Peptides having hair growth promoting or hair loss suppressing properties and their uses

Abstract

The present invention relates to a peptide having hair growth promoting activity and its uses, and provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1, a cosmetic composition for promoting hair growth and improving hair loss containing the peptide, and a pharmaceutical composition for preventing or treating hair loss containing the peptide.

Description

Technical Field

[0001] The present invention relates to a peptide having a hair growth promoting or hair loss suppressing ability and uses thereof.

Background Art

[0002] As society ages, the number of people troubled by hair loss is increasing. Conventionally, it was mainly a problem for middle-aged men, but recently, there has been an increasing interest in hair loss prevention and hair nourishment among the younger generation and women. On the other hand, hair loss has been recognized as a series of aging phenomena, but recently it has become clear that hair loss is promoted by various causes such as stress, Westernized eating habits, nutritional imbalance, and changes in social activities, along with genetic factors.

[0003] Currently, drugs approved by the US FDA that exhibit a hair growth effect include minoxidil (6-Amino-1,2-dihydro-1-hydroxy-2-imino-4-phenoxypyrimidine) (US Patent No. 3,3,82,247) and finasteride (US Patent No. 5,215,894). In the case of minoxidil, it was developed as a vasodilator for the treatment of hypertension in the early 1970s. Although hirsutism was reported as a side effect, it has been used as a hair growth promoter, resulting in thickened pores and increased hair diameter, thus showing the effect of making hair thicker. In the case of finasteride, it was developed as a therapeutic agent for benign prostatic hyperplasia, but currently it is used as a hair loss therapeutic agent, delaying the progression of hair loss and showing a hair growth effect.

[0004] However, in the case of minoxidil, side effects such as weight gain, edema, dermatitis, and increased heart rate have been reported. In the case of finasteride, continuous administration is required, and in men, there are side effects such as sexual dysfunction, and in pregnant women, there are side effects such as the birth of deformed infants. Therefore, the development of a hair loss therapeutic agent without side effects is still required.

[0005] Against this technological backdrop, multifaceted research is underway to improve hair loss through mechanisms such as hormone regulation and metabolic regulation (Korean Patent Publication No. 2002-0005332), but the situation is still inadequate. [Overview of the Initiative] [Problems that the invention aims to solve]

[0006] One embodiment provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0007] Another embodiment provides a cosmetic composition for improving hair loss or promoting hair growth, comprising the peptide.

[0008] Another embodiment provides a pharmaceutical composition for preventing or treating hair loss, comprising the peptide.

[0009] Other objects and advantages of the present invention will become even clearer from the following detailed description, along with the appended claims and drawings. Any matters not described herein are readily apparent and inferable to those skilled in the art of the present invention or similar art, and are therefore omitted from this description. [Means for solving the problem]

[0010] Each description and embodiment disclosed in this invention may also apply to other descriptions and embodiments. That is, all combinations of the various elements disclosed in this invention fall within the scope of this invention. Furthermore, the scope of this invention is not considered to be limited by the following specific descriptions.

[0011] One embodiment provides a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0012] As used herein, the term "peptide" means a linear molecule formed by the bonding of amino acid residues to one another via peptide bonds. Such peptides can be produced by chemical synthesis methods known in the art, particularly by solid-phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85:2149-54 (1963)) or liquid-phase synthesis techniques (US Patent No. 5,516,891).

[0013] The inventors, in their efforts to develop a peptide with biologically effective activity, have identified a peptide consisting of the amino acid sequence of SEQ ID NO: 1. More specifically, the biologically effective activity exhibits one or more of the following characteristics selected from: (a) promotion of human hair follicle dermal papilla cell proliferation, (b) promotion of human hair follicle outer root sheath cell proliferation, (c) promotion of human hair follicle germinal matrix cell proliferation, (d) activation of β-catenin, e.g., increased expression of β-catenin and / or migration of β-catenin from the cytoplasm to the nucleus, (e) increased expression of Lef-1 (Lymphoid enhancer-binding factor-1), Cyclin-D1, or C-Myc, (f) promotion of differentiation of human hair follicle stem cell, and (g) suppression of DKK-1 (Dickkopf-related protein 1) expression.

[0014] Therefore, the peptide can be used for purposes such as promoting hair growth or improving hair loss, preventing or treating hair loss, and improving damaged hair.

[0015] The peptide is fitted with a protecting group at its N or C terminus to acquire chemical stability, enhanced pharmacological properties (half-life, absorption, potency, efficacy, etc.), altered specificity (e.g., broad biological activity spectrum), and reduced antigenicity. Therefore, the peptide can be used as encompassing the peptide itself or its modified derivatives. In one specific example, the N-terminus of the peptide may be bound to any one protecting group selected from the group consisting of acetyl, fluorenylmethoxycarbonyl, formyl, palmitoyl, myristyl, stearyl, butoxycarbonyl, allyloxycarbonyl, and polyethylene glycol (PEG), and / or the C-terminus of the peptide may be bound to any one protecting group selected from the group consisting of amino (-NH2), tertiary alkyl, and azide (-NHNH2). The peptide may also selectively further include targeted sequences, tags, labeled residues, and amino acid sequences manufactured for specific purposes to increase half-life or peptide stability.

[0016] The peptides are artificially synthesized, non-naturally occurring, or engineered, where “non-naturally occurring or engineered” means a state produced by artificial modification rather than being in its natural state. Here, such artificial modification may include artificially synthesizing an amino acid sequence by mimicking a plurality of amino acid structures, or, as described above, being engineered to acquire chemical stability, enhanced pharmacological properties, altered specificity, or reduced antigenicity.

[0017] As used herein, the term "stability" refers not only to in vivo stability, which protects the peptide from attack by endogenous protein-cleaving enzymes, but also to storage stability (e.g., room temperature storage stability).

[0018] Another embodiment provides a cosmetic composition for promoting hair growth or improving hair loss, comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

[0019] Any terms or elements mentioned in the description of the aforementioned peptide that are the same as those already mentioned are as previously stated.

[0020] As used herein, the term "improvement" means all actions that reduce, for example, the degree of symptoms, which are parameters related to the alleviation or treatment of a condition.

[0021] As used herein, the term "hair loss improvement" comprehensively refers to the process or effect of treating, reducing, or alleviating hair loss, and includes all actions that suppress the progression of hair loss, such as promoting the proliferation or activation of hair follicle dermal papilla cells, or suppressing the expression of hair loss inducing factors such as DKK-1. The hair follicle dermal papilla cells are cells that make up the hair follicle, which affects the thickness and growth of hair, and the activation of these cells affects the hair cycle, and through this mechanism, a hair loss improvement effect can be induced.

[0022] As used herein, the term "hair growth promotion" refers to all actions that increase hair growth from hair follicles, including all actions that increase the total amount of hair, such as the proliferation of hair follicle cells, the activation of hair follicle cells, or the effect of promoting the growth of hair follicle cells. Specifically, the hair cycle can be divided into the growth phase (anagen), when hair grows; the degeneration phase (catagen), when growth ends and the hair follicle opening shrinks; the resting phase (telogen), when the dermal papilla ceases activity and the hair remains in the scalp; and the active phase, when the dermal papilla becomes active or generates new hair and causes old hair to fall out. In the hair cycle, dermal papilla cells are connected to capillaries and sensory nerves and supply oxygen and nutrients to matrix cells, which surround the dermal papilla cells and determine the rate of the growth phase through continuous cell division and proliferation. Furthermore, the outer root sheath cells are located in the part of the epidermis that is in contact with the basal layer, and they play a role in protecting the hair until keratinization is completed.

[0023] Conventional functional peptides, despite their effective biological activity, have the disadvantage of not being effectively absorbed into target tissues or cells due to the size of the peptide itself, or being eliminated from the body in a short period due to their short half-life. On the other hand, the cosmetic composition according to one example contains a peptide consisting of 10 or fewer amino acids as an active ingredient, resulting in excellent skin penetration of the active ingredient. For example, when applied topically to the skin, it can promote hair growth and effectively suppress hair loss.

[0024] According to one example, the peptide demonstrated the following effects: promotion of proliferation or activation of hair follicle dermal papilla cells, promotion of proliferation or activation of extrafollicle root sheath cells, promotion of proliferation or activation of hair follicle matrix cells, activation of β-catenin, increased expression of Lef-1, Cyclin-D1, or C-Myc, promotion of differentiation of hair follicle stem cells, and suppression of the expression of hair loss inducing factors such as DKK-1. Therefore, the peptide can be used as an active ingredient in cosmetic compositions for promoting hair growth and improving hair loss.

[0025] The cosmetic composition contains a cosmetically effective amount of a peptide and / or a cosmetically acceptable carrier, but is not limited thereto.

[0026] As used herein, the term "cosmetically effective amount" means an amount sufficient to achieve the hair removal improvement or hair growth promoting efficacy of the cosmetic composition.

[0027] The weight ratio between the peptide and the cosmetically acceptable carrier is, for example, from 500:1 to 1:500. As an example, the weight ratio is from 450:1 to 1:450, from 400:1 to 1:400, from 350:1 to 1:350, from 300:1 to 1:300, from 250:1 to 1:250, from 200:1 to 1:200, from 150:1 to 1:150, from 100 :1 to 1:100, from 80:1 to 1:80, from 60:1 to 1:60, from 40:1 to 1:40, from 20:1 to 1:20, from 10:1 to 1:10, from 8:1 to 1:8, from 6:1 to 1:6, from 4:1 to 1:4, or from 2:1 to 1:2, but is not limited thereto.

[0028] The cosmetic composition is manufactured in any dosage form commonly manufactured in the industry, for example, it is formulated into solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansings, oils, powder foundations, emulsion foundations, wax foundations, and sprays, etc., but is not limited thereto. For example, it can be manufactured into the dosage forms of softening lotions, nourishing lotions, nourishing creams, massage creams, essences, eye creams, cleansing creams, cleansing foams, cleansing waters, packs, sprays or powders.

[0029] When the dosage form of the cosmetic composition is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, tragacanths, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talcs or zinc oxides, etc. can be used as carrier components.

[0030] When the dosage form of the cosmetic composition is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier component, and in the case of a spray, for example, it may additionally contain propellants such as chromolofluorohydrocapone, propane / butane, or dimethyl ether.

[0031] When the dosage form of the cosmetic composition is a solution or emulsion, a solvent, solubilizer, or emulsifier may be used as a carrier component, and may include, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or fatty acid ester of sorbitan.

[0032] When the dosage form of the cosmetic composition is a suspension, the carrier component may be a liquid diluent such as water, ethanol, or propylene glycol; a suspension agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester; or microcrystalline cellulose, aluminum methhydroxyl, bentonite, aga, or tracant.

[0033] When the dosage form of the cosmetic composition is a surfactant-containing cleanser, the carrier component may be an aliphatic alcohol sulfate, an aliphatic alcohol sulfate ether, a sulfosuccinate monoester, isethionate salt, imidazolinium derivative, methyl taurate, sarcosinate salt, fatty acid amide ether sulfate, alkylamide betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester.

[0034] The peptide may be contained in nanosomes or nanoparticles to further improve skin penetration or stability issues. For example, the nanosomes may be produced in a microfluidizer using lecithin as a raw material and contained within lecithin particles. Any known method for producing the nanosomes can be used. The size of the nanosome particles is preferably 30 to 200 nm. If the size of the nanosome particles is less than 30 nm, skin penetration proceeds very quickly, causing skin side effects, and if it exceeds 200 nm, skin penetration is not easy, making it difficult to obtain the benefits of using the nanosome structure.

[0035] The components contained in the cosmetic composition include, in addition to the peptide and carrier components as active ingredients, components commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0036] The content of peptides as active ingredients in the cosmetic composition can be appropriately and non-restrictively selected depending on the shape of the product, the desired use, etc., and can be added in amounts of 0.01 to 15% by weight of the total weight of the cosmetic composition. Alternatively, for example, the cosmetic composition may contain peptides in amounts of 1.0% to 3.0% by weight, preferably 2.0% to 3.0% by weight, based on the total weight.

[0037] Another embodiment provides a method for improving hair loss or promoting hair growth, which includes the step of applying a cosmetic composition containing a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient to the skin of an individual.

[0038] The terms or elements mentioned in the description of the cosmetic composition that are the same as those already mentioned are as described above.

[0039] In this specification, the terms “apply,” “administer,” and “coat” are used interchangeably and may mean bringing a composition according to one embodiment to a desired site at least partially, or placing a composition according to one embodiment into an individual via an administration route.

[0040] Another embodiment provides a pharmaceutical composition for preventing or treating hair loss, comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0041] Any terms or elements mentioned in the description of the aforementioned peptide that are the same as those already mentioned are as previously stated.

[0042] In this specification, the term "prevention" means all actions that suppress or delay the onset of a disease by administering the composition.

[0043] In this specification, the term “treatment” means any form of treatment that provides an effect to an individual suffering from or at risk of developing a disease, including improvement of the individual’s condition (e.g., one or more symptoms), delay of disease progression, delay of symptom onset, or slowing of symptom progression. Accordingly, “treatment” and “prevention” are not intended to mean a cure or complete elimination of symptoms.

[0044] The term "individual" refers to an entity requiring treatment for a disease, and more specifically, to mammals such as humans or non-human primates, mice, dogs, cats, horses, and cattle.

[0045] "Alopecia," a disease targeted for prevention or treatment by pharmaceutical compositions, refers to the absence of hair in areas where hair should normally be present, and specifically, it can mean the shedding of mature hair (thick, dark hair) on the scalp. The causes of alopecia are not limited to genetic factors, but can be triggered by a variety of dietary habits and environmental influences such as exposure to air pollution and consumption of processed foods. For example, alopecia can include hereditary androgenic alopecia (baldness), alopecia areata, tinea capitis due to fungal infection, telogen effluvium, trichotillomania, and hair growth disorders. Scarring alopecia, which leaves a residue, refers to alopecia caused by lupus, bald folliculitis, lichen planus pilaris, burns, and trauma.

[0046] According to one example, the peptide promoted the proliferation or activation of hair follicle dermal papilla cells, hair follicle root sheath cells, hair follicle matrix cells, activated β-catenin, increased expression of Lef-1, Cyclin-D1, or C-Myc, promoted differentiation of hair follicle stem cells, and suppressed the expression of hair loss inducing factors such as DKK-1. Therefore, the peptide can be used as an active ingredient in pharmaceutical compositions for the prevention or treatment of hair loss.

[0047] The pharmaceutical composition comprises, but is not limited to, a pharmaceutical effective amount of the peptide and / or a pharmaceutically acceptable carrier.

[0048] In this specification, the term "pharmaceutical effective amount" means an amount sufficient to achieve the efficacy of the pharmaceutical composition in preventing or treating hair loss.

[0049] The pharmaceutically acceptable carriers mentioned above are those commonly used in formulation and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0050] The weight ratio between the peptide and the pharmaceutically acceptable carrier is, for example, 500:1 to 1:500, and is not limited to, but may be 450:1 to 1:450, 400:1 to 1:400, 350:1 to 1:350, 300:1 to 1:300, 250:1 to 1:250, 200:1 to 1:200, 150:1 to 1:150, 100:1 to 1:100, 80:1 to 1:80, 60:1 to 1:60, 40:1 to 1:40, 20:1 to 1:20, 10:1 to 1:10, 8:1 to 1:8, 6:1 to 1:6, 4:1 to 1:4, or 2:1 to 1:2.

[0051] The aforementioned pharmaceutical composition may further include, but is not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspension agents, preservatives, and the like, in addition to the aforementioned components.

[0052] The pharmaceutical composition described above is administered orally or parenterally, preferably parenterally, and in the case of parenteral administration, by methods such as intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, local administration, or transdermal administration, but is not limited to these methods.

[0053] The dosage of the pharmaceutical composition is 0.0001 to 1000 μg (micrograms), 0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, or 1.0 to 1000 μg per day, but is not limited to these, and can be prescribed in various ways depending on factors such as the formulation method, administration method, patient's age, weight, sex, medical condition, diet, administration time, route of administration, excretion rate, and response sensitivity.

[0054] The pharmaceutical composition may be manufactured in unit dose form or encapsulated in a multi-dose container by formulation using pharmaceutically acceptable carriers and / or excipients, by a method readily implemented by a person with ordinary skill in the art to which the invention pertains.

[0055] The dosage form may be in the form of an oil or aqueous medium, such as a solution, suspension or emulsion, and may be in the form of an extract, powder, granules, tablet or capsule, and may further contain a dispersant and / or stabilizer.

[0056] Another embodiment provides a method for preventing or treating hair loss, comprising the step of administering to an individual a pharmaceutical composition containing a therapeutically effective amount of a peptide comprising the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0057] As stated above, any terms or elements mentioned in the description of the aforementioned pharmaceutical composition that are the same as those already mentioned are as described previously.

[0058] Another embodiment provides a food composition for preventing hair loss or promoting hair growth, comprising a peptide consisting of the amino acid sequence of SEQ ID NO: 1 as an active ingredient.

[0059] As previously stated, any terms or elements mentioned in the description of the peptides that are identical to those already mentioned are as described above.

[0060] The content of the peptide as an active ingredient contained in the food composition can be appropriately and non-restrictively selected depending on the shape of the food, the desired use, etc., and can be added in an amount of 0.01 to 15% by weight of the total food weight. Also, for example, in a health drink composition, it can be added in a ratio of 0.02 to 10 g, preferably 0.3 to 1 g, based on 100 ml. [Effects of the Invention]

[0061] According to one embodiment of the peptide, it can be applied to promoting hair growth by (a) promoting the proliferation of human hair follicle dermal papilla cells (HHFDPC), (b) promoting the proliferation of human hair follicle outer root sheath cells (HHFORSC), (c) promoting the proliferation of human hair follicle germinal matrix cells (HHFGMC), (d) activating β-catenin, (e) increasing the expression of Lef-1, Cyclin-D1, or C-Myc, or (f) promoting the differentiation of human hair follicle stem cells (HHFSC), thereby promoting the proliferation or activity of hair follicle cells or promoting the expression of hair follicle stem cells.

[0062] Peptides in other embodiments exhibit the ability to suppress the expression of hair loss factors and can be applied to hair loss suppression and other related applications. [Brief explanation of the drawing]

[0063] [Figure 1] This is the result of measuring the proliferation-promoting effect of a peptide consisting of the amino acid sequence of Sequence ID No. 1 on human dermal papilla cells. [Figure 2] This is the result of measuring the proliferation-promoting effect of a peptide consisting of the amino acid sequence of Sequence ID No. 1 on human hair follicle extraroot sheath cells. [Figure 3] This is the result of measuring the proliferation-promoting effect of a peptide consisting of the amino acid sequence of Sequence ID No. 1 on human hair follicle matrix cells. [Figure 4] This is the result of measuring the effect of promoting the growth mechanism of hair follicle cells of the peptide consisting of the amino acid sequence of SEQ ID NO: 1. [Figure 5] This is the result of confirming that the expression of Lef-1, Cyclin-D1, and C-Myc, which are factors related to the activity and growth of hair follicle cells of the peptide consisting of the amino acid sequence of SEQ ID NO: 1, is increased. [Figure 6] This is the result of confirming that the expression of the stem cell marker decreases due to the induction of differentiation of the peptide consisting of the amino acid sequence of SEQ ID NO: 1. [Figure 7] This is the result of confirming that the expression of the differentiation marker increases due to the induction of differentiation of the peptide consisting of the amino acid sequence of SEQ ID NO: 1. [Figure 8] This is the result of measuring the effect of suppressing the expression of hair loss-related factors in hair follicle cells of the peptide consisting of the amino acid sequence of SEQ ID NO: 1. [Figure 9] This is the result of a comparative measurement of the effect of promoting the hair follicle cell growth mechanism of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 and the peptide consisting of the amino acid sequence of SEQ ID NO: 26. [Figure 10] This is the result of measuring the expression of the stem cell marker after treatment with the peptide consisting of the amino acid sequence of SEQ ID NO: 1 and the peptide consisting of the amino acid sequence of SEQ ID NO: 26. [Figure 11] This is the result of measuring the expression of the differentiation marker after treatment with the peptide consisting of the amino acid sequence of SEQ ID NO: 1 and the peptide consisting of the amino acid sequence of SEQ ID NO: 26. [Figure 12] This is the result of measuring the effect of suppressing the expression of hair loss-related factors after treatment with the peptide consisting of the amino acid sequence of SEQ ID NO: 1 and the peptide consisting of the amino acid sequence of SEQ ID NO: 26.

Mode for Carrying Out the Invention

[0064] Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes of the present invention, and the scope of the present invention is not limited to these examples.

[0065] Example 1: Peptide synthesis Using an automatic peptide synthesizer (Milligen 9050, Millipore, USA), a peptide having the amino acid sequence of SEQ ID NO: 1 described in Table 1 below was synthesized, and these synthesized peptides were purified by C18 reverse-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). The column used was ACQUITY UPLC BEH300 C18 (2.1 mm × 100 mm 1.7 μM, Waters Co, USA).

[0066]

Table 1

[0067] Example 2: Confirmation of the hair follicle cell proliferation effect We attempted to confirm the effect of promoting the proliferation of hair follicle cells of the peptide consisting of the amino acid sequence of SEQ ID NO: 1. Specifically, each of human hair follicle dermal papilla cells (HHFDPC), human hair follicle outer root sheath cells (HHFORSC), and human hair follicle hair matrix cells (HHFGMC) was seeded in a 96-well plate at a density of 4×10 3 cells / well and cultured for 24 hours. After changing to serum-free mesenchymal stem cell medium, it was cultured for 24 hours. The peptide was treated at each concentration of 10 μM, 50 μM, or 100 μM and cultured at 37°C for 72 hours. Minoxidil (1 μM), human recombinant Wnt-3a protein (100 ng / ml), and human recombinant EGF protein (100 ng / ml), which are used as hair loss treatment agents in the positive control group, were treated. After the culture was completed, the culture supernatant was removed, and the cells were fixed using ethanol. After washing with PBS, SRB solution was treated to proceed with staining. After washing with 1% acetic acid, the cells were observed under a microscope, and 20 mM tris solution was treated to proceed with de-staining. The absorbance at a wavelength of 560 nm was measured using a spectrophotometer (SpectraMax iD3, Molecular Devices (USA, CA)) to measure the survival state of the cells.

[0068] The results are shown in Figures 1 to 3. Figure 1 shows the proliferation results of human hair follicle dermal papilla cells (HHFDPCs), Figure 2 shows the proliferation results of human hair follicle extraroot sheath cells (HHFORSCs), and Figure 3 shows the proliferation results of human hair follicle matrix cells (HHFGMCs). It can be confirmed that proliferation is induced and increased in a concentration-dependent manner in the peptide-treated groups. In the case of HHFDPCs, a maximum increase of approximately 46% (Figure 1) was observed, in the case of HHFORSCs, a maximum increase of approximately 49% (Figure 2), and in the case of HHFGMCs, a maximum increase of approximately 33% (Figure 3).

[0069] Example 3: Confirmation of the effect of promoting the proliferation mechanism of hair follicle cells We attempted to confirm the effect of the peptide consisting of the amino acid sequence of SEQ ID NO: 1 on promoting the proliferation mechanism of hair follicle cells. Specifically, we used 4 × 10⁻¹⁶ HHFDPC. 5 Cells were seeded into 6-well plates at a cell / well density and cultured for 24 hours. After changing to serum-free mesenchymal stem cell medium, they were cultured for another 24 hours. Peptides were treated at concentrations of 10 μM, 50 μM, or 100 μM, respectively, and cultured at 37°C for 24 hours. The positive control group was treated with recombined human Wnt-3a protein at 10 ng / ml. After PBS washing, nucleoproteins were separated using a nucleoprotein extraction kit (Thermo Scientific, USA), and BCA quantification was performed to prepare the same volume of protein sample. Electrophoresis was carried out using a 10% SDS-PAGE gel, and the separated proteins were transferred to a PVDF membrane. Blocking was performed at room temperature for 30 minutes using 5% skim milk. β-catenin (Cell Signaling, USA) and HDAC1 (Santa Cruz, USA) antibodies were diluted 1:1000 in 3% BSA and reacted with the membrane at 4°C for 16 hours. The membrane was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS). The secondary antibody was diluted 1:2000 in 5% skim milk and reacted at room temperature for 1 hour. The membrane was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS). After treatment with ECL solution (GE Healthcare, USA), detection was performed on the membrane.

[0070] The results are shown in Figure 4. From the results in Figure 4, we can confirm that β-catenin expression increases upon peptide treatment, and that peptide treatment increases the nuclear translocation of β-catenin from the cytoplasm to the nucleus by up to 5.34 times.

[0071] Example 4: Confirmation of the effect of promoting the activation of hair follicle cells We attempted to confirm the effect of a peptide consisting of the amino acid sequence of Sequence ID No. 1 on promoting the activity of hair follicle cells. HHFDPC was measured in 4 × 10⁶ units. 5 Cells were seeded into 6-well plates at a cell / well density and cultured for 24 hours. After changing to serum-free mesenchymal stem cell medium, they were cultured for another 24 hours. Peptides were treated at concentrations of 10 μM, 50 μM, or 100 μM, respectively, and cultured at 37°C for 24 hours. The positive control group was treated with 10 ng / ml of recombined human Wnt-3a protein. After PBS washing, RNA was isolated by treatment with 300 μL of easy blue (intron, Korea). After RNA quantification, cDNA synthesis was carried out using a cDNA synthesis kit (enzynomics, Korea). PCR was carried out using the PCR pre-mix (enzynomics, Korea) with Lef-1, Cyclin-D1, and C-Myc primers shown in Table 2. Electrophoresis was performed on 1.5% agaroth gel, and band detection was performed using the Bio-Rad gel image system.

[0072] The results are shown in Figure 5. From the results in Figure 5, it was confirmed that peptide treatment increased the expression of Lef-1, C-Myc, and Cyclin-D1, which are factors associated with the activity and growth of HHFDPC cells. In the case of Lef-1, the expression increased by up to 1.53 times compared to the control group, in the case of Cyclin-D1, it increased by up to 6.13 times, and in the case of C-Myc, it increased by up to 4.86 times.

[0073] [Table 2]

[0074] Example 5: Confirmation of the effect of inducing differentiation of stem cells We attempted to confirm the differentiation-inducing effect of a peptide consisting of the amino acid sequence of SEQ ID NO: 1 on stem cells. HHFSC was divided into 1 × 10⁶ cells. 5 Cells were seeded into 6-well plates at a cell / well density and cultured for 24 hours. After changing to serum-free mesenchymal stem cell medium, they were cultured for another 24 hours. Peptides were treated at concentrations of 10 μM, 50 μM, or 100 μM, respectively, and cultured at 37°C for 72 hours. The positive control group was treated with recombined human Wnt-3a protein at concentrations of 10 ng / ml and 100 ng / ml, respectively. After PBS washing, RNA was separated by treatment with 300 μL of easy blue (intron, Korea). After RNA quantification, cDNA synthesis was carried out using a cDNA synthesis kit (enzynomics, Korea). PCR was carried out using the primers shown in Table 3 and the PCR pre-mix (enzynomics, Korea). Electrophoresis was performed on 1.5% agaroth gel, and band detection was performed using the Bio-Rad gel image system.

[0075] [Table 3]

[0076] Figure 6 shows the results of measuring stem cell markers, and Figure 7 shows the results of measuring differentiation markers. From the results in Figure 6, it can be confirmed that the expression of the stem cell markers CD34, Lhx-2, and Keratin15 is all significantly decreased. On the other hand, from the results in Figure 7, it can be confirmed that the expression of the differentiation markers α-SMA, Corin, Versican, Vimentin, and Nexin-1 is all significantly increased. From this, it was confirmed that the peptide having the amino acid sequence of Sequence ID No. 1 induces the proliferation of stem cells, induces differentiation to go to hair follicles, and generates new hair follicles.

[0077] Example 6: Confirmation of the effect of suppressing the expression of hair loss-related factors in hair follicle cells We attempted to determine whether the expression of DKK-1, a hair loss-related factor, is suppressed in hair follicle cells by using a peptide consisting of the amino acid sequence of SEQ ID NO: 1. HHFDPC was used in 4 × 10⁶ samples.5 Cells were seeded into 6-well plates at a cell / well density and cultured for 24 hours. After changing to serum-free mesenchymal stem cell medium, they were cultured for another 24 hours. Peptides were treated at concentrations of 10 μM, 50 μM, or 100 μM, respectively, with co-treatment with DHT, a DKK-1 hair loss protein inducer, at a concentration of 100 nM. A positive control group was treated with 5 μM of finasteride, a drug that suppresses male hormones. DHT is known to be a hair loss hormone that increases the expression of DKK-1 (Dickkopf-related protein 1), a hair loss inducer, by activating androgen receptors. After culturing at 37°C for 24 hours, cells were washed with PBS, treated with lysis buffer to prepare cell lysates, and BCA quantification was performed to prepare the same volume of protein samples. Electrophoresis was carried out using a 10% SDS-PAGE gel. Proteins separated via SDS-PAGE were transferred to a PVDF membrane. Blocking was performed at room temperature for 30 minutes using 5% skim milk. The DKK-1 (cell signaling, USA) antibody was diluted 1:1000 with 3% BSA and reacted with the membrane at 4°C for 16 hours. The membrane was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS). The secondary antibody was diluted 1:2000 with 5% skim milk and reacted at room temperature for 1 hour. The membrane was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS). After treatment with ECL solution (GE Healthcare, USA), protein band detection and densitometry were performed using an ImageQuant 800 (Amersham, USA).

[0078] The results are shown in Figure 8. From the results in Figure 8, we confirmed the inhibitory effect of peptide treatment on DHT-induced DKK-1 expression. In the high-concentration peptide treatment group, we confirmed that the DKK-1 expression level decreased by approximately 50% compared to the negative control group (DHT-treated group).

[0079] Example 7: Comparison of the effect of promoting the proliferation mechanism of hair follicle cells The effects of a novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 and a peptide consisting of the amino acid sequence of SEQ ID NO: 26 disclosed in KR10-1209117 (Table 4) on promoting the proliferation mechanism of hair follicle cells were compared. In the experiment, as described in Example 3, HHFDPC was treated with the peptide at different concentrations, and then the cell proliferation effect was measured.

[0080] [Table 4]

[0081] The results are shown in Figure 9. From the results in Figure 9, it can be confirmed that both the peptide consisting of the amino acid sequence of SEQ ID NO: 26 and the novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 promote the translocation of β-catenin from the cytoplasm to the nucleus in a concentration-dependent manner. Furthermore, when comparing the activities of the two peptides, it can be confirmed that the novel peptide exhibits significantly higher proliferation mechanism promoting activity than the known peptide at the same concentration.

[0082] Example 8: Comparison of the differentiation-inducing effect of stem cells The differentiation-inducing effects of a novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 and a peptide consisting of the amino acid sequence of SEQ ID NO: 26 on stem cell differentiation were compared. The experiment was conducted as described in Example 5, where HHFSCs were treated with the peptides at different concentrations, and then the differentiation-inducing effect was confirmed.

[0083] The results are shown in Figures 10 and 11. Figure 10 shows the results of measuring stem cell markers, and Figure 11 shows the results of measuring differentiation markers. From the results in Figure 10, it can be confirmed that the expression of the stem cell markers CD34, Lhx-2, and Keratin15 decreased significantly when treated with the two peptides. Furthermore, when comparing the activity of the two peptides, it can be confirmed that the novel peptide significantly reduced the expression of stem cell markers compared to the known peptide when treated at the same concentration. In addition, from the results in Figure 11, it can be confirmed that the expression of the differentiation markers α-SMA, Corin, Versican, Vimentin, and Nexin-1 all increased significantly when treated with the two peptides. Furthermore, when comparing the activity of the two peptides, it can be confirmed that the novel peptide significantly increased the expression of differentiation markers compared to the known peptide when treated at the same concentration.Therefore, it can be confirmed that the novel peptide induces the proliferation of stem cells, induces differentiation to go to hair follicles, and generates new hair follicles compared to the known peptide.

[0084] Example 9: Comparison of the effect of suppressing the expression of hair loss-related factors in hair follicle cells The inhibitory effects of a novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 and a peptide consisting of the amino acid sequence of SEQ ID NO: 26 on the expression of hair loss-related factors were compared. In the experiment, as described in Example 6, HHFDPC was treated with the peptides at different concentrations, and then it was confirmed whether or not the expression of DKK-1, a hair loss-related factor, was suppressed.

[0085] The results are shown in Figure 12. From the results in Figure 12, it can be confirmed that both the peptide consisting of the amino acid sequence of SEQ ID NO: 26 and the novel peptide consisting of the amino acid sequence of SEQ ID NO: 1 exhibit a concentration-dependent inhibitory effect on DKK-1 expression. Furthermore, when comparing the inhibitory effects of the two peptides, it can be confirmed that the novel peptide exhibits significantly higher DKK-1 expression inhibitory ability than the known peptide at the same concentration.

[0086] Dosage Form Example 1: Production of Peptide Nanosomes 50 mg of the peptide from Example 1 was dissolved in 500 ml of distilled water by thorough stirring. The mixture was then mixed with 5 g of lecithin, 0.3 ml of sodium oleate, 50 ml of ethanol, and a small amount of oil. After adjusting the volume with distilled water to a total of 1 L, the mixture was emulsified using a microfluidizer under high pressure to produce peptide nanosomes with a size of approximately 100 nm.

[0087] Dosage form example 2. Lotion A lotion containing a peptide according to one embodiment and having the following composition was manufactured using a method known in the industry.

[0088] [Table 5]

[0089] Dosage form example 3: Nutritional cream A nutritional cream containing a peptide according to one embodiment and having the following composition was manufactured using a method known in the industry.

[0090] [Table 6]

[0091] The above-mentioned description of the present invention is illustrative, and a person with ordinary skill in the art to which the present invention pertains will understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the present invention. Therefore, the above-mentioned embodiments should be understood to be illustrative in all respects and not limiting.

Claims

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

1.

2. The peptide according to claim 1, wherein the N-terminus of the peptide is bonded to one protecting group selected from the group consisting of an acetyl group, a fluoreonylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an allyloxycarbonyl group, and polyethylene glycol (PEG).

3. The C-terminus of the peptide is an amino group (-NH ２ ), tertiary alkyl group and azide (-NHNH) ２ The peptide according to claim 1, which is bonded to any one protecting group selected from the group consisting of ).

4. The peptide according to claim 1, wherein the peptide exhibits one or more of the following characteristics: (a) Promotion of human hair follicle dermal papilla cell proliferation, (b) Promotion of proliferation of human hair follicle outer root sheath cells, (c) Promotion of proliferation of human hair follicle germinal matrix cells, (d) Activation of β-catenin, (e) Increased expression of Lef-1 (Lymphoid enhancer-binding factor-1), Cyclin-D1, or C-Myc. (f) Promotion of differentiation of human hair follicle stem cells (g) Suppression of DKK-1 (Dickkopf-related protein 1) expression.

5. A cosmetic composition for promoting hair growth or improving hair loss, comprising the peptide described in any one of claims 1 to 4 as an active ingredient.

6. A pharmaceutical composition for preventing or treating hair loss, comprising the peptide described in any one of claims 1 to 4 as an active ingredient.

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