Composition comprising hemp stem-derived PDRN and use thereof

Abstract

The present invention relates to a composition comprising hemp stem-derived polydeoxyribonucleotide (PDRN) and a use thereof. The hemp stem-derived PDRN according to the present invention has been confirmed to exhibit wound healing and skin regeneration efficacy, anti-inflammatory efficacy, and hair loss amelioration and hair growth-promoting activity. Accordingly, the composition may be used for treating wounds, reducing inflammation, preventing, treating, and improving hair loss, as well as promoting hair growth.

Description

Composition containing hemp stem PDRN and its use

[0001] The present invention relates to a composition comprising polydeoxyribonucleotide (PDRN) derived from hemp stems and the use thereof.

[0002] Cannabis sativa L. is an annual plant of the genus Cannabis of the family Cannabaceae that has been widely cultivated in tropical and temperate regions, centered in Central Asia, for 12,000 years. It is a collective term for Cannabis chemovars and their variants, including varieties var. indica and var. kafiristanica, as well as Cannabis sativa subspecies sativa, Cannabis sativa subspecies indica, Cannabis sativa subspecies ruderalis, and their genetically crossbred, self-crossbred, or hybrid plants containing various types of cannabinoid compounds known as medicinal and pharmaceutical components.

[0003] In traditional medical texts from China and Korea, hulled cannabis seeds, known as *majain* or *hwamain*, have been used for constipation, diabetes, pain disorders, menstrual irregularities, skin diseases, and dysentery. Additionally, cannabis leaves (*mayeop*) have been used as an anthelmintic, hair protectant, for asthma, for pain relief, anesthesia, and as a diuretic. Furthermore, records indicate that cannabis roots were used to treat difficult childbirth and resolve blood stasis, cannabis bark for bruises and open sores, cannabis flowers for paralysis and itching, and cannabis powder for difficult childbirth, constipation, gout, phlegm, and insomnia, with each part of the plant being used according to the specific ailment.

[0004] Meanwhile, polydeoxyribonucleotide (PDRN) is a DNA polymer primarily manufactured using DNA extracted from salmon sperm. Since PDRN is a biological component, it does not cause allergic reactions or internal rejection reactions typically seen with synthetic substances. Furthermore, as its main component is negatively charged DNA, it possesses hydrophilic properties; these characteristics make it highly suitable for industrial applications.

[0005] Against this backdrop, the present invention was completed by confirming the significantly superior skin improvement efficacy of PDRN derived from cannabis stems, including wound healing, skin regeneration, anti-inflammatory effects, improvement of hair loss, and promotion of hair growth.

[0006] One aspect provides a cosmetic composition for improving skin condition comprising PDRN (polydeoxyribonucleotide) derived from hemp stems.

[0007] Another aspect provides a topical skin composition for improving skin condition, comprising PDRN derived from hemp stems.

[0008] Another aspect provides a pharmaceutical composition for wound healing or skin regeneration comprising PDRN derived from cannabis stems.

[0009] Another aspect provides a pharmaceutical composition for the prevention or treatment of hair loss, comprising PDRN derived from cannabis stems.

[0010] Another aspect provides a food composition for improving hair loss or promoting hair growth, comprising PDRN derived from hemp stems.

[0011] Another aspect provides an anti-inflammatory composition comprising PDRN derived from cannabis stems.

[0012] One aspect provides a cosmetic composition for improving skin condition comprising polydeoxyribonucleotide (PDRN) derived from hemp stems.

[0013] In this specification, the term "cannabis sativa L." refers collectively to annual plants of the genus Cannabis of the family Cannabisaceae, including wild hemp, and plants produced through genetic crossbreeding, self-crossbreeding, or hybridization thereof, including Cannabis chemovars and their variants, varieties var. indica and var. kafiristanica, Cannabis sativa subspecies sativa, Cannabis sativa subspecies indica, Cannabis sativa subspecies ruderalis, and varieties containing various types of cannabinoid compounds known as medicinal or pharmaceutical components. The term "cannabis" may be used interchangeably with "hemp," which refers to all cannabis used for purposes other than hallucinogenic drugs.

[0014] The term "polydeoxyribonucleotide (PDRN)" in this specification refers to a mixture of short deoxyribonucleotides. That is, it may be a low molecular weight DNA complex produced by fractionating DNA chains into specific sizes. The PDRN may be a type of nucleic acid fragment. The method for producing the PDRN is not particularly limited and may be extracted according to methods commonly used in the relevant technical field.

[0015] The size of the above-mentioned hemp stem-derived PDRN may be 2000 bp or less, 1500 bp or less, or 1000 bp or less, specifically 1 bp to 2000 bp, 1 bp to 1700 bp, 1 bp to 1500 bp, 1 bp to 1200 bp, 1 bp to 1000 bp, 10 bp to 2000 bp, 10 bp to 1700 bp, 10 bp to 1500 bp, 10 bp to 1200 bp, 10 bp to 1000 bp, 50 bp to 2000 bp, 50 bp to 1700 bp, 50 bp to 1500 bp, 50 bp to 1200 bp, 50 bp to 1000 bp. It may have a size in the range of 100 bp to 2000 bp, 100 bp to 1700 bp, 100 bp to 1500 bp, 100 bp to 1200 bp, or 100 bp to 1000 bp.

[0016] The above composition may include the hemp stem-derived PDRN as an active ingredient.

[0017] The term "included as an active ingredient" in this specification means that the cannabis stem-derived PDRN of this specification is added to an extent capable of producing the effects mentioned above. Additionally, this may include formulation in various forms by adding various components as auxiliary ingredients for drug delivery and stabilization, etc.

[0018] The above skin condition improvement may include one or more selected from the group consisting of wound healing, skin regeneration, relief of skin inflammation, anti-inflammatory, improvement of hair loss, prevention of hair loss, and hair growth promoting activities.

[0019] The term "wound" in this specification refers to the destruction of the integrity of the skin epithelial tissue due to external pressure, etc., and may also be referred to as a wound. The types of wounds include, but are not limited to, abrasions, contusions, lacerations, incisions made by a blade, stab wounds, slash wounds, gunshot wounds, puncture wounds, and cleavage wounds.

[0020] The terms "wound treatment" or "wound healing" in this specification include all processes in which damaged epithelial tissue of the skin is restored to its wholeness.

[0021] In this specification, the term "alopecia" refers to a condition in which hair is lost in areas where it should normally be present. Hair undergoes a hair cycle consisting of the anagen, catagen, and telogen phases. More specifically, the hair cycle proceeds through the anagen phase, in which keratinocytes stimulated by the dermal papilla actively divide and proliferate to grow hair; the catagen phase, in which blood supply to the hair bulb is cut off and the dermal papilla separates from the hair follicle; and the telogen phase, in which cell proliferation stops and hair does not grow. Subsequently, the hair either returns to the anagen phase or enters the exogen phase, in which it falls out of the scalp. Human hair has independent growth cycles, with some entering the exogen phase and others the anagen phase, thereby maintaining an overall level of hair density. However, in the case of alopecia, this balance shifts toward the exogen phase, resulting in the gradual disappearance of hair. The causes of the aforementioned hair loss are diverse and can be broadly categorized into internal factors such as male hormone degeneration, autoimmune disorders, endocrine diseases, childbirth, menopause, and aging, and external factors such as nutritional deficiencies, medications, stress, scalp contamination, scalp dryness, and demodex mites.

[0022] The terms "hair growth promotion" or "hair growth promotion" in this specification may refer to any act that increases the growth of hair from hair follicles, and may refer to any action that increases the total amount of hair, such as, for example, the proliferation of hair follicle cells, the activation of hair follicle cells, or the effect of promoting the growth of hair follicle cells.

[0023] The above hair loss may include one or more selected from the group consisting of alopecia areata, male pattern baldness, female pattern baldness, neurogenic hair loss, non-scarring hair loss, seborrheic hair loss, middle-aged hair loss, senile hair loss, and postpartum hair loss, but is not limited thereto.

[0024] The above composition may exhibit one or more features selected from the group consisting of promoting the activity of dermal papilla cells, promoting the proliferation of dermal papilla cells, and inhibiting the death of dermal papilla cells.

[0025] The above composition may enhance the expression level and / or activity of factors related to preventing / improving hair loss or promoting hair growth. The factors related to preventing hair loss or promoting hair growth may include one or more selected from the group consisting of Wnt5a (Wnt Family Member 5A), TBR1 (T-Box Brain 1), and VEGF (Vascular Endothelial Growth Factor).

[0026] The above composition may have anti-inflammatory activity.

[0027] The term "anti-inflammatory" in this specification refers to a process of inflammation or a property or efficacy of inhibiting it, and said anti-inflammatory may include the improvement (relief of symptoms), treatment, prevention, inhibition or delay of the onset of inflammatory conditions and / or inflammatory diseases. Specifically, said anti-inflammatory may involve inhibiting or controlling the expression of inflammatory cytokines such as NO, and / or PGE2, IL-1β, and TNF-α, which act on the inflammatory response.

[0028] The formulation of the above cosmetic composition is not particularly limited and may have any one of the formulations selected from skin toners, lotions, essences, creams, packs, foundations, patches, hair tonics, micro-needle patches, and makeup bases, but is not limited thereto.

[0029] The above cosmetic composition may include substances arbitrarily selected to suit the formulation or intended use of the cosmetic, but is not limited thereto. For example, purified water, oils, surfactants, moisturizers, higher alcohols, thickening agents, chelating agents, colorants, fatty acids, antioxidants, preservatives, waxes, pH adjusters, fragrances, etc., may be added, but is not limited thereto.

[0030] When the formulation of the above cosmetic composition is a paste, cream, or gel, the carrier component may include, but is not limited to, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide.

[0031] When the formulation of the above cosmetic composition is a powder or a spray, it may include lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder as a carrier component, and in particular, when it is a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether, but is not limited thereto.

[0032] When the formulation of the above cosmetic composition is a solution or an emulsion, it may include a solvent, a solubilizing agent, or an emulsifying agent as a carrier component, and may include, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan, but is not limited thereto.

[0033] When the formulation of the above cosmetic composition is a suspension, the carrier component may include, but is not limited to, a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracan.

[0034] When the formulation of the above cosmetic composition is a surfactant-containing cleansing agent, the carrier component may include, but is not limited to, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0035]

[0036] Another aspect provides a topical skin composition for improving skin condition comprising PDRN derived from hemp stems. The same parts as described above apply equally to the said composition.

[0037] The above-mentioned topical skin composition may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal patch, drug-containing bandage, lotion, or a combination thereof. For each formulation of the topical skin composition, other ingredients other than those of the composition of the present invention may be appropriately selected and combined by a person skilled in the art without difficulty according to the formulation or purpose of use of other topical skin preparations, and in this case, a synergistic effect may occur when applied simultaneously with other raw materials.

[0038] The above-mentioned topical skin preparation may be appropriately formulated with ingredients commonly used in topical skin preparations such as cosmetics or pharmaceuticals, for example, aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, UV absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, various skin nutrients, or combinations thereof as needed.

[0039] The above topical skin preparation may also appropriately incorporate metal chelating agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid; caffeine, tannin, bellapamil, licorice extract, glablidin, hot water extract of the fruit of *calin*; various herbal medicines; preparations such as tocopherol acetate, glycyrrhizic acid, tranexamic acid and its derivatives or salts; and sugars such as vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, fructose, and trehalose.

[0040]

[0041] Another aspect is to provide a pharmaceutical composition for wound healing or skin regeneration comprising PDRN derived from cannabis stems. The same parts described above apply equally to said composition.

[0042] The above pharmaceutical composition may include a pharmaceutically acceptable carrier. The term "pharmaceuticalally acceptable carrier" may refer to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the injected compound. Here, "pharmaceuticalally acceptable" means that the target of application (prescription) does not possess toxicity beyond an acceptable level without inhibiting the activity of the active ingredient. Any type of carrier that is commonly used in the relevant technical field and is pharmaceutically acceptable may be used in the above pharmaceutical composition. Non-limiting examples of the above carriers include lactose, dextrose, maltodextrin, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, glycerol, ethanol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. These may be used alone or in a mixture of two or more. The above pharmaceutical composition may be prepared into an oral or parenteral formulation according to the route of administration by conventional methods known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The above pharmaceutical compositions may each be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, or sterile injectable solutions according to conventional methods.

[0043] The above pharmaceutical composition may be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, or sterile injectable solutions, each according to conventional methods. When formulating the above pharmaceutical composition, it may be prepared by adding diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, or surfactants.

[0044] When the above pharmaceutical composition is prepared as an oral formulation, it may be prepared in the form of powder, granules, tablets, pills, coated tablets, capsules, liquids, gels, syrups, suspensions, wafers, etc., in accordance with methods known in the art together with a suitable carrier. Examples of pharmaceutically acceptable suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinylpyrrolidone; water; methylhydroxybenzoate; propylhydroxybenzoate; magnesium stearate; mineral oil; malt; gelatin; talc; polyols; vegetable oils, etc. In the case of formulation, the formulation may include diluents and / or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants as needed.

[0045] When the above pharmaceutical composition is prepared as a parenteral formulation, it may be formulated in the form of an injectable, transdermal, nasal inhalant, and suppository according to methods known in the art with a suitable carrier. When formulated as an injectable, suitable carriers may include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof; preferably, Ringer's solution, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, isotonic solutions such as 5% dextrose, etc. When formulated as a transdermal formulation, it may be formulated in the form of an ointment, cream, lotion, gel, topical solution, paste, liniment, aerosol, etc. In the case of nasal inhalers, they can be formulated in the form of an aerosol spray using suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, and carbon dioxide, and when formulated as suppositories, the base may be Witepsol, Tween 61, polyethylene glycols, cocoa starch, laurin starch, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, and sorbitan fatty acid esters.

[0046] The above pharmaceutical composition may be administered in a pharmaceutically effective amount, wherein the term "pharmaceutically effective amount" means an amount sufficient to treat or prevent a disease with a reasonable benefit / risk ratio applicable to medical treatment or prevention, and the effective dose level may be determined based on factors including the severity of the disease, drug activity, patient's age, weight, health, gender, patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and elimination rate, the duration of treatment, drugs combined or used concurrently with the composition of the present invention used, and other factors well known in the medical field. The above pharmaceutical composition may be administered alone or in combination with a component known to exhibit a therapeutic effect for the said disease. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects, taking all of the above factors into consideration.

[0047] The dosage of the above pharmaceutical composition may be determined by a person skilled in the art by taking into consideration the purpose of use, the degree of toxicity of the disease, the patient's age, weight, gender, medical history, or the type of substance used as an active ingredient. For example, the pharmaceutical composition of the present invention may be administered to an adult at a dose of about 0.1 ng to about 1,000 mg / kg, preferably 1 ng to about 100 mg / kg. The frequency of administration of the composition of the present invention is not particularly limited thereto, but may be administered once a day or divided into several doses. The above dosage or frequency of administration does not limit the scope of the present invention in any way.

[0048]

[0049] Another aspect is to provide a pharmaceutical composition for the prevention or treatment of hair loss comprising PDRN derived from cannabis stems. The same parts described above apply equally to the said composition.

[0050]

[0051] Another aspect provides a method for treating wounds and / or hair loss, comprising the step of administering a cannabis stem-derived PDRN or a pharmaceutical composition containing the same to an individual. The same parts as described above apply equally to the method.

[0052] As used herein, the term "entity" may include, without limitation, mammals, birds, reptiles, farmed fish, etc., including dogs, cats, rats, livestock, and humans, that have developed or are at risk of developing wounds and / or hair loss. The said entity may exclude humans.

[0053] The above pharmaceutical composition may be administered as a single or multiple doses in pharmaceutically effective amounts. In this case, the composition may be administered in the form of a liquid, powder, aerosol, injection, intravenous fluid (Ringer), capsule, pill, tablet, suppository, or patch. The route of administration of the above pharmaceutical composition may be any general route as long as it can reach the target tissue.

[0054] The above pharmaceutical composition is not particularly limited thereto, but may be administered via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, nasal administration, pulmonary administration, or rectal administration, depending on the purpose. However, when administered orally, it may be administered in an unformulated form, and since the active ingredient of the above pharmaceutical composition may be denatured or degraded by gastric acid, the oral composition may be administered into the mouth in a form coated with the active agent or formulated to protect it from degradation in the stomach, or in the form of an oral patch. Additionally, the above composition may be administered by any device capable of transporting the active substance to target cells.

[0055]

[0056] Another aspect is to provide a food composition for improving hair loss or promoting hair growth, comprising PDRN derived from hemp stems. The same parts described above apply equally to the said composition.

[0057] In this specification, the terms "hair loss improvement" or "hair growth promotion" may be used interchangeably and have the same meaning as other terms used in the art, such as hair growth or hair growth promotion.

[0058] The term “food” in this specification includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the ordinary sense.

[0059] The above-mentioned food can be manufactured by methods commonly used in the industry, and during such manufacturing, raw materials and ingredients commonly added in the industry may be added. Furthermore, the formulation of the above-mentioned food can be manufactured without restriction as long as it is a formulation recognized as a food. The above-mentioned food composition can be manufactured in various forms of formulations. Unlike general pharmaceuticals, it uses food as a raw material, offering the advantage of not causing side effects that may occur with long-term use, and possesses excellent portability; therefore, the food composition of the present invention can be consumed as an adjuvant to enhance the effects of improving hair loss or promoting hair growth.

[0060] The above food composition may be a health functional food composition.

[0061] The term "health functional food" in this specification refers to a food manufactured and processed using raw materials or ingredients that have functional properties useful to the human body in accordance with Article 6727 of the Health Functional Foods Act, and "functional properties" means obtaining useful effects for health purposes, such as regulating nutrients or physiological actions on the structure and function of the human body.

[0062] The above-mentioned health food refers to a food that has an active effect of maintaining or promoting health compared to general food, and the health supplement food refers to a food intended for the purpose of supplementing health. In some cases, the terms health functional food, health food, and health supplement food may be used interchangeably. Specifically, the above-mentioned health functional food refers to a food prepared by adding a composition to food materials such as beverages, teas, spices, chewing gum, or confectionery, or by encapsulating, powdering, or suspension, and means that consuming it brings about specific health effects. Unlike general medicines, it has the advantage of not having side effects that may occur from long-term use of medicines because it is made from food.

[0063] Since the above food composition can be consumed on a daily basis, it can be expected to have a high effect on improving hair loss or promoting hair growth, and thus can be used very effectively.

[0064] The above food composition may additionally include a physiologically acceptable carrier, the type of carrier is not particularly limited, and any carrier commonly used in the art may be used. Specifically, the above food composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, visual appearance, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). In addition, it may include amino acids such as lysine, tryptophan, cysteine, and valine.

[0065] In addition, the food composition may include food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder and high-grade bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), coloring agents (tar dyes, etc.), colorants (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium hydrogen tartrate, etc.), reinforcing agents, emulsifiers, thickeners (sizing agents), coating agents, gum bases, antifoaming agents, solvents, and improvers. The above additives can be selected according to the type of food and used in appropriate amounts.

[0066] The above food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The amount of the active ingredient can be appropriately determined according to its purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing food or beverages, the food composition of the present invention may be added to the food or beverage in an amount of 50 parts by weight or less, specifically 20 parts by weight or less. However, when consumed over a long period for the purpose of health and hygiene, the content may be below the above range, and since there are no safety issues, the active ingredient may also be used in an amount greater than the above range.

[0067] As an example of the above food composition, it may be used as a health drink composition, in which case it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners may include natural sweeteners such as thaumatin and stevia extract; and synthetic sweeteners such as saccharin and aspartame. The proportion of the above natural carbohydrates may generally be about 0.01 oz, specifically about 0.02 oz, per 100 mL of the health drink composition of the present invention.

[0068] In addition to the above, the health drink composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. Furthermore, it may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, or vegetable beverages. These ingredients may be used independently or in combination. Although the proportion of these additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health drink composition of the present invention.

[0069] The above food composition may contain the active ingredients of the present invention in various weight percent as long as they can exhibit the effect of improving hair loss or promoting hair growth, and specifically, the hemp stem-derived PDRN may be included in an amount of 0.00001 to 100 weight percent or 0.01 to 80 weight percent relative to the total weight of the food composition, but is not limited thereto.

[0070]

[0071] Another aspect is to provide a feed composition for improving hair loss or promoting hair growth, comprising PDRN derived from hemp stems. The same parts described above apply equally to the said composition.

[0072] The term "feed" in this specification may refer to any natural or artificial prescribed food, meal, etc., or the ingredients of said meal, for or suitable for animals to eat, consume, and digest.

[0073] The types of the above feed are not particularly limited, and feeds commonly used in the relevant technical field may be used. Non-limiting examples of the above feed include plant-based feeds such as grains, root vegetables, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, meal or grain by-products; and animal-based feeds such as proteins, inorganic substances, oils and fats, minerals, oils and fats, single-cell proteins, zooplankton, or food waste. These may be used individually or in a mixture of two or more types.

[0074] The above feed composition may further include known additives that can be added for efficacy in improving hair loss or promoting hair growth, depending on the formulation. The above feed composition may be in the form of a highly concentrated liquid, powder, or granules. The above feed composition may further include any protein-containing organic grain flour commonly used to meet the dietary requirements of animals. The above feed composition may be used by adding it to animal feed by immersion, spraying, or mixing.

[0075] The above feed composition may additionally include substances exhibiting various effects, such as supplementing nutrients and preventing weight loss, enhancing the digestibility and utilization of fiber in the feed, improving milk quality, preventing reproductive disorders and improving conception rates, and preventing high-temperature stress during the summer. For example, it may additionally include mineral preparations such as sodium bicarbonate, bentonite, magnesium oxide, and complex minerals; mineral preparations that are trace minerals such as zinc, copper, cobalt, and selenium; vitamin preparations such as carotene, vitamin E, vitamins A, D, E, nicotinic acid, and vitamin B complex; protected amino acid preparations such as methionine and lysic acid; protected fatty acid preparations such as calcium salts of fatty acids; probiotics (lactic acid bacteria preparations); live bacteria such as yeast cultures and mold fermentation products; and yeast preparations.

[0076] The above feed composition can be applied to the diets of a number of animals, including mammals and poultry, i.e., feed and drinking water.

[0077] The above feed composition may include all of the substances added to the feed (i.e., feed additives), feed raw materials, or the feed itself fed to the individual.

[0078]

[0079] Another aspect is to provide an anti-inflammatory composition comprising PDRN derived from cannabis stems. The same parts as described above apply equally to the said composition.

[0080] The above anti-inflammatory composition may comprise one or more selected from the group consisting of pharmaceutical compositions, food compositions, health functional food compositions, cosmetic compositions, feed compositions, and external skin preparation compositions.

[0081]

[0082] Another aspect provides a skin improvement method comprising the step of applying a composition containing the above-mentioned cannabis stem-derived PDRN as an active ingredient to the skin of an individual. The same parts as described above also apply to the above method.

[0083] The term "individual" above refers to all living organisms, including humans, rats, mice, livestock, and cells. Specific examples include mammals, including humans, and encompass not only in vivo but also in vitro states.

[0084] The above method may be a beauty method through skin improvement.

[0085] The above skin improvement may include one or more selected from the group consisting of wound healing, skin regeneration, relief of skin inflammation, anti-inflammatory, hair loss improvement, hair loss prevention, and hair growth promoting activities.

[0086] The term “application” as used herein may mean causing at least partial localization of the composition to a desired area or placing the composition into an individual by a route of administration, and includes methods of administering or applying to the skin of an individual, such as application by finger, brush, cotton ball, pad, spray, sponge, cotton swab, roll-on, etc. A person skilled in the art can easily determine an appropriate method of application. Additionally, the term “application” may be used interchangeably with “administration” or “application.”

[0087] The above composition may be a cosmetic composition or a skin external application composition.

[0088] In the above method, the applying step may be to apply the composition at intervals of 1 minute to 90 minutes, and specifically, may be performed at intervals of 1 minute to 90 minutes, 5 minutes to 90 minutes, 1 minute to 60 minutes, 5 minutes to 60 minutes, 1 minute to 30 minutes, 5 minutes to 30 minutes, 10 minutes to 30 minutes, 10 minutes to 90 minutes, and 10 minutes to 60 minutes.

[0089] In the above method, the application step may be to apply the composition 1 to 30 times, and specifically, may be to apply it 1 to 30 times, 1 to 25 times, 5 to 30 times, 5 to 25 times, 1 to 20 times, 5 to 20 times, 10 to 20 times, 10 to 30 times, or 10 to 25 times.

[0090]

[0091] Another aspect is to provide the use of the above-mentioned hemp stem-derived PDRN or a composition containing it as an active ingredient for improving skin condition. The same parts described above apply equally to the above use.

[0092]

[0093] Another aspect is to provide the use of the above-mentioned cannabis stem-derived PDRN or a composition containing it as an active ingredient for wound healing, anti-inflammation, treatment of hair loss, and / or promotion of hair growth. The same as described above applies equally to the said uses.

[0094] It has been confirmed that the hemp stem-derived PDRN of the present invention possesses wound healing and skin regeneration efficacy, anti-inflammatory efficacy, and hair loss improvement and hair growth promotion activity; therefore, the composition can be used for wound treatment, anti-inflammation and hair loss prevention, hair loss treatment, hair loss improvement and hair growth promotion.

[0095] Figure 1 is a diagram showing the results of size analysis of PDRN derived from hemp stems.

[0096] Figure 2 is a diagram showing the results of confirming the cytotoxicity of PDRN derived from cannabis stems.

[0097] Figure 3 is a diagram showing the results of evaluating the wound healing efficacy of PDRN derived from cannabis stems.

[0098] Figure 4 is a diagram showing the results of evaluating the anti-inflammatory efficacy of PDRN derived from cannabis stems.

[0099] Figure 5 is a figure showing the results of evaluating the efficacy of PDRN derived from cannabis stems in improving, preventing, and / or treating hair loss or enhancing the expression levels of factors related to hair growth promotion.

[0100] The following examples will be explained in more detail. However, these examples are for illustrative purposes only and the scope of the present invention is not limited to these examples.

[0101]

[0102] Example 1: Preparation of Cannabis-Derived PDRN

[0103] The following experiment was conducted to prepare and obtain PDRN (polydeoxyribonucleotide) derived from cannabis, specifically from cannabis stems.

[0104] Specifically, 1.5 mL / g of CTAB buffer was added to the cannabis stem sample and ground using a blender. The ground material (10–150 g) was divided into 10–20 mL portions into conical tubes, 0.2 μl / mL of 100 mg / ml RNase A was added, and the mixture was incubated at 37°C for 1 hour. Subsequently, 1 μl / mL of 10 mg / ml Proteinase K was added, and the mixture was incubated at 65°C for 1–2 hours (vortexing was performed every 15 minutes). After the reaction, the mixture was incubated at room temperature for 10 minutes and on ice for 5 minutes. A chloroform / isoamyl alcohol (24:1) solution was added in a 1:1 ratio and mixed vigorously, followed by centrifugation at 12,000 xg for 15 minutes. After transferring the supernatant to a new tube, half the volume of 5M NaCl was added, and the mixture was incubated on ice for 15 minutes. 1 / 10 of the total volume of 3M Sodium acetate and 2 / 3 of the volume of chilled Isopropanol were added, mixed vigorously, and incubated in a -20°C freezer for at least 1 hour. After centrifuging at 12,000xg for 5 minutes, the supernatant was removed, 10 ml of 70% ethanol was added, and centrifugation was performed at 12,000xg for 5 minutes. After removing the supernatant, the sample was dried at room temperature, dissolved in 5 mL of Tris-EDTA solution (pH 8.0), 5 mL of distilled water was added, and the solution was dissolved by stirring at 37°C for at least 16 hours. After dissolution was complete, sonication was performed, and DNA purity and concentration were measured before (DNA) and after (PDRN) sonication.

[0105] Next, electrophoresis was performed to analyze the size of the cannabis stem PDRN produced through the above process. As a result, it was found that the size ranged from 200 to 10,000 bp before sonication, and that it was cut into sizes of 1,000 bp or smaller through sonication (Fig. 1).

[0106]

[0107] Example 2: Evaluation of Cytotoxicity of Cannabis Stem PDRN

[0108] To evaluate the cytotoxicity of PDRN derived from cannabis stems, the following experiment was performed.

[0109] Specifically, HaCaT cells (keratinocytes) were placed in a 100Φ dish with DMEM (Dulbecoo's Modified Eagle Medium, Gibco) containing 10% FBS (Fetal Bovine Serum) and 1% Antibiotics (Penicillin streptomycin 10,000 U / ml, Gibco), and cultured for 48 hours in an incubator under 5% CO2 conditions at 37°C. Subsequently, cells were treated with 0.25% Trypsin EDTA, collected in 15ml conical tubes, and counted to determine cell concentration. (0.5 ~ 1.0 x 10⁶ 3MTT assay was performed at 100 μl per cell. The required cell concentration was calculated, diluted with medium, and then seeded into a 96-well plate. After culturing for 24 hours, the medium was removed, 100–200 μl of DPBS was added per well and washed, and then completely removed. Subsequently, 100 μl of medium containing cannabis stem PDRN at various concentrations was added. The cells and medium were incubated for 12–48 hours, and the condition of the cells and medium was checked periodically using a microscope. Afterward, the samples were removed and washed with DPBS. Then, 100–200 μl of MTT solution diluted to 0.5 mg / ml was added to the medium, and the samples were incubated for 1–2 hours under conditions of 37°C and 5% CO2. Finally, the samples were washed with DPBS and 100–200 μl of DMSO was added. The assay results were analyzed using a spectrophotometer at wavelengths of 560–650 nm.

[0110] As a result of the above, it was confirmed that cannabis stem PDRN did not exhibit cytotoxicity up to a concentration of 100 μg / ml (Fig. 2), indicating that it does not show toxicity to cells.

[0111]

[0112] Example 3: Evaluation of Wound Healing Efficacy of Cannabis Stem PDRN

[0113] To evaluate the wound healing efficacy of PDRN derived from cannabis stems, a wound healing assay was performed as follows.

[0114] Specifically, HaCaT cells were cultured in the same manner as in Example 2. Subsequently, after securely attaching the mold (25 culture-inserts 2 Wells for self-insertion, ibidi) to a 24-well plate, 4.0 x 10⁴ cells per well were used. 4Cells were seeded at 70 μl / cell and cultured for 12 hours, after which cell density was checked under a microscope. Subsequently, the molds were removed and washed three times with DPBS. Various samples diluted with serum-free medium (FBS X) were seeded at 1 ml per well into the cells. Microscopic images were taken at 0 h and again after 12 hours of culture to analyze the results. The degree of wound healing was calculated using the Image J program based on the microscopic images. Meanwhile, DMEM, a cell medium, was used as a negative control, and madecassoside and salmon testicular PDRN, representative components of wound healing, were used as positive controls.

[0115] As a result of the above experiment, it was confirmed that cannabis stem PDRN showed a healing efficacy of 137.4% compared to DMEM, the negative control (Fig. 3).

[0116] Based on the above results, it can be seen that cannabis stem PDRN has significantly superior wound healing efficacy.

[0117]

[0118] Example 4: Evaluation of Anti-inflammatory Activity of Cannabis Stem PDNR

[0119] To evaluate the anti-inflammatory activity of cannabis stem PDRN, an NO assay was performed.

[0120] Specifically, to perform the NO assay, 1.0 x 10 6 cell / ml (1.0 x 10⁶ per well) 5RAW 264.7 cells were prepared at a cell concentration of 100 µl and seeded into 96-well plates, after which they were cultured for 24 hours. LPS was added to DMEM medium to a concentration of 1 µg / ml, and the samples were added to the medium at various concentrations. After culturing the cells for 24 hours using the medium containing the LPS and samples, 50 µl of the supernatant was separated, mixed with 50 µl of sulfanilamide solution, and subjected to a dark reaction at room temperature for 10 minutes. Subsequently, 50 µl of NED solution was added, and after a dark reaction at room temperature for 10 minutes, absorbance was measured at wavelengths of 520 nm and 550 nm using a microplate reader within 10 minutes. Meanwhile, DMEM, the cell medium, was used as a negative control, and madecassoside and salmon testis PDRN were used as positive controls.

[0121] As a result of the above experiment, compared to the negative control DMEM, the inflammation level of the positive control madecassoside decreased by 34.1%, but the inflammation level of cannabis stem PDRN decreased by 4451%, confirming that it has superior anti-inflammatory efficacy than madecassoside (Fig. 4).

[0122] Based on the above results, it can be seen that cannabis stem PDRN possesses significantly superior anti-inflammatory efficacy.

[0123]

[0124] Example 5: Evaluation of the efficacy of hemp stem PDNR in improving hair loss or promoting hair growth

[0125] To evaluate the hair loss improvement, treatment, or hair growth-promoting activity of PDRN derived from cannabis stems, the following experiments were performed.

[0126] Specifically, to evaluate the expression levels of factors related to hair loss improvement / prevention or hair growth promotion, HaCaT cells were 3X10 5Cells were seeded into each well of a 6-well plate at a concentration of cell / well and cultured in DMEM medium (10% FBS, Penicillin / streptomycin) at 2.5% CO2 and 37°C for 24 hours. Subsequently, the medium was replaced with serum-free DMEM, and the cells were cultured at 5% CO2 and 37°C for 24 hours. Next, serum-free DMEM containing dissolved cannabis stem-derived PDRN (2.5 μg / mL) or minoxidil (10 μM) was added to each well, followed by culture at 5.5% CO2 and 37°C for 24 hours. The cultured cells were collected, and RNA was extracted from the collected cells using the Rneasy®Mini kit. CDNA was synthesized from the extracted RNA using the Revert Aid First strand cDNA Synthesis kit. Next, qRT-PCR analysis was performed on Wnt5a (Wnt Family Member 5A), TBR1 (T-Box Brain 1), and VEGF (Vascular Endothelial Growth Factor), which are factors for improving hair loss or promoting hair growth, using Power SYBR Green master mix according to the guidelines of the AP7500 RT-qPCR System. In addition, relative gene expression between each group was calculated using the 2-△△CT method. Meanwhile, DMEM, a cell culture medium, was used as a negative control, and minoxidil, known for its hair growth efficacy, was used as a positive control.

[0127] As a result of the above experiment, it was confirmed that PDRN derived from cannabis stems increased the expression levels of hair growth-related factors (Wnt5a, TBR1, and VEGF) compared to the negative control group, and it was also confirmed that the expression levels of hair growth-related factors increased significantly when treated with PDRN derived from cannabis stems compared to the positive control group, minoxidil (Fig. 5).

[0128] Based on the above results, it can be seen that PDRN derived from cannabis stems possesses significantly superior efficacy in treating / improving hair loss and promoting hair growth.

[0129]

[0130] The foregoing description of the present invention is for illustrative purposes only, and those skilled in the art will understand that other specific forms can be easily modified without altering the technical spirit or essential features of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

Claims

1. A cosmetic composition for improving skin condition comprising PDRN (polydeoxyribonucleotide) derived from hemp stems.

2. A composition according to claim 1, wherein the PDRN has a size of 2000 bp or less.

3. A composition according to claim 1, wherein the PDRN is ultrasonically treated.

4. A composition according to claim 1, wherein the skin condition improvement comprises one or more selected from the group consisting of wound healing, skin regeneration, relief of skin inflammation, anti-inflammatory, hair loss improvement, hair loss prevention, and hair growth promoting activities.

5. A topical skin composition for improving skin condition, comprising PDRN derived from cannabis stems.

6. A pharmaceutical composition for wound healing or skin regeneration comprising PDRN derived from cannabis stems.

7. A pharmaceutical composition for preventing or treating hair loss, comprising PDRN derived from cannabis stems.

8. The composition of claim 7, wherein the composition enhances the expression level or activity of factors related to hair loss improvement or hair growth promotion.

9. A composition according to claim 8, wherein the hair loss improvement or hair growth promotion related factor comprises one or more selected from the group consisting of Wnt5a (Wnt Family Member 5A), TBR1 (T-Box Brain 1), and VEGF (Vascular Endothelial Growth Factor).

10. A food composition for improving hair loss or promoting hair growth, comprising PDRN derived from hemp stems.

11. A feed composition for improving hair loss or promoting hair growth, comprising PDRN derived from hemp stems.

12. An anti-inflammatory composition comprising PDRN derived from cannabis stems.

Images