Peptides having activity to promote hair growth and inhibit hair loss and uses thereof

Abstract

The present invention relates to peptides having an activity of promoting hair growth or preventing or inhibiting hair loss. The peptides according to the present invention promote proliferation of hair follicle dermal papilla cells (HFDPC) and human hair root sheath cells (HHORSC), upregulate expression of a beta-catenin secondary factor in hair follicle dermal papilla cells (HFDPC), and upregulate expression of a growth factor.

Description

TECHNICAL FIELD

[0001] The present application relates to peptides having an activity of promoting hair growth and inhibiting hair loss and uses thereof. BACKGROUND

[0002] Alopecia is a phenomenon of excessive hair shedding due to an anagen phase becoming longer than normal or an increase in the number of hair follicles in the anagen phase, and means excessive hair loss due to stress, nutritional deficiency, local blood flow disorder, hormonal and genetic factors, etc., and it cannot be recovered.

[0003] As a therapeutic agent for hair loss, 5α-reductase inhibitors, which convert the male hormone testosterone into the androgenic hormone dihydrotestosterone (DHT), such as drugs such as finasteride or dutasteride have been developed and approved for commercial sale, and minoxidil, a drug that promotes hair growth by stimulating hair follicles and increasing blood flow, has also been approved and used.

[0004] However, for 5α-reductase inhibitors, side effects such as sexual dysfunction, depression, anxiety, and breast enlargement are reported to occur relatively frequently; and for minoxidil, side effects such as ECG abnormalities, tachycardia, and endocarditis are reported to occur; in addition, it is reported that once minoxidil administration is stopped, hair loss begins again.

[0005] Peptides have high biocompatibility, and when used as a single active ingredient in a drug or cosmetic, they have fewer side effects than extracts including various ingredients. After inhibiting local hair loss, these peptides are easily broken down and eliminated by proteases in the body; thus, there are fewer concerns about side effects of these peptide drugs compared to existing therapeutic agents.

[0006] PCT International Patent Application WO 2005-082395 discloses a peptide having a hair growth-promoting effect by promoting the progression of the anagen phase to the growth phase in the hair growth cycle, and Korean Patent No. 10-2387764 discloses a peptide derivative in the form of a dimer that promotes the proliferation of hair follicle cells.

[0007] [Prior Art Documents]

[0008] [Patent Documents]

[0009] WO 2005-082395

[0010] Korean Patent No. 10-2387764 SUMMARY

[0011] Technical problem

[0012] The present inventors have conducted research to develop a peptide having improved activity of promoting hair growth and inhibiting hair loss, while effectively promoting hair growth and having fewer side effects. As a result, the present inventors have confirmed through experiments that the new peptide they developed promotes the proliferation of human hair follicle dermal papilla cells (HHFDPC) and human hair outer root sheath cells (HHORSC), improves the expression level of downstream factors of β-catenin (i.e., LEF-1 and cyclin D1), down-regulates the expression level of hair loss protein DKK-1 in HHFDPC, and increases the expression level of growth factors bFGF, HGF, and VEGF, thereby completing the present invention.

[0013] Accordingly, an object of the present invention is to provide a novel peptide having activity of promoting hair growth and inhibiting hair loss.

[0014] Another object of the present invention is to provide a composition for promoting hair growth, or inhibiting, preventing, or improving hair loss, the composition including the peptide having the above-mentioned activity as an active ingredient.

[0015] Still another object of the present invention is to provide a pharmaceutical composition for preventing or treating hair loss.

[0016] Still another object of the present invention is to provide a cosmetic composition for promoting hair growth, or inhibiting or improving hair loss.

[0017] Technical solution

[0018] To achieve the above object,

[0019] One aspect of the present invention provides a peptide including the amino acid sequence of SEQ ID NO: 1.

[0020] Another aspect of the present invention provides a composition for promoting hair growth, or inhibiting, preventing, or improving hair loss, the composition including the peptide as an active ingredient.

[0021] Still another aspect of the present invention provides a pharmaceutical composition for preventing or treating hair loss, the pharmaceutical composition including the peptide as an active ingredient.

[0022] Still another aspect of the present invention provides a cosmetic composition for promoting hair growth, or preventing or improving hair loss, the cosmetic composition including the peptide as an active ingredient.

[0023] Hereinafter, the present invention will be described in detail.

[0024] 1. Peptides and their activity

[0025] According to an aspect of the present application, there is provided a peptide comprising the amino acid sequence disclosed in SEQ ID NO: 1.

[0026] As used herein, the term "peptide" refers to a linear molecule in which amino acid residues are connected to each other by peptide bonds.

[0027] The peptide of the present application comprising the amino acid sequence of SEQ ID NO: 1 can be used without modification, but within the range that does not affect the original activity of the peptide, such as the activity of promoting hair growth or inhibiting hair loss, a variant having a different amino acid sequence or a fragment thereof by deletion, insertion, substitution, or a combination thereof of amino acid residues can also be used.

[0028] The peptide of the present application can be modified by phosphorylation, sulfation, acrylation, glycosylation, methylation, farnesylation, etc., as long as the activity of the peptide is not changed.

[0029] The peptide of the present application includes a peptide comprising an amino acid sequence substantially identical to the peptide comprising the amino acid sequence of SEQ ID NO: 1, and a variant or an active fragment thereof. The substantially identical amino acid sequence refers to an amino acid sequence having at least 75%, for example, at least 80%, at least 85%, at least 90%, at least 95%, at least 97% sequence identity to the amino acid sequence of SEQ ID NO: 1, respectively. In addition, the peptide can additionally include a targeting sequence, a tag, a labeled residue, and an amino acid sequence prepared for a specific purpose of increasing the half-life or stability of the peptide.

[0030] The peptide of the present application can have an induced N-terminal and / or C-terminal modification to select a part of the amino acid sequence and increase the activity thereof. By these N-terminal and / or C-terminal modifications, the stability of the peptide of the present application can be significantly improved, for example, the half-life of the peptide can be increased when administered in vivo. The term "stability" encompasses not only in vivo stability, which protects the peptide of the present application from attack by intracellular protein cleavage enzymes, but also storage stability, for example, storage stability at room temperature.

[0031] The N-terminal modification can be a modification in which a protecting group selected from the group consisting of acetyl, fluorenylmethyloxycarbonyl, formyl, palmitoyl, myristyl, stearyl, and polyethylene glycol (PEG) is bound to the N-terminus of the peptide. The C-terminal modification can be a modification in which a hydroxyl (-OH), an amino (-NH2), a hydrazine (-NHNH2), etc. are bound to the C-terminus of the peptide, but is not limited thereto.

[0032] The peptides of the present application can be prepared by various methods widely known in the art to which the present application pertains. For example, the peptides of the present application can be prepared using chemical synthesis methods known in the art, particularly, solid-phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85: 2149-54 (1963); Stewart, et al., Solid Phase Peptide Synthesis, 2nd. ed., Pierce Chem. Co.: Rockford, Ill (1984)) or liquid-phase synthesis techniques (U.S. Patent No. 5516891).

[0033] The peptides of the present application have an activity of promoting hair growth, or preventing or inhibiting hair loss.

[0034] In an embodiment, the peptides of the present application promote proliferation of hair follicle dermal papilla cells (HFDPCs).

[0035] In an embodiment, the peptides of the present application promote proliferation of human hair outer root sheath cells (HHORSCs).

[0036] In an embodiment, the peptides of the present application upregulate gene expression of one or more factors selected from the group consisting of LEF-1, which is a downstream factor of β-catenin, and Cyclin D1 in hair follicle dermal papilla cells (HFDPCs).

[0037] In an embodiment, the peptides of the present application upregulate expression of a growth factor in hair follicle dermal papilla cells (HFDPCs). The growth factor can be one or more selected from the group consisting of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF).

[0038] By the above-mentioned activity, the above-described peptides of the present application can exhibit an effect of promoting hair growth, or inhibiting, preventing, or ameliorating hair loss.

[0039] 2. Compositions for promoting hair growth, or preventing, treating or ameliorating hair loss

[0040] In another aspect of the present application, there is provided a composition for promoting hair growth, or preventing, treating, or ameliorating hair loss, the composition comprising: a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0041] As described above, the peptide of the present application comprising the amino acid sequence of SEQ ID NO: 1 has an activity of promoting hair growth, or inhibiting or preventing hair loss.

[0042] In another aspect of the present application, the composition of the present application can be a pharmaceutical composition for preventing or treating alopecia, the pharmaceutical composition comprising a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0043] As used herein, the term "hair loss" or "alopecia" refers to a condition in which hair does not exist in an area where hair should normally exist due to abnormal hair loss, or the number of hairs is reduced compared to the normal state. In embodiments, the hair loss or symptoms of hair loss in the present application include hair loss on the scalp.

[0044] In the present application, the hair loss includes genetic androgenetic alopecia (male pattern baldness), female pattern baldness, telogen effluvium, anagen effluvium, alopecia areata, tinea capitis caused by fungal infection, hair loss caused by hypothyroidism or by hyperthyroidism, hair loss caused by inflammation (seborrheic dermatitis), hair loss caused by malnutrition, hair loss caused by diabetes, hair loss caused by lupus, hair loss caused by trichogenesis disorder, hair loss caused by drug treatment (anticoagulants, antidepressants, discontinuation of oral contraceptives, chemotherapy), toxic folliculitis, lichen planopilaris, hair loss caused by burns or trauma, hair loss caused by ultraviolet rays, and hair loss due to contact with fine dust in the air, but is not limited thereto.

[0045] In embodiments, the pharmaceutical composition comprising the peptide of the present application as an active ingredient promotes the proliferation of hair follicle dermal papilla cells (HFDPC).

[0046] In embodiments, the pharmaceutical composition comprising the peptide of the present application as an active ingredient promotes the proliferation of human hair outer root sheath cells (HHORSC).

[0047] In embodiments, the pharmaceutical composition comprising the peptide of the present application as an active ingredient upregulates the gene expression of one or more factors selected from the group consisting of LEF-1, which is a downstream factor of β-catenin, and cyclin D1 in hair follicle dermal papilla cells (HFDPC).

[0048] In embodiments, the pharmaceutical composition comprising the peptide of the present application as an active ingredient upregulates the expression of a growth factor in hair follicle dermal papilla cells (HFDPC). The growth factor can be one or more selected from the group consisting of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF).

[0049] The pharmaceutical composition of the present application can include a therapeutically effective amount of the peptide of the present application.

[0050] The term "therapeutically effective amount" refers to an amount of a peptide (an active ingredient of the pharmaceutical composition of the present application) sufficient to achieve its activity or efficacy, for example, a therapeutically effective amount refers to an amount sufficient to achieve efficacy in treating or preventing hair loss.

[0051] As used herein, the term "preventing" refers to reducing the risk of developing a disease or disorder, and refers to any action that inhibits or delays the onset of a disease by preventing the progression of the disease or one or more clinical symptoms thereof.

[0052] As used herein, the term "treating" refers to alleviating a disease or disorder, and includes all actions that improve or beneficially alter the symptoms of a disease by preventing or reducing the progression of the disease or one or more clinical symptoms thereof.

[0053] In the present application, the prevention or treatment of hair loss can be to eliminate the cause of hair loss or to inhibit the progression of hair loss, or can be to promote hair growth by inhibiting hair loss or promoting hair formation.

[0054] The pharmaceutical composition of the present application can include a pharmaceutically acceptable carrier.

[0055] The pharmaceutically acceptable carriers commonly used in formulations include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginates, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil, etc.; but the carriers are not limited thereto.

[0056] In addition to the above ingredients, the pharmaceutical composition of the present application can further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., but the ingredients are not limited thereto.

[0057] Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington: The Science and Practice of Pharmacy (19th ed., 1995, Williams & Wilkins).

[0058] The pharmaceutical composition of the present application can be administered by any route suitable for treating hair loss, for example, orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, topical application, or transdermal application. Since the pharmaceutical composition of the present application has an activity of preventing or treating hair loss, it can be applied topically, such as to the scalp.

[0059] The dose of the pharmaceutical composition can be 0.0001 μg to 100 mg, 0.001 μg to 100 mg, 0.01 μg to 100 mg, 0.1 μg to 100 mg, or 1.0 μg to 1000 mg per day, but is not limited thereto. The pharmaceutical composition can be prescribed in various ways depending on factors such as the formulation method, the administration method, the age, body weight, sex, pathological condition, food intake, administration time, administration route, excretion rate, and response sensitivity of the patient.

[0060] The pharmaceutical composition of the present application can be formulated in a unit dosage form by using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by one skilled in the art, or can be formulated by placing it in a multiple dose container. Specifically, the dosage form can be in the form of a solution, suspension, or emulsion in an oily or aqueous medium, or can be in the form of an extract, powder, granule, tablet, or capsule, and can additionally include a dispersant or stabilizer.

[0061] The pharmaceutical composition of the present application can be a skin preparation for external use. The skin preparation for external use is a preparation that can be used by applying it to the outside of the skin. When the pharmaceutical composition of the present application is used as a skin preparation for external use, it can be applied to the scalp, particularly the scalp of an area where hair loss has occurred or the scalp of an area where hair growth is to be promoted. The skin preparation for external use can be a cream, a gel, an ointment, a skin emulsifier, a skin suspension, a transdermal delivery patch, a medicated bandage, a lotion, or a combination thereof. The skin preparation for external use can be appropriately mixed with ingredients commonly used in skin preparations for external use (e.g., cosmetics and pharmaceuticals) as needed, such as an aqueous ingredient, an oil-based ingredient, a powdery ingredient, an alcohol, a humectant, a thickening agent, an ultraviolet absorber, a whitening agent, a preservative, an antioxidant, a surfactant, a fragrance, a coloring agent, various skin nutrients, or a combination thereof. The skin preparation for external use can be appropriately mixed with a metal chelating agent (e.g., disodium ethylenediaminetetraacetate, trisodium ethylenediaminetetraacetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, etc.), caffeine, tannin, a grass extract, glabridin, various herbs, a drug (e.g., tocopheryl acetate, glycyrrhizin, tranexamic acid and derivatives or salts thereof, etc.), vitamin C, ascorbic acid phosphate magnesium, ascorbic acid glucoside, arbutin, kojic acid, and a sugar (e.g., glucose, fructose, trehalose, etc.).

[0062] In another aspect of the present application, the composition of the present application can be a cosmetic composition for promoting hair growth, or preventing or improving hair loss, which includes a peptide including the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0063] As used herein, the term "improving" means any action that ameliorates or benefits the symptoms of a disease or disorder.

[0064] In an embodiment, the cosmetic composition including the peptide of the present application as an active ingredient promotes proliferation of hair follicle dermal papilla cells (HFDPC).

[0065] In an embodiment, the cosmetic composition including the peptide of the present application as an active ingredient promotes proliferation of human hair outer root sheath cells (HHORSC).

[0066] In an embodiment, the cosmetic composition including the peptide of the present application as an active ingredient upregulates gene expression of one or more factors selected from the group consisting of LEF-1, which is a downstream factor of β-catenin, and cyclin D1 in hair follicle dermal papilla cells (HFDPC).

[0067] In an embodiment, the cosmetic composition including the peptide of the present application as an active ingredient upregulates expression of a growth factor in hair follicle dermal papilla cells (HFDPC). The growth factor can be one or more selected from the group consisting of basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF).

[0068] The cosmetic composition can be prepared in any dosage form generally prepared in the art to which the present application pertains, and can be a skin preparation for external use. For example, the cosmetic composition can be formulated as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, a spray, etc., but is not limited thereto.

[0069] The cosmetic composition can be prepared in various forms such as a solution (e.g., a skin softening toner, a nutrition toner, a nutrition cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, a loose powder, a hair tonic, a hair paste, a shampoo, a shampoo water, a hair dye, a hair conditioner, a spray-type hair spray, an aerosol-type hair spray, a hair wax, a gel, etc.), a sol-gel, an emulsion, an oil, a wax, an aerosol, etc., but is not limited thereto.

[0070] The cosmetic composition of the present application can include other additives such as excipients and carriers, and can be applied and mixed with common ingredients mixed in general skin cosmetics, as needed.

[0071] When the dosage form of the cosmetic composition is a paste, a cream, or a gel, an animal oil, a vegetable oil, a wax, a paraffin, starch, tragacanth, a cellulose derivative, a polyethylene glycol, a silicone, bentonite, silicon oxide, talc, zinc oxide, etc. can be used as a carrier ingredient.

[0072] When the dosage form of the cosmetic composition is a powder or a spray, lactose, talc, silicon dioxide, aluminum hydroxide, calcium silicate, or polyamide powder can be used as a carrier ingredient, and specifically, when the cosmetic composition is a spray, a propellant such as chlorofluorocarbon, propane / butane, or dimethyl ether can be further included, but is not limited thereto.

[0073] When the cosmetic composition is in the form of a solution or an emulsion, a solvent, a solubilizer, or an emulsifier can be used, and water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol oil, glycerol fatty acid ester, polyethylene glycol, sorbitol fatty acid ester, etc. can be used as a carrier ingredient.

[0074] When the dosage form of the cosmetic composition is a suspension, a liquid diluent (e.g., water, ethanol, and propylene glycol), a suspending agent (e.g., ethoxy isostearyl alcohol, polyoxyethyl sorbitol ester, polyoxyethylene sorbitol ester, etc.), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, tragacanth gum, etc. can be used as a carrier ingredient.

[0075] When the dosage form of the cosmetic composition is a surfactant-containing cleanser, a fatty alcohol sulfate, a fatty alcohol ether sulfate, a sulfosuccinic acid monoester, a hydroxyethyl sulfonate, an imidazoline derivative, a methyl taurate, a sarcosinate, a fatty acid amide ether sulfate, an alkyl amide betaine, a fatty alcohol, a fatty acid glyceride, a fatty acid diethanolamide, a vegetable oil, a lanolin derivative, an ethoxylated glycerol fatty acid ester, etc. can be used as a carrier ingredient.

[0076] When the dosage form of the cosmetic composition is a shampoo, the peptide of the present application can be mixed with basic ingredients for forming a shampoo, such as a thickening agent, a surfactant, a viscosity regulator, a moisturizer, a pH regulator, a preservative, and an essential oil. CDE can be used as a thickening agent; LES (an anionic surfactant) and cocamidopropyl betaine (an amphoteric surfactant) can be used as a surfactant; a polyquaternary ammonium salt can be used as a viscosity regulator; glycerol can be used as a moisturizer; citric acid and sodium hydroxide can be used as a pH regulator. Grapefruit extract, etc. can be used as a preservative; in addition, essential oils of cedar, mint, and rosemary, and silk acid, pentaol, or vitamin E can be added.

[0077] In addition to the peptide of the present application and the carrier ingredient, the ingredients included in the cosmetic composition can include auxiliary ingredients commonly used in cosmetic compositions, such as an antioxidant, a stabilizer, a solubilizer, a vitamin, a pigment, and a fragrance, but are not limited thereto.

[0078] The peptide of the present application can be included in the above composition at a concentration of 0.01 µM to 1000 µM, and specifically, the peptide of the present application can be included in the above composition at a concentration of 0.01 µM to 1000 µM; 0.05 µM to 800 µM, 0.05 µM to 700 µM, 0.05 µM to 600 µM, 0.05 µM to 500 µM, 0.05 µM to 300 µM, 0.05 µM to 200 µM; 0.1 µM to 800 µM, 0.1 µM to 700 µM, 0.1 µM to 600 µM, 0.1 µM to 500 µM, 0.1 µM to 300 µM, 0.1 µM to 200 µM; 1 µM to 800 µM, 1 µM to 700 µM, 1 µM to 600 µM, 1 µM to 500 µM, 1 µM to 300 µM, 1 µM to 200 µM; 5 µM to 800 µM, 5 µM to 700 µM, 5 µM to 600 µM, 5 µM to 500 µM, 5 µM to 300 µM, or 5 µM to 200 µM; but not limited thereto.

[0079] 3. Use of the peptides of the invention

[0080] In another aspect of the present application, there is provided a use of a peptide comprising the amino acid sequence of SEQ ID NO: 1 for promoting hair growth, or inhibiting, preventing, treating or ameliorating alopecia.

[0081] In still another aspect of the present application, there is provided a method for promoting hair growth, or inhibiting, preventing, treating or ameliorating alopecia, the method comprising administering a peptide comprising the amino acid sequence of SEQ ID NO: 1 or a composition comprising the same to a subject in need of promoting hair growth, or preventing, inhibiting, treating or ameliorating alopecia.

[0082] In still another aspect of the present application, there is provided a use of a peptide comprising the amino acid sequence of SEQ ID NO: 1 in the manufacture of a medicament or a cosmetic for promoting hair growth, or preventing, treating or ameliorating alopecia.

[0083] Advantageous effects

[0084] The peptide of the present application has an activity of promoting hair growth, or preventing or inhibiting alopecia. The peptide of the present application promotes proliferation of hair follicle dermal papilla cells (HFDPC) and human hair outer root sheath cells (HHORSC), upregulates expression of LEF-1 and cyclin D1, which are downstream genes of β-catenin, in hair follicle dermal papilla cells (HFDPC), and upregulates expression of growth factors such as bFGF, HGF and VEGF.

[0085] However, the effects of the present invention are not limited to those described above, and those skilled in the art will clearly understand from the following description other effects not mentioned. Attached Figure Description

[0086] Figure 1 Experimental results show that the peptides of the present invention promote the proliferation of HHFDPC cells when treated with 10 μM and 100 μM.

[0087] Figure 2 Experimental results show that the peptides of the present invention promote the proliferation of HHORSC cells when treated with 50 μM and 100 μM.

[0088] Figure 3a Experimental results show that when HHFDPC cells were treated with the peptides of the present invention at 10 μM, 50 μM, and 100 μM, the expression of LEF-1, a downstream factor of β-catenin, and cyclin D1 was increased.

[0089] Figure 3b To measure Figure 3a The intensity of the middle strip is shown in a graph of relative intensity values.

[0090] Figure 4a Experimental results show that when HHFDPC cells were treated with the peptides of the present invention at 10 μM, 50 μM, and 100 μM, the activity of HHFDPC cells increased, thereby increasing the expression of growth factors such as bFGF, HGF, and VEGF.

[0091] Figure 4b To measure Figure 4a The intensity of the middle strip is shown in a graph of relative intensity values. Detailed Implementation

[0092] The present invention will be described in detail below through embodiments. However, the following embodiments are for illustrative purposes only, and the scope of the present invention is not limited to these embodiments.

[0093] Preparation Example 1: Preparation of Peptides and Peptide Complexes

[0094] Peptides having the amino acid sequence SEQ ID NO: 1 shown in Table 1 were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA), and the synthesized peptides were purified using C18 reversed-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). The chromatographic column used was an ACQUITY UPLC BEH300 C18 (2.1 mm × 100 mm, 1.7 μm, Waters Co, USA).

[0095] [Table 1]

[0096]

[0097] The efficacy of the prepared peptide of SEQ ID NO: 1 was evaluated through the following experiments.

[0098] Experimental Example 1: Promoting the proliferation of human follicular dermal papillary cells (HHFDPC)

[0099] The effect of the peptide prepared in Preparation Example 1 above on the proliferation of human follicular dermal papillary cells (HHFDPC) was tested.

[0100] HHFDPC at 4×10 3 Cells were seeded at a density of 10 cells / well in 96-well cell culture plates and cultured for 24 hours in complete mesenchymal stem cell culture medium. The medium was then replaced with serum-free mesenchymal stem cell culture medium and cultured for another 24 hours. The cultured cell cultures were treated with the peptides prepared in Example 1 above at 10 μM and 100 μM, respectively, with 1 μM minoxidil (MNX) as a positive control and no treatment as a negative control (con). These cultures were then cultured at 37°C for another 72 hours. Afterward, 10 μL of 5 mg / mL MTT solution was added, and the mixture was incubated at 37°C in a CO2 incubator for 4 hours. The medium was then removed, and 100 μL of DMSO was added, followed by shaking for 10 minutes. Finally, the absorbance was measured at 540 nm using a spectrophotometer (Molecular Devices (SpectraMax iD3, CA, USA)).

[0101] Experimental results are as follows Figure 1 As shown, treatment of HHFDPC with the peptide from Preparation Example 1 at 10 μM and 100 μM promoted HHFDPC proliferation compared to the negative control (con).

[0102] Experimental Example 2: Promoting the proliferation of human hair outer root sheath cells (HHORSC)

[0103] The effect of the peptide prepared in Preparation Example 1 above on the proliferation of human hair outer root sheath cells (HHORSC) was tested.

[0104] HHORSC at 4×10 3The cells were seeded at a density of 1 cell / well in a 96-well cell culture plate and cultured for 24 hours under mesenchymal stem cell complete medium conditions. Then, the medium was replaced with serum-free mesenchymal stem cell medium and cultured for another 24 hours. The cultured cell culture was treated with the peptide prepared in Preparation Example 1 above at 50 μM and 100 μM, respectively, treated with 100 nM of epidermal growth factor (EGF) as a positive control, and not treated with any substance as a negative control (con), and incubated at 37°C for another 72 hours. Subsequently, 10 μL of 5 mg / mL MTT solution was added thereto, and the mixture was incubated at 37°C in a CO2incubator for 4 hours, the medium was removed, 100 μL of DMSO was added thereto, and shaken for 10 minutes. Finally, the absorbance was measured using a spectrophotometer (Molecular Devices (SpectraMax iD3, CA, USA)) at a wavelength of 540 nm.

[0105] The experimental results, as shown in Table 1, confirmed that the treatment of HHORSC with the peptide of Preparation Example 1 above at 50 μM and 100 μM promoted the proliferation of HHORSC compared to the negative control group (con). Figure 2

[0106] Experimental Example 3: Promotion of expression of β-catenin downstream genes in human hair follicle dermal papilla cells

[0107] The effect of the peptide prepared in Preparation Example 1 above on the expression level of lymphoid enhancer-binding factor 1 (LEF-1) and cyclin D1, which are downstream genes of β-catenin, in human hair follicle dermal papilla cells (HHFDPC) was confirmed by RT-PCR.

[0108] HHFDPC were seeded at a density of 4 x 10 5 ​A density of 1,000 cells / well was seeded in a 6-well cell culture plate and cultured for 24 hours under mesenchymal stem cell complete medium conditions. Then, the medium was replaced with serum-free mesenchymal stem cell medium and cultured for another 24 hours. The cultured cell culture was treated with the peptide prepared in Preparation Example 1 above at 10 μM, 50 μM, and 100 μM, respectively, treated with 10 ng / mL of recombinant human Wnt-3a (rhWnt-3a protein, R&D Systems (MN, USA)) as a positive control, and treated with nothing as a negative control (con), each of which was further cultured at 37°C for 24 hours. After washing each culture with PBS, RNA was isolated by treating with 300 μL of eaay blue (Intron, South Korea). The isolated RNA was quantified using a Nano drop, and then cDNA synthesis was performed using a cDNA synthesis kit (Enzynomics, South Korea). Then, PCR reaction was performed using a PCR premix (Enzynomics, South Korea) and primers shown in Table 2 below. The PCR reaction product was electrophoresed on a 1.5% agarose gel, and the band was detected using a Bio-Rad gel image system.

[0109] [Table 2]

[0110]

[0111] The experimental results, as shown in Figure 3a and Figure 3b , confirmed that when HHFDPC was treated with the peptide of Preparation Example 1 above at 10 μM, 50 μM, and 100 μM, β-catenin, which affects cell proliferation and differentiation, was activated, thereby increasing the expression of LEF-1 and cyclin D1, which are downstream factors of β-catenin.

[0112] Experimental Example 4: Promotion of expression of growth factors in human hair follicle dermal papilla cells

[0113] The effect of the peptide prepared in Preparation Example 1 above on the expression level of growth factors in human hair follicle dermal papilla cells (HHFDPC) was determined by an RT-PCR method.

[0114] HHFDPC was seeded at a density of 4 x 10 5The cells were seeded at a density of 1 cell / well in a 6-well cell culture plate and cultured for 24 hours under mesenchymal stem cell complete medium conditions. Then, the medium was replaced with serum-free mesenchymal stem cell medium and cultured for another 24 hours. The cultured cell culture was treated with the peptide prepared in Preparation Example 1 above at 500 nM, 5 μM, and 50 μM, respectively, treated with minoxidil (MNX) as a positive control, and not treated with any substance as a negative control (con), and each was cultured for another 24 hours at 37°C. After washing each culture with PBS, RNA was isolated by treating the culture with 300 μL of eaay blue (Intron, South Korea). The isolated RNA was quantified using a Nano drop, and then cDNA synthesis was performed using a cDNA synthesis kit (Enzynomics, South Korea). Then, PCR reaction was performed using a PCR premix (Enzynomics, South Korea) and primers shown in Table 3 below. The PCR reaction product was electrophoresed on a 1.5% agarose gel, and the band was detected with a Bio-Rad gel image system.

[0115] [Table 3]

[0116]

[0117] The experimental results, as shown in Figure 4a and Figure 4b , confirmed that when HHFDPC was treated with the peptide of Preparation Example 1 above at 10 μM, 50 μM, and 100 μM, the activity of HHFDPC was increased, thereby increasing the expression of growth factors such as bFGF, HGF, and VEGF.

[0118] Preparation Example 2: Preparation of a pharmaceutical composition

[0119] 2-1. Preparation of a powder

[0120] Peptide of the present application 2 g

[0121] Lactose 1 g

[0122] After mixing the above ingredients, fill into an airtight bubble to prepare a powder.

[0123] 2-2. Preparation of a tablet

[0124] Peptide of the present application 100 mg

[0125] Corn starch 100 mg

[0126] Lactose 100 mg

[0127] Magnesium stearate 2 mg

[0128] The above ingredients are mixed and tableted by compression according to a conventional tablet preparation method.

[0129] 2-3. Preparation of Capsules

[0130] Peptide of the present application 100 mg

[0131] Corn starch 100 mg

[0132] Lactose 100 mg

[0133] Magnesium stearate 2 mg

[0134] The above ingredients are mixed and filled into each gelatin capsule according to a conventional capsule preparation method to prepare capsules.

[0135] 2-4. Preparation of Boluses

[0136] Peptide of the present application 1 g

[0137] Lactose 1.5 g

[0138] Glycerin 1 g

[0139] Xylitol 0.5 g

[0140] The above ingredients are mixed and prepared into boluses in an amount of 4 g per bolus according to a conventional method.

[0141] 2-5. Preparation of Granules

[0142] Peptide of the present application 150 mg

[0143] Soybean extract 50 mg

[0144] Glucose 200 mg

[0145] Starch 600 mg

[0146] The above ingredients are mixed and 100 mg of 30% ethanol is added thereto, and the mixture is dried at 60°C to form granules, which are then filled into a bag.

[0147] Preparation Example 3: Preparation of a cosmetic composition

[0148] 3-1. Preparation of a cream

[0149] Peptide of the present application: 4.6 parts by weight

[0150] Cetyl stearyl alcohol: 2.8 parts by weight

[0151] Beeswax: 2.6 parts by weight

[0152] Stearic acid: 1.4 parts by weight

[0153] Lipophilic glyceryl monostearate: 2 parts by weight

[0154] PEG-100 stearate: 1 part by weight

[0155] Sorbitan sesquioleate: 1.4 parts by weight

[0156] Jojoba oil: 4 parts by weight

[0157] Squalene: 3.8 parts by weight

[0158] Polysorbate 60: 1.1 parts by weight

[0159] Macadamia nut oil: 2 parts by weight

[0160] Tocopheryl acetate: 0.2 parts by weight

[0161] Methyl polysiloxane: 0.4 parts by weight

[0162] Ethyl parahydroxybenzoate: 0.1 parts by weight

[0163] Propyl parahydroxybenzoate: 0.1 parts by weight

[0164] Euxyl K-400: 0.1 parts by weight

[0165] 1,3-Butylene glycol: 7 parts by weight

[0166] Methyl parahydroxybenzoate: 0.05 parts by weight

[0167] Glycerin: 6 parts by weight

[0168] d-Panthenol: 0.2 parts by weight

[0169] Triethanolamine: 0.2 parts by weight

[0170] pt 41891: 0.2 parts by weight

[0171] p-H2O: 46.05 parts by weight

[0172] 3-2. Preparation of the wash

[0173] Peptide of the invention: 3.5 parts by weight

[0174] Cetyl stearyl alcohol: 1.6 parts by weight

[0175] Stearic acid: 1.4 parts by weight

[0176] Lipophilic glyceryl monostearate: 1.8 parts by weight

[0177] PEG-100 stearate: 2.6 parts by weight

[0178] Sorbitol sesquioleate: 0.6 parts by weight

[0179] Squalene: 4.8 parts by weight

[0180] Macadamia nut oil: 2 parts by weight

[0181] Jojoba oil: 2 parts by weight

[0182] Tocopheryl acetate: 0.4 parts by weight

[0183] Methyl polysiloxane: 0.2 parts by weight

[0184] Ethyl paraben: 0.1 parts by weight

[0185] Propyl paraben: 0.1 parts by weight

[0186] 1,3-Butylene glycol: 4 parts by weight

[0187] Methyl paraben: 0.1 parts by weight

[0188] Xanthan gum: 0.1 parts by weight

[0189] Glycerin: 4 parts by weight

[0190] d-Panthenol: 0.15 parts by weight

[0191] Allantoin: 0.1 parts by weight

[0192] Calcium carbonate (2% aq. Sol): 4 parts by weight

[0193] Triethanolamine: 0.15 parts by weight

[0194] Ethanol: 3 parts by weight

[0195] pt 41891: 0.1 parts by weight

[0196] p-H2O: 48.3 parts by weight

[0197] 3-3. Preparation of a skin softening lotion

[0198] Peptide of the invention 0.2 wt%

[0199] Ethanol 10.0 wt%

[0200] Polyoxyethylene sorbitan polylaurate 1.0 wt%

[0201] Methyl paraben 0.2 wt%

[0202] Glycerin 5.0 wt%

[0203] 1,3-Butylene glycol 6.0 wt%

[0204] Fragrance q.s.

[0205] Pigment q.s.

[0206] Purified water q.s.

[0207] Total 100

[0208] 3-4. Preparation of a nutrient toner

[0209] Peptide of the present invention 0.1 wt%

[0210] Petrolatum 2.0 wt%

[0211] Sorbitan sesquioleate 0.8 wt%

[0212] Polyoxyethylene oleyl ether 1.2 wt%

[0213] Methyl paraben q.s.

[0214] Propylene glycol 5.0 wt%

[0215] Ethanol 3.2 wt%

[0216] Carbopol 18.0 wt%

[0217] Pigment q.s.

[0218] Fragrance q.s.

[0219] Purified water q.s.

[0220] Total 100

[0221] 3-5. Preparation of a serum

[0222] Peptide of the present invention 5.0 wt%

[0223] Propylene glycol 10.0 wt%

[0224] Glycerin 10.0 wt%

[0225] Sodium hyaluronate in water (1%) 5.0 wt%

[0226] Ethanol 3.2 wt%

[0227] Polyoxyethylene hydrogenated castor oil 1.0 wt%

[0228] Methyl paraben 0.1 wt%

[0229] Fragrance q.s.

[0230] Purified water q.s.

[0231] Total 100

[0232] 3-6. Preparation of the mask pack

[0233] Peptide of the present application 0.5 wt%

[0234] Glycerin 5.0 wt%

[0235] Propylene glycol 4.0 wt%

[0236] Polyvinyl alcohol 15.0 wt%

[0237] Ethanol 8.0 wt%

[0238] Polyoxyethylene oleyl ether 1.0 wt%

[0239] Methyl paraben 0.2 wt%

[0240] Fragrance q.s.

[0241] Pigment q.s.

[0242] Purified water q.s.

[0243] Total 100

[0244] In the preferred embodiment, the above-mentioned composition of proportions is a mixture of suitable ingredients, but the ingredients or the mixing proportions thereof can be arbitrarily modified according to regional or national preferences, such as demand segments, country demands, and intended uses.

[0245] In the above, representative embodiments of the present application have been described by way of exemplary embodiments; however, the scope of the present application is not limited to the above-described specific embodiments, and those skilled in the art can make appropriate modifications within the scope of the present application as claimed.

Claims

1. A peptide consisting of the amino acid sequence of SEQ ID NO:

1.

2. A composition for promoting hair growth, or preventing or ameliorating hair loss, the composition comprising the peptide of claim 1 as an active ingredient.

3. A pharmaceutical composition for preventing or treating hair loss, the pharmaceutical composition comprising the peptide of claim 1 as an active ingredient.

4. The pharmaceutical composition of claim 3, wherein, The pharmaceutical composition is a preparation for external use on the skin.

5. Use of the peptide according to claim 1 in the manufacture of a pharmaceutical composition for the prevention or treatment of alopecia, wherein, The peptide promotes proliferation of hair follicle dermal papilla cells or outer root sheath cells of hair.

6. Use according to claim 5, wherein, In the hair follicle dermal papilla cells, the peptide: (i) upregulates gene expression of one or more factors selected from the group consisting of LEF-1 and cyclin D1, which are downstream factors of β-catenin; or (ii) upregulates expression of one or more growth factors selected from the group consisting of bFGF, HGF, and VEGF.

7. A cosmetic composition for promoting hair growth, or preventing or ameliorating hair loss, the cosmetic composition comprising the peptide of claim 1 as an active ingredient.

8. The cosmetic composition according to claim 7, wherein, The cosmetic composition is a preparation for external use on the skin.

9. The cosmetic composition according to claim 7, wherein, The cosmetic composition is one or more dosage forms selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, powders, surfactant-containing cleansers, oils, and sprays.

10. The cosmetic composition according to claim 7, wherein, The cosmetic composition is one or more dosage forms selected from the group consisting of lotions, soaps, powdery foundations, emulsion-type foundations, and waxy foundations.

11. The cosmetic composition according to claim 7, wherein, The cosmetic composition is one or more preparations selected from the group consisting of hair tonics, hair creams, shampoos, hair dyes, spray-type hair sprays, aerosol-type hair sprays, hair waxes, and gels.

12. The cosmetic composition of claim 7, wherein, The cosmetic composition is one or more preparations selected from the group consisting of hair conditioners and shampoos.

13. Use of the peptide of claim 1 in the manufacture of a cosmetic composition for promoting hair growth, or preventing or ameliorating hair loss, wherein, The peptide promotes proliferation of hair follicle dermal papilla cells or outer root sheath cells of hair.

14. Use according to claim 13, wherein, In the hair follicle dermal papilla cells, the peptide: (i) upregulates gene expression of one or more factors selected from the group consisting of LEF-1 and cyclin D1, which are downstream factors of β-catenin; or (ii) upregulates expression of one or more growth factors selected from the group consisting of bFGF, HGF, and VEGF. ​

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