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WO2019059512A9 - Composition cosmétique contenant de la purpurogalline pour la protection de la peau contre les espèces réactives de l'oxygène, le rayonnement ultraviolet ou les particules fines - Google Patents

Composition cosmétique contenant de la purpurogalline pour la protection de la peau contre les espèces réactives de l'oxygène, le rayonnement ultraviolet ou les particules fines Download PDF

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Publication number
WO2019059512A9
WO2019059512A9 PCT/KR2018/008350 KR2018008350W WO2019059512A9 WO 2019059512 A9 WO2019059512 A9 WO 2019059512A9 KR 2018008350 W KR2018008350 W KR 2018008350W WO 2019059512 A9 WO2019059512 A9 WO 2019059512A9
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Prior art keywords
purpurogallin
cosmetic composition
oxygen species
skin
skin protection
Prior art date
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Ceased
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PCT/KR2018/008350
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English (en)
Korean (ko)
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WO2019059512A2 (fr
WO2019059512A3 (fr
Inventor
현진원
강희경
고영상
안미정
김태훈
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Industry Academic Cooperation Foundation of Jeju National University
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Industry Academic Cooperation Foundation of Jeju National University
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Priority to CN201880057443.0A priority Critical patent/CN111093607A/zh
Publication of WO2019059512A2 publication Critical patent/WO2019059512A2/fr
Publication of WO2019059512A3 publication Critical patent/WO2019059512A3/fr
Publication of WO2019059512A9 publication Critical patent/WO2019059512A9/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • A61K8/345—Alcohols containing more than one hydroxy group
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/35—Ketones, e.g. benzophenone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00—Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04—Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • 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 cosmetic composition for skin protection against active oxygen species, ultraviolet rays or fine dusts, a quasi-drug composition and a composition for external application for skin, which comprises Purpurogallin or a cosmetically acceptable salt thereof as an active ingredient.
  • Skin is a part of the body that is directly exposed to the external environment. It acts as a protective layer to protect important organs of our body, as well as controlling water evaporation and protecting the body from external infections.
  • skin prevents the penetration of viruses from the outside, excessive stress from the outside such as ultraviolet rays, contaminated environment and the like causes skin irritation and eventually leads to aging of the skin.
  • UVB radiation wavelength: 280-320 nm
  • ROS reactive oxygen species
  • the active oxygen species is oxygen in an unstable state, and oxygen is overproduced due to environmental pollution, chemicals, ultraviolet rays, blood circulation disorder, stress, and the like. This excess oxygen is oxidized in the human body, and cell membranes, DNA, and all other cell structures are damaged, and the cells lose their function or deteriorate depending on the extent of the damage. In addition, reactive oxygen species in skin cells cause oxidative stress and promote skin aging.
  • ROS reactive oxygen species
  • - epigallocatechin-3-gallate
  • resveratrol a wide variety of marine algae or seaweeds containing compounds such as carotenoids, phenolics or antioxidant vitamins can also be found in biological antioxidant systems and free radicals ) Is known to mitigate oxidative stress in skin cells.
  • Fine dust is a very small material that is invisible to the naked eye and refers to a particulate material having a diameter of 10 ⁇ m or less that floats or drifts in the air for a long time. It is an air pollutant that is produced when fossil fuels such as coal and petroleum are burned, or when they come out from exhaust gas such as manufacturing and automobile soot, and are adsorbed to the lungs through the bronchi and cause various kinds of lung diseases. According to the Ministry of Environment, in March 2017, the term "PM10", which is 10 ⁇ m or less in diameter, refers to floating dust and ultra fine dust (PM2. 5) is defined as fine dust.
  • Contaminants such as fine dust, dust, and the like cause stress in the skin, which is produced by active oxygen.
  • Active oxygen acts as an intracellular signaling substance that plays an important role in maintaining normal cellular functions, such as activation of mitochondrial electron transport and leukocyte cells.
  • the active oxygen is unstable and has a high oxidizing power and easily reacts with biomaterials, if it can not be removed in the human body, it causes oxidative stress.
  • the present invention aims to develop a cosmetic composition having skin protection effect against active oxygen species, ultraviolet ray or fine dust.
  • Another object of the present invention is to provide a quasi-drug composition for protecting skin against active oxygen species, ultraviolet rays or fine dusts, which comprises Purpurogallin or a physiologically acceptable salt thereof as an active ingredient.
  • Another object of the present invention is to provide a skin external composition for skin protection against active oxygen species, ultraviolet rays or fine dusts containing Purpurogallin or a physiologically acceptable salt thereof as an active ingredient.
  • Another object of the present invention is to provide the use of Purpurogallin or a physiologically acceptable salt thereof for the preparation of a quasi-drug composition for skin protection against active oxygen species, ultraviolet rays or fine dusts.
  • Another object of the present invention is to provide a use of Purpurogallin or a physiologically acceptable salt thereof for the preparation of a composition for external skin application for skin protection against active oxygen species, ultraviolet rays or fine dusts.
  • Another object of the present invention is to provide a skin protection method against active oxygen species, ultraviolet rays or fine dusts, comprising the step of administering Purpurogallin or a physiologically acceptable salt thereof to a subject.
  • the present invention relates to a cosmetic composition for skin protection against active oxygen species, ultraviolet rays or fine dusts, comprising Purpurogallin or a cosmetically acceptable salt thereof as an active ingredient, wherein the active oxygen species, ultraviolet ray or fine dust Can be used as a cosmetic composition for skin protection.
  • Figures 1 to 3 show the effect of purpurogallin (PG) on ROS production.
  • Figure 1 HaCaT cells were treated with Purpurogallin (PG) (0, 2.5, 5, 10 [mu] M) for 24 hours and cell viability was determined by MTT assay.
  • Fig. 2 The intracellular ROS removal ability of PG (2.5, 5, 10 [mu] M) produced by H2O2 was detected using DCF-DA assay. * Significantly different from control cells of H2O2 (p ⁇ 0.05).
  • Fig. 3 The ability to remove hydroxy radicals from 10 ⁇ M PG was predicted using the pantone reaction. * Significantly different from the control group; # It is significantly different from the UVB group.
  • FIGs 4 to 7 show the effect of UVB-induced cell damage inhibition of Purpurogallin (PG) (10 [mu] M). Confocal microscopy was used to detect intracellular ROS (FIG. 4) and lipid peroxidation (FIG. 5) after DCF-DA (green) and DPPP (blue) staining. (Fig. 6) DNA damage was analyzed by comet analysis. (Fig. 7) Protein oxidation was measured by measuring the amount of carbonyl formation. * Significantly different from control cells (p ⁇ 0.05) and significantly different from # UVB irradiated cells (p ⁇ 0.05).
  • PG Purpurogallin
  • Figures 8 to 10 show the protective effect of Purpurogallin (PG) (10 [mu] M) against UVB induced cell death.
  • PG Purpurogallin
  • Figures 8 to 10 show the protective effect of Purpurogallin (PG) (10 [mu] M) against UVB induced cell death.
  • MTT assay was performed on HaCaT cell viability after UVB irradiation. * Significantly different from the control group; # It is significantly different from the UVB group.
  • Fig. 9 Cells were stained with JC-1 and the mitochondrial membrane potential ([Delta] m) was evaluated by flow cytometry. * Significantly different from control cells (p ⁇ 0.05); # Significantly different from UVB irradiated cells (p ⁇ 0.05).
  • Apoptotic cells (arrows) stained with Hoechst 33342 were detected by fluorescence microscopy. * Significantly different from control cells (p ⁇ 0.05); # Significantly different from UVB irradiated cells (p ⁇ 0.05).
  • Figure 11 stained cells with DCF-DA (green) and detected intracellular ROS induced by particulate matter (PM) using a confocal microscope.
  • Figure 12 detected apoptotic cells (arrows) stained with Hoechst 33342 by fluorescence microscopy. * Significantly different from control cells (p ⁇ 0.05); # PM cells (p ⁇ 0.05).
  • FIG. 13 shows the fluorescence intensity (green) of DCF formed by ROS in the group treated with UVB, PM (particulate matter).
  • FIG. 14 detected apoptotic cells (arrows) stained with Hoechst 33342 by fluorescence microscopy. * Significantly different from control cells (p ⁇ 0.05); # Significantly different from UVB irradiated cells (p ⁇ 0.05).
  • the present invention provides a cosmetic composition for skin protection against active oxygen species, ultraviolet rays or fine dusts, comprising Purpurogallin or a cosmetically acceptable salt thereof as an active ingredient to provide.
  • the cosmetic composition of the present invention contains, as an active ingredient, Purpurogallin or a cosmetically acceptable salt thereof.
  • the cosmetic composition comprising Purpurogallin has a skin protection effect on active oxygen species, ultraviolet rays or fine dusts.
  • purpurogallin was not cytotoxic in the experiment using human keratinocyte cell line (HaCaT), and the concentration of hydrogen peroxide (H2O2) -induced intracellular reactive oxygen species reactive oxygen species (ROS) and hydroxy radicals, and thus can be used as an active ingredient of a cosmetic composition for skin protection against active oxygen species.
  • HaCaT human keratinocyte cell line
  • ROS reactive oxygen species
  • the active oxygen species may be hydrogen peroxide (H2O2) or hydroxyl radical.
  • purpurogallin had UVB-induced reactive oxygen species (ROS) scavenging effect, and DNA damage caused by UVB stimulation, protein Which markedly reduced damage and lipid peroxidation.
  • ROS reactive oxygen species
  • Purpurogallin inhibited increased cell death by UVB irradiation in HaCaT cells, decreased apoptotic body, and showed protection against UVB induced apoptosis.
  • Purpurogallin can be used as an active ingredient of a cosmetic composition for skin protection against ultraviolet rays.
  • the ultraviolet ray may be ultraviolet-B (UVB).
  • the Purpurogallin can exhibit skin protecting activity through scavenging of reactive oxygen species (ROS) generated by ultraviolet ray B.
  • ROS reactive oxygen species
  • the Purpurogallin may exhibit a skin-protecting activity through an action of reducing intracellular lipid peroxidation, DNA damage or protein damage due to UVB.
  • purpurogallin inhibits ROS and apoptosis induced by PM (particulate matter) treatment in human keratinocyte cell line, It was confirmed that the skin was protected against dust. Therefore, Purpurogallin can be used as an active ingredient of a cosmetic composition for skin protection against fine dusts.
  • Purpurogallin is induced in a human keratinocyte cell line by UVB irradiation, particulate matter (PM) or UVB and particulate matter (PM) treatment It was confirmed that ROS and apoptotic cells were inhibited, and skin protection effect against fine dust and ultraviolet rays was confirmed. Therefore, Purpurogallin can be used as an active ingredient of a cosmetic composition for skin protection against fine dust or ultraviolet rays.
  • the Purpurogallin can exhibit skin protecting activity through scavenging of reactive oxygen species (ROS) generated by fine dust.
  • ROS reactive oxygen species
  • the Purpurogallin may exhibit skin-protecting activity through an action of inhibiting apoptosis of cells exposed to ultraviolet B or fine dust.
  • Acid addition salts can be prepared in a conventional manner, for example, by dissolving the compound in an excess amount of an aqueous acid solution and precipitating the salt with a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and acid aqueous solution or alcohol (for example, glycol monomethyl ether) may be heated and then the mixture may be evaporated to dryness or the precipitated salt may be subjected to suction filtration.
  • a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile.
  • organic acids and inorganic acids can be used.
  • inorganic acids hydrochloric acid, phosphoric acid, sulfuric acid, nitric acid, tartaric acid and the like can be used.
  • organic acids include methanesulfonic acid, p- toluenesulfonic acid, acetic acid, trifluoroacetic acid, maleic acid, succinic acid, oxalic acid, benzoic acid, tartaric acid, fumaric acid, mandelic acid, propionic acid, citric acid, lactic acid, glycollic acid, gluconic acid, galacturonic acid, glutamic acid, glutaric acid, glucuronic acid, aspartic acid, ascorbic acid, carbonic acid, vanillic acid and hydroiodic acid may be used. It is not limited.
  • bases can be used to make cosmetically acceptable metal salts.
  • the alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the non-soluble compound salt, and evaporating and drying the filtrate.
  • the corresponding silver salt can also be obtained by reacting an alkali metal or alkaline earth metal salt with a suitable silver salt (e.g., silver nitrate).
  • the cosmetic composition for protecting skin against active oxygen species, ultraviolet ray or fine dust of the present invention may contain 0.001 to 100% by weight of purpurogallin or a cosmetically acceptable salt thereof, based on the total weight of the composition, Of the present invention may include, but is not limited to, 0.01 to 50% by weight of purpurogallin or a cosmetically acceptable salt thereof.
  • the cosmetic composition of the present invention may include, without limitation, commonly accepted ingredients other than the above-mentioned effective ingredients, and includes conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers can do.
  • the cosmetic composition according to the present invention can be used as a cosmetic composition in the form of a solution, an ointment for external use, a cream, a foam, a nutritional lotion, a softening water, a pack, a soft water, an emulsion, a makeup base, May be prepared from one or more formulations selected from the group consisting of emulsions, emulsions, pastes, gels, lotions, powders, soaps, surfactant-containing cleansers, oils, powder foundations, emulsion foundations, wax foundations, patches and sprays But is not limited to.
  • the cosmetic composition of the present invention may further comprise at least one cosmetically acceptable carrier to be incorporated in a general skin cosmetic composition, and examples thereof include oil, water, a surfactant, a moisturizer, A thickening agent, a chelating agent, a coloring matter, an antiseptic, a perfume, and the like may be appropriately compounded, but the present invention is not limited thereto.
  • the cosmetically acceptable carrier to be contained in the cosmetic composition of the present invention varies depending on the formulations.
  • the carrier component may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide Mixtures of these may be used.
  • lactose When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder or a mixture thereof may be used as the carrier component, Propellants such as fluorohydrocarbons, propane / butane or dimethyl ether.
  • a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil may be used, in particular fatty acid esters of cottonseed oil, peanut oil, corn oil, olive oil, castor oil and sesame oil, glycerol aliphatic esters, polyethylene glycols or sorbitan may be used have.
  • the formulation of the present invention is a suspension
  • a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspension such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Crystalline cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.
  • the formulation of the present invention is a soap, an alkali metal salt of a fatty acid, a fatty acid hemiester salt, a fatty acid protein hydrolizate, isethionate, a lanolin derivative, an aliphatic alcohol, a vegetable oil, glycerol, .
  • the carrier component is selected from the group consisting of aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.
  • the present invention provides a quasi-product composition for protecting skin against active oxygen species, ultraviolet rays or fine dusts, comprising Purpurogallin or a physiologically acceptable salt thereof as an active ingredient.
  • physiologically acceptable of the present invention is physiologically acceptable and when administered to an organism, the compound to be administered, without causing an allergic reaction such as gastrointestinal disorder, dizziness, or the like, Quot; means < / RTI >
  • the term "quasi-drug product" used in the present invention means products that are less active than drugs, among the products used for diagnosing, treating, improving, alleviating, treating or preventing diseases of human or animal.
  • the quasi-quasi-drug means a product that is used for the treatment of diseases of humans and animals, except for the products used for medicines, which are slightly or not directly acting on the human body, , And sterilization and insecticides to prevent infectious diseases.
  • the type and formulations of the quasi-drug composition of the present invention are not particularly limited, but may be disinfectant cleaner, shower foam, gagrin, wet tissue, detergent soap, hand wash, humidifier filler, mask, ointment or filter filler.
  • the composition of the present invention When the composition of the present invention is contained in quasi-drugs for skin protection against active oxygen species, ultraviolet rays or fine dusts, the composition may be used as it is or may be used in combination with other quasi-drugs, .
  • the amount of the active ingredient to be mixed can be suitably determined according to the purpose of use.
  • the quasi-drug composition according to the present invention contains the above purpurogallin or a physiologically acceptable salt thereof in an amount of 0.01 to 20 wt% can do.
  • the present invention provides a skin external composition for skin protection against active oxygen species, ultraviolet rays or fine dusts, comprising Purpurogallin or a physiologically acceptable salt thereof as an active ingredient.
  • purpurogallin active oxygen species
  • skin protection against ultraviolet rays or fine dusts and physiologically acceptable salts are as described above.
  • composition for external application for skin according to the present invention includes, for example, ointments, lotions, soluble images, suspensions, emulsions, creams, gels, sprays, papermaking agents, And may be incorporated into any of the bases known in the art.
  • the composition for external application for skin according to the present invention may contain the above purpurogallin or a physiologically acceptable salt thereof in an amount of 0.01 to 20% by weight based on the total weight of the composition.
  • the present invention provides the use of Purpurogallin or a cosmetically acceptable salt thereof for the preparation of cosmetic composition for skin protection against active oxygen species, ultraviolet rays or fine dusts.
  • the present invention provides the use of Purpurogallin or a physiologically acceptable salt thereof for the preparation of a quasi-drug composition for skin protection against active oxygen species, ultraviolet rays or fine dusts.
  • the present invention provides the use of Purpurogallin or a physiologically acceptable salt thereof for the preparation of a composition for external skin application for skin protection against active oxygen species, ultraviolet rays or fine dusts.
  • the present invention provides a method of protecting skin against active oxygen species, ultraviolet light or fine dust, comprising administering to a subject Purpurogallin or a physiologically acceptable salt thereof .
  • composition of the present invention includes, but is not limited to, mammals including humans, such as cows, pigs, horses, rabbits, mice, and humans.
  • the method of skin protection using purpurogallin or a physiologically acceptable salt of the present invention can be carried out by administering Purpurogallin or a physiologically acceptable salt to individual or individual skin.
  • administered means introducing Purpurogallin or a physiologically acceptable salt of the present invention by any appropriate method, and the route of administration of the compound and the salt of the present invention is not particularly limited But can be administered via various routes to reach the desired skin tissue.
  • NAC 1,1-diphenyl-2-picrylhydrazine
  • DCF -DDA 1,1-diphenyl-2-picrylhydrazine
  • MTT [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetololium] bromide
  • DMPO 5,5- -Oxide
  • Human keratinocyte cell line (HaCaT) was purchased from Amore Pacific Company (Yongin, Republic of Korea) and maintained at 37 ° C in an incubator containing 5% carbon dioxide. The cell lines were grown in DMEM medium containing 10% bovine serum, streptomycin (100 ug / ml) and penicillin (100 units / ml) heated at 56 ° C for 30 minutes. (UVP, Upland, CA, USA) was used as a UVB source spectrum having a UVB energy spectrum of 280 nm to 320 nm, and the UVB irradiation amount was 30 mJ / cm 2 in the present invention.
  • UVP Upland, CA, USA
  • Cells were inoculated into 24-well plates at a density of 0.8 x 105 cells / ml. After the cells were completely attached to the bottom of the plate, 2.5, 5, and 10 ⁇ M of Purpurogallin (PG) was added. MTT Stock solution (50 ⁇ l, 2 mg / ml) was added to each well to adjust the total reaction volume to 500 ⁇ l and incubated for 4 hours. In addition, cells were treated with 1 mM NAC or 20 ⁇ M furfurogalline and irradiated with 30 mJ / cm 2 UVB one hour later.
  • PG Purpurogallin
  • the hydroxy radicals produced by the Fenton reaction were reacted with DMPO.
  • the resulting DMPO / ⁇ OH adduct product was detected using an ESR spectrometer.
  • Phosphate buffer pH 7.4 was mixed with 0.02 ml of each of 0.3 M DMPO, 10 mM FeSO4, 10 mM H2O2, and 10 ⁇ M DQA (3,4-dicaffeoylquinic acid) and the ESR spectrum recorded after 2.5 minutes.
  • the ESR spectrometer parameters were set as follows: center magnetic field, 336.8 mT; Power, 1.00 mW; Frequency, 9.4380 GHz; Modulation width, 0.2 mT; Amplitude, 600; Sweep width, 10 mT; Sweep time, 0.5 min; Time constant, 0.03 sec; And temperature, 25 ⁇ .
  • the level of intracellular ROS induced by H2O2 (1 mM) or UVB irradiation (30 mJ / cm2) was measured using the DCF-DA method. After 16 hours plating, the cells were treated with Purpurogallin (PG) at concentrations of 2.5, 5, and 10 [mu] M for 1 hour. Cells were treated with H2O2 or UVB for 30 min and 8 h. After adding 25 ⁇ M DCF-DA for 10 minutes, 2 ', 7'-dichlorofluorescein fluorescence was detected using LS-5B spectrophotometer (PerkinElmer, Waltham, MA, USA).
  • PG Purpurogallin
  • Oxidative DNA damage was assessed by Comet analysis.
  • the cell suspension was mixed with 75 ⁇ l of 0.5% low melting point agarose (LMA) at 39 ° C. and the mixture was spread on a completely frozen microscope slide precoated with 200 ⁇ l of 1% normal molten agarose (NMA) Respectively. After the agarose was solidified, the slides were covered with 170 [mu] l of another 0.5% LMA and then lysed in solution (2.5 M NaCl, 100 mM Na-EDTA, 10 mM Tris, 1% Triton X- 10) at 4 DEG C for 1 hour.
  • LMA low melting point agarose
  • NMA normal molten agarose
  • the slides were then incubated in a gel electrophoresis apparatus containing 300 mM NaOH and 10 mM Na-EDTA (pH 10) for 40 min to induce DNA unwinding and alkaline-labile damage .
  • the electric field 300 mA, 25 V
  • the slides were washed three times for 5 minutes at 4 ° C in neutral buffer (0.4 M Tris, pH 7.5), stained with 80 ⁇ l of 10 ⁇ g / ml ethidium bromide, and analyzed using a fluorescence microscope and an image analyzer Komet 5.5 (Andor Technology, Harbor, UK ).
  • the tail length and the percentage of total fluorescence in the comet tail were recorded in 50 cells per slide.
  • DPPP was used as a probe to evaluate lipid peroxidation.
  • DPPP reacts with lipid hydroperoxide to produce DPPP oxide, a fluorescent material, which provides an indication of membrane damage.
  • Cells were treated with 10 ⁇ M Purpurogallin (PG) for 1 hour and then exposed to UVB (30 mJ / cm 2). After 5 hours, 5 mM DPPP was added and incubated in the dark for 30 minutes.
  • DPPP fluorescence spectra were analyzed with a fluorescence microscope and an image analyzer Komet 5.5 (Andor Technology, Harbor, UK).
  • the MTT assay showed no cytotoxicity to HaCaT cells, regardless of the concentration of purpurogallin (PG) used. Compared to the control group, the cell viability of each sample treated group was higher than 96% (Fig. 1). In addition, after the treatment with H2O2 using DCF-DA analysis, the intracellular ROS elimination activity of PG was determined (Fig. 2). Fluorescence spectroscopy data showed that purpurogallin (PG) had an intracellular ROS removal effect on H2O2-treated cells in a concentration-dependent manner. Specifically, it was 11% at 2.5 ⁇ M and 5 ⁇ M at 2.5 ⁇ M 23%, and 42% at 10 ⁇ M. (Fig. 2).
  • Purpurogallin is not cytotoxic and has excellent H2O2-induced intracellular ROS and hydroxy radical scavenging effect.
  • Lipid peroxidation was also confirmed by examining the fluorescent product DPPP oxide produced from DPPP.
  • the intensity of the DPPP oxide was higher in UVB-exposed cells than in UV-exposed cells (Fig. 5) treated with Purpurogallin (PG) (blue).
  • Purpurogallin (PG) inhibited lipid peroxidation.
  • Purpurogallin has a protective effect against UVB-induced skin cell death.
  • Purpurogallin has protective effects against PM (particulate matter) -induced ROS clearing and skin cell apoptosis.
  • Purpurogallin suppresses ROS and apoptosis induced by UVB irradiation or UVB irradiation and particulate matter (PM) treatment in human keratinocyte cell lines, and ultraviolet light and fine dust It was found that there is a skin protection effect by the skin.

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Abstract

La présente invention concerne une composition cosmétique contenant de la purpurogalline ou un sel de qualité cosmétique de celle-ci, en tant que principe actif, pour la protection de la peau contre les espèces réactives de l'oxygène, le rayonnement ultraviolet ou les particules fines. Le composé peut être utilisé comme composition cosmétique pour la protection de la peau contre les espèces réactives de l'oxygène, le rayonnement ultraviolet ou les particules fines.
PCT/KR2018/008350 2017-09-25 2018-07-24 Composition cosmétique contenant de la purpurogalline pour la protection de la peau contre les espèces réactives de l'oxygène, le rayonnement ultraviolet ou les particules fines Ceased WO2019059512A2 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201880057443.0A CN111093607A (zh) 2017-09-25 2018-07-24 包含红棓酚的针对活性氧簇、紫外线或微尘的用于保护皮肤的化妆品组合物

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020170123603A KR102040230B1 (ko) 2017-09-25 2017-09-25 푸르푸로갈린을 포함하는, 활성산소종, 자외선 또는 미세먼지에 대한 피부 보호용 화장료 조성물
KR10-2017-0123603 2017-09-25

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KR101748806B1 (ko) * 2016-02-29 2017-06-19 대구대학교 산학협력단 Purpurogallin을 유효성분으로 함유하는 멜라닌 생성 억제 화장료 조성물 및 그 제조방법
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