WO2019078463A1 - Composition for treating fine dust-caused skin cell damage, comprising green tea extract - Google Patents

Composition for treating fine dust-caused skin cell damage, comprising green tea extract Download PDF

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Publication number
WO2019078463A1
WO2019078463A1 PCT/KR2018/009261 KR2018009261W WO2019078463A1 WO 2019078463 A1 WO2019078463 A1 WO 2019078463A1 KR 2018009261 W KR2018009261 W KR 2018009261W WO 2019078463 A1 WO2019078463 A1 WO 2019078463A1
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composition
green tea
tea extract
fine dust
skin
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PCT/KR2018/009261
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French (fr)
Korean (ko)
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신승현
안지연
나용주
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(주)아모레퍼시픽
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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
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/82—Theaceae (Tea family), e.g. camellia
    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat

Definitions

  • compositions for treating skin cell damage caused by fine dusts More particularly, the present invention discloses a composition comprising a tea tree extract, wherein the skin cell damage is cared by significantly changing a biomarker, such as a skin cell gene whose degree of expression is changed by fine dust, as compared with a normal skin cell.
  • a biomarker such as a skin cell gene whose degree of expression is changed by fine dust
  • Fine dusts are smaller in size than normal dusts, so they penetrate the human body more and contain harmful substances such as heavy metals, nitrates, ammoniums, and sulfates, and are harmful to human body.
  • harmful substances such as heavy metals, nitrates, ammoniums, and sulfates
  • the cells responsible for the immune function of the human body act to remove dust. These side effects are accompanied by inflammation.
  • the World Health Organization categorizes fine dusts as a group of carcinogens directly related to cancer.
  • Non-patent document 1 discloses that the expression amount of clodin 1 (CLDN 1), which is a major constituent of tight junction, which is a granule layer barrier, is reduced due to fine dust stimulation of PM 2.5.
  • Non-Patent Document 1 Kim, HJ, et al., "Transcriptome analysis of airborne PM 2.5 -induced detrimental effects on human keratinocytes", Toxicology Letters 273, 26-35,
  • the present inventors have found that the effect of fine dusts on the skin is detrimental to the skin, and the skin cell gene expression is affected by such an effect, thereby manifesting symptoms such as skin cell damage, and the green tea extract has an effect of restoring gene expression Respectively.
  • the present invention provides a composition comprising an extract of green tea as an active ingredient, wherein keratin 1 (KRT 1), claudin 1 CLDN1), claudin 4 (CLDN4), and occludin (OCLN) to a normal level.
  • KRT 1 keratin 1
  • CLDN1 claudin 1
  • CLDN4 claudin 4
  • OCLN occludin
  • the amount of gene expression changed by the fine dust can be returned to a normal level and skin cell damage can be cared.
  • Fig. 1 shows the expression of mRNA of KRT 1 gene in skin cells stimulated by ERM-CZ120 fine powder and showed a return to normal level by green tea extract treatment.
  • FIG. 2 shows an increase in mRNA expression of CLDN 1 gene in skin cells stimulated by ERM-CZ120 fine dust, and a return to a normal level by green tea extract treatment.
  • FIG. 3 shows that mRNA expression of CLDN4 gene was increased in skin cells stimulated by ERM-CZ120 fine powder and returned to normal level by green tea extract treatment.
  • Fig. 4 shows the increase in mRNA expression of OCLN gene in ERM-CZ120 micropore-stimulated skin cells and normalization by green tea extract treatment.
  • the composition for care of skin damage due to fine dust may include green tea extract as an active ingredient.
  • Camellia sinensis is a perennial evergreen plant belonging to theaceae. It is distributed in tropical, subtropical and temperate regions.
  • the main ingredients of tea tree are catechins, caffeine, amino acids, vitamins and minerals. Have been reported to exhibit physiological activity and pharmacological effects. Recently, catechins have been reported to have various functions such as antifouling effect, antioxidant effect, antimicrobial effect and anticancer effect.
  • the green tea extract is green tea leaf extract, the green tea leaves are not fermented, and the inner enzyme is inactive.
  • the green tea may be extracted with a specific extraction solvent to form a green tea extract.
  • the green tea extract may be prepared by extracting green tea with water or an organic solvent.
  • the green tea is prepared by dissolving at least one compound selected from the group consisting of water, C 1 -C 6 anhydrous or lower alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform, And extracting it with an extraction solvent.
  • the green tea extract may be extracted at room temperature.
  • the green tea extract may be one obtained by extracting with the extraction solvent, followed by addition of one or more of evaporation, filtration, concentration, separation or drying.
  • the green tea extract may be subjected to one or more filtration processes.
  • the separation process may include a centrifugation process.
  • the extraction can be carried out in the presence of water, a C 1 -C 6 anhydrous or a lower alcohol, a polar solvent comprising acetone and butylene glycol, and a polar solvent including ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane
  • a low polarity solvent may be used as a solvent.
  • the solvent may be 10-90% ethanol aqueous solution, and may be 10-50%, 15-45%, or 20-40% ethanol aqueous solution.
  • the solvent is 20 to 40% aqueous ethanol solution, the effective ingredient can be effectively extracted from the green tea.
  • the solvent is about 30% aqueous ethanol.
  • the extract may be concentrated under reduced pressure to an appropriate temperature in a distillation apparatus equipped with a cooling condenser after extraction.
  • the green tea extract according to the present invention can be extracted by a conventional method in the art, and is not limited by the above-described method.
  • the composition may comprise from 0.000001 to 30% by weight of green tea extract, based on the total weight of the composition.
  • the content thereof is 0.000001 to 30% by weight, the effect of care for skin damage due to fine dust caused by green tea extract is excellent.
  • 0.0000001 wt% or more 0.0000005 wt% or more, 0.0000007 wt% or more, 0.0000009 wt% or more, 0.0000009 wt% or more, 0.000001 wt% or more, 0.000002 wt% or more, 0.000004 wt% or more, 0.000006 wt% or more, 0.000008 wt.
  • % 0.0000007 wt% or less, 0.0000005 wt% or less, 0.0000003 wt% or less, 0.0000002 wt% or less, 0.0000001 wt% or less, or 0.00000009 wt% or less.
  • the active ingredient of the composition according to the present invention may be one which acts on the tight junction of the skin barrier.
  • the tight junction is a kind of cell junction, consisting of claudin, tricellulin, junctional adhesion molecule (JAM) and zona occluden (ZO).
  • the tight junction can be divided into a part contained in the cell membrane and an intracellular part.
  • Occludin and claudin are closely related constitutive proteins contained in cell membranes.
  • Occludin is the first transmembrane protein found in adherent synapses and has four transmembrane domains.
  • Occludin is known to regulate the function of tight junctions as well as constituents of tight junctions.
  • Claudin a close-coupled conformational protein with Occludin, also has four transmembrane domains.
  • the tight junction is a cell-to-cell linkage that fills the gap between neighboring cells, controls the movement of small molecules, controls the permeation of cells between cells, and maintains the polarity of the cells. Recently, it has been reported that tight junctions play an important role in skin barrier function. Tight junctions are a type of intercellular binding. It is a cell-to-cell junction that mainly occurs in epithelial cells and endothelial cells, forming a complex network with a string-like structure. The tight junction has two functions largely from this structure. The first is a barrier function that prevents material from leaking into the cells that make up epithelial cells or endothelial cells. It is mainly treated in blood vessels and digestive tracts.
  • the second is a fence function, which is a function to divide the cell membrane around the membrane fence in the cell membrane lipid. Accordingly, the active ingredient of the composition of the present invention acts on the tight junction of the skin barrier, thereby enhancing the barrier function and the fence function of the skin cells, thereby protecting the skin damaged by the fine dust stimulation.
  • Another aspect of the present invention includes skin care care applications for fine dusts of the composition.
  • a method for skin damage care by fine dust of a subject comprising administering an effective amount of a green tea extract to a subject in need thereof.
  • Another aspect of the present invention provides the use of green tea extract in the preparation of a composition for skin damage care by fine dust.
  • fine dust refers to a particulate matter that is a very small material that is invisible to the naked eye and that floats or drifts in the air for a long time.
  • the particle size of fine dust in the present invention may be not more than 10 ⁇ m (PM 10), specifically not more than 2.5 ⁇ m (PM 2.5).
  • Particulate matter having a particle diameter of not more than 2.5 ⁇ m (PM 2.5) is referred to as "ultrafine dust”.
  • fine dust is also intended to include “ultrafine dust”.
  • the fine dust in the present invention may include one or more of arsenic, cardmium, lead, and nickel.
  • the heavy metal components such as arsenic, cardmium, lead, and nickel are included in the fine dust to attack skin cells and cause skin allergy, pigmentation, inflammation, .
  • the fine dust includes arsenic, cardmium, lead, and nickel.
  • care means to effectively protect skin cells from stimulation and to inhibit, prevent, and restore (restore) the expression level of a specific gene by the stimulation.
  • repair also includes improving the expression level beyond the reference, in addition to changing the expression level closer to the reference level based on the gene expression level in the absence of the stimulus.
  • the present invention provides a composition for inhibiting damage of skin cells caused by fine dusts by controlling the expression level of a specific gene in skin cells damaged by fine dust to a normal level.
  • genes in the skin cells to which the expression amount by the fine dust is affected include keratin 1 (KRT 1), cladin 1 (CLDN 1), cladin 4 (CLDN 4), and occludin And at least one selected from the group. Since keratin 1 (KRT 1), claudin 1 (CLDN 1), claudin 4 (CLDN 4), and occludin (OCLN) are genes whose expression amount is reduced by fine dusts, To inhibit skin cell damage.
  • Analysis of the expression level of the gene or protein can be performed using a microarray, PCR, NGS (Nest Generation Sequencing), Western blot, northern blot, ELISA, radioimmunoassay, radioimmunoassay, tissue immuno staining, Can be analyzed using a variety of analytical methods known in the art.
  • Damage to the skin cells is caused by fine dust, which leads to inflammation and further damage to the skin cells.
  • the skin condition can be improved by taking care of the vicious circle of the skin cell damage by the green tea extract.
  • the composition may be a cosmetic composition, a pharmaceutical composition, or a health functional food composition.
  • cosmetic composition examples include cosmetics such as various creams, lotion creams, lotions, skins, and the like, and cleansing, cleansing agents, soaps, and essences.
  • the cosmetic composition to which the composition containing the green tea extract of the present invention is added may take the form of a solution, an emulsion, a viscous mixture or the like.
  • the cosmetic of the present invention is not particularly limited in its formulation, and examples thereof include an emulsion, cream, lotion, essence, pack, gel, powder, makeup base, foundation, lotion, ointment, patch, Formulations such as foam, cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, shampoo, rinse, body cleanser, soap, hair dye, spray and the like.
  • the components other than the green tea extract may be mixed and selected without difficulty by those skilled in the art depending on the formulation or use of the other cosmetic ingredients.
  • the cosmetic of the present invention may comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.
  • the cosmetic composition of the present invention may contain, in addition to the above essential ingredients, other ingredients usually added to cosmetics, if necessary.
  • Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.
  • organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.
  • the components to be added in addition to these components are not limited thereto, and any of the above components can be compounded within a range that does not impair the objects and effects of the present invention.
  • the pharmaceutical compositions comprising the green tea extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.
  • the pharmaceutical composition containing the green tea extract may be formulated into tablets, capsules, powders or syrups, or external preparations such as ointments, gels, creams, patches or sprays, And may be formulated and used in any form suitable for a clinical formulation.
  • the actual dosage of the active ingredient administered by the pharmaceutical composition should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration selected, the age, sex, weight and health status of the subject.
  • the dosage of the active ingredient may be from 0.0001 mg / kg / day to 3000 mg / kg / day, for example from 10 mg / kg / day to 500 mg / kg / day.
  • the health food is produced by using a raw material or a component (functional raw material) having a function useful for a nutrient or a human body which is likely to be deficient in a daily meal, Or to maintain or improve health through the activation of physiological functions.
  • a raw material or a component having a function useful for a nutrient or a human body which is likely to be deficient in a daily meal, Or to maintain or improve health through the activation of physiological functions.
  • the present invention is not limited thereto.
  • the health food may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles, but is not limited thereto and may be manufactured and processed in any form according to the law.
  • the health beverage composition has no particular limitation on the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages.
  • natural carbohydrates are conventional sugars such as monosaccharide polysaccharides, cyclodextrins and the like, and sugar alcohols such as xylitol, sorbitol and erythritol.
  • Natural flavors tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be used as flavorings other than those described above.
  • the actual dosage of the active ingredient administered by the health functional food composition should be determined in light of various relevant factors such as the severity of the symptoms, the selected route of administration, the age, sex, weight and health status of the subject .
  • the dosage of the active ingredient may be from 0.0001 mg / kg / day to 1000 mg / kg / day, for example from 0.02 mg / kg / day to 6 mg / kg / day.
  • Green tea extracts were prepared using green tea from Amorepacific Group Changwon. The collected green tea leaves were washed with purified water, filtered and dried, and then mixed in a weight ratio of 1: 8 with 30% ethanol (EtOH). The mixture was stirred at 25 ° C for 72 hours and extracted into 250 mesh size.
  • the obtained extract was subjected to three filtration steps, each of which had a size of 3.0 ⁇ m or less, a size of 1.0 ⁇ m or less, and a size of 0.3 ⁇ m or less.
  • the filtered extract was settled at 4 DEG C for 72 hours, and then filtered again to a size of 0.2 mu m or less to prepare a green tea extract.
  • Keratinocyte cell line Human normal epidermal keratinocytes was Lonza ⁇ (Lonza, Inc. Walkersville, Maryland material) was purchased and subcultured in a culture under a CO 2 incubator (CO 2 incubator), 37 °C from, 5% CO 2 conditions Respectively.
  • Bovine pituitary extract (BPE), human epidermal growth factor (hEGF), insulin, hydrocortisone, gentamicin sulfate, and the like were added to 500 ml of KBM-2 (KBMTM-2, CC- (Gentamycin Suflate), Epinephrine, and Transferrin were used.
  • the fine dusts were ERM-CZ120 fine dusts commercially available from Sigma-Aldrich, and PM 10 levels of fine dust and arsenic, cardmium, lead, and nickel ).
  • a 6-well plate was used and 1 ' Cells cultured under the conditions of 3 ⁇ 10 5 cells / well (cells / well) were cultured in the untreated group (control 1), 50 ppm of fine dusts (Control group 2), and a group treated with 50 ppm of fine dust and green tea extract (Example 1), respectively, and then cultured for 3 hours and 6 hours.
  • Control 1 The cells were washed with 2 ml of phosphate buffered saline (PBS) in Control 1, Control 2 and Example 1 obtained in the treatment of fine dust and green tea extract on keratinocyte cell line, Trizol reagent, Invitrogen, Carlsbad, CA, USA).
  • PBS phosphate buffered saline
  • RNA kit QIAGEN RNeasykt, QIAGEN, Valencia, Calif.
  • Agilent Technologies, Inc. Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, ) was used to confirm the quality of the RNA.
  • CDNA was synthesized from RNA using Invitrogen's Reverse Kit (Superscript Reverse Transcriptase (RT) kit, Invitrogen, Carlsbad, Calif.).
  • Q-RT-PCR real-time reverse transcription polymerase chain reaction
  • the control group 2 treated with fine dust at a concentration of 50 ppm contained Keratin 1, Claudin 1, Claudin 1, 4 (Claudin 4), and Occludin (mRNA) of the present invention.
  • the amount of mRNA expression of keratin 1 was 0.19 for 3 hours and 0.13 for 6 hours, indicating that mRNA expression levels were significantly decreased in the keratin 1, 0.44 and 0.24 in the keratin 1 and 0.52 in the keratin 4, respectively , And acrodine decreased to 0.48 and 0.39, respectively. That is, the amount of expression of keratinocyte differentiation markers (ie, keratin 1) and tight junction components (ie, claudin 1, claudin 4, and occludin) in the granule barriers were all reduced by fine dust.
  • Example 1 shows that the green tea extract of the present invention can cure mRNA expression levels of markers reduced by fine dust.
  • FIG. 1 shows that the amount of mRNA expression of keratin 1 is restored by 0.79 for 3 hours and 0.99 for 6 hours
  • FIG. 4 shows the expression level of mucin Was 0.88 for 3 hours and 0.98 for 6 hours, indicating that the amount of mRNA decreased by fine dust was restored.
  • Claudin 1 and Claudin 4 exhibited enhanced mRNA expression levels (1.12 and 1.23, respectively, when treated for 3 hours, and 1.23 and 1.45, respectively, when treated for 6 hours), compared to control group 1 (see FIGS. 2 and 3).
  • the cosmetic composition, the pharmaceutical composition and the health functional food composition can be applied to various formulations, and the present invention is not limited thereto .
  • 100 mg of the green tea extract according to the present invention, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate were mixed and then tableted according to the conventional preparation method.
  • Green tea extract 100 Lactose 400 Corn starch 400 Magnesium stearate 2
  • 100 mg of the green tea extract according to the present invention, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.
  • Green tea extract 100 Lactose 400 Corn starch 400 Magnesium stearate 2
  • green tea extract according to the present invention 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed and granulated into granules using a fluidized bed granulator, and filled in a capsule.
  • Green tea extract 5.00 maintain Suitable amount Sodium hydroxide Suitable amount Sodium chloride Suitable amount Spices Suitable amount Purified water Balance
  • Green tea extract 3.00 Polyethylene glycol monostearate 2.00 Self emulsifying monostearic acid glycerin 5.00 Cetyl alcohol 4.00 Squalene 6.00 Tri-2-ethylhexanoic acid glyceryl 6.00 Sphingoglycolipids 1.00 1,3-butylene glycol 7.00 Purified water Balance
  • Vitamin A acetate 70 ⁇ g Vitamin E 1.0 mg Vitamin B1 0.13 mg Vitamin B2 0.15 mg Vitamin B6 0.5 mg Vitamin B12 0.2 ⁇ g Vitamin C 10 mg Biotin 10 ⁇ g Nicotinic acid amide 1.7 mg Folic acid 50 ⁇ g Calcium pantothenate 0.5 mg Ferrous sulfate 1.75 mg Zinc oxide 0.82 mg Magnesium carbonate 25.3 mg Potassium monophosphate 15 mg Dicalcium phosphate 55 mg Potassium citrate 90 mg Calcium carbonate 100 mg Magnesium chloride 24.8 mg
  • Green tea extract 50 mg Citric acid 1000 mg oligosaccharide 100 g Taurine 1g Purified water Balance

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Abstract

Disclosed in the present specification is a composition comprising a green tea extract as an effective ingredient for treating fine dust-caused skin damage, wherein the expression of at least one selected from the group consisting of KRT 1, CLDN 1, CLDN 4, and OCLN, which are all genes having expression levels affected by fine dust in skin cells, is regulated to a normal level. By using the composition according to the present invention, gene expression changed by fine dust can return to a normal level to treat skin cell damage.

Description

녹차 추출물을 포함하는 미세먼지에 의한 피부 세포 손상 케어용 조성물Composition for care of skin cell damage by fine dust including green tea extract
본 명세서에는 미세먼지에 의한 피부 세포 손상을 케어하는 조성물이 개시된다. 보다 상세하게, 정상 상태의 피부 세포와 비교하여 미세먼지에 의해 발현 정도가 변화되는 피부 세포 유전자인 바이오마커 등을 유의미하게 변화시켜서 피부 세포 손상을 케어하는, 차나무 추출물을 포함하는 조성물이 개시된다.Disclosed herein are compositions for treating skin cell damage caused by fine dusts. More particularly, the present invention discloses a composition comprising a tea tree extract, wherein the skin cell damage is cared by significantly changing a biomarker, such as a skin cell gene whose degree of expression is changed by fine dust, as compared with a normal skin cell.
최근 환경적인 이슈로 미세먼지에 대한 관심이 높아지고 폴루션 시장이 확대되고 있다. 미세먼지는 일반적인 황사 등에 비하여 더 작은 크기의 입자를 갖기 때문에 인체에 더 잘 침투되며, 중금속, 질산염, 암모늄, 황산염 등의 유해물질을 더 많이 포함하고 있어서 인체에 유해하다. 미세먼지가 우리 몸 속으로 들어오면 인체의 면역을 담당하는 세포가 먼지를 제거하는 작용을 한다. 이 때의 부작용으로 염증을 수반하며, 이에 더하여 세계보건기구에서는 미세먼지를 암과 직접적인 관계가 있는 발암물질 1군으로 분류하고 있는 실정이다.Recently, environmental concerns have raised interest in fine dust and the polarization market is expanding. Fine dusts are smaller in size than normal dusts, so they penetrate the human body more and contain harmful substances such as heavy metals, nitrates, ammoniums, and sulfates, and are harmful to human body. When fine dust enters our body, the cells responsible for the immune function of the human body act to remove dust. These side effects are accompanied by inflammation. In addition, the World Health Organization (WHO) categorizes fine dusts as a group of carcinogens directly related to cancer.
미세먼지는 호흡기뿐만 아니라 피부나 모낭으로도 침투되어서 피부트러블이나 탈모를 일으키는 것으로 예상되나, 어떻게 피부와 모발에 부정적 영향을 끼치는지에 대한 기전 및 타겟 연구는 미비한 수준에 그치고 있으며, 이에 따라 문제점을 해결하기 위한 안티폴루션 소재에 대한 연구 또한 진행되고 있지 않다.It is expected that fine dust will penetrate not only the respiratory tract but also the skin or hair follicle, causing skin trouble or hair loss. However, the mechanism and the target research about how the skin and hair are adversely affected are still insufficient. There is also no research on antipolyge materials for this purpose.
비특허문헌 1은 과립층 장벽인 타이트 정션(tight junction)의 주요 구성성분인 클라우딘 1(CLDN 1)의 발현량이 PM 2.5의 미세먼지 자극으로 인하여 감소하였음을 개시하고 있다. Non-patent document 1 discloses that the expression amount of clodin 1 (CLDN 1), which is a major constituent of tight junction, which is a granule layer barrier, is reduced due to fine dust stimulation of PM 2.5.
선행기술문헌Prior art literature
비특허문헌Non-patent literature
(비특허문헌 1) Kim, H.J., et al, "Transcriptome analysis of airborne PM2.5-induced detrimental effects on human keratinocytes", Toxicology Letters 273, 26-35, 2017.(Non-Patent Document 1) Kim, HJ, et al., "Transcriptome analysis of airborne PM 2.5 -induced detrimental effects on human keratinocytes", Toxicology Letters 273, 26-35,
이에 본 발명자는 미세먼지가 피부에 유해한 영향을 미치며, 이러한 영향에 의하여 피부 세포 유전자의 발현에도 영향을 미침으로써 피부 세포 손상 등과 같은 증상이 나타나며, 녹차 추출물이 유전자의 발현을 복구시키는 효과가 있다는 것을 확인 하였다.Accordingly, the present inventors have found that the effect of fine dusts on the skin is detrimental to the skin, and the skin cell gene expression is affected by such an effect, thereby manifesting symptoms such as skin cell damage, and the green tea extract has an effect of restoring gene expression Respectively.
따라서, 일 측면에서 본 발명은 녹차 추출물을 포함하는 미세먼지에 의한 피부 세포의 손상 케어용 조성물을 제공하는 것을 목적으로 한다.Accordingly, it is an aspect of the present invention to provide a composition for care of damaging skin cells caused by fine dust containing a green tea extract.
상기한 목적을 달성하기 위하여, 일 측면에서, 본 발명은 녹차 추출물을 유효성분으로 포함하는 조성물로서, 미세먼지에 의해 발현량이 영향을 받는 피부 세포 내 유전자인 케라틴 1(KRT 1), 클라우딘 1(CLDN 1), 클라우딘 4(CLDN 4), 및 오클루딘(OCLN)으로 이루어진 군에서 선택되는 하나 이상의 발현량을 정상 수준으로 조절하는 미세먼지에 의한 피부 손상 케어용 조성물을 제공한다.In order to achieve the above object, in one aspect, the present invention provides a composition comprising an extract of green tea as an active ingredient, wherein keratin 1 (KRT 1), claudin 1 CLDN1), claudin 4 (CLDN4), and occludin (OCLN) to a normal level.
일 측면에서, 본 발명에 의해 제공되는 미세먼지에 의한 피부 손상 케어용 조성물을 이용함으로써, 미세먼지에 의해 변화되는 유전자 발현량을 정상 수준으로 되돌려 피부 세포의 손상을 케어할 수 있다.In one aspect, by using the composition for care of skin damage due to fine dust provided by the present invention, the amount of gene expression changed by the fine dust can be returned to a normal level and skin cell damage can be cared.
도 1는 ERM-CZ120 미세먼지에 의해 자극된 피부 세포 내에서 KRT 1 유전자의 mRNA 발현량이 증가하였고, 녹차 추출물 처리에 의해 정상 수준으로 되돌아감을 나타낸 것이다.Fig. 1 shows the expression of mRNA of KRT 1 gene in skin cells stimulated by ERM-CZ120 fine powder and showed a return to normal level by green tea extract treatment.
도 2는 ERM-CZ120 미세먼지에 의해 자극된 피부 세포 내에서 CLDN 1 유전자의 mRNA 발현량이 증가하였고, 녹차 추출물 처리에 의해 정상 수준으로 되돌아감을 나타낸 것이다.FIG. 2 shows an increase in mRNA expression of CLDN 1 gene in skin cells stimulated by ERM-CZ120 fine dust, and a return to a normal level by green tea extract treatment.
도 3는 ERM-CZ120 미세먼지에 의해 자극된 피부 세포 내에서 CLDN 4 유전자의 mRNA 발현량이 증가하였고, 녹차 추출물 처리에 의해 정상 수준으로 되돌아감을 나타낸 것이다.FIG. 3 shows that mRNA expression of CLDN4 gene was increased in skin cells stimulated by ERM-CZ120 fine powder and returned to normal level by green tea extract treatment.
도 4는 ERM-CZ120 미세먼지에 의해 자극된 피부 세포 내에서 OCLN 유전자의 mRNA 발현량이 증가하였고, 녹차 추출물 처리에 의해 정상 수준으로 되돌아감을 나타낸 것이다.Fig. 4 shows the increase in mRNA expression of OCLN gene in ERM-CZ120 micropore-stimulated skin cells and normalization by green tea extract treatment.
이하, 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 측면에서, 상기 미세먼지에 의한 피부손상 케어용 조성물은 녹차 추출물을 유효성분으로 포함할 수 있다.In one aspect of the present invention, the composition for care of skin damage due to fine dust may include green tea extract as an active ingredient.
차나무(Camellia sinensis)는　차나무과(theaceae)에 속하는 다년생 상록식물로서 열대, 아열대, 온대지방에 분포되고　차나무의 주요성분으로는 카테킨, 카페인, 아미노산, 비타민 및 무기질 등이 있으며, 이들 화학성분들은 여러 가지 생리활성과 약리효과를 나타내는 것으로 보고되고 있다. 오늘날 건강에 기여하는 생리활성물질로서 활발한 연구가 되고 있으며, 그 중에서도 카테킨류는 충치예방효과, 항산화효과, 항균효과와 항암효과 등의 다양한 기능이 있는 것으로 보고되고 있다. Camellia sinensis is a perennial evergreen plant belonging to theaceae. It is distributed in tropical, subtropical and temperate regions. The main ingredients of tea tree are catechins, caffeine, amino acids, vitamins and minerals. Have been reported to exhibit physiological activity and pharmacological effects. Recently, catechins have been reported to have various functions such as antifouling effect, antioxidant effect, antimicrobial effect and anticancer effect.
일 측면에서, 상기 녹차 추출물은 녹차 잎 추출물이고, 상기 녹차 잎은 발효되지 않은 것으로서, 내부 효소가 불활성된 것일 수 있다.In one aspect, the green tea extract is green tea leaf extract, the green tea leaves are not fermented, and the inner enzyme is inactive.
본 발명의 일 측면에서, 상기 녹차는 특정 추출용매로 추출되어 녹차 추출물을 형성할 수 있다.In one aspect of the present invention, the green tea may be extracted with a specific extraction solvent to form a green tea extract.
본 발명의 일 측면인, 상기 녹차 추출물은 녹차를 물 또는 유기용매로 추출하여 제조될 수 있다. 구체적으로, 녹차를 물, C1 -C6의 무수 또는 함수 저급 알코올, 아세톤, 부틸렌글리콜, 에틸아세테이트, 디에틸아세테이트, 디에틸에테르, 벤젠, 클로로포름, 및 헥산으로 이루어진 군에서 선택된 어느 하나 이상의 추출용매로 추출하여 제조될 수 있다.　In one aspect of the present invention, the green tea extract may be prepared by extracting green tea with water or an organic solvent. Specifically, the green tea is prepared by dissolving at least one compound selected from the group consisting of water, C 1 -C 6 anhydrous or lower alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform, And extracting it with an extraction solvent.
일 측면에서, 상기 녹차 추출물은 상온 추출된 것일 수 있다.In one aspect, the green tea extract may be extracted at room temperature.
일 측면에서, 상기 녹차 추출물은 상기 추출용매로 추출된 이후에, 증발, 여과, 농축, 분리 또는 건조 과정 중 하나 이상을 추가적으로 거쳐서 얻어진 것일 수 있다. 특히, 상기 녹차 추출물은 1회 이상의 여과 과정을 거친 것일 수 있다.In one aspect, the green tea extract may be one obtained by extracting with the extraction solvent, followed by addition of one or more of evaporation, filtration, concentration, separation or drying. In particular, the green tea extract may be subjected to one or more filtration processes.
일 측면에서, 상기 분리 과정은 원심분리 과정을 포함할 수 있다.In one aspect, the separation process may include a centrifugation process.
구체적으로, 상기 추출은 물, C1 - C6의 무수 또는 함수 저급 알코올, 아세톤 및 부틸렌글리콜을 포함하는 극성 용매 및 에틸아세테이트, 디에틸아세테이트, 디에틸에테르, 벤젠, 클로로포름 및 헥산을 포함하는 저극성 용매 중 어느 하나 이상을 용매로 사용하는 것일 수 있다. 더 구체적으로, 상기 용매는 10 - 90%의 에탄올 수용액일 수 있으며, 10 - 50%, 15 - 45%, 또는 20-40%의 에탄올 수용액일 수 있다. 상기 용매가 20 - 40%의 에탄올 수용액인 경우, 녹차로부터 유효성분을 효과적으로 추출할 수 있다. 일 실시예에서, 상기 용매는 약 30%의 에탄올 수용액이다.Specifically, the extraction can be carried out in the presence of water, a C 1 -C 6 anhydrous or a lower alcohol, a polar solvent comprising acetone and butylene glycol, and a polar solvent including ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane And a low polarity solvent may be used as a solvent. More specifically, the solvent may be 10-90% ethanol aqueous solution, and may be 10-50%, 15-45%, or 20-40% ethanol aqueous solution. When the solvent is 20 to 40% aqueous ethanol solution, the effective ingredient can be effectively extracted from the green tea. In one embodiment, the solvent is about 30% aqueous ethanol.
일 측면에서, 상기 추출물은 추출 후 냉각 콘덴서가 달린 증류 장치에서 적정 온도로 감압 농축될 수 있다.In one aspect, the extract may be concentrated under reduced pressure to an appropriate temperature in a distillation apparatus equipped with a cooling condenser after extraction.
다만, 본 발명에 따른 녹차 추출물은 당해 기술 분야의 통상적인 방법으로 추출하여 얻을 수 있으며, 전술한 방법에 의하여 한정되는 것은 아니다.However, the green tea extract according to the present invention can be extracted by a conventional method in the art, and is not limited by the above-described method.
본 발명의 일 관점인, 조성물에 있어서, 상기 조성물은 녹차 추출물을, 조성물 총 중량을 기준으로 0.000001 내지 30중량%로 포함할 수 있다. 그 함량이 0.000001 내지 30중량%인 경우, 녹차 추출물에 의한 미세먼지에 의한 피부 손상 케어 효과가 우수하였다.In one aspect of the invention, the composition may comprise from 0.000001 to 30% by weight of green tea extract, based on the total weight of the composition. When the content thereof is 0.000001 to 30% by weight, the effect of care for skin damage due to fine dust caused by green tea extract is excellent.
구체적으로, 0.0000001 중량% 이상, 0.0000005 중량% 이상, 0.0000007 중량% 이상, 0.0000009 중량% 이상, 0.000001 중량% 이상, 0.000002 중량% 이상, 0.000004 중량% 이상, 0.000006 중량% 이상, 0.000008 중량% 이상, 0.00001 중량% 이상, 0.00003 중량% 이상, 0.00005 중량% 이상, 0.00007 중량% 이상, 0.00009 중량% 이상, 0.0001 중량% 이상, 0.0003 중량% 이상, 0.0005 중량% 이상, 0.0007 중량% 이상, 0.0009 중량% 이상, 0.001 중량% 이상, 0.01 중량% 이상, 0.1 중량% 이상, 1 중량% 이상, 3 중량% 이상, 5 중량% 이상, 7 중량% 이상, 9 중량% 이상, 10 중량% 이상, 13 중량% 이상, 15 중량% 이상, 17 중량% 이상, 19 중량% 이상, 21 중량% 이상, 23 중량% 이상, 25 중량% 이상, 27 중량% 이상, 29 중량% 이상, 30 중량% 이상, 31 중량% 이상일 수 있고, 32 중량% 이하, 31 중량% 이하, 30 중량% 이하, 29 중량% 이하, 28 중량% 이하, 26 중량% 이하, 24 중량% 이하, 22 중량% 이하, 20 중량% 이하, 18 중량% 이하, 16 중량% 이하, 14 중량% 이하, 12 중량% 이하, 10 중량% 이하, 9 중량% 이하, 8 중량% 이하, 6 중량% 이하, 4 중량% 이하, 2 중량% 이하, 1 중량% 이하, 0.1 중량% 이하, 0.09 중량% 이하, 0.04 중량% 이하, 0.01 중량% 이하, 0.006 중량% 이하, 0.001 중량% 이하, 0.0009 중량% 이하, 0.0007 중량% 이하, 0.00005 중량% 이하, 0.00003 중량% 이하, 0.00001 중량% 이하, 0.000009 중량% 이하, 0.000007 중량% 이하, 0.000005 중량% 이하, 0.000003 중량% 이하, 0.000001 중량% 이하, 0.0000009 중량% 이하, 0.0000007 중량% 이하, 0.0000005 중량% 이하, 0.0000003 중량% 이하, 0.0000002 중량% 이하, 0.0000001 중량% 이하, 0.00000009 중량% 이하일 수 있으나, 이에 제한되는 것은 아니다.More specifically, 0.0000001 wt% or more, 0.0000005 wt% or more, 0.0000007 wt% or more, 0.0000009 wt% or more, 0.0000009 wt% or more, 0.000001 wt% or more, 0.000002 wt% or more, 0.000004 wt% or more, 0.000006 wt% or more, 0.000008 wt. At least 0.00003% by weight, at least 0.00005% by weight, at least 0.00007% by weight, at least 0.00009% by weight, at least 0.00009% by weight, at least 0.0001% by weight, at least 0.0003% by weight, at least 0.0005% by weight, at least 0.0007% by weight, % Or more, 0.01 wt% or more, 0.1 wt% or more, 1 wt% or more, 3 wt% or more, 5 wt% or more, 7 wt% or more, 9 wt% or more, 10 wt% or more, 13 wt% or more, Or more, 17 wt% or more, 19 wt% or more, 21 wt% or more, 23 wt% or more, 25 wt% or more, 27 wt% or more, 29 wt% or more, 30 wt% or more, 31 wt% 32 weight% or less, 31 weight% or less, 30 weight% or less, 29 weight% or less, 28 weight% or less, 26 weight% or less, 24 weight % Or less, 22 wt% or less, 20 wt% or less, 18 wt% or less, 16 wt% or less, 14 wt% or less, 12 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% % Or less, 4 wt% or less, 2 wt% or less, 1 wt% or less, 0.1 wt% or less, 0.09 wt% or less, 0.04 wt% or less, 0.01 wt% or less, 0.006 wt% or less, 0.001 wt% 0.000003 wt.% Or less, 0.00003 wt.% Or less, 0.00001 wt.% Or less, 0.00001 wt.% Or less, 0.000009 wt.% Or less, 0.000007 wt.% Or less, 0.000005 wt. %, 0.0000007 wt% or less, 0.0000005 wt% or less, 0.0000003 wt% or less, 0.0000002 wt% or less, 0.0000001 wt% or less, or 0.00000009 wt% or less.
일 측면에서, 본 발명에 따른 조성물의 유효 성분은 피부 장벽의 타이트 정션(tight junction)에 작용하는 것일 수 있다.In one aspect, the active ingredient of the composition according to the present invention may be one which acts on the tight junction of the skin barrier.
밀착연접은 세포연접의 한 종류로 occluding, claudin, tricellulin, junctional adhesion molecule(JAM), zona occluden(ZO) 등으로 구성되어 있다. 밀착연접은 세포막에 포함된 부분과 세포내 부분으로 나눌 수 있다. Occludin과 claudin은 세포막에 포함된 밀착연접 구성 단백질이다. Occludin은 밀착연접에서 처음으로 발견된 막관통 단백질로 4개의 막관통 영역을 가지고 있다. Occludin은 밀착연접의 구성 성분일 뿐만 아니라 밀착연접의 기능을 조절하는 것으로 알려졌다. Occludin과 함께 밀착연접 구성 단백질인 claudin도 4개의 막관통 영역을 가지고 있다. 밀착연접은 세포 간의 연결고리로 이웃한 세포와 세포 사이의 간격을 메워주고 작은 물질들의 이동을 조절하는 기능을 하며, 세포와 세포 간의 물질투과를 조절하고 세포의 극성을 유지하는 역할도 담당한다. 최근, 연구결과를 통해 밀착연접이 피부 장벽 기능에 있어서 중요한 역할을 담당하고 있다고 보고되었다. 타이트 정션(tight junction)은 세포간 결합양식의 일종이다. 주로 상피세포와 내피세포에서 나타나는 세포간 연접으로서, 끈모양 구조를 갖는 복잡한 네트워크(network)를 형성한다. 타이트 정션은 이러한 구조로부터 크게 두가지 기능을 갖는다. 첫째는 격벽(barrier)기능으로, 상피세포나 내피세포를 구성하는 세포 사이로 물질의 누출을 방지한다. 주로 혈관이나 소화관에서 중요하게 다루어진다. 둘째는 울타리기능으로, 세포막 지질에서 끈모양 울타리를 둘러 세포막을 구획하는 기능이다. 따라서, 본 발명에 따른 조성물의 유효 성분은 피부 장벽의 타이트 정션에 작용하여 피부 세포의 격벽 기능 및 울타리 기능을 강화시켜 미세먼지 자극에 의하여 손상된 피부를 케어할 수 있다.The tight junction is a kind of cell junction, consisting of claudin, tricellulin, junctional adhesion molecule (JAM) and zona occluden (ZO). The tight junction can be divided into a part contained in the cell membrane and an intracellular part. Occludin and claudin are closely related constitutive proteins contained in cell membranes. Occludin is the first transmembrane protein found in adherent synapses and has four transmembrane domains. Occludin is known to regulate the function of tight junctions as well as constituents of tight junctions. Claudin, a close-coupled conformational protein with Occludin, also has four transmembrane domains. The tight junction is a cell-to-cell linkage that fills the gap between neighboring cells, controls the movement of small molecules, controls the permeation of cells between cells, and maintains the polarity of the cells. Recently, it has been reported that tight junctions play an important role in skin barrier function. Tight junctions are a type of intercellular binding. It is a cell-to-cell junction that mainly occurs in epithelial cells and endothelial cells, forming a complex network with a string-like structure. The tight junction has two functions largely from this structure. The first is a barrier function that prevents material from leaking into the cells that make up epithelial cells or endothelial cells. It is mainly treated in blood vessels and digestive tracts. The second is a fence function, which is a function to divide the cell membrane around the membrane fence in the cell membrane lipid. Accordingly, the active ingredient of the composition of the present invention acts on the tight junction of the skin barrier, thereby enhancing the barrier function and the fence function of the skin cells, thereby protecting the skin damaged by the fine dust stimulation.
본 발명의 다른 측면은, 상기 조성물의 미세먼지에 의한 피부 손상 케어 용도를 포함한다.Another aspect of the present invention includes skin care care applications for fine dusts of the composition.
본 발명의 다른 측면에서, 대상의 미세먼지에 의한 피부 손상 케어를 위한 방법으로서, 상기 방법은 녹차 추출물의 유효량을 이를 필요로 하는 대상에 투여하는 단계를 포함하는 방법을 제공한다.In another aspect of the present invention there is provided a method for skin damage care by fine dust of a subject comprising administering an effective amount of a green tea extract to a subject in need thereof.
본 발명의 다른 측면에서 미세먼지에 의한 피부 손상 케어를 위한 조성물을 제조하는데 있어서의 녹차 추출물의 용도를 제공한다.Another aspect of the present invention provides the use of green tea extract in the preparation of a composition for skin damage care by fine dust.
본 발명의 다른 측면에서 미세먼지에 의한 피부 손상 케어를 위한 녹차 추출물을 제공한다.In another aspect of the present invention, there is provided a green tea extract for skin damaging treatment by fine dust.
본 명세서에서 사용되는 "미세먼지"라 함은, 우리 눈에 보이지 않는 아주 작은 물질로 대기 중에 오랫동안 떠다니거나 흩날리는 입자상의 물질을 의미한다. 구체적으로, 본 발명에서 미세먼지의 입자 크기는 입경 10μm 이하(PM 10), 구체적으로 입경 2.5μm 이하(PM 2.5)일 수 있다. 특히 입경이 2.5μm(PM 2.5) 이하인 입자상의 물질은 "초미세먼지"라 하는데, 본 명세서에서 "미세먼지"는 "초미세먼지"도 포함하는 것으로 의도된다.As used herein, the term " fine dust " refers to a particulate matter that is a very small material that is invisible to the naked eye and that floats or drifts in the air for a long time. Specifically, the particle size of fine dust in the present invention may be not more than 10 μm (PM 10), specifically not more than 2.5 μm (PM 2.5). Particulate matter having a particle diameter of not more than 2.5 μm (PM 2.5) is referred to as "ultrafine dust". In the present specification, "fine dust" is also intended to include "ultrafine dust".
일 측면에서, 본 발명에서 미세먼지는 비소(arsenic), 카드뮴(cardmium), 납(lead), 및 니켈(nikel) 중 하나 이상을 포함할 수 있다. 상기 비소(arsenic), 카드뮴(cardmium), 납(lead), 및 니켈(nikel) 등의 중금속 성분은 미세먼지에 함께 포함되어 피부 세포를 공격해 피부 알러지, 색소 침착, 염증 등을 일으켜 피부노화를 유발시킬 수 있다. 일 실시예에서, 상기 미세먼지는 비소(arsenic), 카드뮴(cardmium), 납(lead), 및 니켈(nikel)을 포함한다.In one aspect, the fine dust in the present invention may include one or more of arsenic, cardmium, lead, and nickel. The heavy metal components such as arsenic, cardmium, lead, and nickel are included in the fine dust to attack skin cells and cause skin allergy, pigmentation, inflammation, . In one embodiment, the fine dust includes arsenic, cardmium, lead, and nickel.
본 명세서에서 사용되는 용어"케어"라 함은, 자극으로부터 피부 세포를 효과적으로 보호하고, 상기 자극에 의하여 특정 유전자의 발현량이 변화하는 것을 억제, 방지, 복구(복원)시키는 것을 의미한다. 여기서, "복구"는 자극이 없는 경우의 유전자 발현량을 기준으로 할 때, 발현량을 상기 기준에 가깝게 변화시키는 것 외에도 상기 기준 이상으로 발현량을 향상시키는 것을 또한 포함한다.As used herein, the term " care " means to effectively protect skin cells from stimulation and to inhibit, prevent, and restore (restore) the expression level of a specific gene by the stimulation. Here, " repair " also includes improving the expression level beyond the reference, in addition to changing the expression level closer to the reference level based on the gene expression level in the absence of the stimulus.
일 측면에서, 본 발명은 미세먼지에 의해 손상된 피부 세포에서 특정 유전자의 발현량을 정상 수준으로 조절함으로써, 미세먼지에 의한 피부 세포의 손상을 억제하는 조성물을 제공한다.In one aspect, the present invention provides a composition for inhibiting damage of skin cells caused by fine dusts by controlling the expression level of a specific gene in skin cells damaged by fine dust to a normal level.
구체적으로, 본 발명에서 미세먼지에 의해 발현량이 영향을 받는 피부 세포 내 유전자로는 케라틴 1(KRT 1), 클라우딘 1(CLDN 1), 클라우딘 4(CLDN 4), 및 오클루딘(OCLN)으로 이루어진 군에서 선택되는 하나 이상을 포함할 수 있다. 상기 케라틴 1(KRT 1), 클라우딘 1(CLDN 1), 클라우딘 4(CLDN 4), 및 오클루딘(OCLN)은 미세먼지에 의해 발현량이 감소하는 유전자이므로, 이들 유전자의 발현량을 억제하여 정상 수준으로 조절함으로써 피부 세포의 손상을 억제할 수 있다.Specifically, in the present invention, genes in the skin cells to which the expression amount by the fine dust is affected include keratin 1 (KRT 1), cladin 1 (CLDN 1), cladin 4 (CLDN 4), and occludin And at least one selected from the group. Since keratin 1 (KRT 1), claudin 1 (CLDN 1), claudin 4 (CLDN 4), and occludin (OCLN) are genes whose expression amount is reduced by fine dusts, To inhibit skin cell damage.
본 발명에서 사용되는, 미세먼지에 의해 발현량이 변화하는 유전자가 아래 [표 1]에 제시되어 있다. [표 1]은 미세먼지에 의해 발현량이 감소되는 유전자를 나타내는 것이며, 이들 표에서 Gene Symbol은 공식 유전자 심볼을 의미하며, Gene title은 각 유전자의 이름을 의미한다. 이러한 내용은 비특허문헌 1에 기재를 통하여 확인할 수 있다.The genes whose expression levels are changed by fine dusts used in the present invention are shown in Table 1 below. [Table 1] shows a gene whose expression amount is reduced by fine dust. In these tables, Gene Symbol means the official gene symbol and Gene title means the name of each gene. Such contents can be confirmed through the description in Non-Patent Document 1.
감소 유전자Reduction gene
GeneSymbolGeneSymbol Gene title Gene title
KRT 1KRT 1 Keratin 1 Keratin 1
CLDN 1 CLDN 1 Claudin 1 Claudin 1
CLDN 4 CLDN 4 Claudin 4 Claudin 4
OCLNOCLN OccludinOccludin
상기 유전자 또는 단백질의 발현량 분석은 마이크로어레이, PCR, NGS(Nest Generation Sequencing; 차세대 염기서열분석), 웨스턴 블럿, 노던 블럿, ELISA, 방사선 면역 분석, 방사 면역 확산법, 조직면역 염색, 면역침전 분석법 등 당업계에 공지된 다양한 분석 방법을 이용하여 분석될 수 있다.Analysis of the expression level of the gene or protein can be performed using a microarray, PCR, NGS (Nest Generation Sequencing), Western blot, northern blot, ELISA, radioimmunoassay, radioimmunoassay, tissue immuno staining, Can be analyzed using a variety of analytical methods known in the art.
미세먼지에 의해 피부 세포의 손상이 일어나며, 이 때 염증이 유도되어 다시 피부 세포의 손상이 추가로 일어나게 된다. 이렇게 피부 세포 손상의 악순환을 녹차 추출물이 케어함으로써 피부 상태를 개선시킬 수 있게 된다.Damage to the skin cells is caused by fine dust, which leads to inflammation and further damage to the skin cells. Thus, the skin condition can be improved by taking care of the vicious circle of the skin cell damage by the green tea extract.
본 발명의 일 관점인, 상기 조성물은, 화장료 조성물일 수 있고, 약학적 조성물일 수 있으며, 건강기능식품 조성물일 수 있다. In one aspect of the present invention, the composition may be a cosmetic composition, a pharmaceutical composition, or a health functional food composition.
상기 화장료 조성물은, 예컨대, 각종 크림, 로션 각종 크림, 로션, 스킨 등과 같은 화장품 류와 클렌징, 세안제, 비누, 미용액 등이 있다.Examples of the cosmetic composition include cosmetics such as various creams, lotion creams, lotions, skins, and the like, and cleansing, cleansing agents, soaps, and essences.
일 측면에서, 본 발명의 상기 녹차 추출물을 함유하는 조성물이 첨가된 화장료는 용액, 유화물, 점성형 혼합물 등의 형상을 취할 수 있다.In one aspect, the cosmetic composition to which the composition containing the green tea extract of the present invention is added may take the form of a solution, an emulsion, a viscous mixture or the like.
즉, 일 측면에서 본 발명의 화장료는 그 제형에 있어서 특별히 한정되지 않으며, 예를 들어 유액, 크림, 화장수, 에센스, 팩, 젤, 파우더, 메이크업 베이스, 파운데이션, 로션, 연고, 패취, 미용액, 클렌징폼, 클렌징크림, 클렌징워터, 바디로션, 바디크림, 바디오일, 바디에센스, 샴푸, 린스, 바디세정제, 비누, 염모제, 분무제 등과 같은 제형을 들 수 있다.That is, in one aspect, the cosmetic of the present invention is not particularly limited in its formulation, and examples thereof include an emulsion, cream, lotion, essence, pack, gel, powder, makeup base, foundation, lotion, ointment, patch, Formulations such as foam, cleansing cream, cleansing water, body lotion, body cream, body oil, body essence, shampoo, rinse, body cleanser, soap, hair dye, spray and the like.
각 제형의 화장료 조성물에 있어서, 상기 녹차 추출물 이외에 다른 성분들은 기타 화장료의 제형 또는 사용 목적에 따라 당업자가 어려움 없이 적의 선정하여 배합할 수 있다.In the cosmetic composition of each formulation, the components other than the green tea extract may be mixed and selected without difficulty by those skilled in the art depending on the formulation or use of the other cosmetic ingredients.
또한, 일 측면에서 본 발명의 화장료는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어진 군에서 선택된 조성물을 포함할 수 있다.In addition, in one aspect, the cosmetic of the present invention may comprise a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.
일 측면에서, 본 발명의 화장료에는 상기 필수 성분과 더불어 필요에 따라 통상 화장료에 배합되는 다른 성분을 배합해도 된다.In one aspect, the cosmetic composition of the present invention may contain, in addition to the above essential ingredients, other ingredients usually added to cosmetics, if necessary.
이외에 첨가해도 되는 배합 성분으로서는 유지 성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및 무기 안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조정제, 알콜, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다.Examples of the compounding ingredients that may be added include organic solvents such as a preservative component, a moisturizer, an emollient, a surfactant, an organic and inorganic pigment, an organic powder, an ultraviolet absorbent, a preservative, a bactericide, an antioxidant, a plant extract, a pH adjuster, A blood circulation accelerator, a cold agent, an antiperspirant agent, and purified water.
또한, 이외에 첨가해도 되는 배합 성분은 이에 한정되는 것은 아니며, 또, 상기 어느 성분도 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 배합 가능하다.The components to be added in addition to these components are not limited thereto, and any of the above components can be compounded within a range that does not impair the objects and effects of the present invention.
일 측면에서, 본 발명의 녹차 추출물을 포함하는 약학적 조성물은 약학조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.In one aspect, the pharmaceutical compositions comprising the green tea extract of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions.
그 제형으로는, 상기 녹차 추출물을 포함하는 약학적 조성물은 각각 통상의 방법에 따라 정제, 캡슐, 산제 또는 시럽 등의 경구제, 또는 연고, 겔, 크림, 패취 또는 분무제 등의 외용제 등을 비롯하여 약제학적 제제에 적합한 어떠한 형태로든 제형화하여 사용할 수 있다.The pharmaceutical composition containing the green tea extract may be formulated into tablets, capsules, powders or syrups, or external preparations such as ointments, gels, creams, patches or sprays, And may be formulated and used in any form suitable for a clinical formulation.
일반적으로 상기 약학적 조성물에 의해 투여되는 유효성분의 실제 투여량은 증상의 중증도, 선택된 투여 경로, 대상의 연령, 성별, 체중 및 건강상태 등의 여러 관련 인자에 비추어 결정되어야 하는 것으로 이해되어야 한다. 일반적으로 유효성분의 투여량은 0.0001mg/kg/일 내지 3000mg/kg/일, 예를 들어 10 mg/kg/일 내지 500mg/kg/일 일 수 있다.In general, it is to be understood that the actual dosage of the active ingredient administered by the pharmaceutical composition should be determined in light of various relevant factors such as the severity of the symptoms, the route of administration selected, the age, sex, weight and health status of the subject. In general, the dosage of the active ingredient may be from 0.0001 mg / kg / day to 3000 mg / kg / day, for example from 10 mg / kg / day to 500 mg / kg / day.
본 발명의 일 관점인 건강 기능식품 조성물에서, 건강식품은, 일상 식사에서 결핍되기 쉬운 영양소나 인체에 유용한 기능을 가진 원료나 성분(기능성원료)을 사용하여 제조한 것으로, 인체의 정상적인 기능을 유지하거나 생리기능 활성화를 통하여 건강을 유지하고 개선하는 식품을 의미할 수 있으나, 이에 제한되지 않는다. 상기 건강식품은 정제, 캡슐, 분말, 과립, 액상, 환 등의 형태로 제조, 가공될 수 있으나, 이에 한정되지 않으며 법률에 따라 어떤 형태로든지 제조, 가공될 수 있다.In the health functional food composition as one aspect of the present invention, the health food is produced by using a raw material or a component (functional raw material) having a function useful for a nutrient or a human body which is likely to be deficient in a daily meal, Or to maintain or improve health through the activation of physiological functions. However, the present invention is not limited thereto. The health food may be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, and circles, but is not limited thereto and may be manufactured and processed in any form according to the law.
구체적으로, 건강 음료 조성물은 지시된 비율로 필수 성분으로서 상기 화합물을 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 천연 탄수화물의 예는 모노사카라이드 폴리사카라이드, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제 (타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제 (사카린, 아스파르탐 등)를 사용할 수 있다.Specifically, the health beverage composition has no particular limitation on the other ingredients other than the above-mentioned compounds as essential ingredients in the indicated ratios, and may contain various flavors or natural carbohydrates as additional ingredients such as ordinary beverages. Examples of natural carbohydrates are conventional sugars such as monosaccharide polysaccharides, cyclodextrins and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavors (tau martin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.) and synthetic flavors (saccharin, aspartame, etc.) can be used as flavorings other than those described above.
일반적으로 상기 건강 기능식품 조성물에 의해 투여되는 유효성분의 실제 투여량은 증상의 중증도, 선택된 투여 경로, 대상의 연령, 성별, 체중 및 건강상태 등의 여러 관련 인자에 비추어 결정되어야 하는 것으로 이해되어야 한다. 일반적으로 유효성분의 투여량은 0.0001mg/kg/일 내지 1000mg/kg/일, 예를 들어 0.02 mg/kg/일 내지 6mg/kg/일 일 수 있다.In general, it should be understood that the actual dosage of the active ingredient administered by the health functional food composition should be determined in light of various relevant factors such as the severity of the symptoms, the selected route of administration, the age, sex, weight and health status of the subject . In general, the dosage of the active ingredient may be from 0.0001 mg / kg / day to 1000 mg / kg / day, for example from 0.02 mg / kg / day to 6 mg / kg / day.
이하, 실시예를 들어 본 발명의 구성 및 효과를 보다 구체적으로 설명한다. 그러나 이들 실시예는 본 발명에 대한 이해를 돕기 위해 예시의 목적으로만 제공된 것일 뿐 본 발명의 범주 및 범위가 하기 예에 의해 제한되는 것은 아니다.Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to examples. However, these examples are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited by the following examples.
[실시예 1] 녹차 추출물 제조[Example 1] Preparation of green tea extract
녹차 추출물은 아모레퍼시픽 그룹 ㈜ 장원으로부터 분양 받은 녹차 잎(green tea)을 사용하여 제조하였다. 채취한 녹차 잎은 정제수로 세척하고 여과 및 건조시킨 뒤, 30% 에탄올(EtOH)와 1:8의 7중량 비율로 혼합한 뒤 25 ℃에서 72시간 교반하여 250 메쉬(mesh) 사이즈로 추출하였다.Green tea extracts were prepared using green tea from Amorepacific Group Changwon. The collected green tea leaves were washed with purified water, filtered and dried, and then mixed in a weight ratio of 1: 8 with 30% ethanol (EtOH). The mixture was stirred at 25 ° C for 72 hours and extracted into 250 mesh size.
얻어진 추출액은 3차례의 여과과정을 거치는데 각각 3.0 ㎛ 이하 사이즈, 1.0㎛ 이하 사이즈, 및 0.3㎛ 이하 사이즈로 여과하였다. 여과된 추출액을 4 ℃에서 72시간동안 침전시킨 뒤, 다시 0.2 ㎛ 이하 사이즈로 여과시켜 녹차 추출물을 제조하였다.The obtained extract was subjected to three filtration steps, each of which had a size of 3.0 μm or less, a size of 1.0 μm or less, and a size of 0.3 μm or less. The filtered extract was settled at 4 DEG C for 72 hours, and then filtered again to a size of 0.2 mu m or less to prepare a green tea extract.
[실험예 1] 실시간 qPCR 정량을 통한 유전자 발현량 측정 실험[Experimental Example 1] Measurement of gene expression amount by real-time qPCR quantification Experiment
1. 각질형성세포주의 배양1. Culture of keratinocyte cell line
각질형성세포주(Human normal epidermal keratinocytes)는 론자 社(Lonza, Inc. 미국 메릴랜드주 워커스빌 소재)에서 구입하여 계대배양한 후 CO2 배양기(CO2 incubator)에서 37℃, 5% CO2 조건 하에서 배양하였다. 500ml의 KBM-2(KBMTM-2, CC-3103) 배지에 BPE(Bovine pituitary extract), 인간표피 성장인자(human epidermal growth factor, hEGF), 인슐린(Insulin), 하이드로코티손(Hydrocortisone), 젠타마이신 설페이트(Gentamycin Suflate), 에피네프린(Epinephrine), 트랜스페린(Transferrin)을 포함한 배양액을 사용하였다.Keratinocyte cell line (Human normal epidermal keratinocytes) was Lonza社(Lonza, Inc. Walkersville, Maryland material) was purchased and subcultured in a culture under a CO 2 incubator (CO 2 incubator), 37 ℃ from, 5% CO 2 conditions Respectively. (Bovine pituitary extract (BPE), human epidermal growth factor (hEGF), insulin, hydrocortisone, gentamicin sulfate, and the like were added to 500 ml of KBM-2 (KBMTM-2, CC- (Gentamycin Suflate), Epinephrine, and Transferrin were used.
배양시 최대 90% 정도의 컨플루언시(confluency)를 유지하였으며, 그 이후에는 계대배양(subcultivation)하였다.Confluency of up to 90% was maintained during culture, and then subcultivation was performed.
2. 각질형성세포주에 미세먼지 및 녹차 추출물 처리2. Treatment of keratinocyte cell line with fine dust and green tea extract
미세먼지는 시그마-알드리치 社(Sigma-Aldrich)에서 시판하는 ERM-CZ120 미세먼지를 사용하였으며, PM 10 수준의 미세먼지와 비소(arsenic), 카드뮴(cardmium), 납(lead), 및 니켈(nikel)을 포함하고 있다.The fine dusts were ERM-CZ120 fine dusts commercially available from Sigma-Aldrich, and PM 10 levels of fine dust and arsenic, cardmium, lead, and nickel ).
6-웰 플레이트를 사용하였고 '1. 각질형성세포주의 배양'에서의 세포배양조건으로 3 Х 105 세포수/웰(cells/well)인 조건으로 배양한 세포주에 각각 대조군으로 미처리군(대조군 1), 미세먼지를 50 ppm 처리한 군(대조군 2), 및 미세먼지 및 녹차 추출물을 각각 50 ppm 처리한 군(실시예 1)으로 총 3개의 군을 물질 처리를 하여 3 시간 및 6 시간 동안 배양하였다.A 6-well plate was used and 1 ' Cells cultured under the conditions of 3 × 10 5 cells / well (cells / well) were cultured in the untreated group (control 1), 50 ppm of fine dusts (Control group 2), and a group treated with 50 ppm of fine dust and green tea extract (Example 1), respectively, and then cultured for 3 hours and 6 hours.
실험군Experimental group 미세먼지fine dust (( ERMERM -CZ120 Fine Dust)-CZ120 Fine Dust) 녹차 추출물Green tea extract
대조군 1(미세먼지 미처리)Control group 1 (untreated fine dust) -- --
대조군 2(미세먼지 처리)Control group 2 (fine dust treatment) 50 ppm50 ppm --
실시예 1(미세먼지+녹차 추출물 처리)Example 1 (Fine dust + green tea extract treatment) 50 ppm50 ppm 50 ppm50 ppm
3. 실시간 qPCR 정량3. Real-time qPCR quantitation
3-1. RNA 추출 과정3-1. RNA extraction process
'2. 각질형성세포주에 미세먼지 및 녹차 추출물 처리'에서 얻은 대조군 1, 대조군 2 및 실시예 1에서 배양액을 제거하고, 2ml의 인산염 완충액(Phosphate Buffered Saline, PBS)으로 세포를 세척한 다음, 트리졸 시약(Trizol reagent, Invitrogen, Carlsbad, CA, USA)을 사용하여 세포 내의 RNA를 분리하였다.'2. The cells were washed with 2 ml of phosphate buffered saline (PBS) in Control 1, Control 2 and Example 1 obtained in the treatment of fine dust and green tea extract on keratinocyte cell line, Trizol reagent, Invitrogen, Carlsbad, CA, USA).
3-2. cDNA 합성 과정3-2. cDNA synthesis process
분리된 RNA를 키아젠사의 RNA 키트(QIAGEN RNeasy kt, QIAGEN, Valencia, CA)로 한번 더 정제한 후, 애질런트 社의 바이오어낼라이저 2100 모델 기기(Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, CA, USA)를 사용하여 RNA의 질(quality)을 확인하였다. 인비트로젠사의 역전사키트(Superscript Reverse Transcriptase (RT) kit, Invitrogen, Carlsbad, CA)를 이용하여 RNA로부터 cDNA를 합성하였다.The separated RNA was further purified with an RNA kit (QIAGEN RNeasykt, QIAGEN, Valencia, Calif.) From Agilent Technologies, Inc., Agilent 2100 BioAnalyzer, Agilent Technologies, Santa Clara, ) Was used to confirm the quality of the RNA. CDNA was synthesized from RNA using Invitrogen's Reverse Kit (Superscript Reverse Transcriptase (RT) kit, Invitrogen, Carlsbad, Calif.).
3-3. qPCR 수행3-3. Perform qPCR
이를 상기 [표 3]의 프라이머를 이용한 실시간 역전사 중합 효소 연쇄반응(Q-RT-PCR: real time-reverse transcription polymerase chain reaction)을 통해 정량적으로 분석하였다. 유전자의 발현 패턴 변화를 어플라이드바이오시스템사의 택맨 유전자 발현 시스템(TaqMan gene expression assay kit, Applied Biosystems, Foster City, CA)을 이용하여 세포의 유전자 변화를 실시간 PCR로 평가하였으며, 그 결과를 [도 1] 내지 [도 4]에 나타내었다. 이용한 Q-RT-PCR과 실시간 PCR은 모두 라이프테크놀로지에서 배포하는 표준 프로토콜에 따라서 실행하였으며, 구체적으로 95℃에서 20초 동안 처리한 후, 95℃에서 3초 및 60℃에서 30초를 처리하는 공정을 40주기 진행하였다.This was quantitatively analyzed by real-time reverse transcription polymerase chain reaction (Q-RT-PCR) using the primers of Table 3 above. The change in the expression pattern of the gene was evaluated by real-time PCR using the TaqMan gene expression assay kit (Applied Biosystems, Foster City, Calif.) Of Applied Biosystems. To [Figure 4]. Both Q-RT-PCR and real-time PCR were performed according to the standard protocols distributed by Life Technologies. Specifically, the Q-RT-PCR was performed at 95 ° C for 20 seconds, followed by 95 ° C for 3 seconds and 60 ° C for 30 seconds For 40 cycles.
[도 1] 내지 [도 4]에 나타낸 바와 같이, 미세먼지를 50 ppm 농도로 처리한 대조군 2는 미세먼지를 미처리한 대조군 1에 비하여 케라틴 1(Keratin 1), 클라우딘 1(Claudin 1), 클라우딘 4(Claudin 4), 및 오클루딘(Occludin)의 mRNA의 발현량이 감소한 것을 확인할 수 있었다. 구체적으로, 케라틴 1의 mRNA 발현량은 3시간 처리한 경우 0.19, 6시간 처리한 경우는 0.13으로 mRNA 발현량이 현저히 감소한 것을 확인할 수 있었고, 클라우딘 1는 각각 0.44와 0.24로, 클라우딘 4는 모두 0.52로, 오클루딘은 각각 0.48과 0.39로 감소한 것을 확인하였다. 즉, 미세먼지에 의하여 각질세포 분화마커(즉, 케라틴 1)와 과립층 장벽인 타이트 정션(tight junction) 구성성분(즉, 클라우딘 1, 클라우딘 4, 및 오클루딘)의 발현량이 모두 감소하였다.As shown in FIGS. 1 to 4, the control group 2 treated with fine dust at a concentration of 50 ppm contained Keratin 1, Claudin 1, Claudin 1, 4 (Claudin 4), and Occludin (mRNA) of the present invention. Specifically, the amount of mRNA expression of keratin 1 was 0.19 for 3 hours and 0.13 for 6 hours, indicating that mRNA expression levels were significantly decreased in the keratin 1, 0.44 and 0.24 in the keratin 1 and 0.52 in the keratin 4, respectively , And acrodine decreased to 0.48 and 0.39, respectively. That is, the amount of expression of keratinocyte differentiation markers (ie, keratin 1) and tight junction components (ie, claudin 1, claudin 4, and occludin) in the granule barriers were all reduced by fine dust.
실시예 1은 미세먼지에 의하여 감소된 마커들의 mRNA 발현량을 본 발명의 녹차 추출물이 케어할 수 있음을 나타낸다. 구체적으로, 도 1은 케라틴 1의 mRNA 발현량은 3시간 처리한 경우 0.79, 6시간 처리한 경우는 0.99로 미세먼지에 의하여 감소된 mRNA 발현량이 복구됨을 나타내며, 도 4는 오클루딘의 mRNA 발현량은 3시간 처리한 경우 0.88, 6시간 처리한 경우는 0.98로 미세먼지에 의하여 감소된 mRNA 발현량이 복구됨을 나타내다. 특히, 클라우딘 1 및 클라우딘 4는 3시간 처리한 경우 각각 1.12와 1.23을, 6시간 처리한 경우에 각각 1.23과 1.45로 대조군 1에 비하여 향상된 mRNA 발현량을 보인다(도 2 및 도 3 참조).Example 1 shows that the green tea extract of the present invention can cure mRNA expression levels of markers reduced by fine dust. Specifically, FIG. 1 shows that the amount of mRNA expression of keratin 1 is restored by 0.79 for 3 hours and 0.99 for 6 hours, and FIG. 4 shows the expression level of mucin Was 0.88 for 3 hours and 0.98 for 6 hours, indicating that the amount of mRNA decreased by fine dust was restored. In particular, Claudin 1 and Claudin 4 exhibited enhanced mRNA expression levels (1.12 and 1.23, respectively, when treated for 3 hours, and 1.23 and 1.45, respectively, when treated for 6 hours), compared to control group 1 (see FIGS. 2 and 3).
따라서, 미세먼지에 의해 자극된 피부 세포에서 감소한 케라틴 1, 클라우딘 1, 클라우딘 4, 및 오클루딘의 발현량은 녹차 추출물의 처리에 의하여 그 발현량이 복구될 수 있음을 확인할 수 있다.Therefore, it can be confirmed that the expression amount of keratin 1, cladin 1, cladin 4, and occludin decreased in skin cells stimulated by fine dust can be restored by treatment of green tea extract.
이하, 본 발명에 따른 조성물의 제형예를 설명하나, 화장료 조성물, 약학적 조성물 및 건강 기능식품 조성물은 여러 가지 제형으로 응용 가능하며, 이는 본 발명을 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of the composition according to the present invention will be described. However, the cosmetic composition, the pharmaceutical composition and the health functional food composition can be applied to various formulations, and the present invention is not limited thereto .
[제형예 1] 정제 [Formulation Example 1] Tablets
본 발명 실시예에 따른 녹차 추출물 100mg, 락토오스 400mg, 옥수수 전분 400mg 및 스테아린산 마그네슘 2mg을 혼합한 후, 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.100 mg of the green tea extract according to the present invention, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate were mixed and then tableted according to the conventional preparation method.
배합성분Compounding ingredient 함량(mg)Content (mg)
녹차 추출물Green tea extract 100100
락토오스Lactose 400400
옥수수 전분Corn starch 400400
스테아린산 마그네슘Magnesium stearate 22
[제형예 2] 캡슐제[Formulation Example 2]
본 발명 실시예에 따른 녹차 추출물 100mg, 락토오스 400mg, 옥수수 전분 400mg 및 스테아린산 마그네슘 2mg을 혼합한 후, 통상의 캡슐제의 제조 방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.100 mg of the green tea extract according to the present invention, 400 mg of lactose, 400 mg of corn starch and 2 mg of magnesium stearate were mixed and filled in gelatin capsules according to the conventional preparation method of capsules to prepare capsules.
배합성분Compounding ingredient 함량(mg)Content (mg)
녹차 추출물Green tea extract 100100
락토오스Lactose 400400
옥수수 전분Corn starch 400400
스테아린산 마그네슘Magnesium stearate 22
[제형예 3] 과립제[Formulation Example 3]
본 발명 실시예에 따른 녹차 추출물 50mg, 무수결정 포도당 250mg 및 전분 550mg을 혼합하고, 유동층 과립기를 사용하여 과립으로 성형한 후 포에 충진하였다.50 mg of green tea extract according to the present invention, 250 mg of anhydrous crystalline glucose and 550 mg of starch were mixed and granulated into granules using a fluidized bed granulator, and filled in a capsule.
배합성분Compounding ingredient 함량(mg)Content (mg)
녹차 추출물Green tea extract 5050
무수결정 포도당Anhydrous crystalline glucose 250250
전분Starch 550550
[제형예 4] 비누 [Formulation Example 4] Soap
배합성분Compounding ingredient 함량(%)content(%)
녹차 추출물Green tea extract 5.005.00
유지maintain 적량Suitable amount
수산화나트륨Sodium hydroxide 적량Suitable amount
염화나트륨Sodium chloride 적량Suitable amount
향료Spices 적량Suitable amount
정제수Purified water 잔량Balance
[제형예 5] 로션 [Formulation Example 5] Lotion
배합성분Compounding ingredient 함량(%)content(%)
녹차 추출물Green tea extract 5.005.00
L-아스코르빈산-2-인산마그네슘염L-ascorbic acid-2-phosphate magnesium salt 1.001.00
수용성 콜라겐 (1 % 수용액)Water soluble collagen (1% aqueous solution) 1.001.00
시트르산나트륨Sodium citrate 0.100.10
시트르산Citric acid 0.050.05
감초 엑기스Licorice extract 0.200.20
1,3-부틸렌글리콜1,3-butylene glycol 3.003.00
정제수Purified water 잔량Balance
[제형예 6] 크림[Formulation Example 6] Cream
배합성분Compounding ingredient 함량(%)content(%)
녹차 추출물Green tea extract 3.003.00
폴리에틸렌글리콜모노스테알레이트Polyethylene glycol monostearate 2.002.00
자기유화형모노스테아르산글리세린Self emulsifying monostearic acid glycerin 5.005.00
세틸알코올Cetyl alcohol 4.004.00
스쿠알렌Squalene 6.006.00
트리2-에틸헥산산글리세릴Tri-2-ethylhexanoic acid glyceryl 6.006.00
스핑고당지질Sphingoglycolipids 1.001.00
1.3-부틸렌글리콜1,3-butylene glycol 7.007.00
정제수Purified water 잔량Balance
[제형예 7] 연고[Formulation Example 7] ointment
배합성분Compounding ingredient 함량(%)content(%)
녹차 추출물Green tea extract 5.005.00
폴리비닐알코올Polyvinyl alcohol 13.0013.00
L-아스코르빈산-2-인산마그네슘염L-ascorbic acid-2-phosphate magnesium salt 1.001.00
라우로일히드록시프롤린Lauroylhydroxyproline 1.001.00
수용성 콜라겐 (1 % 수용액)Water soluble collagen (1% aqueous solution) 2.002.00
1,3-부틸렌글리콜1,3-butylene glycol 3.003.00
에탄올ethanol 5.005.00
정제수Purified water 잔량Balance
[제형예 8] 미용액 제조 [Formulation Example 8] Preparation of serum
배합성분Compounding ingredient 함량(%)content(%)
녹차 추출물Green tea extract 3.003.00
히드록시에틸렌셀룰로오스(2 % 수용액)Hydroxyethylene cellulose (2% aqueous solution) 12.0012.00
크산탄검(2 % 수용액)Xanthan gum (2% aqueous solution) 2.002.00
1,3-부틸렌글리콜1,3-butylene glycol 6.006.00
진한 글리세린Concentrated glycerin 4.004.00
히알루론산나트륨 (1 % 수용액)Sodium hyaluronate (1% aqueous solution) 2.002.00
정제수Purified water 잔량Balance
[제형예 9] 건강식품[Formulation Example 9] Health food
배합 성분Compounding ingredient 함량content
녹차 추출물Green tea extract 2mg2 mg
비타민 A 아세테이트Vitamin A acetate 70μg70 μg
비타민 EVitamin E 1.0mg1.0 mg
비타민 B1Vitamin B1 0.13mg0.13 mg
비타민 B2Vitamin B2 0.15mg0.15 mg
비타민 B6Vitamin B6 0.5mg0.5 mg
비타민 B12Vitamin B12 0.2μg0.2 μg
비타민 CVitamin C 10mg10 mg
비오틴Biotin 10μg10 μg
니코틴산아미드Nicotinic acid amide 1.7mg1.7 mg
엽산Folic acid 50μg50 μg
판토텐산 칼슘Calcium pantothenate 0.5mg0.5 mg
황산 제1철Ferrous sulfate 1.75mg1.75 mg
산화아연Zinc oxide 0.82mg0.82 mg
탄산마그네슘Magnesium carbonate 25.3mg25.3 mg
제1인산칼륨Potassium monophosphate 15mg15 mg
제2인산칼슘Dicalcium phosphate 55mg55 mg
구연산칼륨Potassium citrate 90mg90 mg
탄산칼슘Calcium carbonate 100mg100 mg
염화마그네슘Magnesium chloride 24.8mg24.8 mg
[제형예 10] 건강음료[Formulation Example 10] Health drinks
배합성분Compounding ingredient 함량content
녹차 추출물Green tea extract 50mg50 mg
구연산Citric acid 1000mg1000 mg
올리고당oligosaccharide 100 g100 g
타우린Taurine 1g1g
정제수Purified water 잔량Balance

Claims (11)

  1. 녹차 추출물을 유효성분으로 포함하는, 미세먼지에 의한 피부 손상 케어용 조성물.A composition for care of skin damage caused by fine dust, comprising green tea extract as an active ingredient.
  2. 제 1 항에 있어서, The method according to claim 1,
    상기 녹차 추출물은 조성물 총 중량에 대하여 0.000001 내지 30중량%로 포함된, 조성물.Wherein the green tea extract is contained in an amount of 0.000001 to 30% by weight based on the total weight of the composition.
  3. 제 1 항에 있어서, The method according to claim 1,
    상기 녹차 추출물은 물, C1 - C6의 무수 또는 함수 저급 알코올, 아세톤, 부틸렌글리콜, 에틸아세테이트, 디에틸아세테이트, 디에틸에테르, 벤젠, 클로로포름 및 헥산으로 이루어진 군에서 선택된 어느 하나 이상의 추출용매로 추출된 것인, 조성물.Wherein the green tea extract is selected from the group consisting of water, C 1 - C 6 anhydrous or lower alcohol, acetone, butylene glycol, ethyl acetate, diethyl acetate, diethyl ether, benzene, chloroform and hexane . ≪ / RTI >
  4. 제 1 항에 있어서, The method according to claim 1,
    상기 조성물의 유효성분은 피부 장벽의 타이트 정션(tight junction)에 작용하는 것인, 조성물.Wherein the active ingredient of the composition acts on the tight junction of the skin barrier.
  5. 제 1 항에 있어서, The method according to claim 1,
    상기 조성물은 케라틴 1(KRT 1), 클라우딘 1(CLDN 1), 클라우딘 4(CLDN 4), 및 오클루딘(OCLN)로 이루어진 군에서 선택되는 하나 이상의 발현을 복구시키는, 조성물.Wherein the composition restores one or more expressions selected from the group consisting of keratin 1 (KRT 1), claudin 1 (CLDN 1), claudin 4 (CLDN 4), and occludin (OCLN).
  6. 제 1 항에 있어서,The method according to claim 1,
    상기 미세먼지는 비소(arsenic), 카드뮴(cardmium), 납(lead), 및 니켈(nikel) 중 하나 이상을 포함하는, 조성물.Wherein the fine dust comprises at least one of arsenic, cardmium, lead, and nickel.
  7. 제 1 항에 있어서,The method according to claim 1,
    상기 미세먼지는 입자크기가 PM 10 이하인, 조성물.Wherein the fine dust has a particle size of PM 10 or less.
  8. 제 1 항에 있어서,The method according to claim 1,
    상기 녹차 추출물은 10 내지 500 mg/kg/일의 투여량으로 투여되는, 조성물.Wherein the green tea extract is administered at a dose of 10 to 500 mg / kg / day.
  9. 제 1 항에 있어서, The method according to claim 1,
    상기 조성물은 화장료 조성물인, 조성물.Wherein the composition is a cosmetic composition.
  10. 제 1 항에 있어서, The method according to claim 1,
    상기 조성물은 약학적 조성물인, 조성물.Wherein the composition is a pharmaceutical composition.
  11. 제 1 항에 있어서, The method according to claim 1,
    상기 조성물은 건강 기능식품 조성물인, 조성물.Wherein the composition is a health functional food composition.
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