WO2022191339A1 - 송어에서 분리된 pdrn과 난포액을 포함하는 화장료 조성물 - Google Patents
송어에서 분리된 pdrn과 난포액을 포함하는 화장료 조성물 Download PDFInfo
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- WO2022191339A1 WO2022191339A1 PCT/KR2021/002798 KR2021002798W WO2022191339A1 WO 2022191339 A1 WO2022191339 A1 WO 2022191339A1 KR 2021002798 W KR2021002798 W KR 2021002798W WO 2022191339 A1 WO2022191339 A1 WO 2022191339A1
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- Prior art keywords
- trout
- skin
- pdrn
- follicular fluid
- present
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- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
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Images
Classifications
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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
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
- A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/60—Fish, e.g. seahorses; Fish eggs
-
- 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
- A61K8/60—Sugars; Derivatives thereof
-
- 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
- A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- 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
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- 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
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/52—Stabilizers
- A61K2800/522—Antioxidants; Radical scavengers
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
Definitions
- the present invention relates to a cosmetic composition
- a cosmetic composition comprising PDRN isolated from trout and follicular fluid.
- the skin is a part of the body that is directly exposed to the external environment, and it not only acts as a protective film to protect important organs of our body, but also regulates water evaporation and protects the body from external infections.
- collagen is present in the dermal layer of the skin, and is known as a major component that gives elasticity to the skin together with elastin, which accounts for about 70 to 80% of the total dry weight of the skin.
- Collagen is reduced by internal factors such as decreased cell activity due to natural aging, and biosynthesis is reduced or decomposition is promoted by external factors such as increased stress in various harmful environments or increased reactive oxygen species caused by sunlight. have.
- the breakdown of collagen and elastin in aged skin is mainly caused by increased expression of its degrading enzymes, such as the MMP protein.
- a person's skin color largely depends on the amount of melanin pigment contained in skin cells. People with a lot of melanin have brown or dark skin, while people with very little melanin cause vitiligo.
- Melanin pigment is produced from tyrosine by tyrosinase in pigment cells called melanocytes present in the basement membrane of the skin. Melanin pigment plays an important role in preventing skin damage and skin cancer by protecting the skin from excessive UV rays. However, when melanin is excessively produced by sunlight, hormonal changes, inflammation, drugs, etc., pigmentation of the skin is deposited, which causes various cosmetic problems.
- a whitening ingredient substances that inhibit tyrosinase enzyme activity such as kojic acid, arbutin, etc., hydroquinone, L-ascorbic acid, derivatives thereof, and various There are plant extracts.
- melanin pigment By inhibiting the synthesis of melanin pigment, it is possible not only to realize skin whitening by brightening the skin tone, but also to improve skin hyperpigmentation such as spots or freckles caused by ultraviolet rays, hormones, or heredity.
- the amount of use is limited due to safety issues such as irritation and redness, or the actual effect cannot be expected because the effect is insignificant.
- An object of the present invention is to provide a cosmetic composition for whitening comprising PDRN and/or follicular fluid isolated from trout sperm and ovary as an active ingredient.
- Another object of the present invention is to provide a pharmaceutical composition for preventing or treating pigmentation diseases comprising PDRN and/or follicular fluid isolated from trout as an active ingredient.
- Another object of the present invention is to provide a quasi-drug composition for whitening comprising PDRN and/or follicular fluid isolated from trout as an active ingredient.
- Another object of the present invention is to provide a composition for external application for skin whitening comprising PDRN and/or follicular fluid isolated from trout as an active ingredient.
- Another object of the present invention is to process the papain enzyme in trout eggs; degumming by treating with phosphate; Deoxidation by treating sodium hydroxide; And to provide a method for separating the follicular fluid from the trout comprising the step of treating the acid clay.
- Another object of the present invention is to provide a method for skin whitening, a method for regenerating the skin, a method for improving skin wrinkles, or a method for preventing skin aging, comprising administering to a subject follicular fluid or/and PDRN isolated from trout. .
- Another object of the present invention is to provide a method for treating pigmentation diseases, wound regeneration or skin regeneration comprising administering to damaged skin a pharmaceutical composition comprising follicular fluid isolated from trout and/or PDRN as an active ingredient. .
- Another object of the present invention is to provide a use of follicular fluid and/or PDRN isolated from trout for the preparation of a composition for skin whitening, skin aging prevention, skin regeneration, antioxidant or skin wrinkle improvement.
- Another object of the present invention is to provide a skin whitening use, skin aging prevention use, skin regeneration use, antioxidant use, or skin wrinkle improvement use of follicular fluid and/or PDRN isolated from trout.
- the present invention relates to a cosmetic composition for whitening comprising follicular fluid isolated from trout, wherein the cosmetic composition has antioxidant activity in addition to whitening.
- the cosmetic composition of the present invention further contains PDRN in addition to the follicular fluid isolated from trout, and has whitening and antioxidant effects, as well as wrinkle improvement, elasticity enhancement, and cell regeneration effects. Accordingly, it can be usefully used as a cosmetic composition, a quasi-drug composition, a composition for external application to the skin, and a therapeutic agent for a skin pigmentation disease.
- the left view of FIG. 1 is the result of confirming highly polymerized DNA extracted from semen and testes by gel electrophoresis.
- Lane 1 DNA ladder
- Lane 2 Lane 3: PDRN extracted from semen (SEMEN)
- Lane 4 Lane 5: PDRN extracted from Testis (testis).
- the right side view of FIG. 1 shows a low-molecular-weight DNA polymer made by segmenting a DNA chain to a predetermined size.
- Figure 10 shows that PDRN suppresses the expression of MMP-1 and MMP-3 increased by the treatment with reactive oxygen species.
- 11 is a result showing that PDRN increases collagen gene expression reduced by reactive oxygen species treatment.
- the present invention provides a cosmetic composition for whitening comprising follicular fluid isolated from trout as an active ingredient.
- Trout has a body similar to salmon, but is rounder and smaller than salmon. The body color is dark blue on the back, silvery white on the belly, and dark brown spots on the sides. During the breeding season, the male's snout is extended, and irregular peach-colored patterns appear on the side of the body, and the length of the body reaches 60 cm.
- the trout may be specifically, rainbow trout.
- the follicular fluid is isolated from the ovaries of trout, specifically rainbow trout.
- follicular fluid is an albumin-like liquid in the cyst follicle surrounding an egg, and the liquid component is alkaline. During ovulation, follicular fluid is expelled out of the ovary at the same time as the egg.
- the follicular fluid is treated with a papain enzyme in trout eggs; degumming by treating with phosphate; Deoxidation by treating sodium hydroxide; and treating the acid clay.
- the step of treating the papain enzyme in trout roe is 50-60 ° C, specifically 55 After the reaction at °C, to obtain a primary extract by filtration.
- the step of degumming by treating the ginseng salt is a process of degumming by treating the primary extract with 0.05 to 0.15%, specifically 0.1% phosphate. Thereafter, the process of washing the degummed primary extract with DW may be additionally included.
- degumming is one of refining to remove impurities, and the refining includes degumming, deoxidation (refining), bleaching, deodorization, and the like.
- the degumming is a process of removing protein degradation products or phospholipids using phosphoric acid or sulfuric acid.
- the step of deoxidation by treating sodium hydroxide is a process of deoxidation by treating 1-5%, specifically 2.5% sodium hydroxide in the primary extract that has undergone the degumming and washing process. Thereafter, the process of washing the deoxidized primary extract with DW may be additionally included.
- Deoxidation removes free fatty acids, also called alkali refining.
- sodium hydroxide is used to neutralize free fatty acids, sodium fatty acid is produced. This sodium fatty acid adsorbs impurities such as pigments and precipitates and removes them.
- the extract that has undergone the deoxidation and washing process is treated with 1 to 10% (w/v), specifically 5% (w/v) of the acid clay to dehydration, decolorization, and deodorization process. is a process through Thereafter, a second purified extract is obtained by stirring and heating, and this is filtered to obtain a purified trout follicle fluid.
- skin whitening refers to the function of preventing the increase of melanocytes in the skin and thereby preventing excessive deposition of melanin on the skin, or thinning the previously deposited melanin. Accordingly, it is possible to suppress the generation of spots or freckles caused by the deposition of excessive melanin pigment.
- the follicular fluid isolated from the trout exhibits a whitening effect by inhibiting tyrosinase activity (FIG. 4).
- the cosmetic composition since the cosmetic composition has antioxidant activity, it can be used as a cosmetic composition for antioxidant in addition to whitening.
- the follicular fluid isolated from the trout exhibited DPPH radical scavenging activity and had an antioxidant effect through the DDPH assay (active oxygen) scavenging effect measurement experiment (FIG. 5).
- antioxidant refers to the action of inhibiting oxidation, and active oxygen oxidizes and destroys cells in the body, and thus is exposed to various diseases.
- the antioxidant refers to the function of inhibiting oxidation of cells by highly reactive free radicals or reactive oxygen species (ROS) depending on intracellular metabolism or oxidative stress caused by the influence of ultraviolet rays. and removing free radicals or reactive oxygen species, thereby reducing cell damage.
- ROS reactive oxygen species
- the cosmetic composition of the present invention may further include a polydeoxyribonucleotide (PDRN).
- PDRN polydeoxyribonucleotide
- PDRN refers to a polydeoxyribonucleotide, and is a mixture of short deoxyribonucleotides. That is, it is a low molecular weight DNA complex made by fractionating a DNA chain into a certain size.
- the PDRN may be extracted from human placenta, trout sperm, soft sperm, or other fish tissue, but is not limited thereto.
- PDRN had no cytotoxicity (FIG. 6), and inhibited the expression of MMP (Matrix metalloproteinases)-1 and MMP-3, which are genes related to skin aging increased by treatment with reactive oxygen species.
- MMP Microx metalloproteinases
- MMP-3 genes related to skin aging increased by treatment with reactive oxygen species.
- FIG. 10 it was confirmed that there is an anti-aging effect.
- PDRN increased the collagen gene expression decreased by the treatment with reactive oxygen species, thereby preventing skin wrinkles and increasing skin elasticity to prevent skin aging (FIG. 11).
- PDRN promotes the proliferation and migration of skin cells, confirming that it can induce skin cell regeneration and heal wounds.
- the cosmetic composition containing PDRN may induce or have these effects, in addition to whitening and antioxidant, improving skin wrinkles, enhancing skin elasticity, or regenerating skin cells.
- skin cell regeneration refers to a process of repairing skin tissue against damage caused by external and internal causes of the skin.
- the damage caused by the external cause may include ultraviolet rays, external pollutants, wounds, trauma, and the like, and the damage caused by the internal cause may include stress.
- the term “improving skin wrinkles” or “enhancing skin elasticity” refers to maintaining or enhancing skin wrinkles and elasticity.
- active ingredient means a component that can exhibit the desired activity alone or can exhibit activity together with a carrier that is not active by itself.
- the cosmetic composition for whitening of the present invention may contain, in addition to the follicular fluid isolated from trout as an active ingredient, a compound or natural extract known to have a skin whitening effect so as to enhance or reinforce the skin whitening effect.
- compounds known to have a whitening effect or natural extracts include mercaptosuccinic acid, mercaptodextran, teprenone, dihydroxy-isoquinoline, indomethacin, 3-hydroxymannul, vitamin K, thiazolidone, and kinu. renin, lemon extract, cucumber extract, cucumber extract, rosemary extract, acerola extract, cherry extract, ginkgo extract, geranium extract, and the like, but are not limited thereto.
- the active ingredient may be included in any amount (effective amount) according to the use, formulation, purpose of blending, etc. as long as it can exhibit skin whitening activity. It may be included in the range of 0.001 weight % to 99.99 weight % based on the weight.
- effective amount refers to the amount of an active ingredient capable of inducing a whitening effect. Such effective amounts can be determined empirically within the ordinary ability of one of ordinary skill in the art.
- the trout follicle fluid 1 to 15% by weight of the trout follicle fluid, and 0.05 to 5% by weight of PDRN may be included.
- the cosmetic composition for whitening of the present invention may be prepared in various forms, for example, the cosmetic composition of the present invention may be prepared in any formulation conventionally prepared in the art, for example, a solution, suspension, emulsion, It may be formulated as a paste, gel, cream, lotion, powder, soap, cleansing, oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto.
- skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisture cream, hand cream, essence, nourishing essence, pack, soap, shampoo It may have a formulation selected from the group consisting of cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, lipstick, makeup base, foundation, press powder, and loose powder, but is not limited thereto.
- the cosmetic composition for whitening of the present invention may include an acceptable carrier in the cosmetic preparation in addition to the active ingredient thereof.
- acceptable carrier for cosmetic preparations refers to compounds or compositions that are already known and used that can be included in cosmetic preparations, or compounds or compositions to be developed in the future that do not have toxicity, instability, or irritation beyond what the human body can adapt to when in contact with the skin. say that
- alcohol, oil, surfactant, fatty acid, silicone oil, humectant, humectant, viscosity modifier, emulsion, stabilizer, sunscreen, color developer, fragrance, and the like may be exemplified.
- Alcohols, oils, surfactants, fatty acids, silicone oils, humectants, humectants, viscosity modifiers, emulsifiers, stabilizers, sunscreens, coloring agents, perfumes, etc. that can be used as the carrier are already known in the art. Therefore, those skilled in the art can select and use an appropriate material/composition.
- the present invention provides a pharmaceutical composition for the prevention or treatment of pigmentation diseases comprising follicular fluid isolated from trout as an active ingredient.
- trout and follicular fluid in the present invention is the same as described above.
- the follicular fluid isolated from trout had a whitening effect within a range that did not cause toxicity to cells.
- the pharmaceutical composition of the present invention may have a function of helping to lighten the color of the melanin pigment deposited on the skin, and inhibit the occurrence of skin pigmentation diseases by preventing the melanin pigment from being excessively deposited on the skin. It may have a function to help whitening of
- the prevention or treatment of pigmentation diseases in the present invention does not stop at preventing the deposition of melanin pigment on the skin by applying a composition containing the follicular fluid isolated from trout as an active ingredient to hyperpigmented skin, but more Furthermore, it includes the act of removing melanin already deposited on the skin.
- the pigmentation disease is a skin pigmentation disease, freckles, senile spots, liver spots, melasma, brown or black spots, solar pigmentation, cyanic melasma, hyperpigmentation after drug use, gestational brown spots (gravidic chloasma), or wounds including abrasions and burns, or post-inflammatory hyperpigmentation due to dermatitis, etc., but are not limited thereto.
- the pharmaceutical composition according to the present invention may contain the follicular fluid isolated from the trout of the present invention alone or may additionally contain one or more pharmaceutically acceptable carriers, excipients or diluents.
- the pharmaceutical composition of the present invention may be administered orally or parenterally during clinical administration, but may be administered directly to the skin topically as an external formulation in general.
- composition of the present invention when formulated, it may be formulated using a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant.
- a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, and a surfactant.
- Solid preparations for oral administration include tablets, pills, powders, granules, and capsules, and such solid preparations include at least one excipient in the active ingredient of the present invention, for example, starch, calcium carbonate, sucrose, lactose and It is prepared by mixing gelatin, etc. In addition to simple excipients, lubricants may also be used.
- Liquid preparations for oral administration include suspensions, solutions, emulsions and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances and preservatives are used. may be included.
- Formulations for parenteral administration include topical formulations such as creams, lotions, ointments (semi-solid external preparations), microemulsions, gels, pastes, transdermal preparations (TTS), and the like.
- the daily dosage of the pharmaceutical composition of the present invention is usually in the range of 0.001 to 150 mg/kg body weight, and may be administered once or divided into several doses. However, since the actual dosage of the pharmaceutical composition of the present invention is determined in light of several related factors such as the route of administration, the age, sex, weight, and severity of the patient, the dosage is within the scope of the present invention in any aspect. should not be construed as limiting
- the present invention provides a quasi-drug composition for whitening comprising the follicular fluid isolated from trout as an active ingredient.
- quasi-drug used in the present invention refers to articles with a milder action than pharmaceuticals among articles used for the purpose of diagnosing, treating, improving, alleviating, treating or preventing diseases of humans or animals, for example, in the Pharmaceutical Affairs Act. According to the report, quasi-drugs exclude products used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products with minor or no direct action on the human body.
- the quasi-drug composition of the present invention is a body cleanser, disinfectant cleaner, detergent, kitchen cleaner, cleaning agent, toothpaste, gargle, wet tissue, detergent, soap, hand wash, hair cleaner, hair softener, humidifier filler, mask, ointment and filter filler. It may be prepared in a formulation selected from the group consisting of, but is not limited thereto.
- the present invention provides a composition for external application for skin whitening comprising follicular fluid isolated from trout as an active ingredient.
- external preparation is a preparation provided for external use, and there are external acid preparations, external tablets, external preparations, ointments, warning preparations, suppositories, and the like.
- the external preparation for skin may be a parenteral preparation formulated in solid, semi-solid or liquid form by adding commercially available inorganic or organic carriers, excipients and diluents.
- the preparation for parenteral administration may be a transdermal dosage form selected from the group consisting of drops, ointments, lotions, gels, creams, patches, sprays, suspensions and emulsions, but is not limited thereto.
- Carriers, excipients and diluents that may be included in the external preparation include lactose, dextrose, sucrose, oligosaccharide, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
- composition for external application for skin can be formulated by those skilled in the art without difficulty depending on the formulation or purpose of use of other external preparations for skin other than the follicular fluid isolated from the trout of the present invention, and in this case, it is applied simultaneously with other raw materials. If so, a synergistic effect may occur.
- the composition for external application for skin of the present invention may contain 0.0001 to 30% (w/v) of active ingredients based on the weight of the total composition. If it is included in less than 0.0001% (w/v), the skin whitening effect cannot be substantially expected. On the other hand, if it is included in excess of 30% (w/v), the overall processability of the composition, such as solubility in solvents, may deteriorate, and thus its use for various purposes such as formulations may be limited.
- the present invention comprises the steps of treating trout eggs with papain enzyme; degumming by treating with phosphate; Deoxidation by treating sodium hydroxide; And it provides a method for separating the follicular fluid from the trout comprising the step of treating the acid clay.
- the method for isolating follicular fluid from trout includes treating trout roe with papain enzyme. Specifically, after shredding trout eggs, 1-10% (w/v), specifically 5% (w/v) of papain enzyme was treated to react at 50-60 ° C., specifically 55 ° C., and filtered to 1 Tea extract is obtained.
- the papain enzyme is a vegetable proteolytic enzyme contained in papaya (Carica papaya), and serves to hydrolyze proteins.
- the method of separating the follicular fluid from the trout in the present invention includes the step of degumming by treating with phosphate. Specifically, it is a process of degumming by treating the primary extract with 0.05-0.15%, specifically, 0.1% phosphate. Thereafter, the process of washing the degummed primary extract with DW may be additionally included.
- degumming is one of refining to remove impurities, and the refining includes degumming, deoxidation (refining), bleaching, deodorization, and the like.
- the degumming is a process of removing protein degradation products or phospholipids using phosphoric acid or sulfuric acid.
- the method of separating the follicle fluid from the trout in the present invention includes the step of deoxidation by treating sodium hydroxide. Specifically, it is a process of deoxidizing the primary extract that has undergone the degumming and washing process by treating 1-5%, specifically 2.5% sodium hydroxide. Thereafter, the process of washing the deoxidized primary extract with DW may be additionally included.
- the deoxidation is to remove free fatty acids, also called alkali purification.
- sodium hydroxide is used to neutralize free fatty acids, sodium fatty acid is produced. This sodium fatty acid adsorbs impurities such as pigments and precipitates and removes them.
- the method of separating the follicular fluid from the trout in the present invention includes the step of treating the acid clay.
- the step of treating the acid clay is a process of dehydration, decolorization, and deodorization by treating the acid clay of 1 to 10% (w/v), specifically 5% (w/v) of the extract that has undergone the deoxidation and washing process. is a process through Thereafter, a second purified extract is obtained by stirring and heating, and this is filtered to obtain a purified trout follicle fluid.
- the present invention provides a skin whitening method, a skin regenerating method, a method for improving skin wrinkles, or a method for preventing skin aging, comprising administering to an individual follicular fluid and / and PDRN isolated from trout. to provide.
- the term "administration” means introducing the composition of the present invention to a patient by any suitable method, and the administration route of the composition may be administered through any general route as long as it can reach the target tissue.
- Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration may be administered intranasally, but is not limited thereto.
- composition of the present invention is effective in antioxidant, whitening, skin regeneration, skin aging prevention or wrinkle improvement
- route of administration of the composition may be applied to the skin and administered.
- the present invention provides a method for treating pigmentation diseases, wound regeneration or skin regeneration comprising administering to the damaged skin a pharmaceutical composition comprising follicular fluid isolated from trout and / and PDRN as an active ingredient. to provide.
- treatment refers to any action in which the pigmentation, wound or skin regeneration is improved or changed advantageously by administration of the composition.
- the treatment method using the follicular fluid and/or PDRN isolated from trout of the present invention comprises administering the follicular fluid and/or PDRN isolated from trout to the damaged skin. It is apparent to those skilled in the art that a suitable total daily amount can be determined by a treating physician within the scope of sound medical judgment. In addition, it can be administered once or divided into several doses.
- a specific therapeutically effective amount for a particular patient depends on the type and extent of the response to be achieved, the specific composition, including whether other agents are used, if necessary, the specific composition, the patient's age, weight, general health, It is preferable to apply differently depending on various factors including sex and diet, administration time, administration route and secretion rate of the composition, treatment period, drugs used together or concurrently with a specific composition, and similar factors well known in the pharmaceutical field.
- composition of the present invention includes, without limitation, mammals including humans, and examples thereof may be cattle, pigs, horses, rabbits, mice, and humans.
- the present invention provides the use of follicular fluid and/or PDRN isolated from trout for the preparation of a composition for skin whitening, skin aging prevention, skin regeneration, antioxidant or skin wrinkle improvement.
- composition for skin whitening, skin aging prevention, skin regeneration, antioxidant or skin wrinkle improvement is not particularly limited thereto, but may be a pharmaceutical composition or a cosmetic composition.
- the terms “trout”, “follicular fluid”, “PDRN”, “whitening”, “anti-aging”, “skin regeneration”, “antioxidant”, and “wrinkle improvement” are the same as described above.
- it provides a skin whitening use, skin aging prevention use, skin regeneration use, antioxidant use, or skin wrinkle improvement use of follicular fluid and/or PDRN isolated from trout.
- the eggs were cut into pieces of 0.2 cm or less. Then, the minced eggs were treated with papain enzyme 5% (w/v) and reacted at 55° C. overnight. After that, the enzyme was inactivated by reacting at 95° C. for 2 hours. Thereafter, the primary extract was obtained by filtration with 0.45 ⁇ m filter paper.
- the primary extract was degummed by treatment with 0.1% phosphate and washed with DW at 70° C., and 2.5% NaOH was treated with the washed primary extract at a ratio of 1:1 (v/w) to deoxidize After the process, it was washed again with 70°C DW.
- the washed extract was treated with 5% (w/v) acid clay to undergo dehydration, decolorization, and deodorization processes, and then stirred and heated at 100° C. to obtain a second purified extract.
- a purified trout follicle fluid was obtained by using 0.45 ⁇ m filter paper or filtration under reduced pressure for the second purified extract.
- the fatty acid composition of the purified trout follicle fluid prepared above is shown in Table 1.
- 3X volume TE buffer (10mM Tris-HCl, 1mM EDTA pH 8.0
- a lysis buffer was mixed with the sperm pellet obtained in 2.1 above at a ratio of 1:100 (w/v).
- composition of the lysis buffer (pH 7.5) is as follows.
- Protease K + RNase A (10 mg/ml) was added to the sperm pellet, and the cell lysate was reacted overnight at 55° C. to remove proteins and RNA in addition to nucleic acids.
- the cell lysate cultured in 2.2 above was cooled to 23°C. Thereafter, the sperm cell lysate was filtered through a filter (autoclaved cheese clothes) to collect DNA fibers. Thereafter, after centrifugation at 10,000 ⁇ g for 10 minutes, the supernatant was discarded. Thereafter, cold ethanol (95%) of 2-2.5 times the volume of the sample was added and refrigerated for 1 hour. DNA was collected using a glass rod and washed twice with cold ethanol (70%). Ethanol was evaporated and DNA was stored in liquid form using TE buffer or freeze-dried for long-term storage.
- the extracted DNA was confirmed by 0.4% agarose gel. Pure double-stranded DNA has 1.0 at OD 260 and 1.8 at OD 260 /OD 280 . When contaminated with protein, the OD 260 /OD 280 value is much lower than 1.8, and when contaminated with RNA, the ratio is greater than 1.8.
- FIG. 1 The results of checking the characteristics of DNA extracted from trout sperm in the present invention are shown in Table 1 below. Therefore, it was found that relatively pure DNA was extracted from the trout sperm.
- electrophoresis results of the extracted DNA are shown in FIG. 1 .
- the left view of FIG. 1 is the result of confirming highly polymerized DNA extracted from semen and testes by gel electrophoresis. Lane 1: DNA ladder, Lane 2, Lane 3: PDRN extracted from semen (SEMEN), Lane 4, Lane 5: PDRN extracted from Testis (testis).
- the right side view of FIG. 1 shows a low molecular weight DNA polymer made by slicing a DNA chain to a predetermined size.
- a cream containing the trout follicle fluid prepared in Example 1 and PDRN was prepared.
- Carbopol 940 and Sodium Hyaluronate were stirred in purified water at 75-80° C. for 5 minutes using an azimuth mixer. Then, purified water and glycerin were added, and the mixture was stirred at 75-80°C for 5 minutes using a homomixer. Next, Bees wax, Montanov L, GCC 300kc (INCI Name: Caprylic/Capric Triglyceride), and CEH 100kc (INCI Name: Cetyl Ethylhexanoate) were added and stirred at 75-80°C for 5 minutes using a homomixer.
- NIH3T3 cells were used as fibroblasts. After dispensing the NIH3T3 cells so as to become 10,000 cells/ml, 1 ⁇ L, 5 ⁇ L, and 10 ⁇ L of trout follicle was treated, and it was checked whether the growth of fibroblasts was inhibited.
- FIG. 3 is a result of evaluating the cytotoxicity of trout follicle fluid. As a result, as shown in FIG. 3, the growth of fibroblasts was promoted when 1 ⁇ L and 5 ⁇ L of trout follicle was treated. However, it was confirmed that the cells were toxic at high concentrations by inhibiting proliferation or inducing apoptosis when treated with 10 ⁇ L.
- the whitening effect of the trout follicle fluid isolated in Example 1 was evaluated using a tyrosinase enzyme activity inhibition test. After dispensing NIH3T3 cells so as to become 10,000 cells/ml, 1 ⁇ L to 50 ⁇ L of trout follicle was treated. After removing the medium and washing with PBS, the cells were lysed using a lysis buffer, and then centrifuged to separate the protein supernatant. As the blank sample solution, 0.1M phosphate buffer (phosphate buffer, pH 6.5) was used.
- a 96-well plate 220 ⁇ l of 0.1M phosphate buffer (phosphate buffer, pH 6.5), 20 ⁇ l of protein supernatant, 20 ⁇ l of 1,500 U/ml tyrosinase, and 40 ⁇ l of 1.5 mM L-tyrosine were mixed and reacted at 37° C. for 10 minutes. Then, the absorbance was measured at 490 nm. The activity inhibition rate was converted to the following formula.
- Tyrosinase activity inhibition rate (%) 100-(b-b'/a-a')*100
- a', b' Absorbance measured by substituting a buffer for tyrosinase
- the antioxidant effect of the trout follicle fluid isolated in Example 1 was evaluated using the DPPH assay (active oxygen) scavenging effect measurement experiment.
- the DPPH radical inhibitory effect was prepared by dissolving 0.5 mL of the trout follicle fluid sample in DMSO, and the DPPH solution was dissolved in 95% ethanol to make 120 ⁇ M. After mixing 10 ⁇ L of follicle extract and 195 ⁇ L of DPPH solution and vortexing, it was left at room temperature without light passing for 15 minutes, and then absorbance was measured at 517 nm.
- Example 2 In order to determine whether the PDRN isolated in Example 2 is suitable for use as a cosmetic composition, it was checked whether it contains toxic elements including heavy metals such as mercury, lead, arsenic, antimony, cadmium, and nickel. Prosecutors were commissioned by Soonchunhyang University for objectivity and fairness. 2 is a test test report of PDRN. As a result, as shown in FIG. 2 , it was found that heavy metals such as mercury, lead, arsenic, antimony, cadmium, and nickel were not included, so that it was suitable for use as a cosmetic composition.
- toxic elements including heavy metals such as mercury, lead, arsenic, antimony, cadmium, and nickel.
- HaCaT cells which are human dermal keratinocytes
- HDF-1 cells which are human dermal fibroblasts
- PDRN and EGF were treated by concentration and cytotoxicity was evaluated.
- FIG. 6 is a result of evaluating the cytotoxicity of PDRN.
- 7 is a result of evaluating the cytotoxicity of EGF.
- FIG. 6 it was confirmed that there was no toxicity to HaCaT cells and HDF-1 cells even when PDRN was treated with 0.01 to 100 ⁇ g/ml.
- EGF was treated with 0.01 to 100 ⁇ g/ml, there was no toxicity to these skin cells.
- the purpose of this study was to evaluate whether PDRN and EGF exist in the dermis of the skin and inhibit the activity of collagenase that promotes skin aging.
- the activity of collagen degrading enzyme was evaluated by treatment with 50-200 ⁇ g/ml of PDRN and 50-200 ⁇ g/ml of EGF.
- FIG. 8 is a result of evaluation of inhibition of collagenase activity of PDRN and EGF. As shown in FIG. 8 , it was confirmed that epigallocatechin gallate (EGCG) used as a positive control showed about 33% collagenase inhibitory ability, whereas PDRN and EGF had no collagenase inhibitory ability at all concentrations.
- EGCG epigallocatechin gallate
- PDRN and EGF inhibit the activity of elastin-degrading enzyme (elastase), which is present in the skin dermis and promotes skin aging.
- elastin-degrading enzyme elastase
- the activity of elastin-degrading enzyme was evaluated by treating 50-200 ⁇ g/ml of PDRN and 50-200 ⁇ g/ml of EGF.
- FIG. 9 is a result of evaluation of inhibition of elastin-degrading enzyme activity of PDRN and EGF. As shown in FIG. 9 , it was confirmed that when PDRN was treated with 200 ⁇ g/ml, the activity of elastin degrading enzyme was inhibited by 12% compared to the control. However, in the EGF-treated group, elastin-degrading enzyme inhibitory ability did not appear at any concentration.
- Photoaging which is extrinsic aging, is caused by ultraviolet rays, and ultraviolet rays absorbed into the skin increase the activity of MMPs (Matrix metalloproteinases) through induction of reactive oxygen species (ROS) and decrease collagen synthesis to accelerate aging.
- MMPs Microx metalloproteinases
- ROS reactive oxygen species
- Figure 10 shows that PDRN suppresses the expression of MMP-1 and MMP-3 increased by the treatment with reactive oxygen species.
- FIG. 10 it was confirmed through RT-PCR that PDRN suppressed the expression of MMP-1 and MMP-3 increased by H 2 O 2 , and EGCG (Epigallocatechin gallate) used as a positive control. ) was also confirmed to inhibit the expression of MMP-1 and MMP-3.
- PDRN has anti-aging effects such as inhibiting the process of MMPs decomposing collagen by suppressing the expression of MMPs induced by reactive oxygen species, suppressing skin wrinkles and improving skin elasticity.
- cell regeneration efficacy evaluation was performed using HaCaT cells, which are human skin keratinocytes.
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Abstract
Description
지방산 종류 | 평균함량(중량%) | 표준편차 |
Myristic acid | 1.55 | 0.34 |
Myristoleic acid | 0.02 | 0.01 |
Pentadecanoic acid | 0.22 | 0.03 |
Palmitic acid | 7.38 | 0.88 |
Palmitoleic acid | 2.06 | 0.25 |
Heptadecanoic acid | 0.31 | 0.04 |
cis-10-heptadecanoic acid | 0.16 | 0.03 |
Stearic acid | 1.85 | 0.21 |
Elaidic acid | 0.07 | 0.01 |
Oleic acid | 4.34 | 0.98 |
Linoleic acid | 1.25 | 0.23 |
Arachidic acid | 0.08 | 0.01 |
c-linolenic acid | 0.15 | 0.02 |
Eicosenoic acid | 0.27 | 0.09 |
a-linolenic acid | 1.33 | 0.29 |
cis-11,14-eicosadienoic acid | 1.09 | 0.23 |
Behenic acid | 0.03 | 0.01 |
Homo-c-linolenic acid | 0.11 | 0.02 |
Erucic acid | 0.06 | 0.02 |
Eicosatrienoic acid | 0.16 | 0.03 |
Arachidonic acid | 2.47 | 0.28 |
cis-13,16-docosadienoic acid | 0.42 | 0.11 |
Eicosapentaenoic acid | 2.81 | 0.34 |
Nervonic acid | 0.18 | 0.02 |
Docosapentaenoic acid | 0.64 | 0.07 |
Docosahexaenoic acid | 5.59 | 0.70 |
omega-3 지방산 | 14.10 | 0.33 |
omega-6 지방산 | 5.32 | 0.18 |
포화지방산 | 15.51 | 0.19 |
단일 불포화 지방산 | 9.17 | 0.42 |
다중 불포화 지방산 | 21.30 | 0.28 |
총합 | 100 | 6.65 |
Controls | Specifications | Results |
Appearance of Solution S (1.0% (w/v) in water) | Clear | Complies |
Absorbance at 600nm of the S solution | A 600nm≤ 0.010 | 0.002 |
Absorbance ratio 260nm/280nm | 1.6 to 2.0 | 1.78 |
Ribonucleic acid | ≤3.0% | 0.6% |
Protein | ≤1.0% | 0.5% |
Sodium | 4.5~5.5% | 2.1% |
Calcium | ≤ 500ppm | 60 ppm |
Magnesium | ≤ 500ppm | 5 ppm |
Sulfate | ≤ 0.3% | 0.2% |
Loss on Drying | ≤ 13% | 8.1% |
Microbiological Quality: Bacteria | ≤ 100CFU/g | 0 CFU/g |
성분 | 함량(중량%) |
정제수 | 38.67 |
Glycerin | 8.00 |
정제수 | 30.00 |
Carbopol 940 | 0.40 |
Sodium Hyaluronate | 0.03 |
Bees wax | 1.00 |
Montanov L | 2.50 |
GCC 300kc | 3.00 |
CEH 100kc | 3.00 |
정제수 | 3.00 |
L-Arginine | 0.40 |
1,2-헥산다이올 | 2.00 |
송어알(정제 난포액) | 7.00 |
PDRN | 1.00 |
총합 | 100.00 |
Claims (9)
- 송어에서 분리된 난포액을 유효성분으로 포함하는 미백용 화장료 조성물.
- 제1항에 있어서,상기 화장료 조성물은 항산화 활성을 가지는 것인, 화장료 조성물.
- 제1항에 있어서,상기 난포액은 송어 알에 파파인 효소를 처리하는 단계; 인산염을 처리하여 탈검하는 단계; 수산화나트륨을 처리하여 탈산하는 단계; 및 산성백토를 처리하는 단계에 의해 제조되는 것인, 화장료 조성물.
- 제1항에 있어서,상기 화장료 조성물은 PDRN(polydeoxyribonucleotide)을 더 포함하는 것인, 화장료 조성물.
- 제4항에 있어서,상기 화장료 조성물은 피부 주름 개선, 피부 탄력 증진 또는 피부 세포 재생을 유도하는 것인, 화장료 조성물.
- 송어에서 분리된 난포액을 유효성분으로 포함하는 색소 침착 질환의 예방 또는 치료용 약학적 조성물.
- 송어에서 분리된 난포액을 유효성분으로 포함하는 미백용 의약외품 조성물.
- 송어에서 분리된 난포액을 유효성분으로 포함하는 미백용 피부 외용제 조성물.
- 송어 알에 파파인 효소를 처리하는 단계;인산염을 처리하여 탈검하는 단계;수산화나트륨을 처리하여 탈산하는 단계; 및산성백토를 처리하는 단계를 포함하는 송어에서 난포액을 분리하는 방법.
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PCT/KR2021/002798 WO2022191339A1 (ko) | 2021-03-08 | 2021-03-08 | 송어에서 분리된 pdrn과 난포액을 포함하는 화장료 조성물 |
KR1020217043109A KR20220127738A (ko) | 2021-03-08 | 2021-03-08 | 송어에서 분리된 pdrn과 난포액을 포함하는 화장료 조성물 |
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Citations (4)
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KR20090079413A (ko) * | 2008-01-17 | 2009-07-22 | 정래준 | 어류 정액 또는 알로부터 분리된 dna 중합체단편복합체 및 그의 제조방법 |
KR101353628B1 (ko) * | 2006-03-31 | 2014-01-20 | 닛세이 바이오 카부시키 가이샤 | 기초화장품용 배합제 및 기초화장품 |
KR20140012274A (ko) * | 2012-07-19 | 2014-02-03 | (주)아모레퍼시픽 | 간 x 수용체 활성제를 포함하는 피부 미백제 |
KR20190139633A (ko) * | 2018-06-08 | 2019-12-18 | (주) 비앤에프솔루션 | 어류 정액 및 정소에서 피디알엔을 추출하는 방법 |
-
2021
- 2021-03-08 KR KR1020217043109A patent/KR20220127738A/ko not_active Application Discontinuation
- 2021-03-08 WO PCT/KR2021/002798 patent/WO2022191339A1/ko active Application Filing
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KR101353628B1 (ko) * | 2006-03-31 | 2014-01-20 | 닛세이 바이오 카부시키 가이샤 | 기초화장품용 배합제 및 기초화장품 |
KR20090079413A (ko) * | 2008-01-17 | 2009-07-22 | 정래준 | 어류 정액 또는 알로부터 분리된 dna 중합체단편복합체 및 그의 제조방법 |
KR20140012274A (ko) * | 2012-07-19 | 2014-02-03 | (주)아모레퍼시픽 | 간 x 수용체 활성제를 포함하는 피부 미백제 |
KR20190139633A (ko) * | 2018-06-08 | 2019-12-18 | (주) 비앤에프솔루션 | 어류 정액 및 정소에서 피디알엔을 추출하는 방법 |
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Title |
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NOH TAI, CHUNG BO, KIM SU, LEE MI, KIM MOON, YOUN CHOON, CHANG SUNG: "Novel Anti-Melanogenesis Properties of Polydeoxyribonucleotide, a Popular Wound Healing Booster", INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 17, no. 9, pages 1448, XP055965501, DOI: 10.3390/ijms17091448 * |
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