG AGA TGC TGA AGC GGA ACGAG AGA TGC TGA AGC GGA AC 서열번호 5SEQ ID NO: 5
SPT 역방향SPT reverse TGG TAT GAG CTG CTG ACA GGTGG TAT GAG CTG CTG ACA GG 서열번호 6SEQ ID NO: 6
β-액틴 정방향β-actin forward GGA CTT CGA GCA AGA GAT GGGGA CTT CGA GCA AGA GAT GG 서열번호 7SEQ ID NO: 7
β-액틴 역방향β-actin reverse AGC ACT GTG TTG GCG TAC AGAGC ACT GTG TTG GCG TAC AG 서열번호 8SEQ ID NO: 8
그리고 나서, 모든 PCR 결과물은 1.5%의 아가로스 젤에 전기영동을 수행하고, BioRad Molecular Imager® GelDoc™ XR(BioRad, USA)로 이미지를 확인하였으며, 음성 대조군의 IL-6, 필라그린 및 세린 팔미토일트렌스퍼레이즈 발현량을 기준으로 하여 농도별로 DMEM/F12를 처리한 군의 발현량을 비교한 결과를 도 3 내지 5에 나타내었다.Then, all PCR results were subjected to electrophoresis on 1.5% agarose gel and images were verified with BioRad Molecular Imager® GelDoc ™ XR (BioRad, USA), IL-6, pilaggrin and serine palmi of negative controls. 3 to 5 show the results of comparing the expression levels of the groups treated with DMEM / F12 for each concentration based on the expression of the toy trancease.
도 3 내지 5에서 보는 바와 같이, 염증반응 관련물질인 IL-6의 경우 DMEM/F12 배지를 처리한 농도에 의존적으로 발현이 억제된 반면, 피부장벽 관련물질인 필라그린 및 세린 팔미토일트렌스퍼레이즈는 DMEM/F12 배지의 처리 농도 의존적으로 발현이 증가함으로써, 상기 DMEM/F12 배지가 염증반응을 억제하고, 피부 상태를 개선시키는 효과가 있음을 알 수 있다.As shown in FIGS. 3 to 5, the expression of IL-6, which is an inflammation-associated substance, was suppressed depending on the concentration of DMEM / F12 medium, whereas the skin barrier-related substance, filaggrin and serine palmitoyltransferase By increasing the expression-dependent expression of DMEM / F12 medium, it can be seen that the DMEM / F12 medium has an effect of inhibiting the inflammatory response and improving the skin condition.
실험예Experimental Example 4: 세포 증식률 확인 4: cell proliferation rate
상기 실시예 2에서 제조된 조성물이 피부 세포의 증식에 어떠한 영향을 미치는지 확인하기 위하여 라우릴황산나트륨(sodium lauryl sulfate, SLS) 또는 UV에 의한 손상으로부터 회복률을 확인하였다.In order to confirm how the composition prepared in Example 2 affects the proliferation of skin cells, recovery rate from damage by sodium lauryl sulfate (SLS) or UV was confirmed.
먼저, 배양 접시에서 배양 중인 인간 피부 섬유아세포(human dermal fibroblast cell, 한국세포주은행)를 트립신 0.5%(w/v)로 처리하여 배양 접시로부터 분리시켰다. 상기 분리된 섬유아세포 4×103개를 96 웰 플레이트의 각 웰에 넣어 주었다. 하루 동안 37℃ 및 5% CO2 조건의 인큐베이터에서 무혈청 배지로 굶주리게 한 뒤, 0.005%(v/v)의 SLS 또는 30 mJ의 UVA를 처리하고, 실시예 2의 혼합 조성물을 각각의 농도로 첨가하여 24시간 동안 반응시켰다. 이때, 대조군으로는 아무것도 처리하지 않은 무처리 대조군 및 상기 비교예 1에서 제조된 마치현 추출물 처리군을 사용하였다.First, human dermal fibroblast cells (Korean Cell Line Bank) being cultured in a culture dish were treated with trypsin 0.5% (w / v) and separated from the culture dish. The separated fibroblasts 4 × 10 3 were put in each well of a 96 well plate. After starving with serum-free medium in a 37 ° C. and 5% CO 2 condition incubator for one day, treated with 0.005% (v / v) of SLS or 30 mJ of UVA and the mixed composition of Example 2 at each concentration It was added and reacted for 24 hours. At this time, as a control, a non-treated control, which was not treated with anything, and the gusset extract treatment group prepared in Comparative Example 1 were used.
상기 24시간 동안 반응한 세포의 증식율을 분석하기 위해 각각의 웰에 CCK 용액(Dojindo, Japan)을 10 ㎕ 첨가한 뒤, 2시간 동안 반응시키고, 모델 680 마이크로플레이트 리더기를 이용하여 450 nm에서 흡광도를 측정하였다.In order to analyze the proliferation rate of the cells reacted for 24 hours, 10 μl of CCK solution (Dojindo, Japan) was added to each well, followed by reaction for 2 hours, and absorbance at 450 nm using a model 680 microplate reader. Measured.
아무것도 처리하지 않은 무처리 대조군의 흡광도 값을 기준으로 하여, 실시예 2 또는 비교예 1의 조성물을 처리한 군의 세포 증식율을 비교한 결과를 도 6 및 7에 나타내었다.6 and 7 show the results of comparing the cell proliferation rates of the groups treated with the composition of Example 2 or Comparative Example 1, based on the absorbance values of the untreated control treated with nothing.
도 6 및 7에서 보는 바와 같이, 마치현 추출물을 처리한 군에 있어서는 세포 증식이 억제되는 것을 알 수 있으나, 실시예 2의 혼합 조성물은 농도 의존적으로 세포의 증식을 촉진시킴을 확인함으로써, 본 발명의 혼합 조성물 또는 상기 조성물을 구성성분으로 포함하는 DMEM/F12 배지가 피부재생 또는 피부자극 완화 용도로 사용될 수 있음을 알 수 있다.As shown in Figures 6 and 7, it can be seen that the cell proliferation is inhibited in the group treated with gut extract, but by confirming that the mixed composition of Example 2 promotes the proliferation of cells in a concentration-dependent manner, It can be seen that the mixed composition or DMEM / F12 medium containing the composition as a component can be used for skin regeneration or skin irritation relief.
실험예Experimental Example 5:  5: UVA에To UVA 대한 민감도 완화 확인 Sensitivity Mitigation
상기 실시예 2에서 제조된 조성물의 인공피부에서 UVA에 대한 민감도 완화 효과를 확인하였다.In the artificial skin of the composition prepared in Example 2 was confirmed the effect of reducing the sensitivity to UVA.
먼저, 인공피부는 테고사이언스㈜에 표피부분의 제조를 의뢰하여 구입하였다. 상기 인공피부 6개를 한천에서 떼어내어 6웰 플레이트로 옮긴 뒤, 함께 제공받은 보존 배지(maintenance medium, TA-MM001, Tego science, Korea)를 3 ㎖ 첨가하여 하루 동안 37℃ 및 5% CO2 조건의 인큐베이터에서 안정화시켰다. 상기 안정화된 인공피부 중 5개를 1 ㎖의 PBS가 첨가된 새로운 6웰 플레이트로 옮겨 1 J의 UVA(ultraviolet-A, 방사스펙트럼 315 내지 400 ㎚)에 노출시킨 뒤, 이를 PBS로 2회 세척하고, UVA를 처리하지 않은 대조군을 포함하는 6개의 인공피부를 새 6 웰 플레이트에 넣어주었다. 그 후, UVA에 노출되지 않은 대조군 및 UVA에 노출된 1개의 인공피부에 3 ㎖의 상기 보존 배지를 넣어주고, 나머지 4개의 인공피부에는 실시예 2의 조성물의 농도가 2%, 및 비교예 1의 마치현 추출물의 농도가 0.5%가 되도록 보존 배지와 섞어 3 ㎖씩 넣어주고 3일간 배양하였다.First, artificial skin was purchased by Tego Science Co., Ltd. for the preparation of the epidermal part. 6 artificial skins were removed from the agar and transferred to a 6-well plate, followed by adding 3 ml of the preservation medium (maintenance medium, TA-MM001, Tego science, Korea) provided at 37 ° C. and 5% CO 2 conditions for one day. Stabilized in an incubator. Five of the stabilized artificial skins were transferred to a new 6-well plate with 1 ml of PBS, exposed to 1 J of UVA (ultraviolet-A, radiation spectrum 315-400 nm) and washed twice with PBS. Six artificial skins, including controls not treated with UVA, were placed in a new 6 well plate. Thereafter, 3 ml of the preservation medium was added to the control group not exposed to UVA and one artificial skin exposed to UVA, and the remaining four artificial skins had a concentration of the composition of Example 2 at 2%, and Comparative Example 1 Mix with 3 ml of preservation medium so that the concentration of the extract of Machi Prefecture was 0.5% and incubated for 3 days.
상기 배양된 인공피부를 웰에서 꺼내 카세트에 넣은 뒤, 5% 포름알데히드 용액에 넣어 하루 동안 고정하고, 이를 H&E로 염색하여 피부조직을 관찰 및 촬영한 결과를 도 8에 나타내었다.The cultured artificial skin was removed from the well, placed in a cassette, and then fixed in a 5% formaldehyde solution for one day, and stained with H & E to observe and photograph skin tissue.
도 8에서 보는 바와 같이, UVA만 처리한 군은 이를 처리하지 않은 군과 비교하여 인공피부가 손상되었고, 여기에 비교예 1의 마치현 추출물을 처리한 경우에는 손상된 인공피부에 변화가 없었으나, 실시예 2의 혼합 조성물을 처리한 경우에는 UVA에 의해 손상된 인공 피부가 정상에 가깝게 회복됨을 확인함으로써, 상기 실시예 2의 혼합 조성물 또는 상기 조성물을 구성성분으로 포함하는 DMEM/F12 배지가 피부 민감성 완화 용도로 사용될 수 있음을 알 수 있다.As shown in Figure 8, the group treated with UVA only damaged the artificial skin compared to the group not treated, and when treated with the gut extract of Comparative Example 1 there was no change in the damaged artificial skin, but When the mixed composition of Example 2 was treated, it was confirmed that artificial skin damaged by UVA was restored to normal, so that the mixed composition of Example 2 or the DMEM / F12 medium containing the composition as a component reduced skin sensitivity. It can be seen that it can be used as.
실험예Experimental Example 6:  6: UVA에To UVA 노출된 인공피부에서 염증반응 관련 물질에 대한 영향 Influence of Inflammatory Response Substances on Exposed Artificial Skin
상기 실시예 2에서 제조된 조성물이 UVA에 노출된 인공피부에서 염증성 사이토카인인 IL-1α(interleukin-1α) 및 TNF-α(tumor necrosis factor-α)의 발현에 어떠한 영향을 주는지 확인하였다.It was confirmed that the composition prepared in Example 2 affects the expression of inflammatory cytokines IL-1α (interleukin-1α) and TNF-α (tumor necrosis factor-α) in artificial skin exposed to UVA.
먼저, 상기 실험예 5에 기재된 바와 같이 인공피부에 실시예 2의 조성물 및 비교예 1의 마치현 추출물을 처리하여 3일 동안 배양하였다. 상기 배양된 배양액을 수득하여 IL-1α 및 TNF-α의 양을 각각 IL-1α ELISA 키트(Koma biotech, Korea) 및 TNF-a ELISA 키트(R&D System, USA)를 사용하여 제조사의 가이드라인을 따라 측정하여 그 결과를 도 9 및 10에 나타내었다.First, as described in Experimental Example 5, the artificial skin was treated with the composition of Example 2 and the extract of Portico of Comparative Example 1 and cultured for 3 days. Obtain the cultured cultures and use the IL-1α ELISA kit (Koma biotech, Korea) and the TNF-a ELISA kit (R & D System, USA), respectively, to obtain the amount of IL-1α and TNF-α, respectively. The measurement results are shown in FIGS. 9 and 10.
도 9 및 10에서 보는 바와 같이, UVA만 처리한 군은 이를 처리하지 않은 군과 비교하여 IL-1α 및 TNF-α의 양이 증가하였고, 여기에 비교예 1의 마치현 추출물을 처리한 경우와 비교하여 실시예 2의 혼합 조성물을 처리한 경우에 상기 IL-1α 및 TNF-α의 양을 더욱 현저하게 저해하는 것을 확인함으로써, 본 발명의 실시예 2의 혼합 조성물 또는 상기 조성물을 구성성분으로 포함하는 DMEM/F12 배지가 아토피 피부염과 같은 피부질환 치료 용도로 사용될 수 있음을 알 수 있다.As shown in Figures 9 and 10, the group treated with UVA only increased the amount of IL-1α and TNF-α compared to the group not treated, compared to the case treated with the gusset extract of Comparative Example 1 When the mixed composition of Example 2 was treated to further inhibit the amount of IL-1α and TNF-α, the mixed composition of Example 2 of the present invention or the composition as a component It can be seen that DMEM / F12 medium can be used for the treatment of skin diseases such as atopic dermatitis.

Claims (23)

  1. 세포 배양배지를 유효성분으로 포함하는 피부 질환 예방 및 치료용 약학 조성물.Pharmaceutical composition for preventing and treating skin diseases comprising a cell culture medium as an active ingredient.
  2. 제1항에 있어서, 상기 세포 배양배지는 DMEM/F12, DMEM, MEMα 및 이의 조합으로 구성된 군으로부터 선택된 어느 하나인, 약학 조성물.The pharmaceutical composition of claim 1, wherein the cell culture medium is any one selected from the group consisting of DMEM / F12, DMEM, MEMα, and combinations thereof.
  3. 제1항에 있어서, 상기 세포 배양배지는 무혈청(serum-free) 배지인, 약학 조성물.The pharmaceutical composition of claim 1, wherein the cell culture medium is a serum-free medium.
  4. 제1항에 있어서, 상기 세포 배양배지는 아미노산 성분, 비타민 성분, 무기염 성분 및 기타 성분으로 구성되며,The method of claim 1, wherein the cell culture medium is composed of an amino acid component, vitamin component, inorganic salt component and other components,
    a) 상기 아미노산 성분은 글리세린, L-알라닌, L-아르기닌 하이드로클로라이드, L-시스테인 하이드로클로라이드-모노하이드레이트, L-글루타민, L-히스티딘 하이드로클로라이드-모노하이드레이트, L-리신 하이드로클로라이드, L-메티오닌, L-프롤린, L-세린, L-트레오닌 및 L-발린으로 구성된 군으로부터 선택되는 적어도 하나의 아미노산 또는 이의 조합이고,a) the amino acid component is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L-lysine hydrochloride, L-methionine, At least one amino acid selected from the group consisting of L-proline, L-serine, L-threonine and L-valine, or a combination thereof,
    b) 상기 비타민 성분은 i-이노시톨, 티아민 하이드로클로라이드, 나이아신아미드 및 피리독신 하이드로클로라이드로 구성된 군으로부터 선택되는 적어도 하나의 비타민 또는 이의 조합이며,b) said vitamin component is at least one vitamin selected from the group consisting of i-inositol, thiamine hydrochloride, niacinamide and pyridoxine hydrochloride or combinations thereof,
    c) 상기 무기염 성분은 염화나트륨(NaCl), 탄산수소나트륨(NaHCO3), 염화칼륨(KCl), 염화칼슘(CaCl2)(무수) 및 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O)로 구성된 군으로부터 선택되는 적어도 하나의 무기염 또는 이의 조합이고,c) The inorganic salt component is sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), potassium chloride (KCl), calcium chloride (CaCl 2 ) (anhydrous) and sodium hydrogen phosphate monohydrate (NaH 2 PO 4 -H 2 O) At least one inorganic salt selected from the group consisting of or a combination thereof,
    d) 상기 기타 성분은 D-글루코즈(덱스트로즈) 또는 소듐 피루베이트인, 약학 조성물.d) The other component is D-glucose (dextrose) or sodium pyruvate.
  5. 제4항에 있어서, 상기 세포 배양배지는 글리세린, L-알라닌, L-아르기닌 하이드로클로라이드, L-시스테인 하이드로클로라이드-모노하이드레이트, L-글루타민, L-히스티딘 하이드로클로라이드-모노하이드레이트, L-리신 하이드로클로라이드, L-메티오닌, L-프롤린, L-세린, L-트레오닌, L-발린, i-이노시톨, 티아민 하이드로클로라이드, 나이아신아미드, 피리독신 하이드로클로라이드, 염화나트륨(NaCl), 탄산수소나트륨(NaHCO3), 염화칼륨(KCl), 염화칼슘(CaCl2)(무수), 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O), D-글루코즈(덱스트로즈) 및 소듐 피루베이트로 구성되는, 약학 조성물.The cell culture medium of claim 4, wherein the cell culture medium is glycerin, L-alanine, L-arginine hydrochloride, L-cysteine hydrochloride-monohydrate, L-glutamine, L-histidine hydrochloride-monohydrate, L-lysine hydrochloride. , L-methionine, L-proline, L-serine, L-threonine, L-valine, i-inositol, thiamine hydrochloride, niacinamide, pyridoxine hydrochloride, sodium chloride (NaCl), sodium bicarbonate (NaHCO 3 ), potassium chloride Pharmaceutical composition, consisting of (KCl), calcium chloride (CaCl 2 ) (anhydrous), sodium hydrogen phosphate monohydrate (NaH 2 PO 4 —H 2 O), D-glucose (dextrose) and sodium pyruvate.
  6. 제4항에 있어서, 상기 아미노산 성분은 0.1 내지 0.2 중량%의 글리세린, 0.01 내지 0.1 중량%의 L-알라닌, 0.5 내지 3 중량%의 L-아르기닌 하이드로클로라이드, 0.1 내지 0.2 중량%의 L-시스테인 하이드로클로라이드-모노하이드레이트, 2 내지 3 중량%의 L-글루타민, 0.1 내지 0.5 중량%의 L-히스티딘 하이드로클로라이드-모노하이드레이트, 0.4 내지 1.0 중량%의 L-리신 하이드로클로라이드, 0.1 내지 0.5 중량%의 L-메티오닌, 0.1 내지 0.5 중량%의 L-프롤린, 0.1 내지 0.3 중량%의 L-세린, 0.3 내지 0.6 중량%의 L-트레오닌 및 0.3 내지 0.6 중량%의 L-발린을 포함하는, 약학 조성물.The method according to claim 4, wherein the amino acid component is 0.1 to 0.2 wt% glycerin, 0.01 to 0.1 wt% L-alanine, 0.5 to 3 wt% L-arginine hydrochloride, 0.1 to 0.2 wt% L-cysteine hydro Chloride-monohydrate, 2-3 wt% L-glutamine, 0.1-0.5 wt% L-histidine hydrochloride-monohydrate, 0.4-1.0 wt% L-lysine hydrochloride, 0.1-0.5 wt% L- A pharmaceutical composition comprising methionine, 0.1-0.5 wt% L-proline, 0.1-0.3 wt% L-serine, 0.3-0.6 wt% L-threonine and 0.3-0.6 wt% L-valine.
  7. 제4항 있어서, 상기 비타민 성분은 0.01 내지 0.1 중량%의 i-이노시톨, 0.01 내지 0.1 중량%의 티아민클로라이드, 0.005 내지 0.03 중량%의 나이아신아미드 및 0.01 내지 0.1 중량%의 피리독신 하이드로클로라이드를 포함하는, 약학 조성물.The method of claim 4, wherein the vitamin component comprises 0.01 to 0.1% by weight of i-inositol, 0.01 to 0.1% by weight of thiaminchloride, 0.005 to 0.03% by weight of niacinamide and 0.01 to 0.1% by weight of pyridoxine hydrochloride. Pharmaceutical composition.
  8. 제4항에 있어서, 상기 무기염 성분은 45 내지 50 중량%의 염화나트륨(NaCl), 15 내지 18 중량%의 탄산수소나트륨(NaHCO3), 2.0 내지 2.5 중량%의 염화칼륨(KCl), 0.8 내지 0.9 중량%의 염화칼슘(CaCl2)(무수) 및 0.4 내지 0.5 중량%의 인산수소나트륨 모노하이드레이트(NaH2PO4-H2O)를 포함하는, 약학 조성물.The method according to claim 4, wherein the inorganic salt component is 45 to 50% by weight sodium chloride (NaCl), 15 to 18% by weight sodium hydrogen carbonate (NaHCO 3 ), 2.0 to 2.5% by weight potassium chloride (KCl), 0.8 to 0.9 A pharmaceutical composition comprising weight percent calcium chloride (CaCl 2 ) (anhydrous) and 0.4 to 0.5 weight percent sodium hydrogen phosphate monohydrate (NaH 2 PO 4 —H 2 O).
  9. 제4항에 있어서, 상기 기타 성분은 20 내지 25 중량%의 D-글루코즈(덱스트로즈) 또는 0.35 내지 0.4 중량%의 소듐 피루베이트를 포함하는, 약학 조성물.The pharmaceutical composition of claim 4, wherein the other component comprises 20 to 25 wt% D-glucose (dextrose) or 0.35 to 0.4 wt% sodium pyruvate.
  10. 제1항에 있어서, 상기 세포 배양배지가 1 내지 10 중량%인, 약학 조성물.The pharmaceutical composition of claim 1, wherein the cell culture medium is 1 to 10 wt%.
  11. 제1항에 있어서, 상기 피부 질환이 아토피성 피부염, 알레르기, 피부 습진, 여드름, 건선, 가려움증 및 이의 조합으로 이루어진 군에서 선택되는, 약학 조성물.The pharmaceutical composition of claim 1, wherein the skin disease is selected from the group consisting of atopic dermatitis, allergy, skin eczema, acne, psoriasis, itching, and combinations thereof.
  12. 제4항에 있어서, 상기 세포 배양배지에 아미노산 성분, 비타민 성분, 무기염 성분, 기타 성분 및 이의 조합으로 이루어진 군에서 선택되는 성분을 더 포함하는, 약학 조성물.The pharmaceutical composition of claim 4, further comprising a component selected from the group consisting of an amino acid component, a vitamin component, an inorganic salt component, other components, and combinations thereof in the cell culture medium.
  13. 제12항에 있어서, 상기 아미노산 성분이 L-아스파라긴-모노하이드레이트, L-아스파르트산, L-시스틴 2HCl, L-글루탐산, L-이소류신, L-류신, L-페닐알라닌, L-트립토판, L-티로신 디소듐염 디하이드레이트 및 이의 조합으로 이루어진 군에서 선택되는, 약학 조성물.The method according to claim 12, wherein the amino acid component is L-asparagine-monohydrate, L-aspartic acid, L-cystine 2HCl, L-glutamic acid, L-isoleucine, L-leucine, L-phenylalanine, L-tryptophan, L-tyrosine Disodium salt dihydrate, and combinations thereof.
  14. 제12항에 있어서, 상기 비타민 성분이 바이오틴, D-판토텐산칼슘, 엽산, 리보플라빈, 비타민 이의 조합으로 이루어진 군에서 선택되는, 약학 조성물.The pharmaceutical composition of claim 12, wherein the vitamin component is selected from the group consisting of biotin, calcium D-pantothenate, folic acid, riboflavin, vitamins, and combinations thereof.
  15. 제12항에 있어서, 상기 무기염 성분이 황산동 펜타하이드레이트(CuSO4-5H2O), 황산제이철 헵타하이드레이트(FeSO4-7H2O), 염화마그네슘(무수), 황산마그네슘(MgSO4)(무수), 인산수소이나트륨(Na2HPO4) 무수, 황산아연 헵타하이드레이트(ZnSO4-7H2O) 및 이의 조합으로 이루어진 군에서 선택되는, 약학 조성물.The method of claim 12, wherein the inorganic salt components are copper sulfate pentahydrate (CuSO 4 -5H 2 O), ferric sulfate hepta-hydrate (FeSO 4 -7H 2 O), magnesium chloride (anhydrous), magnesium sulfate (MgSO 4), (anhydrous ), phosphoric acid susoyi sodium (Na 2 HPO 4) anhydrous, zinc sulfate hepta-hydrate (ZnSO 4 -7H 2 O), and, a pharmaceutical composition is selected from the group consisting of a combination thereof.
  16. 제12항에 있어서, 상기 기타 성분이 히포크산틴 Na, 리놀렌산, 리포산, 푸트레신 2HCl, 티미딘 및 이의 조합으로 이루어진 군에서 선택되는, 약학 조성물.The pharmaceutical composition of claim 12, wherein the other component is selected from the group consisting of hypoxanthine Na, linolenic acid, lipoic acid, putrescine 2HCl, thymidine, and combinations thereof.
  17. 제1항에 따른 약학 조성물을 이를 필요로 하는 대상의 피부에 도포하는 단계를 포함하는, 피부 질환의 예방 또는 치료 방법.A method of preventing or treating skin diseases comprising applying the pharmaceutical composition according to claim 1 to the skin of a subject in need thereof.
  18. 제17항에 있어서, 상기 대상은 포유동물인, 방법.The method of claim 17, wherein the subject is a mammal.
  19. 피부 질환의 예방 또는 치료를 위한 약제의 제조에 사용하기 위한 제1항에 따른 약학 조성물의 용도.Use of the pharmaceutical composition according to claim 1 for use in the manufacture of a medicament for the prevention or treatment of skin diseases.
  20. 세포 배양배지를 유효성분으로 포함하는 피부 상태 개선용 화장료 조성물.Cosmetic composition for improving skin conditions comprising a cell culture medium as an active ingredient.
  21. 제20항에 있어서, 상기 피부 상태 개선이 주름의 발생 억제, 피부 노화 억제, 피부 탄력 개선, 피부 재생, 상처 또는 창상 회복(wound healing), 각막 재생, 피부자극완화 및 이의 조합으로 이루어진 군에서 선택되는, 피부 세포의 기능 저하 또는 손실로부터 피부를 보호 또는 피부 상태를 개선시키는, 조성물.The method of claim 20, wherein the improvement in skin condition is selected from the group consisting of inhibition of wrinkles, inhibition of skin aging, improvement of skin elasticity, skin regeneration, wound or wound healing, corneal regeneration, skin irritation relief, and combinations thereof. Which protects the skin from deterioration or loss of skin cells or improves the skin condition.
  22. 제20항에 따른 화장료 조성물을 이용하여 피부 상태를 개선시키는 방법.A method for improving skin condition using the cosmetic composition according to claim 20.
  23. 피부 상태를 개선시키기 위한 화장료의 제조에 사용하기 위한 제20항에 따른 화장료 조성물의 용도.Use of a cosmetic composition according to claim 20 for use in the preparation of a cosmetic for improving skin condition.
PCT/KR2016/000276 2015-01-26 2016-01-12 Pharmaceutical composition for preventing or treating skin diseases, containing cell culture medium WO2016122140A1 (en)

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