WO2016122140A1 - Composition pharmaceutique pour la prévention ou le traitement de maladies de la peau, contenant un milieu de culture cellulaire - Google Patents

Composition pharmaceutique pour la prévention ou le traitement de maladies de la peau, contenant un milieu de culture cellulaire 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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English (en)
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/ko
Publication of WO2016122140A1 publication Critical patent/WO2016122140A1/fr

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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

La présente invention concerne : une composition pour la prévention ou le traitement de maladies de la peau ou pour améliorer l'état de la peau, contenant un milieu de culture cellulaire; et un procédé pour la prévention ou le traitement de maladies de la peau ou pour améliorer l'état de la peau par l'utilisation de la composition. La composition selon la présente invention inhibe la dégranulation de la peau et l'expression de substances liées à des réactions inflammatoires, favorise l'expression de substances liées à la barrière cutanée, et présente un effet facilitant la guérison de la peau endommagée; de ce fait, la composition contenant un milieu de culture cellulaire peut être utile dans la prévention ou le traitement de maladies de la peau ou l'amélioration de l'état de la peau.
PCT/KR2016/000276 2015-01-26 2016-01-12 Composition pharmaceutique pour la prévention ou le traitement de maladies de la peau, contenant un milieu de culture cellulaire WO2016122140A1 (fr)

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KR102216302B1 (ko) * 2018-09-21 2021-02-17 메디포스트(주) 피부 세포 활성화용 조성물 및 이의 용도

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WO2010107286A2 (fr) * 2009-03-20 2010-09-23 주식회사 스템메디언스 Composition utilisée dans le traitement des affections cutanées et utilisant des cellules souches foetales du liquide amniotique
KR20120027440A (ko) * 2012-01-19 2012-03-21 바이오스펙트럼 주식회사 천연추출물 또는 이로부터 단리된 화합물을 포함하는 동물세포 배양용 무혈청 배지 및 피부개선조성물
KR20130061950A (ko) * 2011-12-02 2013-06-12 서경대학교 산학협력단 피부줄기세포 배양방법 및 이를 이용한 피부상태 개선용 조성물
KR20140103759A (ko) * 2013-02-19 2014-08-27 주식회사 엘지생활건강 동물세포 배양용 무혈청 배지 및 이를 이용한 피부상태 개선용 조성물

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KR20130061950A (ko) * 2011-12-02 2013-06-12 서경대학교 산학협력단 피부줄기세포 배양방법 및 이를 이용한 피부상태 개선용 조성물
KR20120027440A (ko) * 2012-01-19 2012-03-21 바이오스펙트럼 주식회사 천연추출물 또는 이로부터 단리된 화합물을 포함하는 동물세포 배양용 무혈청 배지 및 피부개선조성물
KR20140103759A (ko) * 2013-02-19 2014-08-27 주식회사 엘지생활건강 동물세포 배양용 무혈청 배지 및 이를 이용한 피부상태 개선용 조성물

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