Hemp-derived biomaterial for preventing and treating hair loss

Abstract

The present invention relates to a hemp-derived extract or exosomes for preventing and treating hair loss. A composition comprising a hemp-derived extract or exosomes for preventing and treating hair loss according to the present invention can be used to effectively prevent and treat alopecia in a subject.

Description

Hemp-derived biomaterial for hair loss prevention and treatment

[0001] The present invention was made under the support of the Ministry of Science and ICT of the Republic of Korea under the research project number 2022R1G1A1008848, and the research management specialized organization of the project is the National Research Foundation of Korea, the research project name is "Individual Basic Research (Ministry of Science and ICT)", the research project name is "Biological material for preventing hair loss using hemp germinated seeds", the main organization is Everbio Co., Ltd., and the research period is from 20224.03.01 to 2023.12.31.

[0002] This patent application claims priority to Republic of Korea Patent Application No. 10-2024-0002448, filed with the Korean Intellectual Property Office on January 5, 2024, the disclosure of which is incorporated herein by reference.

[0003] The present invention relates to a hemp-derived extract or exosome for preventing and treating hair loss.

[0004]

[0005] Hair is one of the body's major structures, originating from the outermost layer of the skin. Along with sweat glands, sebaceous glands, and nails, it is considered a binding structure of the outer skin. Hair protects the head from various external stimuli, including physical shock, UV rays, cold, and heat. While not essential for life, it is a part of the body that serves as a barometer of health and influences appearance. Hair is produced from the hair follicle and undergoes a hair growth cycle, including growth phases such as anagen, resting phase, and catagen. The anagen phase is the period during which hair grows and accounts for 90% of normal hair. The telogen phase is the period when the hair bulb dries and becomes a club, while the catagen phase is the period when hair growth ceases and the hair root atrophies. Hair repeats growth and shedding according to these three stages of the growth cycle. Hair formation is a cyclical pattern of growth and shedding through the anagen, resting, and catagen phases. This cycle repeats over 3 to 6 years, with an average of 50 to 100 hairs falling out per day. Alopecia occurs when many hair follicles undergo apoptosis during the catagen phase and enter the resting phase, reducing the size of the hair follicles and causing an abnormal increase in the number of hairs lost. Causes of hair loss include the effect of androgens on organs such as hair roots and sebaceous glands, decreased blood flow to the hair follicles, scalp abnormalities caused by bacterial growth or excessive sebum secretion, aging, and genetic factors. Recently, the number of hair loss patients has been increasing due to various reasons such as excessive stress, westernized eating habits, environmental pollution, side effects from drug abuse, nutritional imbalances, changes in social activities, perms, and dyeing. In the case of male pattern baldness, testosterone, a type of androgen, is activated into dihydrotestosterone (DHT) by the enzyme 5α-reductase, and this DHT binds to a specific receptor, causing hair loss.Treatments for alopecia include oral medications, topical applications, and hair transplants. FDA-approved medications include minoxidil and finasteride. Finasteride is used to treat male pattern baldness by inhibiting the activity of the 5α-reductase enzyme, thereby reducing DHT levels. Minoxidil was originally developed as a vasodilator for high blood pressure, but due to reports of hirsutism as a side effect, it has been used as a hair growth agent. While the exact mechanism of action of minoxidil is not yet fully understood, it is believed to promote hair growth by increasing blood flow to hair follicles through vasodilation. These treatments, relying on medications or steroids, are associated with side effects and inconsistent efficacy across individuals.

[0006] Therefore, there is a need to develop an effective hair loss treatment.

[0007]

[0008] The inventors of the present invention have conducted extensive research to develop an effective hair loss treatment. As a result, they have discovered that hemp seed extract or hemp seed-derived exosomes can effectively prevent hair loss and promote hair growth, thereby completing the present invention.

[0009] Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating alopecia comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0010] Another object of the present invention is to provide a cosmetic composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0011] Another object of the present invention is to provide a food composition for preventing hair loss or promoting hair growth, comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0012] Another object of the present invention is to provide a feed composition for preventing hair loss or promoting hair growth, comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0013] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0014]

[0015] The present invention provides the following inventions 1 to 17.

[0016] 1. A pharmaceutical composition for preventing or treating alopecia comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0017] 2. A cosmetic composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0018] 3. A food composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0019] 4. A feed composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0020] 5. A method for preventing or treating alopecia, comprising administering a hemp seed extract, hemp seed-derived exosomes, or a combination thereof to a subject in need of prevention or treatment.

[0021] 6. A composition or method according to any one of 1 to 5, wherein the hemp seed is a germinated hemp seed.

[0022] 7. A composition or method according to any one of 1 to 6, wherein the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

[0023] 8. A composition or method according to any one of 1 to 7, wherein the alcohol is 30 to 70% by volume of ethanol.

[0024] 9. A composition or method according to any one of 1 to 8, wherein the concentration of the hemp seed extract is 500 to 3000 μg / mL.

[0025] 10. A composition or method according to any one of 1 to 9, wherein the hemp seed-derived exosome is an exosome isolated from hemp seed callus.

[0026] 11. A composition or method according to any one of 1 to 10, wherein the concentration of the hemp seed-derived exosome is 10 to 100 μg / mL.

[0027] 12. A composition or method according to any one of 1 to 11, wherein the size of the hemp seed-derived exosome is 50 to 300 nm.

[0028] 13. A composition or method according to any one of 1 to 12, comprising a pharmaceutically acceptable carrier, excipient or diluent.

[0029] 14. A composition or method according to any one of 1 to 13, wherein the formulation of the composition is a hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, powder, gel, hair pack, hair treatment, eyebrow hair tonic, eyelash hair tonic, eyelash nutrient or ointment.

[0030] 15. A composition or method according to any one of 1 to 14, wherein the composition is prepared in any one of the following dosage forms: powder, granules, pills, tablets, capsules, candies, syrups, and beverages.

[0031] 16. A composition or method according to any one of 1 to 15, wherein the alopecia is selected from the group consisting of androgenetic alopecia, female pattern alopecia, inflammatory alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania alopecia, compression alopecia, anagen alopecia, pityriasis versicolor, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, non-scarring alopecia, scarring alopecia, and congenital alopecia, and combinations thereof.

[0032] 17. A composition or method according to any one of 1 to 16, wherein the alopecia is alopecia caused by dihydrotestosterone.

[0033] According to one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating alopecia comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0034] The inventors of this study have conducted extensive research to develop an effective hair loss treatment. As a result, they have discovered that hemp seed extract or hemp seed-derived exosomes can effectively prevent hair loss and promote hair growth.

[0035] As used herein, the term "hemp seed" refers to the edible portion of the cannabis (Cannabis sativa) plant. Hemp seed contains essential fatty acids such as omega-3 and omega-6 fatty acids, various essential amino acids required for keratin synthesis, and various vitamins and minerals, including vitamin E, magnesium, calcium, iron, and zinc.

[0036] As used herein, the term "prevention" refers to the prevention or protective treatment of a disease or disease state. As used herein, the term "treatment" refers to the reduction, suppression, alleviation, or eradication of a disease state.

[0037] The pharmaceutical composition of the present invention can be manufactured in a unit dose form or can be manufactured by placing it in a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains, and the like. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or in the form of an extract, powder, suppository, powder, granule, tablet or capsule, and may additionally include a dispersing agent or stabilizer.

[0038] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are those commonly used in formulations, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweetening agents, flavoring agents, emulsifiers, suspending agents, preservatives, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0039] The pharmaceutical composition of the present invention can be administered orally or parenterally, and can be administered by, for example, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, intrasternal injection, intratumoral injection, local administration, intranasal administration, intrapulmonary administration, and rectal administration.

[0040] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. A skilled physician can easily determine and prescribe a dosage effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the anticancer composition of the present invention is 0.0001-100 mg / kg.

[0041] In one embodiment of the present invention, the alopecia is selected from the group consisting of androgenetic alopecia, female pattern alopecia, inflammatory alopecia, alopecia areata, telogen effluvium, traumatic alopecia, trichotillomania alopecia, compression alopecia, anagen alopecia, pityriasis alopecia, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, non-scarring alopecia, scarring alopecia, and congenital alopecia, and combinations thereof.

[0042] In one embodiment of the present invention, the alopecia is alopecia caused by dihydrotestosterone.

[0043] In one embodiment of the present invention, the composition has the properties of promoting differentiation of follicular dermal papilla cells into papilla cells, promoting germ cell differentiation, inhibiting secretion of hair loss activating factors by CD8+ T cells, promoting hair growth, reducing hair loss, or a combination thereof.

[0044] In one embodiment of the present invention, the composition is administered in combination with a drug for preventing or treating hair loss.

[0045] In one specific example of the present invention, the drug for preventing or treating hair loss may be, for example, minoxidil, finpecia, dutasteride, spironolactone, ketoconazole, or a combination thereof, but is not limited thereto.

[0046] The pharmaceutical composition according to the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents. It can be administered sequentially or simultaneously with conventional therapeutic agents, or in single or multiple doses. Taking all of the above factors into account, it is important to administer an amount that achieves maximum efficacy with minimal side effects. This amount can be readily determined by those skilled in the art to which the present invention pertains.

[0047] The hemp seed extract used in the present invention can be purchased or obtained through direct extraction. When the hemp seed extract used in the composition of the present invention is obtained by treating it with an extraction solvent, various extraction solvents, such as polar solvents or nonpolar solvents, can be used.

[0048] Suitable polar solvents include, but are not limited to, (i) water, (ii) alcohols (preferably, methanol, ethanol, propanol, butanol, n-propanol, iso-propanol, n-butanol, 1-pentanol, 2-butoxyethanol or ethylene glycol), (iii) acetic acid, (iv) dimethyl-formamide (DMFO) and (v) dimethylsulfoxide (DMSO).

[0049] Suitable non-polar solvents include, but are not limited to, acetone, acetonitrile, ethyl acetate, methyl acetate, fluoroalkanes, pentane, hexane, 2,2,4-trimethylpentane, decane, cyclohexane, cyclopentane, diisobutylene, 1-pentene, 1-chlorobutane, 1-chloropentane, o-xylene, diisopropyl ether, 2-chloropropane, toluene, 1-chloropropane, chlorobenzene, benzene, diethyl ether, diethyl sulfide, chloroform, dichloromethane, 1,2-dichloroethane, aniline, diethylamine, ether, carbon tetrachloride, and THF (Tetrahydrofuran).

[0050] In one embodiment of the present invention, the hemp seed is a germinated hemp seed.

[0051] Hemp seeds may exhibit some differences in composition depending on whether they are sprouted or not. The sprouting process is known to increase the availability and activity of nutrients by inducing biochemical changes within the seed. While unsprouted hemp seeds already contain beneficial fatty acids, protein, essential amino acids, fiber, vitamins, and minerals, sprouting alters these nutrients to make them more absorbable. For example, sprouting increases enzyme activity, converting nutrients within the seed into more digestible forms and converting insoluble nutrients into soluble forms, making them more readily available to the body. Furthermore, sprouting can improve mineral absorption by reducing levels of the antinutrient phytic acid.

[0052] In one embodiment of the present invention, the extract is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

[0053] In one specific example of the present invention, the alcohol is selected from the group consisting of methanol, ethanol, propanol, butanol, normal-propanol, iso-propanol, normal-butanol, and mixed solvents thereof.

[0054] In the present invention, the "extract" has the meaning commonly used in the art as a crude extract as described above, but in a broad sense, it also includes a fraction obtained by further fractionating the extract. That is, the buckwheat sprout extract includes not only the one obtained using the above-described extraction solvent, but also the one obtained by additionally applying a purification process thereto. For example, the fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, the fraction obtained through various additional purification methods such as separation by various chromatographies (designed for separation according to size, charge, hydrophobicity, or affinity) may also be included in the hemp seed extract of the present invention.

[0055] In the present invention, hemp seeds may be extracted using a hot water extraction method, a pressurized hot water extraction method, a cold extraction method, a reflux extraction method, a room temperature extraction method, or an ultrasonic extraction method, but is not limited thereto.

[0056] In order to prepare an extract according to one embodiment of the present invention, the amount of solvent used for extraction may be appropriately selected depending on the amount of hemp seeds used. Specifically, the volume may be 1 to 20 times the weight of the hemp seeds used for preparing the extract, more specifically 1 to 15 times the volume, 1 to 12 times the volume, 1 to 10 times the volume, and even more specifically 4, 5, 6, 7, 8, 9, or 10 times the weight of the hemp seeds, but is not limited thereto.

[0057] In one specific embodiment of the present invention, the alcohol is 30 to 70 vol% ethanol. The alcohol may be, for example, 30 to 70 vol%, 30 to 60 vol%, 30 to 50 vol%, 30 to 40 vol%, 40 to 70 vol%, 50 to 70 vol%, 60 to 70 vol%, 35 to 65 vol%, 40 to 60 vol%, or 45 to 55 vol%, but is not limited thereto.

[0058] The extraction temperature of the water or ethanol extract according to one embodiment of the present invention is not particularly limited and may be, for example, 10°C to 110°C, and specifically, 20°C to 100°C, 30°C to 100°C, 40°C to 100°C, 50°C to 100°C, 60°C to 100°C, 20°C to 90°C, 30°C to 90°C, 40°C to 90°C, 50°C to 90°C, 60°C to 90°C, 20°C to 80°C, 30°C to 80°C, 40°C to 80°C, 50°C to 80°C, 60°C to 80°C, 20°C to 80°C, 30°C to 80°C, 40°C to 80°C, 50°C to 80°C, or 60°C to It can be 80℃, but is not limited to this.

[0059] The extraction time of the water or ethanol extract according to one embodiment of the present invention is not particularly limited, and may be, for example, 1 hour to 20 hours, specifically, 1 hour to 20 hours, 1 hour to 18 hours, 1 hour to 16 hours, 1 hour to 14 hours, 1 hour to 12 hours, 1 hour to 10 hours, 1 hour to 8 hours, 1 hour to 6 hours, 1 hour to 4 hours, 1 hour to 2 hours, 2 hours to 20 hours, 4 hours to 20 hours, 6 hours to 20 hours, 8 hours to 20 hours, 10 hours to 20 hours, 12 hours to 20 hours, 14 hours to 20 hours, 16 hours to 20 hours, 18 hours to 20 hours, 2 hours to 18 hours, 4 hours to 16 hours, 6 hours to 14 hours, 8 hours to It may be, but is not limited to, 14 hours, 10 to 14 hours, or 11 to 13 hours.

[0060] The hemp seed extract used in the present invention can be manufactured into a powder form by additional processes such as reduced pressure distillation and freeze drying or spray drying.

[0061] In one embodiment of the present invention, the concentration of the hemp seed extract is 500 to 3000 μg / mL. The concentration of the hemp seed extract may be, for example, but is not limited to, 500 to 3000, 500 to 2700, 500 to 2400, 500 to 2100, 500 to 1800, 500 to 1500, 500 to 1200, 500 to 900, 800 to 3000, 1100 to 3000, 1400 to 3000, 1700 to 3000, 2000 to 3000, 2300 to 3000, 2600 to 3000, 1000 to 2500, 1500 to 2500, or 1800 to 2200 μg / mL.

[0062] In one embodiment of the present invention, the hemp seed-derived exosome is an exosome isolated from hemp seed callus.

[0063] In one specific example of the present invention, the hemp seed callus is a callus derived from germinated hemp seed meristem.

[0064] The term "exosome" in this specification refers to a small membrane-structured vesicle secreted from various cells, and refers to a type of extracellular vesicle. All cells secrete extracellular vesicles to exchange information with other cells or the external environment. Exosomes are approximately 50 to 200 nm in size and contain physiologically active substances such as proteins, lipids, and nucleic acids. Exosomes exist in all living things, including mammals, bacteria, and plants, and reflect the state of the organisms they originate from, so they can be used for diagnosis and treatment. Exosomes are nano-sized phospholipid bilayer structures that easily penetrate cells and have various pathophysiological functions, such as immune response, signaling, anti-allergy, antibacterial and anti-inflammatory action, skin cell regeneration, wound healing, skin immune protection from UV rays, and skin moisturizing effect.

[0065] The term "exosomes" in this specification refers to nano-sized vesicles having a membrane structure secreted or released from plant cells into the extracellular space, and are also defined as exosome-like vesicles or exosome-like particles.

[0066] The term "hemp seed callus" as used herein refers to a tissue that normally functions similarly to plant tissue, but proliferates in a form similar to plant cancer tissue due to physical stress such as plant growth hormones. Unlike general plant tissue, it is a cell tissue that is differentiating into callus, which has rapid cell or tissue differentiation and is rich in mitochondria, vacuoles, and endoplasmic reticulum, and has rapid metabolism and proliferation. The "cell tissue differentiating into callus" refers to a cell or tissue that is in the process of differentiating into callus from a tissue section of hemp seed. From this, exosomes of hemp seed-derived callus can be extracted.

[0067] The above callus is an amorphous tissue or cell mass derived from plant tissue that has lost the ability to induce normal organ formation or tissue differentiation. It is produced by culturing excised plant tissue in a medium containing auxin and cytokines, by wounding the plant, or by treating plant sections with auxin. This callus can be derived from any part of the plant, including leaves, stems, roots, rhizomes, and fruits, using plant tissue culture technology, and has the unique characteristic of being continuously produced through subculture. The basic principle of plant tissue culture technology utilizes the biological fact that all plant tissues have the ability to induce the entire plant from which they are derived. This totipotency is comparable to the pluripotency of animal embryonic stem cells. Therefore, if callus culture, which is a mass of undifferentiated cells and parenchyma that utilizes this plant property, is utilized, it can be considered a valuable raw material with excellent physiological activity. In particular, callus is a mass of soft cells and parenchyma tissues with less well-developed cell walls than plants, so it is easy to remove the cell walls and grows quickly. In addition, it can easily release secondary metabolic products, physiologically active substances, and even exosomes from the cells, thereby showing much better yield and productivity than plants with thick cell walls.

[0068] Specifically, the hemp seed-derived exosome may be produced through a production method including the steps of isolating a meristem from a hemp seed; culturing the meristem in a callus induction medium; and purifying an exosome from the callus.

[0069] In a specific example, the exosomes may be isolated from the lysate of the hemp seed callus. In one specific example, the lysate of the hemp seed callus may be a lysate of a mixed culture of hemp seed callus. In another specific example, the lysate of the hemp seed callus may be a lysate of a callus recovered by removing the culture supernatant from a mixed culture of hemp seed callus. In yet another specific example, the lysate of the hemp seed callus may be a lysate of a culture supernatant recovered by removing the callus from a mixed culture of callus. A standard sieve and mesh net can be used to separate callus or culture supernatant from the above hemp seed callus mixed culture, and the pore size may be suitably 50 ㎛ to 300 ㎛, but is not limited thereto, and a conventional method used in the art to separate callus and culture supernatant, such as a centrifuge, a filter device, or a qualitative filter paper, can be used.

[0070] The term “callus mixed culture” as used herein may refer to the entire medium including callus, its metabolites, and extra nutrients, etc., obtained by culturing callus for a certain period of time in a medium that can supply nutrients so that callus can grow and survive in vitro. For example, it may be obtained by culturing callus in a medium at a temperature of more than 10°C or less than 40°C for a certain period of time, for example, 1 to 40 days. Those skilled in the art may appropriately select or modify the culture medium and culture conditions for culturing the callus.

[0071] In this specification, the term “culture supernatant” may mean only the liquid in the upper layer after leaving the culture solution still for a certain period of time, excluding the portion that has settled in the lower layer, or may mean only the liquid in the upper layer after centrifuging the culture solution, excluding the sediment in the lower layer.

[0072] The term “crushed material” in this specification may mean a product obtained by crushing with chemical or physical force, and may be, for example, a crushed material obtained by using a high-pressure homogenizer.

[0073] In one embodiment of the present invention, the concentration of the hemp seed-derived exosome is 10 to 100 μg / mL. The concentration of the hemp seed derived exosomes can be, for example, but is not limited to, 10 to 100, 10 to 90, 10 to 80, 10 to 70, 10 to 60, 10 to 50, 10 to 40, 10 to 30, 10 to 20, 20 to 100, 30 to 100, 40 to 100, 50 to 100, 60 to 100, 70 to 100, 80 to 100, 90 to 100, 20 to 80, 30 to 60, 30 to 50, or 35 to 45 μg / mL.

[0074] In one embodiment of the present invention, the size of the hemp seed-derived exosome is 50 to 300 nm. The size of the exosome may be, for example, 50 to 300, 50 to 270, 50 to 240, 50 to 210, 50 to 180, 50 to 150, 50 to 120, 50 to 90, 80 to 300, 110 to 300, 140 to 300, 170 to 300, 200 to 300, 230 to 300, 260 to 300, or 100 to 200 nm, but is not limited thereto.

[0075] In one embodiment of the present invention, the hemp seed-derived exosome expresses CD68 protein on its surface.

[0076] In one embodiment of the present invention, the composition comprises a pharmaceutically acceptable carrier, excipient or diluent.

[0077] In one aspect of the present invention, the present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0078] The cosmetic composition for preventing hair loss or promoting hair growth of the present invention can be manufactured in any formulation commonly manufactured in the art, and for example, can be formulated as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. More specifically, it can be manufactured in the form of a flexible toner, a nourishing toner, a lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

[0079] The cosmetic composition of the present invention may include, in addition to the active ingredient, a carrier acceptable for use in cosmetic formulations. The term "acceptable carrier for use in cosmetic formulations" refers to an additional ingredient known and used to be capable of being included in cosmetic formulations, and which can improve the application of the active ingredient to the skin, user convenience, and preference without significantly reducing the primary efficacy of the active ingredient or causing adverse effects on the human body.

[0080] The carrier may be included in an amount of about 1 wt% to about 99.99 wt%, preferably about 50 wt% to about 99 wt%, of the total weight of the composition of the present invention. However, the content of the carrier may be appropriately adjusted depending on the formulation of the cosmetic, the specific application site, the desired application amount, etc., and is not particularly limited.

[0081] When the formulation of the present invention is a paste, cream, lotion, or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as a carrier component.

[0082] When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0083] When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0084] When the formulation of the present invention is a suspension, 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 tragacanth may be used as a carrier component.

[0085] When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0086] The components included in the cosmetic composition of the present invention include, in addition to the active ingredient and the carrier component, components commonly used in cosmetic compositions, and may include conventional auxiliary agents such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0087] In one embodiment of the present invention, the formulation of the composition is a hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, powder, gel, hair pack, hair treatment, eyebrow hair growth agent, eyelash hair growth agent, eyelash nutrient or ointment.

[0088] The subject of administration of the cosmetic composition of the present invention is an individual in need of prevention or improvement of alopecia, and the individual may include all animals, including humans, that have developed or may develop alopecia, and the animal may include, but is not limited to, non-human primates, such as chimpanzees, other apes, or monkey species; livestock animals, such as cows, horses, sheep, goats, and pigs; domesticated animals, such as rabbits, dogs, or cats; laboratory animals, such as rodents, such as rats, mice, or guinea pigs; and non-mammals, such as birds or fish.

[0089] In one aspect of the present invention, the present invention provides a food composition for preventing hair loss or promoting hair growth, comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0090] The above food may be a health functional food, a health food, or a food additive, and in this case, there is no particular limitation on the type of food. Examples of the above food include dairy products including drinks, meat, sausages, bread, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, alcoholic beverages, and vitamin complexes, dairy products, and dairy products, and includes all health functional foods and health food compositions in the conventional sense, such as tablets, capsules, pills, or liquids.

[0091] The health functional food, health food and food additive composition comprising the hemp seed extract, hemp seed-derived exosome, or a combination thereof according to the present invention can be added to food as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method.

[0092] In the present invention, the amount of the composition in the health functional food, health food, or food additive may be 0.1 to 90 parts by weight based on the total food weight. However, in the case of long-term intake for the purpose of maintaining or regulating health, the amount may be below the above range.

[0093] In the present invention, there is no particular limitation on other ingredients added in addition to those containing the hemp seed extract, hemp seed-derived exosomes, or a combination thereof, and various flavoring agents or natural carbohydrates, etc., may be contained as additional ingredients, like in a typical beverage. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents such as thaumatin, stevia extracts (such as rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the above natural carbohydrates may generally be about 1 g to 20 g, preferably about 5 g to 12 g, per 100 g of the health functional food, health food or food additive composition of the present invention.

[0094] In the present invention, the food composition containing the hemp seed extract, hemp seed-derived exosomes, or a combination thereof may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the health functional food and health food composition of the present invention may contain fruit pulp for producing natural fruit juice and fruit juice drinks and vegetable drinks.

[0095] In one embodiment of the present invention, the composition is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages.

[0096]

[0097] In one aspect of the present invention, the present invention provides a feed composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0098] The above feed composition may include feed or feed additives for feeding a diet to an animal.

[0099] When the hemp seed extract, hemp seed-derived exosome, or a combination thereof of the present invention is contained and used in a feed composition or feed additive composition, the composition may be a highly concentrated solution of 20 to 90% or may be manufactured in the form of a powder or granules. The feed additive may further include one or more of organic acids such as citric acid, fumaric acid, adipic acid, lactic acid, and malic acid; phosphates such as sodium phosphate, potassium phosphate, and acid pyrophosphate; and natural antioxidants such as polyphenols, catechins, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid.

[0100] In the present invention, when the hemp seed extract, hemp seed-derived exosomes, or a combination thereof are used as feed, the composition can be formulated in the form of a conventional feed and can contain conventional feed ingredients together. The feed additives and feed can further include grains such as ground or crushed wheat, oats, barley, corn, and rice; plant-based protein feeds such as feeds mainly composed of rapeseed, soybeans, and sunflower; animal-based protein feeds such as blood meal, meat meal, bone meal, and fish meal; dry ingredients composed of sugars and dairy products such as various types of milk powder and whey powder, and in addition, nutritional supplements, digestion and absorption enhancers, growth promoters, etc.

[0101] In the present invention, when the hemp seed extract, hemp seed-derived exosomes, or a combination thereof are used as the feed additive, they may be administered to animals alone or in combination with other feed additives in an edible carrier. In addition, the feed additive can be easily administered to animals as a top dressing, by mixing them directly into animal feed, or in an oral formulation separate from the feed. When the feed additive is administered separately from animal feed, it can be prepared as an immediate-release or sustained-release formulation by combining it with a pharmaceutically acceptable edible carrier, as is well known in the art. Such edible carriers can be solid or liquid, for example, corn starch, lactose, sucrose, soybean flakes, peanut oil, olive oil, sesame oil, and propylene glycol. When a solid carrier is used, the feed additive can be a top dressing in the form of a tablet, capsule, powder, troche, sugar tablet, or microdispersible form. When a liquid carrier is used, the feed additive may be in the form of a gelatin soft capsule, or a syrup, suspension, emulsion, or solution. The feed may include any protein-containing organic grain meal commonly used to satisfy the dietary needs of animals. Such protein-containing grain meal typically consists of corn, soybean meal, or a corn / soybean meal mix. The feed composition may also contain, for example, preservatives, stabilizers, wetting or emulsifying agents, and solution promoters. The feed additive composition may also be used by adding it to animal feed by immersion, spraying, or mixing.

[0102] The subject of the feed or feed additive administration of the present invention is an individual in need of prevention or improvement of alopecia, and the individual may include any animal other than humans that has developed or may develop alopecia, and the animal may include, but is not limited to, non-human primates such as chimpanzees, other apes, or monkey species; livestock animals such as cows, horses, sheep, goats, and pigs; domestic animals such as rabbits, dogs, or cats; laboratory animals such as rodents such as rats, mice, or guinea pigs; and non-mammals such as birds or fish.

[0103]

[0104] In one aspect of the present invention, the present invention provides a method for preventing or treating alopecia, comprising administering to a subject in need of prevention or treatment a hemp seed extract, hemp seed-derived exosome, or a combination thereof; or a composition comprising the same.

[0105] Since the above method for preventing or treating alopecia overlaps with the content of the above composition in that it administers a hemp seed extract, hemp seed-derived exosome, or a combination thereof, the content described in relation to the composition may also be applied to the method for preventing or treating alopecia, and the description thereof is omitted to avoid excessive complexity of the specification.

[0106]

[0107] The features and advantages of the present invention are summarized as follows:

[0108] (a) The present invention provides a pharmaceutical composition for preventing or treating alopecia comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0109] (b) The present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0110] (c) Provides a food composition for preventing hair loss or promoting hair growth, comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0111] (d) Provides a feed composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

[0112] (e) When the composition of the present invention is used, hair loss of the subject can be effectively prevented and treated.

[0113]

[0114] Figure 1 shows an overview of the action of the hemp seed extract and hemp seed-derived exosomes of the present invention.

[0115] Figure 2 shows the effective concentration analysis results of HFSC using sprouted hemp seed extract and callus exosomes. Figure 2a shows the results of comparing the effects of hot water extract (HE) and alcohol extract (AE) on sprouted hemp seed, Figure 2b shows the results of isolation and purification of exosomes extracted from the meristematic callus of sprouted hemp seed, and Figures 2c, 2d, 2e, and 2f show the results of treatment dose analysis of sprouted hemp seed extract, exosomes, and dihydrotestosterone (DT): Con; control, ns; not significant, HFSC: human hair follicle dermal papilla stem cell. (* p < 0.05, ** p < 0.01, *** p < 0.001).

[0116] Figure 3 shows the results of analyzing the role of hemp seed callus exosomes and sprouted hemp seed extract (GE) as metabolic regulators in HFSCs. Figures 3a and 3b show the levels of metabolic activation genes and alopecia factor genes, respectively, in HFSCs: control, GE2000; sprouted hemp seed extract 2000 μg / mL, E40; callus exosomes 40 μg / mL, DT; (dihydrotestosterone, 1 ng / mL), DT+GE2000; DT after exposure to GE2000, E40+DT; DT after exposure to E40 (* p < 0.05, ** p < 0.01, *** p < 0.001). Figure 4 shows the activation of differentiation of HFSCs into DPCs and germ cells according to treatment with hemp seed callus exosomes or sprouted hemp seed extract. Figures 4a and 4b show marker gene levels of dermal papilla cells (DPCs) and germ cells, respectively, in HFSCs: control, GE2000; sprouted hemp seed extract 2000 μg / mL, E40; callus exosomes 40 μg / mL, DT; (dihydrotestosterone, 1 ng / mL), DT+GE2000; DT after GE2000 exposure, E40+DT; DT after E40 exposure (* p < 0.05, ** p < 0.01, *** p < 0.001). Figure 5 shows the image results of differentiated DPCs and germ cells upon treatment with hemp seed callus exosomes or sprouted hemp seed extract. Green and red represent FITC-anti-cytokeratin 15 (germ cell marker) antibody and APC-anti-Sox (dermal papilla cell, DPC marker) antibody, respectively: Control, GE2000; sprouted hemp seed extract 2000 μg / mL, E40; callus exosomes 40 μg / mL, DT; (dihydrotestosterone, 1 ng / mL), DT+GE2000; DT after GE2000 exposure, E40+DT; DT after E40 exposure (* p < 0.05, ** p < 0.01, *** p < 0.001). Figure 6 shows the CD8 expression levels according to treatment with hemp seed callus exosomes or sprouted hemp seed extract. +Flow cytometry analysis results for the inactivation of T cells by alopecia factors are shown: control, GE2000; sprouted hemp seed extract 2000 μg / mL, E40; callus exosomes 40 μg / mL, DT; (dihydrotestosterone, 1 ng / mL), DT+GE2000; DT after exposure to GE2000, E40+DT; DT after exposure to E40 (* p < 0.05, ** p < 0.01, *** p < 0.001). Figure 7 shows the CD8 + The results of T cell regulatory ability analysis are shown. A is the standard curve of IFNγ, and b and c represent the IFNγ regulation of CD8+ T cells by two substances: control, GE2000; sprouted hemp seed extract 2000 μg / mL, E40; callus exosome 40 μg / mL, DT; (dihydrotestosterone, 1 ng / mL), DT+GE2000; DT after GE2000 exposure, E40+DT; DT after E40 exposure (* p < 0.05, ** p < 0.01, *** p < 0.001). Figure 8 shows the results of the hair loss prevention effect analysis of hemp seed callus exosome or sprouted hemp seed extract compared to hemp seed oil.

[0117]

[0118] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0119]

[0120] Example

[0121] Example 1: Preparation of extracts and exosomes derived from sprouted hemp seeds.

[0122] Sterilized hemp seeds (Cheongsam cultured in Andong, Korea) were soaked in 1% H2O2 for 1 day at 5 g, and the hydrated seeds were germinated at 24℃ for 3 days. The seeds were then germinated on MS (Murashige & Skoog Medium; KisanBio, Seoul, Korea) containing GA3 (Gibberellic Acid; KisanBio, Seoul, Korea), IAA (Indole-3-Acetic Acid; KisanBio, Seoul, Korea), and sucrose (KisanBio, Seoul, Korea). Meristematic tissue was isolated from the germinated hemp seeds and cultured on MS (Murashige & Skoog Medium; KisanBio, Seoul, Korea) callus inducing medium containing 6BAP (6-Benzylaminopurine; KisanBio, Seoul, Korea), IAA, and sucrose. After culturing calli for 5 weeks in callus transfer medium (MS medium supplemented with 0.3% agar, 2.3% sucrose, 6BAP, and IAA), exosomes were isolated and purified from calli using exosome standard kits, exoEasy Maxi Kit (QIAGEN, Hilden, DE), and CD68 Exo-Flow Capture Kit (SBI, CA, USA). Germinated hemp seed extract (GE) was prepared by treating germinated hemp seeds with alcohol (AE) or hydrothermal extract (HE). Sprouted hemp seeds were extracted with distilled water or ethanol (30 to 70%) at 70°C using a rotary concentrator for 12 hours, and then concentrated and sterilized by filtration.

[0123]

[0124] Example 2: Establishment of therapeutic capacity of hemp seed-derived extract and exosomes.

[0125] Proliferated hair follicle dermal papilla stem cells (HFSC) (SKU: M36007-08S, Celprogen, CA, USA) were cultured in specific growth medium (SKU: 36007-08, Celprogen) at 37°C and 5% CO under various conditions (HE, AE30–70, exosomes and extracts) for 1 day to establish the therapeutic dose. To compare the effects of water and alcohol extraction, total RNA of the exposed cells was isolated using RiboEx reagent (GeneAll, Seoul, Korea), and cDNA was synthesized from the isolated RNA using Maxime RT PreMix (iNtRON, Seongnam, Korea). cDNA was amplified with primers (SEQ ID NOs: 13, 14, 15, and 16) using the following cycling parameters: 95°C for 1 min, 35 cycles of 59°C for 35 s each, and 72°C for 1 min. The amplified DNA was analyzed using iBright FL1000 and iBright Analysis Software 4.0.0 (Invitrogen).

[0126] The results are shown in Figs. 2a, 2b, 2c, 2d, 2e and 2f.

[0127] As shown in Fig. 2a, the results of the experiment on the promotion of mRNA synthesis of the target substance BMP6 gene by hydrothermal extract (HE) and ethanol extract showed that the 50% ethanol extract showed the best effect. In addition, as shown in Fig. 2c, the treating dosage of the 50% ethanol extract for hair follicle dermal papilla stem cells (HFSC) was determined to be 2000 μg / mL based on the expression of β-catenin. Exosomes extracted from germinated hemp seed meristem callus were isolated and purified (Fig. 2b) to confirm the therapeutic dosage of the isolated exosomes. As shown in Fig. 2d, the appropriate therapeutic dosage of the isolated exosomes was determined to be 40 μg / mL based on the analysis of β-catenin expression. As shown in Figures 2e and eh 2f, the concentration of DT was set to 1 ng / mL in HFDPSCs.

[0128]

[0129] Example 3: Confirmation of the metabolic regulatory function of hemp seed-derived extracts and exosomes.

[0130] After exposure to hemp seed-derived extracts and exosomes (GE2000, E40) for one day, human hair follicle papilla stem cells (HFSCs) (Sku: M36007-08S, Celprogen, CA, USA) were isolated using RiboEx reagent (GeneAll, Seoul, Korea), and cDNA was synthesized from the isolated RNA using Maxime RT PreMix (iNtRON, Seongnam, Korea). cDNA was amplified using primers (SEQ ID NOs: 5 to 8, 11 to 24) at the following cycling parameters: 35 cycles of 95°C for 1 min, 59°C for 35 s, and 72°C for 1 min. The amplified DNA was analyzed using iBright FL1000 and iBright Analysis Software 4.0.0 (Invitrogen).

[0131] The results are shown in Figures 3a and 3b.

[0132] As shown in Fig. 3a, the transcript (RNA) levels of metabolic activator genes BMP6, β-catenin, and Wnt in HFSCs were examined, and the expression levels were 2.4 times higher on average in GE2000 and 3.2 times higher in E40 compared to the control group. In addition, even when exposed to DT1, the two substances (GE2000 and E40) prevented hair loss by inhibiting the activation of TCF (T cell factor), β-catenin, and Wnt (Wingless and Int-1) protein genes in HFDPSCs. In particular, E40 significantly upregulated related genes in HFDPSCs.

[0133] On the other hand, as shown in Fig. 3b, the expression levels of hair loss-promoting genes STAT1, IL-15R, 5α-reductase type 1, and NKG2DL were analyzed according to treatment with hemp seed-derived extracts and exosomes. GE2000 showed a 0.7-fold increase in expression compared to the control group, and E40 showed a 0.5-fold increase in expression. In summary, it was confirmed that both hemp seed-derived extracts and exosomes have the functionality of inhibiting hair loss.

[0134]

[0135] Example 4: Confirmation of the ability of hemp seed-derived extracts and exosomes to induce differentiation of HFSCs into dermal papilla cells (DPCs) and germ cells.

[0136] After exposure to hemp seed-derived extracts and exosomes (GE2000, E40) for 1 day, total RNA from human hair follicle papilla stem cells (HFSCs) (Sku: M36007-08S, Celprogen, CA, USA) was isolated from cells using RiboEx reagent (GeneAll, Seoul, Korea), and cDNA was synthesized from the isolated RNA using Maxime RT PreMix (iNtRON, Seongnam, Korea). cDNA was amplified using primers (SEQ ID NOs: 1 to 4) at the following cycling parameters: 95°C for 1 min, 59°C for 35 s, and 72°C for 1 min, for 35 cycles. The amplified DNA was analyzed using iBright FL1000 and iBright Analysis Software 4.0.0 (Invitrogen). To evaluate the results of flow cytometry, cultured cells were fixed with 2% paraformaldehyde for 4 h and treated with 0.02% Tween 20 for 5 min. The treated cells were incubated with three fluorescently conjugated immunoglobulins, FITC-anti-K15 (Novous, CA, USA) and APC-anti-Sox2, at 37°C for 2 days. The stained cells were evaluated using a flow cytometer (BD FACScalibur), FlowJo 10.6.1 (BD science), and Prism 7 (GraphPad, CA, USA).

[0137] The results are shown in Figures 4a and 4b.

[0138] As shown in Fig. 4a and Fig. 4b, the expression levels of DPC marker genes Sox2 and Itga9 were 1.8 times higher on average for GE2000 and 2.8 times higher for E40 compared to the control group when differentiation into DPCs and germ cells of HFSCs was confirmed. In addition, the differentiation into germ cells was 1.6 times higher for GE2000 and 3.3 times higher for E40. From the above results, it was confirmed that both GE2000 and E40 materials promote differentiation into cells that form hair follicles.

[0139]

[0140] To evaluate immunocytochemical results, cultured cells were fixed with 2% paraformaldehyde for 4 h and treated with 0.02% Tween 20 for 5 min. The treated cells were incubated with three fluorescently conjugated immunoglobulins, FITC-anti-K15 (Novous, CA, USA) and APC-anti-Sox2, at 37°C for 2 days. The stained cells were evaluated using a flow cytometer (BD FACScalibur), FlowJo 10.6.1 (BD science), and Prism 7 (GraphPad, CA, USA).

[0141] The results are shown in Fig. 5.

[0142] As shown in Fig. 5, the differentiation of HFSCs into DPCs and germ cells was confirmed by ICC (immunocytochemistry). The number of cells stained with the Sox2 fluorescent antibody, a DPC marker, was approximately 3.1 times higher in GE2000 and approximately 5.3 times higher in E40 than the control group. In addition, when the K14 fluorescent antibody, a germ cell marker, was used, GE2000 and E40 were 2.4 times and 4.1 times higher, respectively. Therefore, as with the results using flow cytometry, both materials were effective in promoting differentiation, and the effect was particularly excellent in the case of E40.

[0143]

[0144] Example 5: Confirmation of CD8+ T cell regulatory function of hemp seed-derived extracts and exosomes.

[0145] To assess downregulation of inactivating factors, proliferated T cells (MOLT-4, ATCC) were exposed to conditioned medium (GE2000CM, E40CM), fixed with 2% paraformaldehyde for 4 h, and treated with 0.02% Tween 20 for 5 min. The treated cells were incubated with three fluorescently conjugated immunoglobulins: FITC-anti-CD8 (BD biosciences, CA, USA), PE-anti-IL-2Rβ (Biolegend, CA, USA), PerCP-anti-JAK1 (ThermoFisher, MA, USA), and APC-anti-STAT1 (Abcam, CA, UK) at 37°C for 2 days. The stained cells were analyzed using a flow cytometer (BD biosciences FACScalibur), FlowJo 10.6.1 (BD biosciences), and Prism 7 (GraphPad, CA, USA).

[0146] The results are shown in Fig. 6.

[0147] As shown in Figure 6, CD8 + As a result of confirming the effect of suppressing the hair loss activator of T cells, the expression level of IL-2Rβ was 0.5 times in GE2000CM and 0.3 times in E40CM compared to the control group. Jak1 was 0.5 times in GE2000CM and 0.3 times in E40CM compared to the control group, and STAT1 was 0.5 times in GE2000CM and 0.2 times in E40CM compared to the control group. In summary, it was confirmed that both materials showed the effect of suppressing the hair loss activator.

[0148]

[0149] To assess IFNγ regulation, proliferated T cells (MOLT-4, ATCC) were exposed to conditioned media (GE2000CM, E40CM) and the supernatant was isolated. Stimulated CD8 +IFNγ secreted from T cells was analyzed using an IFNγ ELISA kit (ThermoFisher), AMR-100 (ALLSHENG, HZ, CN), and Prism 7 (GraphPad, CA, USA).

[0150] The results are shown in Fig. 7.

[0151] As shown in Figure 7, CD8 + As a result of checking the amount of IFNγ secretion by T cells, it was 0.43 times higher in GE2000CM and 0.12 times higher in E40CM compared to the control group. When calculated as secretion amount, the control group was 341.1 pg / mL, GE2000CM was 172.6 pg / mL, and E40 was 56.3 pg / mL. In summary, both materials showed the effect of suppressing hair loss activating factors, and it was found that the effect of E40 was particularly excellent.

[0152]

[0153] Example 6: Comparison of the effects of hemp seed oil (HSO) and two substances.

[0154] The hair loss prevention and treatment effects of hemp seed-derived extracts and exosomes were compared with those of hemp seed oil (HSO). The results are shown in Fig. 8. As shown in Fig. 8, GE2000 and E40 more strongly upregulated hair loss prevention genes, such as TCF, β-catenin, and Wnt, in HFDPSCs even when exposed to DT1. The preventive effect of GE2000 was slightly higher than that of HSO, but the effect of E40 was approximately 2.2 times higher than that of hemp seed oil (HSO).

[0155] This indicates that the hemp seed-derived extract and exosome of the present invention are more effective against DT, a hair loss-inducing substance, than HSO, which has been reported to be effective in preventing hair loss.

[0156]

[0157] In summary, both germinated hemp seed extract and callus exosome showed excellent hair loss prevention effects, and the exosome material showed a higher effect. Both materials promoted the differentiation of dermal papilla cells (DPCs), which are important for hair growth and hair follicle cycle control, and germ cells, which are important for hair, sebaceous gland, and apocrine gland formation, and CD8, which are immune cells that affect hair loss. + The expression of hair loss-promoting markers in T cells was effectively suppressed. Therefore, the two materials are considered to have potential for use as hair care products and drugs related to hair loss prevention in humans and dogs.

Claims

1. A pharmaceutical composition for preventing or treating alopecia comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

2. A pharmaceutical composition for preventing or treating alopecia, wherein the hemp seed in claim 1 is a sprouted hemp seed.

3. A pharmaceutical composition for preventing or treating alopecia, wherein the extract in paragraph 1 is extracted with water, an alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof.

4. A pharmaceutical composition for preventing or treating alopecia, wherein in the third paragraph, the alcohol is ethanol at a volume content of 30 to 70%.

5. A pharmaceutical composition for preventing or treating alopecia, wherein the concentration of the hemp seed extract in paragraph 1 is 500 to 3000 ㎍ / mL.

6. A pharmaceutical composition for preventing or treating alopecia, wherein the hemp seed-derived exosome in paragraph 1 is an exosome isolated from hemp seed callus.

7. A pharmaceutical composition for preventing or treating alopecia, wherein the concentration of the hemp seed-derived exosome in paragraph 1 is 10 to 100 ㎍ / mL.

8. A pharmaceutical composition for preventing or treating alopecia, wherein the size of the hemp seed-derived exosome in paragraph 1 is 50 to 300 nm.

9. A pharmaceutical composition for preventing or treating alopecia, wherein the composition comprises a pharmaceutically acceptable carrier, excipient or diluent in paragraph 1.

10. A cosmetic composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

11. A cosmetic composition for preventing hair loss or promoting hair growth, wherein the formulation of the composition in paragraph 10 is a hair tonic, hair cream, hair lotion, hair shampoo, hair rinse, hair conditioner, hair spray, hair aerosol, pomade, powder, gel, hair pack, hair treatment, eyebrow hair tonic, eyelash hair tonic, eyelash nutrient or ointment.

12. A food composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

13. A food composition for preventing hair loss or promoting hair growth, wherein the composition is manufactured in any one formulation selected from powder, granules, pills, tablets, capsules, candy, syrup, and beverage in the 12th paragraph.

14. A feed composition for preventing hair loss or promoting hair growth comprising a hemp seed extract, hemp seed-derived exosomes, or a combination thereof.

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