Composition for preventing hair loss and promoting hair growth, comprising connarus conchocarpus extract

Abstract

The present invention relates to a composition for promoting hair growth, the composition comprising a Connarus conchocarpus extract as an active ingredient, and relates to a composition for preventing hair loss and promoting hair growth, the composition being for promoting hair growth and containing ingredients for inhibiting male pattern hair loss, inhibiting alopecia areata, and inducing new growth or growth promotion of hair. When the Connarus conchocarpus extract according to the present invention is used, hair loss can be effectively prevented or treated, and hair growth can be promoted.

Description

Composition for preventing hair loss and promoting hair growth containing extract of Cornarus conchocapis

[0001] The present invention relates to a composition for promoting hair growth comprising an extract of Connarus conchocarpus as an active ingredient, and to a composition for preventing hair loss and promoting hair growth containing a component that suppresses male pattern baldness, suppresses alopecia areata, and induces new hair growth or hair growth promotion.

[0002] This patent application claims priority to Republic of Korea Patent Application No. 10-2024-0092487, filed with the Korean Intellectual Property Office on July 12, 2024, the disclosure of which is incorporated herein by reference.

[0003]

[0004] In modern society, hair loss is a common cosmetic and psychological problem for both men and women, and demand for its prevention and treatment is steadily increasing. Hair loss is caused by a variety of factors, including genetics, hormonal imbalances, stress, medications, aging, and environmental pollution, acting singly or in combination. Its pathophysiological mechanisms are also highly complex.

[0005] Hair grows through a cycle of growth (anagen), regression (catagen), and resting (telogen) phases. However, an imbalance or shortening of this cycle can lead to miniaturization of hair follicles and hair loss, resulting in alopecia. Alopecia is classified into congenital, acquired, and pathological alopecia based on its cause. Among the acquired alopecia types, male pattern baldness and alopecia areata are known to have a high incidence and relatively clear pathogenesis.

[0006] Male pattern baldness is caused by dihydrotestosterone (DHT) binding to the androgen receptor (AR) on hair papilla cells, which induces the expression of hair growth inhibitors such as DKK1, TGF-β, and IL-6. This suppresses the Wnt / β-catenin signaling pathway, leading to apoptosis and shortened growth phase of hair follicle cells. Furthermore, the conversion of testosterone to DHT by 5α-reductase further accelerates this process.

[0007] Alopecia areata is an autoimmune disease in which the immune privilege of hair follicles is disrupted and the JAK-STAT signaling pathway is overactivated. Increased secretion of cytokines such as IFN-γ and IL-15 promotes inflammatory gene expression through phosphorylation of JAK1 / 2 / 3 and STAT1 / 3 / 5, and CD8 + Hair follicle cells are destroyed by T cell invasion and excessive granzyme B and perforin secretion. Furthermore, activation of the JAK-STAT pathway inhibits the transition of hair follicle stem cells from the resting phase to the growth phase, contributing to persistent hair loss.

[0008] To date, Minoxidil, Finasteride, steroids, immunosuppressants, and phototherapy have been suggested as hair loss treatments. However, their effectiveness is limited due to the reported side effects of long-term use, such as increased body hair, skin irritation, decreased sexual function, depression, and Cushing's syndrome.

[0009] Therefore, there is an urgent need to develop a safe and effective hair loss treatment material of natural origin that has few side effects and can comprehensively regulate hair follicle and papilla cells functions.

[0010] Connarus conchocarpus F.Muell. is an evergreen shrub of the legume family, native to tropical regions such as New Guinea and Queensland. Its main components, flavonoids and phenolic compounds, are known to possess antioxidant, anti-inflammatory, and antibacterial properties. While it has traditionally been used to treat digestive and respiratory ailments, scientific research on its hair loss prevention and hair growth promotion effects is limited.

[0011]

[0012] The present inventors have diligently researched and developed an effective hair loss treatment. As a result, they have discovered that Connarus conchocarpus extract exhibits excellent hair loss prevention and treatment effects, as well as hair growth promotion, due to its physiological effects, including promoting dermal papilla cell proliferation, activating the Wnt / β-catenin pathway, suppressing JAK and AR expression, and inhibiting 5α-reductase activity. This has led to the completion of the present invention.

[0013] Accordingly, an object of the present invention is to provide a pharmaceutical composition for preventing or treating alopecia comprising an extract of Connarus conchocarpus.

[0014] Another object of the present invention is to provide a method for producing a Cornarus conchocapis extract.

[0015] Another object of the present invention is to provide a cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0016] Another object of the present invention is to provide a food composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0017] Another object of the present invention is to provide a feed composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0018] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0019]

[0020] The present invention provides inventions 1 to 17 below.

[0021] 1. A pharmaceutical composition for preventing or treating alopecia comprising an extract of Connarus conchocarpus.

[0022] 2. A pharmaceutical composition for preventing or treating alopecia, wherein the extract of Cornarus conchocapis in 1 is a solvent extract selected from the group consisting of glycol solvents, water, alcohol, or a combination thereof.

[0023] 3. In 1 or 2, the extract is a pharmaceutical composition for preventing or treating alopecia, which is as follows:

[0024] (a) the glycol solvent is a glycol solvent having a concentration of 20% (v / v) to 80% (v / v); or

[0025] (b) The alcohol has a concentration of 20% (v / v) to 80% (v / v).

[0026] 4. A pharmaceutical composition for preventing or treating alopecia, wherein the Connarus conchocarpus extract in any one of 1 to 3 is a hot water extract of Connarus conchocarpus residue extracted with a glycol solvent at a concentration of 20% (v / v) to 80% (v / v).

[0027] 5. A pharmaceutical composition for preventing or treating alopecia, wherein the extract has the properties of inhibiting expression of 5-α-reductase, inhibiting expression of androgen receptor, inhibiting activity of JAK (Janus kinase), activating Wnt / β-catenin signaling pathway, or a combination thereof, in any one of 1 to 4.

[0028] 6. A pharmaceutical composition for preventing or treating alopecia, wherein in any one of 1 to 5, 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, congenital alopecia, and combinations thereof.

[0029] 7. A pharmaceutical composition for preventing or treating alopecia, wherein the extract is obtained by any one of the manufacturing methods of 10 to 13 below, in any one of 1 to 6.

[0030] 8. Use for preparing any one of the pharmaceutical compositions or agents of 1 to 7 for preventing or treating alopecia.

[0031] 9. A method for preventing or treating alopecia, comprising administering to a subject any one of the pharmaceutical compositions of 1 to 7.

[0032] 10. A method for producing a Connarus conchocarpus extract, comprising a step of extracting Connarus conchocarpus by adding a solvent selected from the group consisting of a glycol solvent, water, alcohol, or a combination thereof.

[0033] 11. A method for producing a Cornarus conconapis extract, wherein in 10, the glycol solvent has a concentration of 20% (v / v) to 80% (v / v); or the alcohol has a concentration of 20% (v / v) to 80% (v / v).

[0034] 12. A method for producing a Cornarus conchocapis extract, wherein the method comprises a step of hot water extraction of the residue remaining after the glycol solvent extraction step at a concentration of 20% (v / v) to 80% (v / v) in 10 or 11.

[0035] 13. A method for producing a Cornarus conchocapis extract, wherein the method further comprises a step of concentrating under reduced pressure after extraction, in any one of 10 to 12.

[0036] 14. A cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0037] 15. 14, a cosmetic composition for preventing hair loss or promoting hair growth, 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 growth agent, eyelash hair growth agent, eyelash nutrient or ointment.

[0038] 16. A food composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0039] 17. A feed composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0040]

[0041] In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating alopecia comprising an extract of Connarus conchocarpus.

[0042] The present inventors have conducted extensive research to develop an effective hair loss treatment. As a result, we have discovered that Connarus conchocarpus extract exhibits excellent hair loss prevention and treatment effects and hair growth promotion, due to its physiological effects, including promoting hair papilla cell proliferation, activating the Wnt / β-catenin pathway, suppressing JAK and AR expression, and inhibiting 5α-reductase activity.

[0043] The term "Connarus conchocarpus" used herein refers to an evergreen shrub or small vine belonging to the Connaraceae family, native to tropical regions such as New Guinea and northeastern Australia (Queensland). This plant has traditionally been used in folk medicine to treat digestive or respiratory diseases, and has recently attracted attention as a medicinal plant containing various physiologically active substances such as flavonoids and phenolic compounds.

[0044] The above-mentioned Cornarus concocapis may be the entire plant, above-ground parts (leaves, stems, etc.), or underground parts (roots, rhizomes, rhizomes, etc.). Furthermore, it may be manufactured into a powder form through processes such as washing, drying, and grinding, but is not limited thereto. The powder may be used as a starting material for the solvent extraction process described below.

[0045] 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.

[0046] 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.

[0047] 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).

[0048] 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.

[0049] 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.

[0050] In one embodiment of the present invention, the composition is administered in combination with a drug for preventing or treating hair loss.

[0051] 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.

[0052] 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.

[0053] The Cornulus concornapis extract used in the present invention can be purchased or obtained through direct extraction. When the Cornulus concornapis 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 non-polar solvents, can be used.

[0054] Suitable polar solvents include, but are not limited to, (i) water, (ii) low molecular weight alcohols and polyhydric alcohols (e.g., methanol, ethanol, n-propanol, isopropanol, n-butanol, 1-pentanol, 2-butoxyethanol, ethylene glycol, butylene glycol, glycerol, etc.), (iii) acetic acid, (iv) dimethylformamide (DMF), and (v) dimethyl sulfoxide (DMSO).

[0055] 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).

[0056] 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 extract of Cornarus conchocapis includes not only one obtained using the above-described extraction solvent, but also one obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, a 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 Cornarus conchocapis extract of the present invention.

[0057] In the present invention, when extracting Cornarus conchocapis, 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 may be used, but is not limited thereto.

[0058] To prepare an extract according to one embodiment of the present invention, the amount of solvent used for extraction can be appropriately selected depending on the amount of Cornarus conchocapis used. Specifically, a solvent may be used in an amount of 1 to 20 times, 1 to 18 times, 1 to 16 times, 1 to 14 times, 1 to 12 times, 1 to 10 times, 1 to 8 times, 1 to 6 times, 1 to 4 times, 1 to 2 times, 2 to 20 times, 4 to 20 times, 6 to 20 times, 8 to 20 times, 10 to 20 times, 12 to 20 times, 14 to 20 times, 16 to 20 times, 18 to 20 times, 5 to 15 times, or 10 to 15 times the weight or volume of Cornarus conchocapis used in the preparation of the extract, but is not limited thereto.

[0059] The extraction temperature of the extract according to one embodiment of the present invention is, for example, 10°C to 110°C, 20°C to 110°C, 30°C to 110°C, 40°C to 110°C, 50°C to 110°C, 60°C to 110°C, 70°C to 110°C, 80°C to 110°C, 90°C to 110°C, 100°C to 110°C, 10°C to 100°C, 10°C to 90°C, 10°C to 80°C, 10°C to 70°C, 10°C to 60°C, 10°C to 50°C, 10°C to 40°C, 10°C to 30°C, 10°C to 20°C, 20°C to 30°C, 60°C to 90°C, 70°C to It may be, but is not limited to, 90℃, 75℃ to 85℃, 90℃ to 100℃, 90℃ to 105℃.

[0060] The extraction time of the extract according to one embodiment of the present invention is, for example, 1 hour to 100 hours, 1 hour to 90 hours, 1 hour to 80 hours, 1 hour to 72 hours, 1 hour to 70 hours, 1 hour to 60 hours, 1 hour to 50 hours, 1 hour to 40 hours, 1 hour to 30 hours, 1 hour to 20 hours, 1 hour to 10 hours, 10 hours to 100 hours, 20 hours to 100 hours, 30 hours to 100 hours, 40 hours to 100 hours, 50 hours to 100 hours, 60 hours to 100 hours, 70 hours to 100 hours, 80 hours to 100 hours, 90 hours to 100 hours, 10 hours to 80 hours, 20 hours to 60 hours, 40 hours to It may be, but is not limited to, 60 hours, 2 to 10 hours, 3 to 10 hours, 3 to 5 hours, 60 to 80 hours, or 65 to 75 hours.

[0061] The Cornarus conchocapis 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.

[0062] In one embodiment of the present invention, the Cornarus conchocapis extract is a solvent extract selected from the group consisting of glycol solvents, water, alcohol, or a combination thereof.

[0063] The above solvent is a means for effectively extracting effective ingredients from the Cornarus conchocapis plant, and the type and physiological activity of the extracted ingredients may vary depending on the characteristics of each solvent.

[0064] For example, glycol solvents may include, but are not limited to, butylene glycol, propylene glycol, ethylene glycol, and dipropylene glycol. These solvents have excellent water solubility and skin penetration ability, and thus can provide extracts suitable for use in cosmetic and over-the-counter drug compositions.

[0065] Water or purified water can be used as a solvent to extract mainly water-soluble polysaccharides, flavonoids, phenolic compounds, etc. through high-temperature hot water extraction or leaching methods.

[0066] For example, alcohol solvents can be low molecular weight alcohols such as, but not limited to, methanol, ethanol, n-propanol, isopropanol, n-butanol, n-propanol, n-butanol, 1-pentanol, 2-butoxyethanol, etc. These are effective for extracting not only polar compounds but also some non-polar bioactive substances. These solvents can be used alone or as aqueous solutions at a concentration of 20 to 80% (v / v), and can also be applied in composite solvent compositions.

[0067] In one embodiment of the present invention, the extract is as follows:

[0068] (a) the glycol solvent is a glycol solvent having a concentration of 20% (v / v) to 80% (v / v); or

[0069] (b) The alcohol has a concentration of 20% (v / v) to 80% (v / v).

[0070] In one embodiment of the present invention, the Connarus conchocarpus extract is a hot water extract of Connarus conchocarpus residue extracted with a glycol solvent at a concentration of 20% (v / v) to 80% (v / v).

[0071] In one specific embodiment of the present invention, the Connarus conchocarpus extract can be prepared in various ways depending on the type and concentration of the solvent used. In particular, the concentration of the extraction solvent affects the dissolution efficiency, stability, and physiological activity of the active ingredient, so it is important to set it within an appropriate concentration range.

[0072] Specifically, the glycol solvent extract uses a solution prepared by diluting a glycol solvent such as butylene glycol, propylene glycol, etc. in purified water at a concentration of 20% (v / v) to 80% (v / v) as an extraction solvent, and can be obtained by soaking or stirring and extracting the plant body of Cornarus conchocapis with this solution.

[0073] Additionally, water extracts are prepared by first extracting with a glycol solvent (20-80% v / v) and then secondary extracting the remaining solid residue with hot water (purified water at around 100°C). This hot water extraction can additionally secure water-soluble components (e.g., polysaccharides, water-soluble phenols, flavonoids, etc.) that were not extracted during the first solvent extraction, making it advantageous for developing compositions exhibiting complex physiological activities.

[0074] Meanwhile, alcohol extracts are obtained by steeping or stirring Cornarus conchocapis in an extraction solution containing low-molecular-weight alcohols such as methanol, ethanol, propanol, and butanol diluted in purified water at a concentration of 20% (v / v) to 80% (v / v), enabling the extraction of not only polar components but also some semi-polar components. Ethanol is a preferred solvent due to its high safety, especially for applications in food and pharmaceuticals.

[0075] In this way, the Cornarus conchocapis extract of the present invention can be prepared using a single solvent such as a glycol solvent, water, alcohol, etc. or a combination of stepwise extractions, and by setting the concentration of the extraction solvent in the range of 20% (v / v) to 80% (v / v), it is possible to stably secure an effective ingredient suitable for the purpose.

[0076] The concentration of the glycol solvent or alcohol is 20% (v / v) to 80% (v / v), 20% (v / v) to 75% (v / v), 20% (v / v) to 70% (v / v), 20% (v / v) to 60% (v / v), 20% (v / v) to 55% (v / v), 20% (v / v) to 50% (v / v), 20% (v / v) to 45% (v / v), 20% (v / v) to 40% (v / v), 20% (v / v) to 35% (v / v), 20% (v / v) to 30% (v / v), 20% (v / v) to 25% (v / v), 25% (v / v) to 80% (v / v), 30% (v / v) to 80% (v / v), 35% (v / v) to 80% (v / v), 40% (v / v) to 80% (v / v), 45% (v / v) to 80% (v / v), 50% (v / v) to 80% (v / v), 55% (v / v) to 80% (v / v), 60% (v / v) to 80% (v / v), 65% (v / v) to 80% (v / v), 70% (v / v) to 80% (v / v), 75% (v / v) to 80% (v / v), 30% (v / v) to 70% (v / v), 40% (v / v) to 60% (v / v), 25% (v / v) to It may be, but is not limited to, 35% (v / v), 40% (v / v) to 60% (v / v), 45% (v / v) to 55% (v / v), or 65% (v / v) to 75% (v / v).

[0077] In one embodiment of the present invention, the extract has the properties of inhibiting the expression of 5-α-reductase, inhibiting the expression of androgen receptor, inhibiting the activity of JAK (Janus kinase), activating the Wnt / β-catenin signaling pathway, or a combination thereof.

[0078] The Cornarus conchocapis extract according to the present invention can exhibit hair loss prevention or treatment effects by acting on various molecular biological mechanisms related to hair loss, such as 5-α-reductase inhibition, androgen receptor inhibition, JAK signaling pathway inhibition, and Wnt / β-catenin signaling pathway activation. Each of the above mechanisms is a scientifically established target in the pathophysiology of hair loss, and it has already been proven through various literature and clinical studies that when acting alone or in combination, it exhibits a hair loss improvement effect.

[0079] First, 5-alpha-reductase is an enzyme that converts testosterone, a male hormone, into dihydrotestosterone (DHT), a more potent androgen. DHT binds to androgen receptors in hair papilla cells, inducing the expression of hair loss-related genes, resulting in hair follicle atrophy and shortened growth phase. Accordingly, finasteride and dutasteride, which are 5-alpha-reductase inhibitors, have been approved by the FDA as treatments for male pattern baldness, and have been reported to increase hair count and improve hair thickness when taken long-term. Inhibiting this enzyme, as in the extract of the present invention, can scientifically induce hair loss improvement effects.

[0080] Furthermore, the androgen receptor (AR) is a key target protein that binds to DHT and induces transcription of hair loss genes. It is known that increased AR expression accelerates the progression of male pattern baldness. Some studies have reported that AR inhibition also affects the restoration of Wnt / β-catenin signaling and the activation of hair follicle stem cells. RNA-based candidate substances that indirectly regulate AR are actively being studied in the preclinical stage. Therefore, a composition that inhibits AR expression, such as the extract according to the present invention, can effectively alleviate hair loss progression.

[0081] The JAK (Janus kinase) signaling pathway is a key pathway driving the pathogenesis of autoimmune hair loss, particularly in alopecia areata. Inflammatory cytokines such as IFN-γ and IL-15 induce phosphorylation of JAK1 / 2 / 3 and STAT1 / 3 / 5, which leads to CD8+ T cell invasion into hair follicles and destruction of hair follicle cells. Accordingly, JAK inhibitors such as tofacitinib and ruxolitinib have clinically demonstrated hair regrowth effects in patients with alopecia areata, and in 2022, the JAK inhibitor baricitinib was approved by the FDA as a treatment for alopecia areata. Therefore, the extract of the present invention, which inhibits JAK activity, can be effectively applied to inflammatory alopecias such as alopecia areata.

[0082] The Wnt / β-catenin signaling pathway is an essential pathway that induces the survival and anagen entry of hair follicle stem cells. When the activity of this pathway is suppressed, hair regeneration is inhibited and telogen effluvium is induced. There are research results that show that while hair loss is induced in a mouse model in which the β-catenin gene is deleted, inducing the expression of Wnt3a or β-catenin promotes hair follicle regeneration and hair growth. In fact, minoxidil has also been reported to exhibit a hair growth effect by indirectly activating the Wnt / β-catenin pathway. The extract of the present invention can induce the survival of hair follicle cells and recovery of the growth cycle by showing activity that increases the expression of Wnt3a and β-catenin.

[0083] Thus, the Cornulus conchocapis extract of the present invention can contribute to the treatment of hair loss through each mechanism alone. When combined, it can provide even more effective treatment by blocking various causes of hair loss or promoting hair growth. This scientific and clinical background supports the claim that the composition of the present invention is a technology sufficiently effective in the prevention or treatment of hair loss.

[0084] 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, congenital alopecia, and combinations thereof.

[0085] In particular, the Cornarus conchocapis extract of the present invention can effectively suppress the progression of hormonal alopecia such as androgenetic alopecia through inhibition of 5-α-reductase and androgen receptor expression, and can also have a significant effect on autoimmune alopecia such as alopecia areata through inhibition of the JAK pathway. In addition, activation of the Wnt / β-catenin pathway can contribute to the recovery of telogen effluvium or seborrheic or inflammatory alopecia through induction of regeneration and growth phase of hair follicle cells.

[0086] Therefore, the extract of the present invention can be widely applied to alopecia caused by various etiologies, and can be utilized as a natural therapeutic material that shows excellent efficacy, especially in types of hair loss involving hormonal, autoimmune, and inflammatory mechanisms.

[0087] In one aspect of the present invention, the present invention provides a pharmaceutical composition or use for preparing a medicament comprising a Connarus conchocarpus extract for the prevention or treatment of alopecia. Since the use has in common with the pharmaceutical composition in that it comprises a Connarus conchocarpus extract, description of the common portions is omitted to avoid excessive redundancy.

[0088] In one aspect of the present invention, there is provided a method for preventing or treating alopecia, comprising administering to a subject a pharmaceutical composition comprising a Connarus conchocarpus extract. Since the method for preventing or treating alopecia shares similarity with the pharmaceutical composition in that it comprises a Connarus conchocarpus extract, description of the common portions is omitted to avoid excessive duplication.

[0089]

[0090] In one aspect of the present invention, the present invention provides a method for preparing a Connarus conchocarpus extract, comprising the step of extracting Connarus conchocarpus by adding a solvent selected from the group consisting of a glycol solvent, water, alcohol, or a combination thereof.

[0091] The method for producing the Cornarus conchocapis extract of the present invention has something in common in that it obtains the Cornarus conchocapis extract described above, and therefore, it is common in the pretreatment or state of Cornarus conchocapis, the type, amount or concentration of the solvent, extraction conditions such as extraction time or temperature, extraction method, etc., and in order to prevent excessive redundant description in the specification, the description of the common parts is omitted.

[0092] In one embodiment of the present invention, the glycol solvent has a concentration of 20% (v / v) to 80% (v / v); or the alcohol has a concentration of 20% (v / v) to 80% (v / v).

[0093] In one embodiment of the present invention, the manufacturing method includes a step of hot water extraction of the residue remaining after the glycol solvent extraction step at a concentration of 20% (v / v) to 80% (v / v).

[0094] In one aspect of the present invention, a Cornarus conchocapis extract can be prepared through a two-step sequential extraction process. In particular, by additionally performing hot water extraction on the plant residue remaining after the first extraction step using a glycol solvent, the effective ingredients contained in Cornarus conchocapis can be more comprehensively secured.

[0095] More specifically, first, a 20% (v / v) to 80% (v / v) glycol solvent, such as butylene glycol, propylene glycol, etc., diluted in purified water is used to perform immersion or stirring extraction on the dried powder of Cornarus conchocapis. The extraction can be performed at a temperature of 25 to 60°C for 10 to 100 hours, and then the glycol solvent extract is separated through filtration.

[0096] The solid residue remaining after the above filtration may contain water-soluble physiologically active substances (e.g., polysaccharides, phenols, water-soluble flavonoids, etc.) that were not recovered in the first extraction. Therefore, by performing a second hot water extraction on this residue, a complex extract containing a wider variety of components can be obtained.

[0097] Secondary extraction is performed by adding purified water to the residue in an amount 10 to 20 times the weight of the residue, followed by heating, stirring, or immersion extraction at a temperature of approximately 100°C for 2 to 5 hours or more. The final hot water extract is then obtained through filtration and reduced pressure concentration. This hot water extract can be combined with the primary glycol solvent extract, fractionated separately, or used as a complementary agent within the composition.

[0098] In this way, the extraction method of the present invention has the advantage of being able to more quantitatively and efficiently secure polar and water-soluble active ingredients present in the Cornarus conchocapis plant compared to a single solvent extraction method, and by expanding the spectrum of physiologically active ingredients through two-step extraction, the hair loss inhibition and hair growth promotion effects of the final composition can also be improved.

[0099] In one embodiment of the present invention, the manufacturing method additionally includes a reduced pressure concentration step performed after extraction.

[0100] In the above manufacturing method, a vacuum concentration step can be added after the extraction process to remove the solvent and increase the concentration of the active ingredient. Vacuum concentration is typically performed using a rotary evaporator or vacuum concentrator, evaporating the solvent under low pressure at a temperature of 40 to 60°C to concentrate the extract. This process has the advantage of increasing concentration efficiency while minimizing the decomposition of heat-sensitive active ingredients, ultimately securing a high-concentration extract in a form suitable for composition manufacturing.

[0101]

[0102] In one aspect of the present invention, the present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0103] 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.

[0104] 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.

[0105] 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.

[0106] 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.

[0107] 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.

[0108] 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.

[0109] 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.

[0110] 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.

[0111] 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.

[0112] 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.

[0113] 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.

[0114]

[0115] In one aspect of the present invention, the present invention provides a food composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0116] 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.

[0117] The health functional food, health food and food additive composition containing the above-described Cornarus conchocapis extract 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.

[0118] 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.

[0119] In the present invention, there is no particular limitation on other ingredients added in addition to the above-mentioned Conarus conchocapis extract, and various flavoring agents or natural carbohydrates, etc. can be contained as additional ingredients like in a typical beverage. Examples of the above-mentioned natural carbohydrates are monosaccharides such as glucose, fructose, etc.; disaccharides such as maltose, sucrose, etc.; and polysaccharides such as dextrin, cyclodextrin, etc., and typical sugars and sugar alcohols such as xylitol, sorbitol, erythritol, etc. In addition to the above-mentioned flavoring agents, 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-mentioned natural carbohydrates can 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.

[0120] In the present invention, the food composition containing the extract of Cornarus conchocapis 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 the production of natural fruit juice and fruit juice drinks and vegetable drinks.

[0121] 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.

[0122]

[0123] In one aspect of the present invention, the present invention provides a feed composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0124] The above feed composition may include feed or feed additives for feeding a diet to an animal.

[0125] When the above-described Cornarus conchocapis extract 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 additionally include any 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 polyphenol, catechin, alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, and phytic acid.

[0126] In the present invention, when the Cornarus conchocapis extract is used as a 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.

[0127] In the present invention, when the Cornarus conchocapis extract is used as the feed additive, it 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 it 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, such as 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-containing tablet, or microdispersible. 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.

[0128] 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.

[0129] The features and advantages of the present invention are summarized as follows:

[0130] (a) The present invention provides a pharmaceutical composition for preventing or treating alopecia comprising an extract of Connarus conchocarpus.

[0131] (b) The present invention provides a method for preparing a Cornarus conchocapis extract.

[0132] (c) The present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0133] (d) The present invention provides a food composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0134] (e) The present invention provides a feed composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

[0135] (f) When the Connarus conchocarpus extract of the present invention is used, hair loss can be effectively prevented or treated, and hair growth can be promoted.

[0136] Figure 1 shows the results of the Janus kinase (JAK)1 inhibitory activity analysis of the Cornarus conchocapis extract.

[0137] Figure 2 shows the results of the 5-α-reductase inhibition assay of the Cornarus conchocapis extract.

[0138] Figure 3 shows the results of an analysis of the androgen receptor production inhibition ability of the Cornarus conchocapis extract.

[0139] Figure 4 shows the results of the analysis of the ability of Cornarus conchocapis extract to activate the Wnt / β-Catenin signaling pathway.

[0140]

[0141] 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.

[0142]

[0143] Example

[0144] Example 1: Preparation of Connarus conchocarpus extract

[0145] 1-1. Preparation of Connarus conchocarpus butylene glycol extract

[0146] Butylene glycol (BG) was mixed with purified water to prepare 30% (v / v), 50% (v / v), and 70% (v / v) aqueous solutions of butylene glycol, each prepared by mixing BG with purified water, and added 10 times the weight of BG to 200 g of dried Connarus conchocarpus, followed by extraction at 25°C for 3 days. Each extract was concentrated in a vacuum evaporator to obtain more than 10 g of Connarus conchocarpus extract. These were designated CC-BG30, CC-BG50, and CC-BG70.

[0147]

[0148] 1-2. Preparation of Connarus conchocarpus hot water extract

[0149] After the first extraction with 30% (v / v), 50% (v / v), and 70% (v / v) butylene glycol aqueous solutions, purified water was added 15 times to each remaining residue, extracted at 100°C for 4 hours, and each extract was concentrated in a vacuum evaporator to obtain more than 10 g of Connarus conchocarpus extract, respectively. These were named CC-W30, CC-W50, and CC-W70.

[0150]

[0151] 1-3. Preparation of Connarus conchocarpus ethanol extract

[0152] Ethanol 30% (v / v), 50% (v / v), and 70% (v / v) aqueous solutions, prepared by mixing ethanol with purified water, were added to 200 g of dried Connarus conchocarpus in an amount 10 times their weight, and extracted at 80°C for 4 hours. Each extract was concentrated in a reduced pressure evaporator to obtain more than 10 g of Connarus extract, which were named CC-EE30, CC-EE50, and CC-EE70.

[0153]

[0154] The method for preparing the above extract is summarized in Table 1.

[0155] Sample number Name Manufacturing method 1 CC-BG30 Extracted at 25°C for 3 days with 30% (v / v) butylene glycol 2 CC-BG50 Extracted at 25°C for 3 days with 50% (v / v) butylene glycol 3 CC-BG70 Extracted at 25°C for 3 days with 70% (v / v) butylene glycol 4 CC-W30 Add 15 times purified water to the CC-BG30 extraction residue and extract at 100°C for 4 hours 5 CC-W50 Add 15 times purified water to the CC-BG50 extraction residue and extract at 100°C for 4 hours 6 CC-W70 Add 15 times purified water to the CC-BG70 extraction residue and extract at 100°C for 4 hours 7 CC-EE30 Extracted at 80°C with 30% (v / v) ethanol for 4 hours Extraction8CC-EE50Extracted with 50% (v / v) ethanol at 80℃ for 4 hours9CC-EE70Extracted with 70% (v / v) ethanol at 80℃ for 4 hours

[0156] Example 2: Evaluation of Janus kinase (JAK) 1 inhibition ability In order to evaluate the JAK1 (Janus kinase 1) inhibition ability, a major biomarker of alopecia areata, of each sample prepared in Example 1, an experiment was performed using HEK293T cells with high transfection efficiency. JAK1 overexpression conditions were established by transfecting HEK293T cells with FLAG-tagged JAK1 (FALG-JAK1), and 500 μL of each test sample was treated according to the concentration.

[0157] After sample treatment, cells were harvested in 1 mL of phosphate-buffered saline (PBS), transferred to a microtube (e-tube), and centrifuged at 12,000 rpm for 3 minutes to remove the supernatant, leaving only the pellet. Subsequently, a lysis buffer containing the following components was used to lyse the cells: 50 mM Tris-HCl (pH 7.4), 120 mM NaCl, 25 mM β-glycerol phosphate (pH 7.5), 20 mM sodium fluoride (NaF), 2% NP-40 (Nonidet P-40), and protease inhibitors.

[0158] After dissolution, sonication was performed, and then the supernatant was obtained by re-centrifugation at 12,000 rpm at 4°C for 3 minutes. The obtained supernatant was separated through 10-15% concentration SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and Western blotting analysis was performed using each antibody for p-JAK1, JAK1, p-STAT3, STAT3, and β-actin.

[0159] The results are shown in Figure 1.

[0160] As shown in Fig. 1, in the FLAG-JAK1 transfection group, the band intensities of FLAG and JAK1 proteins were confirmed to increase compared to the control group (Normal). As a result of comparing the JAK1 inhibitory effect of each extract, the 50% ethanol extract (sample 8, CC-EE30) showed the best inhibitory activity, and the 30% and 70% ethanol extracts (sample 7, CC-EE30; sample 9, CC-EE70) also showed a significant decrease in JAK1 phosphorylation. In addition, the 70% butylene glycol extract (sample 3, CC-BG70) and the hot water extract (sample 6, CC-W70) were also confirmed to show a certain level of inhibitory effect.

[0161] These results suggest that Connarus conchocarpus ethanol extract is a natural material with significant inhibitory activity against JAK1, a major target of alopecia areata.

[0162]

[0163] Example 3: Evaluation of 5-α-reductase inhibition ability

[0164] 5-alpha-reductase, a key target of male pattern baldness, converts the male hormone testosterone into dihydrotestosterone (DHT). Increased expression of this enzyme leads to increased DHT production, and DHT is known to induce androgen receptor (AR) activity more strongly than testosterone, promoting hair follicle shrinkage and hair loss.

[0165] The 5-α-reductase inhibitory activity of the Connarus conchocarpus extract prepared in Example 1 was evaluated. 10-week-old male Sprague-Dawley rats were used for the experiment, and the entire liver of the rat was extracted, homogenized with a lysis buffer containing the following composition, and the enzyme was extracted: 7.5 nM K2HPO₄, 3.25 nM KH₂PO₄, 1 mM dithiothreitol (DTT), 32 mM sucrose, 0.2 mM phenylmethylsulfonyl fluoride (PMSF), and additional protease inhibitors.

[0166] For 4 μL of the extracted enzyme, 34 mM NADPH (nicotinamide adenine dinucleotide phosphate) and 0.4 mM testosterone were added to the reaction system in which the sample to be evaluated was treated at a concentration of 20 μM, and then McIlvaine buffer (pH 5.0) was immediately added. The reaction was terminated by heat treatment at 80°C for 5 minutes, and the oxidation of NADPH was measured as a change in absorbance at a wavelength of 340 nm to evaluate the change in enzyme activity.

[0167] The results are shown in Fig. 2.

[0168] As shown in Fig. 2, butylene glycol (BG) extract and hot water extract showed no inhibitory effect or rather a tendency to increase enzyme activity, whereas ethanol extracts (CC-EE30, sample 7; CC-EE50, sample 8; CC-EE70, sample 9) all showed excellent inhibitory activity. In particular, the 30% ethanol extract (CC-EE30, sample 7) showed higher inhibitory activity than the positive control, finasteride, and the 50% and 70% ethanol extracts (CC-EE50, sample 8; CC-EE70, sample 9) also showed strong inhibitory effects.

[0169] In summary, these results show that the ethanol extract among Connarus conchocarpus extracts effectively inhibits 5-α-reductase activity, and in particular, the 30% and 50% ethanol extracts (CC-EE30, sample 7; CC-EE50, sample 8) showed excellent inhibitory effects on the main causative mechanism of male pattern baldness. In addition, the ethanol extract also consistently exhibited the previously confirmed JAK1 inhibitory effect, suggesting that it has dual efficacy that can be simultaneously applied to alopecia areata. On the other hand, the butylene glycol extract and the hot water extract showed limited effects only on specific targets.

[0170]

[0171] Example 4: Evaluation of androgen receptor (AR) production inhibition ability

[0172] Male pattern baldness is caused by increased blood dihydrotestosterone (DHT) concentrations, which promote the expression of androgen receptors (ARs), ultimately leading to hair follicle shrinkage and hair loss. This is one of the major pathological mechanisms underlying hair loss. Therefore, the Connarus conchocarpus extracts prepared in Example 1 were evaluated for their ability to inhibit androgen receptor production.

[0173] In this experiment, HEK293T cells, which have high transfection efficiency, were used and transfected with the Myc-DDK-SRD5A2 plasmid to establish an androgen receptor overexpression model. The cells were then treated with testosterone to create conditions for androgen receptor overexpression, and 500 μL of each test sample was administered.

[0174] Cells were washed with 1 mL of phosphate-buffered saline (PBS), transferred to an e-tube, and centrifuged at 12,000 rpm for 3 minutes to remove the supernatant, leaving only the pellet. Cells were then lysed using a lysis buffer composed of the following composition: 50 mM Tris-HCl (pH 7.4), 120 mM NaCl, 25 mM β-glycerol phosphate (pH 7.5), 20 mM NaF (sodium fluoride), 2% NP-40 (Nonidet P-40), and protease inhibitors.

[0175] After dissolution, sonication was performed, and the supernatant was obtained by centrifugation at 12,000 rpm for 3 minutes at 4°C. The obtained sample was subjected to 10-15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE) to separate proteins, and Western blot analysis was performed using antibodies against Myc, androgen receptor (AR), and β-actin.

[0176] The results are shown in Fig. 3.

[0177] As shown in Fig. 3, among the nine samples, the 30% butylene glycol extract (CC-BG30, sample 1), the hot water extract (CC-W50, sample 5), and the 50% ethanol extract (CC-EE50, sample 8) showed excellent inhibitory activity against androgen receptor production, and exhibited even stronger inhibitory activity than dutasteride, which was used as a positive control. In addition, CC-BG50 (sample 2), CC-W70 (sample 6), and CC-EE30 (sample 7) also showed a certain level of inhibitory activity.

[0178] These results suggest that extracts derived from Connarus conchocarpus have physiological activity that can effectively inhibit DHT-induced androgen receptor expression, and in particular, it was confirmed that not only ethanol extracts but also some BG and water extracts can act target-specifically.

[0179]

[0180] Example 5. Evaluation of the effect of activating the Wnt / β-Catenin signaling pathway.

[0181] During the progression of male pattern baldness, increased dihydrotestosterone (DHT) concentrations promote androgen receptor (AR) expression, thereby inhibiting the Wnt / β-Catenin signaling pathway, thereby interfering with hair follicle anagen induction and inducing hair loss. Therefore, in this example, we evaluated whether Connarus conchocarpus extract could activate the Wnt / β-Catenin signaling pathway.

[0182] The experiment was conducted using HEK293T cells with high transfection efficiency. After establishing experimental conditions by transfecting HEK293T cells with the SRD5A1-GFP plasmid, 500 μL of each sample prepared in Example 1 was treated according to the concentration.

[0183] Cells were washed with 1 mL of phosphate-buffered saline (PBS), transferred to an e-tube, and centrifuged at 12,000 rpm for 3 minutes to remove the supernatant, leaving only the pellet. Cells were then lysed using a lysis buffer containing the following components: 50 mM Tris-HCl (pH 7.4), 120 mM NaCl, 25 mM β-glycerol phosphate (pH 7.5), 20 mM NaF (sodium fluoride), 2% NP-40 (Nonidet P-40), and protease inhibitors.

[0184] After dissolution, sonication was performed, and the supernatant was obtained by centrifugation at 12,000 rpm for 3 minutes at 4°C. The obtained proteins were separated through 10-15% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), and Western blot analysis was performed using each antibody against GFP, Wnt3a, β-catenin, and β-actin.

[0185] The results are shown in Fig. 4.

[0186] As shown in Fig. 4, among the samples with increased expression of Wnt3a and β-catenin, the 50% ethanol extract (CC-EE50, sample 8) showed the most excellent activation effect, and the 4th and 6th water extracts (CC-W30, sample 4; CC-W70, sample 6) also showed excellent Wnt3a expression induction effects. In addition, the 50% and 70% butylene glycol extracts (CC-W50, sample 5; CC-W70, sample 6) were also observed to have the effect of inducing a certain level of signal pathway activation.

[0187] According to the results of the above examples, when ethanol was used as a solvent in the preparation of Connarus conchocarpus extract, it exhibited various physiological activities, and in particular, 30% and 50% ethanol extracts (CC-EE30, sample 7; CC-EE50, sample 8) showed excellent inhibitory effects on major biomarkers related to hair loss. On the other hand, in the evaluation of Wnt3a and β-catenin expression, not only the 50% ethanol extract (CC-EE50, sample 8) but also the hot water extracts (CC-W30, sample 4; CC-W70, sample 6) showed excellent expression induction effects, confirming that the hot water extract is also effective in promoting hair growth.

[0188] Therefore, these results confirmed that Connarus conchocarpus extracts, especially the 30% or 50% ethanol extracts (CC-EE30, sample 7; CC-EE50, sample 8) and the hot water extracts prepared from the residue after 30% or 70% butylene glycol extraction (CC-W30, sample 4; CC-W70, sample 6), activated the Wnt / β-Catenin pathway. Meanwhile, Wnt signaling activity is known to be closely related to the proliferation of dermal papilla cells. The Wnt / β-catenin signaling pathway regulates the survival and proliferation of dermal papilla cells and plays an important role in the hair growth cycle. When Wnt signaling is activated, β-catenin is not degraded in the cytoplasm but translocates to the nucleus, inducing the expression of genes that promote cell proliferation and differentiation. Therefore, it is suggested that the Cornarus conchocapis extract of the present invention can effectively act to promote hair follicle growth and induce hair growth by activating the Wnt / β-catenin pathway to increase the proliferation of hair papilla cells and promote hair growth.

Claims

1. A pharmaceutical composition for preventing or treating alopecia comprising an extract of Connarus conchocarpus.

2. A pharmaceutical composition for preventing or treating alopecia, wherein the extract of Cornarus conchocapis in paragraph 1 is a solvent extract selected from the group consisting of glycol solvents, water, alcohol, or a combination thereof.

3. In the second paragraph, the extract is a pharmaceutical composition for preventing or treating alopecia, which is as follows: (a) the glycol solvent is a glycol solvent having a concentration of 20% (v / v) to 80% (v / v); or (b) The alcohol has a concentration of 20% (v / v) to 80% (v / v).

4. A pharmaceutical composition for preventing or treating alopecia, wherein the Connarus conchocarpus extract in paragraph 1 is a hot water extract of Connarus conchocarpus residue extracted with a glycol solvent at a concentration of 20% (v / v) to 80% (v / v).

5. A pharmaceutical composition for preventing or treating alopecia, wherein the extract has the properties of inhibiting the expression of 5-α-reductase, inhibiting the expression of androgen receptor, inhibiting the activity of JAK (Janus kinase), activating the Wnt / β-catenin signaling pathway, or a combination thereof, in the first paragraph.

6. A pharmaceutical composition for preventing or treating alopecia, wherein the alopecia in paragraph 1 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, congenital alopecia, and combinations thereof.

7. A method for producing a Connarus conchocarpus extract, comprising the step of extracting Connarus conchocarpus by adding a solvent selected from the group consisting of a glycol solvent, water, alcohol, or a combination thereof.

8. A method for producing a Cornarus conconapis extract, wherein in the 7th paragraph, the glycol solvent has a concentration of 20% (v / v) to 80% (v / v); or the alcohol has a concentration of 20% (v / v) to 80% (v / v).

9. A method for producing a Cornarus conchocapis extract, wherein the method comprises a step of hot water extraction of the residue remaining after the glycol solvent extraction step at a concentration of 20% (v / v) to 80% (v / v) in the 7th paragraph.

10. A method for producing a Cornarus conchocapis extract, wherein the method further comprises a step of concentrating under reduced pressure after extraction, according to any one of claims 7 to 9.

11. A cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

12. A cosmetic composition for preventing hair loss or promoting hair growth, wherein the formulation of the composition in claim 11 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.

13. A food composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

14. A feed composition for preventing hair loss or promoting hair growth comprising an extract of Connarus conchocarpus.

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