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WO2025037843A1 - Procédé de production de pdrn à partir d'une plante - Google Patents

Procédé de production de pdrn à partir d'une plante Download PDF

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Publication number
WO2025037843A1
WO2025037843A1 PCT/KR2024/011785 KR2024011785W WO2025037843A1 WO 2025037843 A1 WO2025037843 A1 WO 2025037843A1 KR 2024011785 W KR2024011785 W KR 2024011785W WO 2025037843 A1 WO2025037843 A1 WO 2025037843A1
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WO
WIPO (PCT)
Prior art keywords
pdrn
skin
composition
present
plant
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/KR2024/011785
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English (en)
Korean (ko)
Inventor
모상현
서효현
최선미
권석현
오경언
이나영
김유아
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BIO-FD&C Co Ltd
BIO FD&C CO Ltd
Kolmar Korea Co Ltd
Original Assignee
BIO-FD&C Co Ltd
BIO FD&C CO Ltd
Kolmar Korea Co Ltd
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Filing date
Publication date
Application filed by BIO-FD&C Co Ltd, BIO FD&C CO Ltd, Kolmar Korea Co Ltd filed Critical BIO-FD&C Co Ltd
Publication of WO2025037843A1 publication Critical patent/WO2025037843A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/153—Nucleic acids; Hydrolysis products or derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • A23L33/13—Nucleic acids or derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/711—Natural deoxyribonucleic acids, i.e. containing only 2'-deoxyriboses attached to adenine, guanine, cytosine or thymine and having 3'-5' phosphodiester links
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • 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
    • A61P17/06—Antipsoriatics
    • 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
    • A61P17/16—Emollients or protectives, e.g. against radiation
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/04—Plant cells or tissues
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/26—Preparation of nitrogen-containing carbohydrates
    • C12P19/28—N-glycosides
    • C12P19/30—Nucleotides
    • C12P19/34—Polynucleotides, e.g. nucleic acids, oligoribonucleotides

Definitions

  • the present invention relates to a method for producing PDRN (polydeoxyribonucleotide) from a plant, PDRN produced by the method, a cosmetic composition for improving skin, a pharmaceutical composition, a quasi-drug composition, a food composition, a health functional food, and a feed composition containing the PDRN as an active ingredient.
  • PDRN polydeoxyribonucleotide
  • Polydeoxyribonucleotide (PDRN), recently used as a key material for cosmeceuticals and bio-cosmetics, is a DNA-derived polymeric substance that promotes self-regeneration of damaged bodies and tissues as a tissue regeneration active substance.
  • PDRN is a DNA fragment that is cut into small sizes by various physical or chemical methods. When administered into the body, it is known to stimulate the adenosine A2A receptor on the surface of the tissue, promote body regeneration, and have the effect of quickly recovering from wounds and reducing pain.
  • PDRN is mainly manufactured from DNA extracted from salmon semen, and is mainly used for the treatment of wounds caused by skin grafting and tissue repair, but it is also used for some tissue regeneration or inflammation treatment for which there is no proper treatment depending on the medical professional's judgment (Korean Patent No. 10-1722181). It is known to be used for the treatment of a wide range of disorders such as diabetic ulcers, scars, vascular insufficiency, and female pattern hair loss.
  • PDRN is an animal-based raw material extracted from salmon semen, and its effectiveness has been proven, but it is not suitable for vegan products that do not contain animal ingredients and pest-free cosmetics that have not been tested on animals.
  • vegan certification is granted only to products that do not use any raw materials that can be obtained from animals, it has the advantage of not only protecting animals but also being less irritating to the skin. Therefore, methods to extract PDRN using plants as raw materials rather than animals are being attempted, but there have been few visible results so far.
  • the inventors of the present invention have conducted extensive research efforts to develop a method for producing PDRN from plant materials, and have developed a method for producing high-purity PDRN from plants, and have confirmed that PDRN produced by this method has the effects of improving wrinkles, healing wounds, protecting the skin from ultraviolet rays, strengthening the skin barrier, moisturizing, and anti-aging, thereby completing the present invention.
  • the present invention aims to solve the above-mentioned problems and other problems related thereto.
  • An exemplary object of the present invention is to provide a method for producing PDRN (polydeoxyribonucleotide) from a plant, comprising the following steps.
  • Another exemplary object of the present invention is to provide PDRN produced by the above production method.
  • Another exemplary object of the present invention is to provide a cosmetic composition for skin improvement, comprising PDRN as an effective ingredient.
  • Another exemplary object of the present invention is to provide a pharmaceutical composition for preventing or treating any skin disease selected from the group consisting of skin diseases caused by ultraviolet rays, psoriasis, skin wounds, skin scars, and skin inflammation, comprising the PDRN as an active ingredient.
  • Another exemplary object of the present invention is to provide a pharmaceutical composition for skin improvement, comprising PDRN as an effective ingredient.
  • Another exemplary object of the present invention is to provide a food composition for skin improvement, comprising PDRN as an effective ingredient.
  • Another exemplary object of the present invention is to provide a skin improvement feed composition comprising the PDRN as an effective ingredient.
  • the present invention provides a method for producing PDRN (polydeoxyribonucleotide) from a plant, comprising the following steps.
  • PDRN refers to polydeoxyribonucleotide, which 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, and the PDRN used in the present invention may have a size of 2000 bp or less and a molecular weight of 74 kDa or less.
  • the term “PDRN” may be a type of the nucleic acid fragment.
  • the plant body is a concept including the whole plant or a part of the plant body, such as a leaf, stem, root or a part thereof, and includes a culture solution in which the whole or a part of the plant body is cultured.
  • the culturing in step (b) can be performed at 60°C to 70°C, and specifically, can be performed at 65°C, but is not limited thereto.
  • the freezing may be performed at -90°C to -70°C, specifically at -85°C to -75°C, and more specifically at -80°C.
  • the crushing may be performed by i) first crushing the frozen plant body by thawing it at room temperature, and ii) secondarily crushing it through physical crushing.
  • Plants contain 70-80% of moisture inside, so cells can be naturally destroyed during the thawing process after the freezing step as described above.
  • the concentration of the sodium chloride may be 0.1 M to 1 M, specifically, 0.3 M to 0.8 M, more specifically, 0.4 M to 0.6 M, and even more specifically, 0.5 M, but is not limited thereto.
  • the PDRN production method can be applied to all plants regardless of type.
  • PDRN was produced by applying the production method to hibiscus ( Hibiscus sabdariffa ) and Houttuynia Cordata , which are of different genus and species.
  • the present invention provides PDRN produced by the above production method.
  • the present invention provides a cosmetic composition for skin improvement comprising the PDRN as an effective ingredient.
  • skin improvement means any act of improving or benefiting the skin by using PDRN produced by the production method of the present invention.
  • the skin improvement may be, for example, skin protection from ultraviolet rays, strengthening of the skin barrier, moisturizing, wrinkle improvement, skin regeneration, wound healing, or anti-aging.
  • the cosmetic composition of the present invention can be manufactured in various forms, for example, the cosmetic composition can be manufactured in any formulation commonly manufactured in the art, and for example, can be formulated as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray, but is not limited thereto.
  • the cosmetic composition can have a formulation selected from the group consisting of a skin lotion, a skin softener, a skin toner, an astringent, a lotion, a milk lotion, a moisture lotion, a nutrition lotion, a massage cream, a nutrition cream, a moisture cream, a hand cream, an essence, a nutrition essence, a pack, a soap, a shampoo, a cleansing foam, a cleansing lotion, a cleansing cream, a body lotion, a body cleanser, an emulsion, a lipstick, a makeup base, a foundation, a pressed powder, and a loose powder, but is not limited thereto.
  • the cosmetic composition of the present invention may contain, in addition to its effective ingredient, a carrier acceptable for cosmetic formulations.
  • a carrier acceptable for cosmetic formulations refers to a compound or composition already known and used or a compound or composition to be developed in the future that can be included in cosmetic formulations, which does not have toxicity, instability or irritation beyond what the human body can adapt to when in contact with the skin.
  • the carrier may be included in the cosmetic composition of the present invention in an amount of about 1 wt % to about 99.99 wt % based on the total weight of the composition, preferably about 5 wt % to about 99 wt % of the composition weight.
  • the carrier may include alcohol, oil, surfactant, fatty acid, silicone oil, humectant, moisturizer, viscosity modifier, emulsion, stabilizer, UV blocker, colorant, fragrance, etc.
  • Compounds or compositions that can be used as the carrier, such as alcohol, oil, surfactant, fatty acid, silicone oil, humectant, moisturizer, viscosity modifier, emulsion, stabilizer, UV blocker, colorant, fragrance, etc. are already known in the art, and therefore, a person skilled in the art can select and use an appropriate corresponding material or composition.
  • the present invention provides a pharmaceutical composition for preventing or treating any skin disease selected from the group consisting of skin diseases caused by ultraviolet rays, psoriasis, skin wounds, skin scars, and skin inflammation, comprising the PDRN as an active ingredient.
  • the skin disease caused by ultraviolet rays may be at least one selected from the group consisting of sunburn, erythema, edema, pigmentation, skin inflammation, skin keratinization, photoallergy, phototoxicity, photosensitivity, and photoaging.
  • prevention means any act of improving a skin disease by administering the pharmaceutical composition of the present invention
  • treatment means any act of improving, alleviating, or beneficially changing a skin disease caused by administering the pharmaceutical composition of the present invention.
  • Immprovement in the present invention means any act of improving a skin disease by administering the composition.
  • the "pharmaceutical composition” means a composition manufactured for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to conventional methods.
  • it may be formulated in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, and syrups, depending on the route of administration, and may be formulated and used in the form of external preparations and sterile injectable solutions.
  • the administration route may be any appropriate route including a topical route, an oral route, an intravenous route, an intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be combined and used.
  • An example of a combination of two or more routes is a case where two or more drug formulations are combined according to the route of administration, for example, a case where one drug is first administered via the intravenous route and another drug is secondarily administered via the topical route.
  • the pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent according to a conventional method.
  • Pharmaceutically acceptable carriers are well known in the art depending on the administration route or formulation, and specifically, reference can be made to the pharmacopoeias of each country including the 'Korean Pharmacopoeia'.
  • Carriers, excipients and diluents that may be included in the composition of the present invention include, but are not limited to, lactose, dextrose, sucrose, 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. Additionally, carriers, excipients and diluents that may be included in the composition of the present invention may be non-natural carriers, but are not limited thereto.
  • the pharmaceutical composition of the present invention can be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions according to conventional methods. Specifically, when formulating, it can be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants that are commonly used.
  • Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations can be prepared by mixing the compound with at least one excipient, for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc.
  • excipients for example, starch, calcium carbonate, sucrose, lactose, gelatin, etc.
  • lubricants such as magnesium stearate and talc can also be used.
  • Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives may be included.
  • Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories.
  • Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.
  • Suppository bases may include withepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin, etc.
  • the pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount.
  • the pharmaceutically effective amount means an amount sufficient to treat a disease with a reasonable benefit/risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined based on factors including the patient's health condition, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the method of administration, the time of administration, the route of administration, and the excretion rate, the treatment period, the drugs used in combination or simultaneously, and other factors well known in the medical field.
  • the dosage and frequency of administration do not limit the scope of the present invention in any way.
  • the pharmaceutical composition of the present invention can be administered to mammals such as rats, dogs, cats, cows, horses, pigs, and humans through various routes, and humans may be preferred. All modes of administration are conceivable, and include, but are not limited to, oral, intravenous, intramuscular, or subcutaneous injection.
  • the meaning of “comprising as an effective ingredient” means including an effective amount capable of exhibiting a preventive or therapeutic effect on a skin disease as a pharmaceutical composition.
  • the present invention provides a pharmaceutical composition for skin improvement comprising the PDRN as an effective ingredient.
  • PDRN and “skin improvement” of the present invention are as described above.
  • the quasi-drug composition of the present invention may contain, in addition to the PDRN, ingredients commonly used in quasi-drug compositions, and may include, for example, an abrasive, a humectant, a binder, a foaming agent, a sweetener, a preservative, a medicinal ingredient, a flavoring agent, a pigment, a solvent, a whitening agent, a solubilizer, or a pH adjuster, but is not limited thereto.
  • ingredients commonly used in quasi-drug compositions may include, for example, an abrasive, a humectant, a binder, a foaming agent, a sweetener, a preservative, a medicinal ingredient, a flavoring agent, a pigment, a solvent, a whitening agent, a solubilizer, or a pH adjuster, but is not limited thereto.
  • the pharmaceutical composition of the present invention may be, but is not particularly limited to, an external preparation, a powder, a sterilizing disinfectant, a toothpaste, an ointment, a lotion, an internal preparation (vitamin/mineral preparation, a tonic), a wet tissue, a spray patch, a bandage, or a patch, and the like.
  • the pharmaceutical composition may be appropriately selected by a person skilled in the art from conventional techniques known in the art.
  • the present invention provides a food composition for skin improvement comprising the PDRN as an effective ingredient.
  • PDRN and “skin improvement” of the present invention are as described above.
  • the food composition of the present invention can be manufactured into various dosage forms, and unlike general drugs, has the advantage of having no side effects that may occur when taking drugs for a long period of time since it uses food as a raw material.
  • the food composition of the present invention can be manufactured into any form, and specifically, it can be at least one dosage form selected from the group consisting of health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, beverages, gums, and candies, but is not particularly limited thereto.
  • the food composition of the present invention may contain food additives in addition to the effective ingredients.
  • Food additives can be generally understood as substances that are added to, mixed in, or infiltrated into food during the manufacturing, processing, or preservation of food, and their safety must be guaranteed because they are consumed daily and for a long period of time with food.
  • the food additives are classified into sweeteners, flavoring agents, preservatives, emulsifiers, acidulants, thickeners, etc. in terms of function, and are not particularly limited as long as they are consistent with the purpose to be achieved by the food composition of the present invention.
  • the food composition of the present invention may contain, in addition to the food additives, physiologically active substances or minerals known in the art for the purpose of functionality and nutritional supplementation and whose safety as a food additive is guaranteed.
  • physiologically active substances or minerals are not particularly limited as long as they are consistent with the purpose to be achieved by the food composition of the present invention.
  • the food composition of the present invention may contain the aforementioned food additives in an effective amount that can achieve the purpose of addition depending on the product type, and with respect to other food additives that may be contained in the food composition of the present invention, reference may be made to the food codes or food additive codes of each country.
  • active ingredient includes an ingredient that exhibits the desired activity alone or can exhibit the activity together with a carrier that is inactive by itself.
  • the composition of the present invention may contain a compound or natural extract known to have a skin improvement effect so as to increase or enhance the skin improvement effect.
  • the effective ingredient may be included in any amount (effective amount) depending on the intended use, formulation, mixing purpose, etc., as long as it can exhibit skin improvement activity.
  • a typical effective amount may be included within a range of 0.001 wt % to 99.99 wt % based on the total weight of the composition.
  • the "effective amount” refers to the amount of the effective ingredient that can induce a skin improvement effect. This effective amount can be experimentally determined within the normal ability range of a person skilled in the art.
  • the present invention provides a health functional food for skin improvement containing the PDRN as an effective ingredient.
  • PDRN and “skin improvement” of the present invention are as described above.
  • health functional food in the present invention refers to a food manufactured and processed using raw materials or ingredients having functionality useful to the human body.
  • the “functionality” above means obtaining a useful effect for health purposes such as regulating nutrients or physiological effects for the structure and function of the human body.
  • the health functional food of the present invention can be manufactured by a method commonly used in the art, and can be manufactured by adding raw materials and ingredients commonly added in the art during manufacturing.
  • the formulation of the above health functional food can be manufactured without limitation as long as it is a formulation recognized as a health functional food, and non-limiting examples of the formulation include one or more formulations selected from the group consisting of health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, beverages, gums, and candies, but are not particularly limited thereto.
  • health functional food preparations such as tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, bars, beverages, gums, and candies, but are not particularly limited thereto.
  • the present invention provides a skin improvement feed composition comprising the PDRN as an effective ingredient.
  • PDRN and “skin improvement” of the present invention are as described above.
  • the above feed composition may include a feed additive.
  • the feed additive of the present invention corresponds to an auxiliary feed under the Feed Management Act.
  • the term "feed” may mean any natural or artificial diet, meal, etc., or a component of said meal, especially for eating, ingesting, and digesting by an animal or suitable therefor.
  • the type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used.
  • Non-limiting examples of the above feed include plant feed such as grains, roots, food processing by-products, algae, fibers, pharmaceutical by-products, fats, starches, meal, or grain by-products; and animal feed such as proteins, inorganic substances, fats, minerals, fats, simple proteins, zooplankton, or food. These may be used alone or in combination of two or more.
  • the present invention provides a method for improving skin, comprising a step of administering or applying a composition containing the PDRN in an effective amount for improving skin to a subject in need thereof.
  • PDRN and “skin improvement” of the present invention are as described above.
  • “effective amount” means an amount sufficient for the composition of the present invention described above to achieve skin improvement efficacy.
  • subject includes, but is not particularly limited to, a human, a monkey, a cow, a horse, a sheep, a pig, a chicken, a turkey, a quail, a cat, a dog, a mouse, a rat, a rabbit or a guinea pig, and is preferably a mammal, and more preferably a human.
  • the term “administration” means providing a given substance to a subject by any suitable method.
  • the term “application” means any method of bringing a composition according to the present invention into contact with the skin of a subject by any suitable method, thereby aiming at allowing the composition to be absorbed into the skin.
  • the route of administration of the composition of the present invention may be oral or parenteral administration through any common route as long as it can reach the target tissue.
  • the composition of the present invention may be administered using any device capable of delivering the active ingredient to the target cell, tissue or organ.
  • Another aspect of the present invention to achieve the above object provides a skin improvement use of PDRN of the present invention.
  • Another aspect of the present invention for achieving the above object provides the use of PDRN for the manufacture of a cosmetic composition for improving skin.
  • Another aspect of the present invention for achieving the above object provides the use of PDRN for the manufacture of a pharmaceutical composition for preventing or treating skin diseases.
  • PDRN and “skin improvement” of the present invention are as described above.
  • high purity and high concentration PDRN can be produced from a plant, and the plant-derived PDRN thus produced has the effects of protecting the skin from ultraviolet irradiation, strengthening the skin barrier, moisturizing, improving wrinkles, regenerating the skin, healing wounds, or anti-aging, and can be usefully used as a cosmetic composition for improving the skin, a pharmaceutical composition, a food composition, a health functional food, a quasi-drug composition, and a feed composition.
  • Figure 1 shows the results of confirming PDRN obtained from the plants of Aster japonica and Hibiscus.
  • lysis buffer contained 37.5 mL of distilled water, 1.16 g of sodium chloride (NaCl, final concentration 0.5 M), and 1.75 mL of surfactant (Alpicare gl 612).
  • the frozen samples were thawed at room temperature with the lysis buffer to induce cell rupture, and the cells were ground finely in a blender to perform secondary cell disruption.
  • the ethanol was discarded, the pellet was completely dried in the air, and purified water was added to obtain genomic DNA.
  • the obtained genomic DNA was treated 4 or 8 times with a sonicator to obtain PDRN with a small molecular weight of 100-300 bp (Fig. 1).
  • the purity of the obtained PDRN was measured to be 1.8 or more and less than 2.0 (Table 1).
  • the above purity was determined by the ratio of the absorbance value at 260 nm/the absorbance value at 280 nm (A260/280 ratio). If the A260/280 ratio is less than 1.8, the DNA concentration is considered to be very low or there is protein contamination. If it is 1.8 to 1.9, the purity is considered to be good, and if it is 2.0 or higher, the DNA concentration is considered to be high, or there is RNA contamination or DNA fragmentation.
  • Example 1 An experiment was conducted to determine whether the PDRN obtained in Example 1 causes irritation to skin cells.
  • HaCaT cells keratinocytes
  • Detroit cells fibroblasts
  • DMEM Dubelcco's Modified Eagle's Medium
  • FBS Fetal Bovine Serum
  • Antibiotic-Antimycotic 1%
  • control group to which purified water was added and the experimental group to which PDRN derived from the Eoseongcho plant was treated at 0.5%, 1%, and 5% weight parts of each cell were cultured for an additional 24 hours, respectively.
  • MTT 3-(4, 5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide
  • Control group PDRN 0.5% PDRN 1% PDRN 5% Cell viability (%) HaCaT cells 100 103 112 119 Detroit cell 100 106 110 113
  • Detroit551 cells were seeded on a 48-well plate and cultured for 24 hours under the same conditions as in Example 3-1. Thereafter, the control group to which purified water was added, the experimental group treated with 0.5%, 1%, and 5% by weight of PDRN derived from the plant of E. coli, and the positive control group treated with 5 ng/ml TGF- ⁇ 1 were additionally cultured for 48 hours. Then, 20 ⁇ L of the diluted supernatant of the cultured medium was placed into each antibody-coated microtiter plate along with 20 ⁇ L of the PIP standard substance in the PIP EIA Kit (Takara, Cat.
  • Example 1 In order to examine the skin cell regeneration effect of PDRN obtained in Example 1, a wound healing assay was used to observe whether cell proliferation and migration were promoted by the test substance, thereby evaluating the anti-aging efficacy.
  • HaCaT cells were seeded on a 24-well plate and cultured for 24 hours under the same conditions as in Example 2-1. Thereafter, the control group to which purified water was added, the experimental group treated with 0.5%, 1%, or 2% by weight of PDRN derived from the plant of E. coli, and the positive control group treated with 100 ng/ml EGF were additionally cultured for 18 hours at 37°C and 5% CO2 .
  • Cell images were taken under a microscope before the start of the additional culture and cultured. After 16 hours of culture, the medium was removed, a fixing solution (4% paraformaldehyde) was added, and the cells were incubated at room temperature for 15 minutes and washed three times with PBS. The fixed cells could be stored at 4°C for one month.
  • the degree of wound healing was calculated using a program called Image J by taking photographs under a microscope.
  • the light source for UV irradiation was Ultraviolet crosslinker CL-1000 (Ultra-Violet Products, CA), which emits ultraviolet rays with a wavelength of 302 nm.
  • Human keratinocytes (HaCaT) were placed in a 24-well plate at a concentration of 1x105 cells/ml and cultured in an incubator for 24 hours. The medium was removed and washed with phosphate buffer. Again, 500 ⁇ L of phosphate buffer was added, and then irradiated with UV rays at 10 mJ/cm2.
  • the medium was replaced with fresh cell culture medium without FBS, and PDRN derived from the plant obtained in Example 1 was treated at various concentrations (0.5%, 1%, and 5%), and then cultured for an additional 24 hours. After that, 5 mg/ml of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma) was added and cultured in an incubator for 3 hours.
  • the formed fomazan was dissolved in DMSO and transferred to a 96-well plate, and the absorbance was measured at 595 nm using an ELISA reader to confirm the cell viability.
  • HaCaT cells negative control
  • a normal control group that was not treated with UV or PDRN in HaCaT cells were prepared.
  • Example 1 cell viability increased in a concentration-dependent manner when PDRN derived from the plant was treated compared to the negative control group (Table 5). Therefore, it was confirmed that the PDRN obtained in Example 1 can protect skin cells from ultraviolet rays.
  • Example 1 The skin moisturizing effect of PDRN obtained in Example 1 was evaluated.
  • human keratinocyte cell line HaCaT was cultured in a 100 mm/60.1 cm2 culture dish with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS), and 1% Antibiotic-Antimycotic (GIBCO, Cat.# 15240-062) at 37°C, 5% CO2 .
  • DMEM Dulbecco's Modified Eagle's Medium
  • FBS Fatal bovine serum
  • GEBCO Antibiotic-Antimycotic
  • the medium was replaced with DMEM free medium (without FBS), and then treated with PDRN derived from the plant of Haematology japonica obtained in Example 1 at various concentrations (0.5%, 1%, 5%). Human keratinocytes not treated with PDRN were prepared as a control. After additional culture for 24 hours, the effect on the expression of genes related to moisturizing (Aquaporin-3, AQP-3) was confirmed.
  • Gene expression rates were measured using the following Real-time PCR method.
  • RNA isolation was performed using the FastLane Cell one-step buffer set (QIAGEN Cat.# 216213).
  • FCW cell wash buffer
  • 50 ul of cell treatment mixture a mixture of gDAN Wipeout buffer added to FCPW buffer containing FCPL buffer
  • the extracted RNA was used as a template and the PCR was performed using a Rotor Gene Q Real-time PCR Machine (Qiagen) according to the manual of 2X QuantiTect SYBR Green RT-PCR Master mix.
  • the primers used in the experiment were normalized with QuantiTect® primerassays (AQP-3, Cat.# QT00212996) from Qiagen.
  • PDRN Treatment concentration (%) TERT gene expression rate (% of control) Negative control 0 1.0 Positive control (1 mM SA) 0 5.2 Experimental group 0.5 1.4 1.0 4.2 5.0 3.8
  • filaggrin a gene related to the skin barrier function of human keratinocytes, was measured to evaluate the skin barrier strengthening effect of PDRN derived from the plant of ...

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Abstract

La présente invention concerne un procédé de production de polydésoxyribonucléotide (PDRN) à partir d'une plante, le PDRN produit par le procédé, et une composition cosmétique, une composition pharmaceutique, une composition de quasi-médicament, une composition alimentaire, un aliment fonctionnel pour la santé, et une composition d'aliments pour animaux, chacun comportant le PDRN comme principe actif pour le traitement des affections cutanées. La présente invention permet de produire du PDRN de haute pureté et de haute concentration à partir d'une plante, et le PDRN dérivé de la plante ainsi produit présente une protection de la peau contre les rayons ultraviolets, une amélioration de la barrière cutanée, une hydratation, une atténuation des rides, une régénération de la peau, une cicatrisation des plaies ou des effets antivieillissement, et peut donc être avantageusement utilisé comme composition cosmétique pour le traitement des affections cutanées, comme composition pharmaceutique, comme composition alimentaire, comme aliment fonctionnel pour la santé, comme composition quasi-médicamenteuse et comme composition pour l'alimentation animale.
PCT/KR2024/011785 2023-08-11 2024-08-08 Procédé de production de pdrn à partir d'une plante Pending WO2025037843A1 (fr)

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KR102682938B1 (ko) * 2023-08-11 2024-07-09 주식회사 바이오에프디엔씨 식물체로부터 pdrn 생산 방법
KR102838969B1 (ko) * 2024-07-31 2025-07-28 주식회사 래디안 새싹 보리 유래 pdrn의 추출 방법과 이를 유효성분으로 포함하는 피부개선용 화장료 조성물
KR102838970B1 (ko) * 2024-08-12 2025-07-28 주식회사 래디안 인진쑥 유래 pdrn의 추출 방법과 이를 유효성분으로 포함하는 피부개선용 화장료 조성물
KR102843806B1 (ko) * 2024-12-13 2025-08-12 주식회사 유니솔브 엘더베리 유래 pdrn을 유효성분으로 포함하는 화장료 조성물
KR102843808B1 (ko) * 2024-12-13 2025-08-12 주식회사 유니솔브 파바빈 유래 pdrn을 유효성분으로 포함하는 화장료 조성물
KR102843810B1 (ko) * 2024-12-13 2025-08-12 주식회사 유니솔브 감초 유래 pdrn을 유효성분으로 포함하는 화장료 조성물

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