Peptides having hair growth promoting and damaged hair repair activity and their uses

Abstract

The present application relates to peptides having the activity of promoting hair growth and improving damaged hair, and uses thereof, and provides a peptide consisting of an amino acid sequence represented by Arg-Cys-Cys-Gly or Glu-Glu, a cosmetic composition containing the peptide for improving damaged hair, a cosmetic composition containing the peptide for improving hair loss or promoting hair growth, and a pharmaceutical composition containing the peptide for preventing or treating hair loss.

Description

Technical Field

[0001] This application relates to a peptide having hair growth promoting and damaged hair improving activities and its uses.

Background Art

[0002] Human hair consists of about 150,000 hairs, and each hair repeats the processes of growth and hair loss through different hair cycles. The hair growth cycle is roughly divided into three stages: the anagen stage where hair grows, the catagen stage where growth stops and the hair is maintained, and the telogen stage where the hair papilla contracts and the hair follicle shrinks, resulting in hair loss. These three stages are cyclically repeated. The hair growth cycle varies depending on internal factors such as family strength, genetics, constitution, and hormonal regulation, and external factors such as nutritional status, aging, the surrounding environment, and stress.

[0003] On the other hand, the ends of the hair are exposed to the external environment for a longer time than the scalp side part, so hair damage caused by the external environment is more severe. The weathering effect due to exposure to the external environment induces the loss of the hair cuticle, causing the hair tip to branch and eventually break. When high heat is continuously applied to the hair with a hair dryer, etc., the phenomenon of the hair becoming weak with a decrease in hair tension is a typical example of such damage. Also, various physical, chemical, or environmental factors such as frequent perms and dyeing cause the proteins and lipids inside the hair to elute, promoting the porosity of the hair. As a result, the hair becomes limp, loses its luster, has an increased frictional force, making hair loss difficult, and phenomena such as split ends and branched hairs occur. Moreover, such porosity of the hair causes a decrease in the elasticity of the hair, resulting in a lack of firmness and volume in the hair or the problem of thinning hair.

[0004] Furthermore, with the development of an aging society, the number of people suffering from hair loss is increasing. While it was traditionally a major problem for middle-aged men, recently, interest in hair loss prevention and hair care has been increasing among young people and women as well. On the other hand, while hair loss has been recognized as a series of aging processes, it has recently become clear that hair loss is driven by a variety of causes, including genetic factors, stress, Westernized eating habits, nutritional imbalances, and changes in social activities. Currently, drugs that show hair growth effects are minoxidil (6-Amino-1,2-dihydro-1-hydroxy-2-imino-4-phenoxypyrimidine) (US Patent No. 3,382,247) and finasteride (US Patent No. 5,215,894), both approved by the US FDA. In the case of minoxidil, it was developed in the early 1970s as a vasodilator for the treatment of hypertension, but despite reports of hirsutism as a side effect, it has been used as a hair growth stimulant, showing the effect of widening hair follicles and expanding and thickening hair. Furthermore, finasteride, originally developed as a treatment for benign prostatic hyperplasia, is now used as a hair loss treatment, slowing the progression of hair loss and promoting hair growth. However, minoxidil has been reported to cause side effects such as weight gain, edema, dermatitis, and increased heart rate. Finasteride requires continuous use and can cause side effects such as sexual dysfunction in men and birth defects in pregnant women. Therefore, there is still a need for the development of a hair loss treatment without side effects.

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

[0006] One embodiment provides a peptide consisting of an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0007] Another embodiment provides a cosmetic composition for improving damaged hair, comprising as an active ingredient a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0008] Another embodiment provides a cosmetic composition for improving hair loss or promoting hair growth, which contains as an active ingredient a peptide comprising an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0009] Another embodiment provides a pharmaceutical composition for preventing or treating hair loss, comprising as an active ingredient a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0010] Other purposes and advantages of this application will be further clarified by the following detailed description, along with the claims and drawings. Any matters not described herein are readily apparent and inferable to those skilled in the art of this application or similar art, and are therefore omitted from this description. [Means for solving the problem]

[0011] Each description and embodiment disclosed in this application may also apply to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of this application. Furthermore, the scope of this application is not considered to be limited by the specific descriptions described below.

[0012] One embodiment provides a peptide consisting of an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0013] As used herein, the term "peptide" may mean a linear molecule formed by the bonding of amino acid residues to each other via peptide bonds. Such peptides can be produced by chemical synthesis methods known to those skilled in the art, particularly by 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, 111(1984)) or liquid synthesis techniques (US Patent No. 5,516,891). The inventors, through diligent efforts to develop a peptide with biologically effective activity, have identified a peptide consisting of an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E). Here, the biologically effective activity is also one or more selected from the following characteristics: (a) promotion of proliferation or activation of hair follicle dermal papilla cells; (b) promotion of activation of outer root sheath cells; (c) promotion of activation of hair follicle cells; and (d) suppression of DKK-1 (Dickkopf-related protein 1) expression. More specifically, the biologically effective activities include (aa) improved phosphorylation of ERK and AKT, factors involved in the proliferation of hair follicle dermal papilla cells; (bb) activation of β-catenin, LEF-1 (Lymphoid enhancer-binding factor-1), c-Myc, and cyclin D1 in hair follicle dermal papilla cells; (cc) enhanced expression of Ha3-II (type I cuticular Ha3-Keratin 5, Keratin 14, and Keratin 19) in outer root sheath cells; and (dd) enhanced expression of HOXC13 (Homeobox protein Hox-C13), MSX2 (Msh homeobox 2), and FOXN1 (Forkhead box protein N1) in hair follicle cells. Therefore, the peptide can be used for applications in improving hair loss or promoting hair growth, preventing or treating hair loss, and improving damaged hair.

[0014] The peptide may also have a protecting group attached to its N-terminus or C-terminus to acquire chemical stability, enhanced pharmacological properties (such as half-life, water absorption, potency, and efficacy), altered specificity (e.g., a broad biological activity spectrum), or reduced antigenicity. Therefore, the peptide may be used to include either the peptide itself or a protected derivative thereof. In one specific example, the N-terminus of the peptide is bonded to one protecting group selected from the group consisting of acetyl, fluorenylmethoxycarbonyl, formyl, palmitoyl, myristyl, stearyl, butoxycarbonyl (Boc), allyloxycarbonyl (Alloc), and polyethylene glycol (PEG); and / or, the C-terminus of the peptide may be bonded to one protecting group selected from the group consisting of amino (-NH2), tertiary alkyl (-NH2), 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.

[0015] 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 the state of existence that occurs naturally. Here, the artificial modification may include artificially synthesizing an amino acid sequence by mimicking multiple amino acid structures, or being engineered to acquire chemical stability, enhanced pharmacological properties, altered specificity, or reduced antigenicity, as described above.

[0016] As used herein, the term "stability" may refer 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).

[0017] Another embodiment provides a cosmetic composition for improving damaged hair, comprising a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient.

[0018] As used herein, the term "improvement" may mean any action that alleviates or treats a condition, such as any action that at least reduces the severity of the symptoms.

[0019] As used herein, the term "damaged hair" may refer to hair that has been deformed or weakened due to physical, chemical, or environmental factors. Damaged hair may also include, for example, hair in which the spacing between the cuticles that make up the hair is widened or in which part of the cuticle has peeled off. Damaged hair may also mean hair in which the overall texture is rougher and the elasticity and volume of the hair are reduced, with decreased tensile strength and shine compared to normal hair, while roughness and friction are increased. Damaged hair may also refer to hair in which the bonds between cysteine ​​molecules within the hair are broken due to physical or chemical external stimuli.

[0020] As used herein, the term "damaged hair improvement" may refer to restoring the aforementioned damaged hair condition to a normal hair condition. Such damage improvement may include, for example, improved hair elasticity (tensile strength) and shine, reduced hair roughness and friction, and restoration of the hair cuticle. Furthermore, such damage improvement may mean reconnecting or restoring cysteine ​​bonds that have been broken by external stimuli. This term may also be used interchangeably with terms such as "promoting damaged hair regeneration" and "improving hair protection function."

[0021] Conventional functional peptides, despite their effective biological activity, suffer from disadvantages such as not being effectively absorbed into target tissues or cells due to their size, or being eliminated from the body quickly 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 or hair penetration of the active ingredient. For example, when applied locally to a specific area, it can improve the condition of damaged hair.

[0022] According to one embodiment, the peptide exhibited efficacy such as promoting the proliferation or activation of hair follicle dermal papilla cells, promoting the activation of outer root sheath cells, and promoting the activation of hair follicle cells. Furthermore, in experiments using human hair or human hair wigs, the peptide showed an effect of improving tensile strength, shine, roughness, and frictional force of damaged hair, and demonstrated excellent skin safety. In other words, the aforementioned efficacy demonstrates the effect of improving / promoting the regeneration of damaged hair and enhancing hair protection function, and the peptide can be utilized as an active ingredient in cosmetic compositions for improving damaged hair.

[0023] In one specific example, the active ingredient of the cosmetic composition, for example, a functional cosmetic composition, may include the biologically active peptide itself, a derivative thereof into which a protecting group has been introduced, or a peptide in which the terminal region has been extended based on the amino acid sequence of the peptide (for example, 10-mer, 9-mer, 8-mer, 7-mer, 6-mer, 5-mer peptides).

[0024] The cosmetic composition contains a cosmetically effective amount of the peptide; and / or a cosmetically acceptable carrier, but is not limited thereto.

[0025] As used herein, the term "cosmetically effective amount" may mean an amount sufficient to achieve the hair removal improvement or hair growth promotion efficacy of the cosmetic composition.

[0026] The weight ratio between the peptide and the cosmetically acceptable carrier is, for example, also 500:1 to 1:500. As an example, the weight ratio is 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:! 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, but is not limited thereto.

[0027] The cosmetic composition can be manufactured into any dosage form commonly manufactured in the industry, for example, it can be formulated into solutions, suspensions, emulsions, pastes, gels, creams, emulsions, lotions, mists, powders, soaps, oils, powder foundations, emulsion foundations, wax foundations, and sprays, etc., but is not limited thereto. The cosmetic composition can be commercialized into any one selected from the group consisting of, for example, scalp clinic agents, scalp scaling agents, scalp massage agents, scalp care products, detergents, shampoos, hair tonics, hair conditioners, hair lotions, hair gels, hair packs, hair masks, hair essences, ointments, hair styling agents, hair dyes, and hair perm agents, but is not limited thereto.

[0028] When the dosage form of the cosmetic composition is a paste, cream, or gel, animal oils, vegetable oils, waxes, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, or zinc oxide may be used as the carrier component.

[0029] 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. For example, in the case of a spray, it may further contain propellants such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether.

[0030] 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, benzylbenzoic acid, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or sorbitan fatty acid ester.

[0031] 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.

[0032] The peptide may be contained in nanosomes or nanoparticles to further improve skin penetration or stability issues. For example, the nanosomes may be produced by a microfluidizer using lecithin as a raw material and contained within lecithin particles. Any known method for producing the nanosomes may 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.

[0033] In addition to the peptide and carrier components as active ingredients, the components contained in the cosmetic composition may include ingredients commonly used in cosmetic compositions, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0034] The content of peptides as active ingredients in the cosmetic composition can be appropriately and non-restrictively selected depending on the product form, desired use, etc., and can be added in amounts of, for example, 0.01 to 15% by weight of the total cosmetic composition weight. Alternatively, for example, the cosmetic composition may contain peptides in amounts of 0.2% by weight, 0.5% by weight, 1.0% to 3.0% by weight, preferably 2.0% to 3.0% by weight, based on the total weight. The cosmetic composition may contain, for example, 0.1 to 1.5% by weight, for example, 0.2 to 1.0% by weight, 0.2 to 0.8% by weight, or 0.2 to 0.5% by weight of peptides, 0.2 to 0.5% by weight of surfactants, 0.01 to 0.1% by weight of pH adjusters, and / or 0.2 to 0.5% by weight of preservatives, and selectively 4.0 to 6.0% by weight of emulsifiers, based on the total weight of the cosmetic composition.

[0035] Another embodiment provides a cosmetic composition for improving hair loss or promoting hair growth, comprising a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient.

[0036] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0037] As used herein, the term "hair loss improvement" comprehensively refers to the process or effect of treating, reducing, or alleviating hair loss, and may include all actions that suppress the progression of hair loss, such as promoting the proliferation or activation of hair follicle dermal papilla cells, promoting the activation of outer root sheath cells, promoting the activation of hair follicle cells, and suppressing the expression of hair loss inducing factors such as DKK-1. The hair follicle dermal papilla cells, outer root sheath cells, and hair follicle cells are cells that constitute the hair follicle, which affects the thickness and growth of hair, and the activation of these cells affects the hair cycle, and through such a mechanism, a hair loss improvement effect can be induced.

[0038] As used herein, the term "hair growth promotion" refers to all actions that increase hair growth in hair follicles, and may include 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 is divided into the growth phase (anagen), when hair grows; the degeneration phase (catagen), when growth ends and the hair bulb shrinks; the resting phase (telogen), when the activity of the dermal papilla stops and the hair remains in the scalp; and the active phase, when the activity of the dermal papilla begins or when new hair is generated and old hair falls out. In the hair cycle, dermal papilla cells are connected to capillaries and sensory nerves and play a role in supplying oxygen and nutrients to hair follicle cells, while hair follicle cells surround the dermal papilla cells and play a role in determining the rate of the growth phase through continuous cell division and proliferation. Furthermore, the outer root sheath cells are located at the point of contact with the basal layer of the epidermis and play a role in protecting the hair until keratinization is complete.

[0039] According to one embodiment, the peptide exhibited effects such as promoting the proliferation or activation of hair follicle dermal papilla cells, promoting the activation of outer root sheath cells, promoting the activation of hair follicle cells, and suppressing 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 improving hair loss or promoting hair growth.

[0040] Another embodiment provides a method for improving damaged hair, improving hair loss, or promoting hair growth, comprising the step of applying a cosmetic composition containing a peptide comprising an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient to the skin or hair of an individual.

[0041] As stated above, any terms or elements mentioned in the description relating to the cosmetic composition that are the same as those already mentioned are as described above.

[0042] As used herein, the terms “apply,” “administer,” and “coat” are interchangeable 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 within an individual via an administration route.

[0043] Another embodiment provides a pharmaceutical composition for preventing or treating hair loss, comprising as an active ingredient a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0044] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0045] In this specification, the term "prevention" means all actions that suppress or delay the onset of a disease by administering the composition.

[0046] 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.

[0047] 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.

[0048] "Hair loss," which is a disease targeted for prevention or treatment by the aforementioned pharmaceutical compositions, refers to a condition in which hair is absent in areas where hair should normally be present, and specifically, it may mean the shedding of mature hair (thick, black hair) on the scalp. The causes of hair loss are not limited, but can be triggered by a variety of factors including genetic causes, hormonal imbalances, mental stress, exposure to air pollution, and environmental influences such as the consumption of processed foods. For example, hair loss can include hereditary androgenic alopecia (baldness), alopecia areata, tinea capitis due to fungal infection, telogen effluvium, trichotillomania, and hair growth disorders. Scarring alopecia, which involves the formation of scars, may include lupus, bald folliculitis, lichen planus pilaris, burns, and trauma.

[0049] According to one embodiment, the peptide exhibited effects such as promoting the proliferation or activation of hair follicle dermal papilla cells, promoting the activation of outer root sheath cells, promoting the activation of hair follicle cells, and suppressing the expression of hair loss inducing factors such as DKK-1. Therefore, the peptide can be utilized as an active ingredient in pharmaceutical compositions for the prevention or treatment of hair loss.

[0050] The active ingredient of the pharmaceutical composition may include the biologically active peptide itself, a derivative thereof into which a protecting group has been introduced, or a peptide in which the terminal region has been extended based on the amino acid sequence of the peptide (for example, 10-mer, 9-mer, 8-mer, 7-mer, 6-mer, and 5-mer peptides).

[0051] The pharmaceutical composition may, but is not limited to, contain a pharmaceutically effective amount of the peptide and / or a pharmaceutically acceptable carrier.

[0052] As used herein, the term "pharmaceutical effective amount" means an amount sufficient to achieve the efficacy of the pharmaceutical composition in preventing or treating hair loss.

[0053] The pharmaceutically acceptable carriers mentioned above are those commonly used in formulation and include, but are not limited to, lactose, dextrose, saccharose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoic acid, propylhydroxybenzoic acid, talc, magnesium stearate, and mineral oil. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0054] The weight ratio between the peptide and the pharmaceutically acceptable carrier may be, for example, 500:1 to 1:500, and may also be, but is not limited to, 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.

[0055] The aforementioned pharmaceutical composition may further contain, but is not limited to, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspension agents, preservatives, and the like, in addition to the aforementioned components.

[0056] The aforementioned pharmaceutical composition may be administered orally or parenterally, preferably parenterally, and in the case of parenteral administration, it may be administered by intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, local administration, transdermal administration, etc., but is not limited to these.

[0057] The dosage of the aforementioned pharmaceutical composition may be 0.0001 to 1000 μg per day (0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, or 1.0 to 1000 μg), but is not limited to these values ​​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.

[0058] The pharmaceutical composition may be manufactured in unit dose form or encapsulated in a multi-dose container by formulating it with pharmaceutically acceptable carriers and / or excipients using a method readily available to a person with ordinary skill in the art to which the invention pertains.

[0059] The dosage form 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, granules, tablet or capsule, and may additionally contain a dispersant and / or stabilizer.

[0060] 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 an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient.

[0061] As stated above, any terms or elements mentioned in the description of the pharmaceutical composition that are the same as those already mentioned are as described above.

[0062] Another embodiment provides a food composition for improving damaged hair, preventing hair loss, or promoting hair growth, comprising a peptide containing an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient.

[0063] As stated above, any terms or elements mentioned in the description of the peptides that are the same as those already mentioned are as described above.

[0064] The content of the peptide as an active ingredient contained in the food composition can be appropriately and non-restrictively selected depending on the form 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 beverage 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]

[0065] According to one embodiment of the peptide, it can be applied to improve damaged hair, improve hair loss, promote hair growth, and promote hair development by promoting the proliferation or activity of dermal papilla cells, hair follicle cells, and outer root sheath cells that constitute hair follicles, and by suppressing the expression of DKK-1, a hair loss-related factor.

[0066] According to one embodiment of the peptide, the peptide may be used as an active ingredient in a composition for improving damaged hair, improving hair loss, or promoting hair growth. [Brief explanation of the drawing]

[0067] [Figure 1] This is the result of checking the cell proliferation level after adding Peptide-1 according to one example to hair follicle dermal papilla cells. [Figure 2] This is the result of checking the cell proliferation level after adding Peptide-2 according to one example to hair follicle dermal papilla cells. [Figure 3]This is the result of adding Peptide-1 according to one example to hair follicle dermal papilla cells and confirming the phosphorylation of ERK and AKT, factors involved in the proliferation of hair follicle dermal papilla cells. [Figure 4] This is the result of adding Peptide-2 according to one example to hair follicle dermal papilla cells and confirming the phosphorylation of ERK and AKT, factors involved in the proliferation of hair follicle dermal papilla cells. [Figure 5] This is the result of adding Peptide-1 according to one example to hair follicle dermal papilla cells and confirming the migration of β-catenin, a factor involved in the proliferation of hair follicle dermal papilla cells, from the cytoplasm to the nucleus. [Figure 6] This is the result of adding Peptide-2 according to one example to hair follicle dermal papilla cells and confirming the migration of β-catenin, a factor involved in the proliferation of hair follicle dermal papilla cells, from the cytoplasm to the nucleus. [Figure 7] This is the result of adding Peptide-1 according to one example to hair follicle dermal papilla cells and then checking the expression levels of c-Myc and cyclin D1, which are downstream target genes of the β-catenin activation mechanism. [Figure 8] This is the result of adding Peptide-2 according to one example to hair follicle dermal papilla cells and then checking the expression levels of LEF-1, c-Myc, and cyclin D1, which are downstream target genes of the β-catenin activation mechanism. [Figure 9] This is the result of adding Peptide-1 according to one example to hair follicle dermal papilla cells and then checking the expression level of DKK-1, a hair loss inducing factor. [Figure 10] This is the result of adding Peptide-2 according to one example to hair follicle dermal papilla cells and then checking the expression level of DKK-1, a hair loss inducing factor. [Figure 11] This is the result of adding Peptide-1 according to one example to outer root sheath cells and then checking the expression levels of Ha3-II, Keratin 14, and Keratin 19, which are activators of outer root sheath cells. [Figure 12]This is the result of adding Peptide-2 according to one example to outer root sheath cells and then checking the expression levels of Ha3-II, Keratin 5, Keratin 14, and Keratin 19, which are activators of outer root sheath cells. [Figure 13] This is the result of adding Peptide-1 according to one example to hair follicle cells and then checking the expression levels of HOXC13, MSX2, and FOXN1, which are activators of hair follicle cells. [Figure 14] This is the result of adding Peptide-2 according to one example to hair follicle cells and then checking the expression levels of MSX2 and FOXN1, which are activators of hair follicle cells. [Figure 15] This shows the results of measuring the hair tensile strength over time after applying the composition according to one embodiment (Example 1) to human hair. [Figure 16] The following are the results of measuring the hair tensile strength over time after applying the composition according to one embodiment (Example 2) to human hair. [Figure 17] This is the result of applying the composition according to one embodiment (Example 1) to a human hair wig and then photographing it using a digital camera. [Figure 18] This is the result of applying the composition according to one embodiment (Example 2) to a human hair wig and then photographing it using a digital camera. [Figure 19] This shows the results of observing the roughness (cuticle) of the hair over time using an electron microscope after applying the composition according to one embodiment (Example 1) to human hair. [Figure 20] This is the result of observing the roughness (cuticle) of the hair over time using an electron microscope after applying the composition according to one embodiment (Example 2) to human hair. [Modes for carrying out the invention]

[0068] The present invention will be described in more detail below based on examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

[0069] Experimental Example 1: Peptide Synthesis The peptides listed in Table 1 below were synthesized using an automated peptide synthesizer (Milligen 9050, Millipore, USA), and these synthesized peptides were separated into pure molecules using C18 reversed-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA). The column used was ACQUITY UPLC BEH300 C18 (2.1 mm x 100 mm, 1.7 μm, Waters Co., USA).

[0070] [Table 1]

[0071] Experimental Example 2. Confirmation of the effect of promoting the proliferation or activity of hair follicle dermal papilla cells. 2-1. Confirmation of the effect of promoting the proliferation of hair follicle dermal papilla cells. In this experiment, we evaluated the changes in the survival rate of human hair follicle dermal papilla cells (HFDPCs) to confirm the effect of the peptide from one example on the proliferation of human hair follicle dermal papilla cells. Specifically, human hair follicle dermal papilla cells were placed in a 96-well plate in a 4 × 10⁶ format. 3After seeding at a density of cells / well, the cells were cultured in mesenchymal stem cell complete media for 24 hours. Subsequently, the culture medium was replaced with serum-free mesenchymal stem cell complete media, and the cells were cultured again for 24 hours. Then, the peptide from one example was dispensed in 500 mM, 5 μM, or 50 μM concentrations and cultured at 37°C for 72 hours. After washing the cultures with PBS, 10 μL of 5 mg / ml MTT solution was dispensed into each well. The cultures were then cultured in a CO2 incubator for 4 hours, the medium was removed, and 100 μL of DMSO (Dimethyl sulfoxide) was added and stirred for 10 minutes. The absorbance at a wavelength of 540 nm was then measured using a spectrophotometer. A control group (untreated group) was used, and a group treated with 1 μM EGF was used as the positive control group.

[0072] As a result, as shown in Figures 1 and 2, it was confirmed that Peptide-1 or Peptide-2 promotes the proliferation of dermal papilla cells.

[0073] 2-2. Confirmation of the activation effect of proliferation-related factors of hair follicle and dermal papilla cells. In this experiment, we confirmed the effect of the peptide described in one example on the activation of proliferation-related factors in human hair follicle papilla cells. Specifically, human hair follicle papilla cells were divided into 4 × 10⁶ cells. 5After seeding a 6-well plate at a cell / well density, the cells were cultured in mesenchymal stem cell complete media for 24 hours. Subsequently, the peptide according to one example was dispensed in 500 mM, 5 μM, or 50 μM concentrations and cultured at 37°C for 24 hours. After washing the culture with PBS, cell lysates were prepared by processing with 100 μL of lysis buffer. Subsequently, BCA (Bicinchoninic Acid) was quantified to prepare the same volume of protein sample, and electrophoresis was performed using a 10% SDS-PAGE gel. Subsequently, the proteins separated via SDS-PAGE were transferred through a PVDF membrane, and the reaction was blocked at room temperature for 30 minutes using 5% skim milk. Subsequently, phospho-AKT and phospho-ERK (cell signaling, USA) antibodies were diluted 1:1000 in 3% BSA and reacted with the blocked PVDF membrane at 4°C for 16 hours. Subsequently, the reaction product was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS), and the secondary antibody was diluted 1:2000 with 5% skim milk and reacted with the reaction product for 1 hour. After that, the reaction product was washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS), treated with ECL solution (GE Healthcare, USA), and the protein expression level was checked. On the other hand, an untreated group was used as the control group, and a group treated with 1 μM Minoxidil was used as the positive control group.

[0074] As a result, as shown in Figures 3 and 4, it was confirmed that Peptide-1 or Peptide-2 induces phosphorylation of ERK and AKT, factors involved in the proliferation of hair follicle dermal papilla cells.

[0075] 2-3. Confirmation of the effect of promoting the proliferation mechanism of hair follicle dermal papilla cells. In this experiment, we investigated the effect of the peptide from Example 1 on the proliferation mechanism of human hair follicle papilla cells. Specifically, human hair follicle papilla cells were cultured in the same manner as in Experimental Example 2-2, treated with the peptide from Example 1, and then nuclear protein was isolated from the culture using a nuclear protein extraction kit (Thermo Scientific, USA). Subsequently, the protein expression levels were confirmed using beta-catenin (Cell Signaling, USA) and HDAC1 (Santa Cruz, USA) antibodies in the same manner as in Experimental Example 2-2. On the other hand, an untreated group was used as the control group, and a group treated with 10 ng / mL of rhWnt-3a (recombinant human Wnt-3a protein) (R&D Systems (USA, MN)) was used as the positive control group.

[0076] Furthermore, we confirmed the level changes of downstream target genes LEF-1 (Lymphoid enhancer-binding factor-1), c-Myc, or cyclin D1 due to the β-catenin activation mechanism. To this end, human hair follicle papilla cells were cultured in the same manner as in Experimental Example 2-2 and treated with the peptide according to Example 1. Subsequently, the culture was washed with PBS and then treated with 300 μL of easy blue (intron, Korea) to separate the RNA. After quantifying the separated RNA with a nano drop, cDNA synthesis was carried out using the RNA template with a cDNA synthesis kit (enzynomics, Korea). Subsequently, a PCR reaction was carried out using the primers and PCR premix (enzynomics, Korea) shown in Table 2 below, followed by electrophoresis on 1.5% agarose gel, and gene expression at the RNA level was confirmed through the Bio-Rad gel image system.

[0077] [Table 2]

[0078] As a result, as shown in Figures 5 and 6, we confirmed that Peptide-1 or Peptide-2 increased the translocation of β-catenin from the cytoplasm to the nucleus. Furthermore, as shown in Figures 7 and 8, we confirmed that Peptide-1 or Peptide-2 activated β-catenin, thereby increasing the expression of its downstream target genes, LEF-1, c-Myc, and / or cyclin D1.

[0079] Experimental Example 3. Confirmation of the effect of suppressing the expression of hair loss-related factors. In this experiment, we investigated the effect of the peptide from Example 1 on the expression of DKK-1 (Dickkopf-1), a hair loss inducing factor. Specifically, human hair follicle dermal papilla cells were cultured in the same manner as in Experimental Example 2-2, and the peptide from Example 1 was dispensed in 500 mM, 5 μM, or 50 μM doses while dihydrotestosterone (DHT), a DKK-1 expression inducer, was added. Subsequently, the protein expression level was confirmed using the DKK-1 (cell signaling, USA) antibody in the same manner as in Experimental Example 2-2. On the other hand, an untreated group was used as the control group, and a group treated with 5 μM finasteride was used as the positive control group.

[0080] As a result, as shown in Figures 9 and 10, it was confirmed that Peptide-1 or Peptide-2 may suppress the expression of DKK-1, a hair loss inducing factor.

[0081] Experimental Example 4. Confirmation of the effect of promoting the activation of outer root sheath cells. In this experiment, we confirmed the effect of the peptide from one example on the expression of cytokines associated with the activation of human hair outer root sheath cells (HHORSCs): Ha3-II (type I cuticular Ha3-II), Keratin 5, Keratin 14, and Keratin 19. Specifically, 4 × 10⁶ human hair outer root sheath cells were used. 5 After seeding cells in a 6-well plate at a cell / well density, the cells were cultured in mesenchymal stem cell complete media for 24 hours. Subsequently, the culture medium was replaced with serum-free mesenchymal stem cell complete media, and the cells were cultured again for 24 hours. Thereafter, the peptide from Example 1 was dispensed in 500 mM, 5 μM, or 50 μM concentrations and cultured at 37°C for 24 hours. Subsequently, gene expression at the RNA level was confirmed using the primers shown in Table 3 below, in the same manner as in Experimental Examples 2-3. As a control group, an untreated group was used, and as a positive control group, a group treated with 50 nM EGF was used.

[0082] [Table 3]

[0083] As a result, as shown in Figures 11 and 12, it was confirmed that Peptide-1 or Peptide-2 increased the expression of Ha3-II, Keratin 14, and Keratin 19, which are activators of outer root sheath cells.

[0084] Experimental Example 5. Confirmation of the effect of promoting the activation of hair follicle cells. In this experiment, we confirmed the effect of the peptide described in one example on the expression of HOXC13 (Homeobox protein Hox-C13), MSX2 (Msh homeobox 2), and FOXN1 (Forkhead box protein N1), transcription factors associated with the activation of human hair follicle cells (HHGMCs). Specifically, human hair follicle cells were subjected to 4 × 10⁶ 5 After seeding cells in a 6-well plate at a cell / well density, the cells were cultured in mesenchymal stem cell complete media for 24 hours. Subsequently, the culture medium was replaced with serum-free mesenchymal stem cell complete media, and the cells were cultured again for 24 hours. Thereafter, the peptide from Example 1 was dispensed in 500 mM, 5 μM, or 50 μM concentrations and cultured at 37°C for 24 hours. Subsequently, gene expression at the RNA level was confirmed using the primers shown in Table 4 below, in the same manner as in Experimental Examples 2-3. A non-treated group was used as the control group, and a group treated with 50 nM EGF was used as the positive control group.

[0085] [Table 4]

[0086] As a result, as shown in Figures 13 and 14, it was confirmed that Peptide-1 or Peptide-2 increased the expression of HOXC13, MSX2, and FOXN1, which are activators of hair follicle cells.

[0087] Experimental Example 6. Production of a composition for improving damaged hair. In this experiment, a composition for improving damaged hair containing a peptide (Peptide-1) as an active ingredient was prepared. Specifically, the mist-type composition of Example 1 was prepared using the components and concentrations listed in Table 5 below, employing a method known in the industry. Furthermore, the emulsion-type composition of Example 2 was prepared using the components and concentrations listed in Table 6 below, employing a method known in the industry.

[0088] [Table 5]

[0089] [Table 6]

[0090] Experimental Example 7. Confirmation of the effect on improving the tensile strength of damaged hair. This experiment investigated the effect of a peptide from one example on the tensile strength (elasticity) of damaged hair. Specifically, 23 subjects participated in the experiment. For three days, each subject shampooed once a day, dried their hair, and then applied either the composition from Example 1 or the composition from Example 2 evenly to their hair. After massaging and brushing, the hair was left to dry for 5 minutes. Subsequently, hair samples were taken from the back of each subject's head, and the Break Load (gmf) value over time was measured using an MTT175 (Miniature Tensile Tester, Diastron Ltd., UK).

[0091] The results of Examples 1 and 2 are shown in Tables 7 and 8 below, respectively, and the results for one of the subjects in Examples 1 and 2 are shown in Figures 15 and 16, respectively. An increase in the Break Load value indicates an improvement in the tensile strength of the hair.

[0092] [Table 7]

[0093] [Table 8]

[0094] As a result, as shown in Tables 7 and 8, when the composition of Example 1 was used, the average Break Load increased by 15.81% immediately after one use and by 33.86% after three days of use. When the composition of Example 2 was used, the average Break Load increased by 18.22% immediately after one use and by 38.96% after three days of use. Statistically significant differences were observed in both the immediate post-use and three-day post-use stages compared to the pre-use stage for both Example 1 and Example 2 (p<0.001).

[0095] Through the results described above, it was confirmed that the peptide according to one example is effective in improving the tensile strength (elasticity) of damaged hair.

[0096] Experiment Example 8. Confirmation of the effect on improving hair shine. In this experiment, we confirmed the effect of the peptide from one example on the shine (angel ring) of damaged hair. Specifically, 20 human hair wigs were used as subjects. After shampooing and drying the wigs, either the composition from Example 1 or the composition from Example 2 was uniformly applied to the wigs, massaged, and brushed, and left for 5 minutes. Subsequently, the gloss unit value at 60° was measured using a gloss meter (Multi Gloss 268 PLUS, Konica Minolta, Japan).

[0097] The results for Examples 1 and 2 are shown in Tables 9 and 10 below, respectively, indicating that an increase in the Gloss Unit value signifies an improvement in hair shine. Furthermore, the results of digital camera photography after applying Examples 1 and 2 to one human hair wig are shown in Figures 17 and 18, respectively.

[0098] [Table 9]

[0099] [Table 10]

[0100] As a result, as shown in Tables 9 and 10, it was confirmed that when the composition of Example 1 was treated, the average Gloss Unit value increased by 78.61% immediately after one use, and when the composition of Example 2 was treated, it increased by 82.15% immediately after one use. A statistically significant difference was observed immediately after one use compared to before use for both Example 1 and Example 2 (p<0.001).

[0101] Through the results described above, it was confirmed that the peptide according to one embodiment is effective in improving the shine (angel ring) of damaged hair.

[0102] Experiment Example 9. Confirmation of the effect on improving the roughness of damaged hair. In this experiment, we confirmed the effect of the peptide from Example 1 on the roughness (cuticle) of damaged hair. Specifically, the composition of Example 1 or the composition of Example 2 was applied to 23 subjects using the same method as in Experimental Example 7. Subsequently, hair from the back of the subjects' heads was photographed using a scanning electron microscope (S-4700, Hitachi, Japan), and the variable value Ra (μm), which indicates the roughness of the hair surface over time, was measured using an image analysis program (Image J, National Institutes of Health, USA) at 700x magnification.

[0103] The results of Examples 1 and 2 are shown in Tables 11 and 12 below, respectively. A decrease in the Ra value indicates an improvement in the hair cuticle and hair roughness. Furthermore, the electron microscope images obtained from one of the subjects after treatments in Examples 1 and 2 are shown in Figures 19 and 20, respectively.

[0104] [Table 11]

[0105] [Table 12]

[0106] As a result, as shown in Tables 11 and 12, when the composition of Example 1 was used, the average Ra value decreased by 20.04% immediately after one use and by 35.64% after three days of use. When the composition of Example 2 was used, the average Ra value decreased by 23.59% immediately after one use and by 37.94% after three days of use. Statistically significant differences were observed in both the immediate post-use and three-day post-use stages compared to the pre-use stage for both Example 1 and Example 2 (p<0.001).

[0107] Through the results described above, it was confirmed that the peptide according to one embodiment is effective in improving the roughness (cuticle) of damaged hair.

[0108] Experimental Example 10. Confirmation of the effect of improving the frictional force of damaged hair. In this experiment, we confirmed the effect of the peptide from one example on the frictional force of damaged hair. Specifically, the composition of Example 1 or the composition of Example 2 was applied to the human hair pieces (bleached hair) in the same manner as in Experimental Example 8, except that 20 human hair pieces (bleached hair) were used. Subsequently, the Mean Horizontal Force Out value (gmf / mm), which indicates the average horizontal frictional force of the hair at the hair root area of ​​the human hair pieces (bleached hair), was measured using an MTT175 (Miniature Tensile Tester, Dia-stron Ltd., UK).

[0109] The results for Examples 1 and 2 are shown in Tables 13 and 14 below, respectively. A decrease in the Mean Horizontal Force Out value indicates an improvement in hair friction.

[0110] [Table 13]

[0111] [Table 14]

[0112] As a result, as shown in Tables 13 and 14, when the composition of Example 1 was used, the average Mean Horizontal Force Out value decreased by 26.94% immediately after one use, and when the composition of Example 2 was used, it decreased by 31.06% immediately after one use. A statistically significant difference was observed immediately after one use compared to before use for both Example 1 and Example 2 (p<0.001).

[0113] Through the results described above, it was confirmed that the peptide according to one embodiment is effective in improving the frictional force of damaged hair.

[0114] Experimental Example 11. Evaluation of abnormal skin reactions The 23 subjects in Experimental Example 7 were observed for the occurrence of abnormal skin reactions after application of the composition. As a result, it was confirmed that no abnormal reactions to allergic contact dermatitis or irritant contact dermatitis were observed. In addition, the subjects were asked to report on erythema (redness), edema (swelling), scaling (dead skin cells), itching, stinging (pain), burning sensation, stiffness, and tingling, and it was confirmed that no abnormal skin reactions were reported.

[0115] Through the results described above, it was confirmed that the peptide-containing composition according to one example is suitable for skin application.

[0116] 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.

[0117] 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.

[0118] [Table 15]

[0119] 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.

[0120] [Table 16]

[0121] The above description of the present invention is illustrative, and a person with ordinary skill in the art to which the invention pertains will understand that it can be easily modified into other specific forms without altering the technical idea or essential features of the invention. Therefore, the embodiments described above should be understood to be illustrative in all respects and not limiting.

Claims

1. A peptide consisting of an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G).

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 fluorenylmethoxycarbonyl 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 (-NH₂). 2 ), tertiary alkyl group and hydrazino group (-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) Promoting the proliferation or activation of hair follicle dermal papilla cells; (b) Promoting activation of outer root sheath cells; (c) Promoting the activation of hair follicle cells; and (d) Suppression of DKK-1 (Dickkopf-related protein 1) expression.

5. A cosmetic composition for improving damaged hair, containing as an active ingredient a peptide consisting of an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G).

6. A cosmetic composition for improving hair loss or promoting hair growth, comprising a peptide consisting of an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E) as an active ingredient.

7. A pharmaceutical composition for preventing or treating hair loss, comprising a peptide as an active ingredient, consisting of an amino acid sequence represented as Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

8. The cosmetic composition according to claim 5 or 6, or the pharmaceutical composition according to claim 7, wherein the N-terminus of the peptide is bonded to one protecting group selected from the group consisting of an acetyl group, a fluorenylmethoxycarbonyl group, a formyl group, a palmitoyl group, a myristyl group, a stearyl group, a butoxycarbonyl group, an allyloxycarbonyl group, and polyethylene glycol (PEG).

9. The cosmetic composition according to claim 5 or 6, or the pharmaceutical composition according to claim 7, wherein the C-terminus of the peptide is bonded to one protecting group selected from the group consisting of an amino group (-NH₂), a tertiary alkyl group, and a hydrazino group (-NHNH₂).

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