Peptides with hair growth promoting and damaged hair improving 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] The present application relates to a peptide having hair growth promoting and damaged hair improving activity, and uses thereof. [Background technology]

[0002] Humans are made up of approximately 150,000 hairs, each of which goes through a different hair cycle, repeating the process of growth and shedding. The hair growth cycle is broadly divided into three stages: the anagen phase, in which hair grows; the catagen phase, in which growth stops and hair is maintained; and the telogen phase, in which the hair papilla shrinks, the hair follicle shrinks, and hair loss occurs. The hair growth cycle varies depending on internal factors such as family strength, genetics, constitution, and hormone regulation, as well as external factors such as nutritional status, aging, surrounding environment, and stress.

[0003] On the other hand, hair ends are exposed to the external environment for a longer period of time than the scalp, and therefore are more susceptible to damage from the external environment. Weathering caused by exposure to the external environment causes the hair cuticle to break, resulting in split ends and ultimately breakage. A typical example of such damage is the phenomenon of hair weakening and loss of tension when high heat is continuously applied to hair using a hair dryer, etc. In addition, various physical, chemical, and environmental factors such as frequent perming and dyeing can leach proteins and lipids from within the hair, promoting hair porosity. As a result, hair becomes dry and loses luster, and friction increases, making hair loss more difficult and resulting in breakage and split ends. Furthermore, such hair porosity can reduce hair elasticity, leading to problems such as loss of firmness and volume, or thinning.

[0004] Additionally, with the aging society, the number of people suffering from hair loss is increasing. While traditionally a problem primarily affecting middle-aged men, interest in hair loss prevention and hair restoration has recently been growing among young people and women. While hair loss has long been recognized as a symptom of aging, it has recently become clear that it is caused by a variety of factors, including genetic factors, stress, Westernized eating habits, nutritional imbalances, and changes in social activities. Currently, drugs with hair growth effects, approved by the U.S. FDA, include minoxidil (6-Amino-1,2-dihydro-1-hydroxy-2-imino-4-phenoxypyrimidine) (U.S. Patent No. 3,382,247) and finasteride (U.S. Patent No. 5,215,894), are used as hair growth promoters. Minoxidil was developed as a vasodilator for the treatment of hypertension in the early 1970s, but hirsutism has been reported as a side effect. It has been used as a hair growth promoter, increasing the diameter of hair follicles and thickening hair. In addition, finasteride was developed as a treatment for benign prostatic hyperplasia, but is currently used as a hair loss treatment, delaying the progression of hair loss and promoting hair growth. However, minoxidil has been reported to have side effects such as weight gain, edema, dermatitis, and increased heart rate, and finasteride requires continuous administration 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] Under such technical background, various researches are being conducted to improve damaged hair and prevent hair loss through mechanisms such as hormone regulation and metabolic regulation (Korean Patent Publication No. 2002-0005332), but the current situation is still incomplete. Summary of the Invention [Problem to be solved by the invention]

[0006] One embodiment is to provide a peptide consisting of the amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0007] Another aspect of the present invention is to provide a cosmetic composition for improving damaged hair, which comprises, as an active ingredient, a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0008] Yet another aspect is to provide a cosmetic composition for improving hair loss or promoting hair growth, which comprises, as an active ingredient, a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0009] Yet another aspect is to provide a pharmaceutical composition for preventing or treating hair loss, which comprises, as an active ingredient, a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0010] Other objects and advantages of the present application will become more apparent from the following detailed description taken in conjunction with the claims and drawings. The contents not described in this specification are fully understood and can be inferred by those skilled in the art of the present application or a similar art, and therefore, the description thereof will be omitted. [Means for solving the problem]

[0011] Each description and embodiment disclosed in this application may also be applied to each other description and embodiment. In other words, all combinations of various elements disclosed in this application belong to the scope of this application. In addition, the specific descriptions described below are not intended to limit the scope of this application.

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

[0013] As used herein, the term "peptide" refers to a linear molecule formed by linking amino acid residues together via peptide bonds. The peptide may be prepared by chemical synthesis methods known to those skilled in the art, particularly solid-phase synthesis techniques (Merrifield, J. Amer. Chem. Soc. 85:2149-54 (1963); Stewart, et al., Solid Phase Peptide Synthesis, 2nd ed., Pierce Chem. Co.: Rockford, 111 (1984)) or liquid synthesis techniques (U.S. Patent No. 5,516,891). The present inventors have made extensive efforts to develop peptides with biologically effective activity and have identified peptides consisting of the amino acid sequence Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E). Here, the biologically effective activity may be any one or more of the following properties: (a) promoting the proliferation or activation of hair follicle dermal papilla cells; (b) promoting the activation of outer root sheath cells; (c) promoting the activation of hair follicle cells; and (d) suppressing the expression of DKK-1 (Dickkopf-related protein 1). More specifically, the biologically effective activities include (aa) increased phosphorylation of ERK and AKT, which are 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) increased expression of Ha3-II (type I cuticular Ha3-Keratin 5, Keratin 14, and Keratin 19) in outer root sheath cells; and (dd) increased 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 improving hair loss or promoting hair growth, preventing or treating hair loss, and improving damaged hair.

[0014] The peptides may also have a protecting group attached to their N- or C-terminus to achieve chemical stability, enhanced pharmacological properties (e.g., half-life, absorbency, potency, efficacy), altered specificity (e.g., a broader spectrum of biological activity), or reduced antigenicity. Thus, the peptides may be used as either the peptide itself or a protected derivative thereof. In one embodiment, the N-terminus of the peptide may be conjugated with any 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 conjugated with any one protecting group selected from the group consisting of amino (-NH), tertiary alkyl, and azide (-NHNH). The peptide may also optionally further comprise a targeting sequence, a tag, a labeled residue, or a specifically engineered amino acid sequence to increase half-life or peptide stability.

[0015] The peptides are artificially synthesized or non-naturally occurring or engineered, and the term "non-naturally occurring or engineered" refers to a state in which an artificial modification has been added, rather than a state in which the peptide exists in nature. Here, the artificial modification can include artificially synthesizing an amino acid sequence by mimicking multiple amino acid structures, or engineering to obtain 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 in vivo proteolytic enzymes, but also to storage stability (eg, storage stability at room temperature).

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

[0018] As used herein, the term "amelioration" can refer to any action that at least reduces the parameters of the alleviation or treatment of a condition, for example, the severity of symptoms.

[0019] As used herein, the term "damaged hair" refers to hair that has been deformed or weakened due to physical, chemical, or environmental factors. Damaged hair can also refer to hair in which the cuticle spacing has widened or a portion of the cuticle has peeled off. Damaged hair can also refer to hair that has a coarse overall texture and reduced elasticity and volume due to reduced tensile strength and luster compared to normal hair, while having increased roughness and friction. Damaged hair can also refer to hair in which the bonds between cysteines in the hair have been broken due to physical or chemical external stimuli.

[0020] As used herein, the term "damaged hair improvement" refers to restoring the state of damaged hair to a normal hair state. The improvement of damaged hair may refer to, for example, improving hair elasticity (tensile strength) and shine, reducing hair roughness and friction, and restoring the cuticle within the hair. The improvement of damaged hair may also refer to reconnecting or restoring cysteine ​​bonds that have been broken by external stimuli. The term may also be used interchangeably with terms such as "promoting damaged hair regeneration" and "improving hair protection function."

[0021] Although conventional functional peptides have effective biological activities, they have drawbacks such as ineffective delivery to target tissues or cells due to their size, or a short half-life that causes them to disappear in the body within a short period of time. In contrast, a cosmetic composition according to one embodiment contains a peptide consisting of 10 or fewer amino acids as an active ingredient, which allows for excellent penetration of the active ingredient into the skin or hair. For example, when applied topically to a specific area, the composition can improve the condition of damaged hair.

[0022] According to one embodiment, the peptide has been shown to promote the proliferation or activation of hair follicle dermal papilla cells, promote the activation of outer root sheath cells, and promote the activation of hair follicle cells. Furthermore, in experiments using human hair or a human hair wig, the peptide has been shown to improve the tensile strength, gloss, roughness, and friction of damaged hair, and to be highly safe for the skin. In other words, the above-mentioned efficacy proves the effects of improving / promoting the regeneration of damaged hair and improving hair protection function, and the peptide can be used as an active ingredient in a cosmetic composition for improving damaged hair.

[0023] In one embodiment, the active ingredient of the cosmetic composition, e.g., functional cosmetic composition, may include the above-mentioned biologically active peptide itself, a derivative thereof into which a protecting group has been introduced, or a peptide having the amino acid sequence of the above-mentioned peptide as a basic skeleton and an extended terminal region (e.g., a 10-mer, 9-mer, 8-mer, 7-mer, 6-mer, or 5-mer peptide).

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

[0025] The term "cosmetically effective amount" as used herein may mean an amount sufficient to achieve the efficacy of the cosmetic composition in improving hair loss or promoting hair growth.

[0026] The weight ratio between the peptide and the cosmetically acceptable carrier may be, for example, 500:1 to 1:500. By way of example, the weight ratio may 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.

[0027] The cosmetic composition may be prepared in any formulation commonly prepared in the art, such as, but not limited to, a solution, suspension, emulsion, paste, gel, cream, emulsion, lotion, mist, powder, soap, oil, powder foundation, emulsion foundation, wax foundation, spray, etc. The cosmetic composition may be prepared in, but not limited to, any one product selected from the group consisting of a scalp clinic agent, a scalp scaling agent, a scalp massage agent, a scalp care product, a cleanser, a shampoo, a hair tonic, a hair conditioner, a hair lotion, a hair gel, a hair pack, a hair mask, a hair essence, an ointment, a hair styling agent, a hair dye, and a hair perm agent.

[0028] When the cosmetic composition is in the form of a paste, cream, or gel, the carrier component may be animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, or the like.

[0029] When the cosmetic composition is in the form of a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder is used as a carrier component, and in the case of a spray, for example, a propellant such as chlorofluorohydrocarbon, propane / butane, or dimethyl ether may be further included.

[0030] When the cosmetic composition is in the form of a solution or emulsion, a solvent, solubilizer, or emulsifier is 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 fatty esters, polyethylene glycol, or sorbitan fatty acid esters.

[0031] When the cosmetic composition is in the form of a suspension, a liquid diluent such as water, ethanol, or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used as a carrier component.

[0032] To further improve skin penetration or stability, the peptide can be incorporated into nanosomes or nanoparticles. For example, the nanosomes can be prepared using lecithin as a raw material using a microfluidizer, and then incorporated into lecithin particles. Any known method can be used to prepare the nanosomes. The nanosome particle size is preferably 30 to 200 nm. Nanosome particle sizes less than 30 nm can penetrate the skin too quickly, resulting in side effects. Nanosome particle sizes greater than 200 nm can penetrate the skin too quickly, making it difficult to achieve the benefits of using the nanosome structure.

[0033] The components contained in the cosmetic composition include, in addition to the peptide as an active ingredient and a carrier component, components commonly used in cosmetic compositions, such as common adjuvants such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

[0034] The content of the peptide as an active ingredient contained in the cosmetic composition is appropriately selected without limitation depending on the product form, desired use, etc., and may be added in an amount of, for example, 0.01 to 15 wt% of the total cosmetic composition weight. Furthermore, for example, the cosmetic composition may contain 0.2 wt%, 0.5 wt%, 1.0 wt% to 3.0 wt%, and preferably 2.0 wt% to 3.0 wt% of the peptide based on the total weight of the cosmetic composition. For example, the cosmetic composition may contain 0.1 to 1.5 wt%, e.g., 0.2 to 1.0 wt%, 0.2 to 0.8 wt%, or 0.2 to 0.5 wt% of the peptide, 0.2 to 0.5 wt% of a surfactant, 0.01 to 0.1 wt% of a pH adjuster, and / or 0.2 to 0.5 wt% of a preservative, and optionally 4.0 to 6.0 wt% of an emulsifier, based on the total weight of the cosmetic composition.

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

[0036] Among the terms or elements mentioned in the description of the peptide, the same as those already mentioned are as described above.

[0037] The term "improving hair loss" as used herein comprehensively refers to the process or effect of treating, alleviating, or alleviating hair loss conditions, and may refer to all actions that inhibit 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 make up hair follicles and affect the thickness and growth of hair, and activation of these cells affects the hair cycle, and through this mechanism, a hair loss improvement effect can be induced.

[0038] The term "hair growth promotion" as used herein refers to any action that increases hair growth in hair follicles, and may refer to any action that increases the total amount of hair, such as the effect of promoting hair follicle cell proliferation, hair follicle cell activity, or hair follicle cell growth. Specifically, the hair cycle is divided into the anagen phase, in which hair grows; the catagen phase, in which growth ends and the hair bulb shrinks; the telogen phase, in which the activity of the dermal papilla stops and the hair remains on the scalp; and the anagen phase, in which the activity of the dermal papilla begins or new hair is generated and old hair is shed. During the hair cycle, dermal papilla cells are connected to capillaries and sensory nerves and supply oxygen and nutrients to hair follicle cells, while hair follicle cells surround the dermal papilla cells and determine the rate of the anagen phase through continuous cell division and proliferation. In addition, outer root sheath cells are cells located at the contact area 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 exhibits effects of 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 a cosmetic composition for improving hair loss or promoting hair growth.

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

[0041] Among the terms and elements mentioned in the description of the cosmetic composition, the same as those already mentioned are as described above.

[0042] As used herein, the terms "apply," "administer," and "apply" are used interchangeably and may refer to causing at least partial localization of a composition according to an embodiment to a desired site, or to the placement of a composition according to an embodiment within an individual by a route of administration.

[0043] Yet another aspect provides a pharmaceutical composition for preventing or treating hair loss, which comprises, as an active ingredient, a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0044] Among the terms or elements mentioned in the description of the peptide, the same as those already mentioned are as described above.

[0045] As used herein, the term "prevention" refers to any action that inhibits or delays the onset of a disease by administering the composition.

[0046] As used herein, the term "treatment" refers to any form of care that provides a benefit to an individual suffering from or susceptible to a disease, including improvement of the individual's condition (e.g., one or more symptoms), delay in disease progression, delay in symptom onset, or slowing of symptom progression, etc. Thus, the terms "treatment" and "prevention" are not intended to mean a cure or complete elimination of symptoms.

[0047] The term "individual" refers to a subject in need of treatment for a disease, and more specifically refers to mammals such as human or non-human primates, mice, dogs, cats, horses, and cows.

[0048] "Hair loss," a disease to be prevented or treated by the pharmaceutical composition, refers to the absence of hair in areas where hair normally exists, specifically the loss of adult hair (thick, dark hair) on the scalp. Hair loss can be caused by, but is not limited to, genetic factors, hormonal imbalance, psychological stress, exposure to air pollution, various dietary habits such as the consumption of processed foods, and environmental influences. Examples of hair loss include hereditary androgenetic alopecia (baldness), alopecia areata, tinea capitis due to fungal infection, telogen effluvium, trichotillomania, and hair production disorders. Examples of cicatricial alopecia, which leaves scars, include hair loss due to lupus, folliculitis barbae, lichen planus pilaris, and hair loss due to burns and trauma.

[0049] According to one embodiment, the peptide exhibits effects of 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 inhibiting the expression of hair loss-inducing factors such as DKK-1. Therefore, the peptide can be used as an active ingredient in a pharmaceutical composition for preventing or treating hair loss.

[0050] The active ingredient of the pharmaceutical composition may include the peptide itself having the above-mentioned biological activity, a derivative thereof into which a protecting group has been introduced, or a peptide having the amino acid sequence of the peptide as a basic skeleton and an extended terminal region (e.g., 10-mer, 9-mer, 8-mer, 7-mer, 6-mer, or 5-mer peptide).

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

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

[0053] The pharmaceutically acceptable carriers are those commonly used in formulations, 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, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. 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. By way of example, the weight ratio may 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 pharmaceutical composition may further contain, in addition to the above ingredients, lubricants, wetting agents, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, etc., but is not limited to these.

[0056] The pharmaceutical composition may be administered orally or parenterally, preferably parenterally. In the case of parenteral administration, it may be administered by intramuscular injection, intravenous injection, subcutaneous injection, intraperitoneal injection, topical administration, transdermal administration, etc., but is not limited thereto.

[0057] The dosage of the pharmaceutical composition may be, but is not limited to, 0.0001 to 1000 μg (0.001 to 1000 μg, 0.01 to 1000 μg, 0.1 to 1000 μg, or 1.0 to 1000 μg) per day, and may be variously prescribed depending on factors such as formulation method, administration route, age, weight, sex, pathological condition, diet of the patient, administration time, administration route, excretion rate, and reaction sensitivity.

[0058] The pharmaceutical composition may be prepared in a unit dose form or in a multi-dose container by formulating it with pharmaceutically acceptable carriers and / or excipients in a manner that can be easily carried out by a person skilled 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 oily or aqueous medium, or in the form of an extract, powder, granules, tablet or capsule, and may additionally contain dispersing agents and / or stabilizers.

[0060] Yet another embodiment provides a method for preventing or treating hair loss, comprising administering to an individual a pharmaceutical composition containing, as an active ingredient, a therapeutically effective amount of a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

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

[0062] Yet another aspect provides a food composition for improving damaged hair, preventing hair loss, or promoting hair growth, which comprises, as an active ingredient, a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

[0063] Among the terms or elements mentioned in the description of the peptide, 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 is appropriately selected without limitation depending on the food form, desired use, etc., and may be added at, for example, 0.01 to 15 wt% of the total food weight. For example, in the case of a health drink composition, the peptide may be added at 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, the peptide promotes the proliferation or activity of hair papilla cells, hair follicle cells, and outer root sheath cells that make up hair follicles, and suppresses the expression of DKK-1, a factor associated with hair loss, and can therefore be used to improve damaged hair, improve hair loss, promote hair growth, and promote hair growth.

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

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

[0068] The present invention will be described in more detail below with reference to 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 the synthesized peptides were purified and separated using C18 reverse-phase high-performance liquid chromatography (HPLC) (Waters Associates, USA) on an ACQUITY UPLC BEH300 C18 column (2.1 mm ∘ 100 mm, 1.7 μm, Waters Co., USA).

[0070] [Table 1]

[0071] Experimental Example 2: Confirmation of the effect of promoting proliferation or activity of hair follicle dermal papilla cells 2-1. Confirmation of the effect of promoting proliferation of hair follicle dermal papilla cells In this experimental example, the effect of the peptide according to one embodiment on the proliferation of human hair follicle dermal papilla cells (HFDPC) was confirmed by evaluating the change in viability of the cells. Specifically, human hair follicle dermal papilla cells were cultured in a 96-well plate at 4 × 10 3After seeding at a density of 1000 cells / well, the cells were cultured in mesenchymal stem cell complete media for 24 hours. The cell culture medium was then replaced with serum-free mesenchymal stem cell complete media and cultured again for 24 hours. A peptide according to one embodiment was then dispensed at 500 mM, 5 μM, or 50 μM and cultured at 37°C for 72 hours. The cultures were then washed with PBS, and 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. After removing the medium, 100 μL of DMSO (dimethyl sulfoxide) was added and stirred for 10 minutes. The absorbance at 540 nm was measured using a spectrophotometer. An untreated group served as a control, and a group treated with 1 μM EGF served as a positive control.

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

[0073] 2-2. Confirmation of the activation effect of factors related to hair follicle dermal papilla cell proliferation In this experiment, the effect of the peptide according to one embodiment on the activation of proliferation-related factors in human hair follicle dermal papilla cells was confirmed. Specifically, 4×10 human hair follicle dermal papilla cells were cultured in a 200-well culture medium. 5After seeding at a density of 100 cells / well into a 6-well plate, the cells were cultured in mesenchymal stem cell complete media for 24 hours. Then, a peptide according to one embodiment was added at 500 mM, 5 μM, or 50 μM and cultured at 37°C for 24 hours. The culture was then washed with PBS and treated with 100 μL of lysis buffer to prepare cell lysates. BCA (bicinchoninic acid) was measured to prepare equal amounts of protein samples, which were then electrophoresed on a 10% SDS-PAGE gel. Proteins separated by SDS-PAGE were transferred to a PVDF membrane and blocked with 5% skim milk for 30 minutes at room temperature. 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. The reaction mixture was then washed three times for 15 minutes each with 0.1% PBS-T (0.1% Tween-20 in PBS). A secondary antibody was diluted 1:2000 with 5% skim milk and incubated with the reaction mixture for 1 hour. The reaction mixture was then 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 expression levels were confirmed. An untreated control group was used, and a positive control group was treated with 1 μM minoxidil.

[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, which are 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 experimental example, the effect of a peptide according to one embodiment on the proliferation mechanism of human hair follicle dermal papilla cells was examined. Specifically, human hair follicle dermal papilla cells were cultured in the same manner as in Experimental Example 2-2, treated with a peptide according to one embodiment, and then nuclear proteins were isolated from the culture using a nuclear protein extraction kit (Thermo Scientific, USA). Protein expression levels were then determined in the same manner as in Experimental Example 2-2 using antibodies against beta-catenin (Cell Signaling, USA) and HDAC1 (Santa Cruz, USA). An untreated control group was used, and a positive control group was treated with 10 ng / mL of rhWnt-3a (recombinant human Wnt-3a protein) (R&D Systems, USA, MN).

[0076] We also investigated changes in the levels of lymphoid enhancer-binding factor-1 (LEF-1), c-Myc, and cyclin D1, downstream target genes of β-catenin activation. To this end, human hair follicle dermal papilla cells were cultured and treated with a peptide according to one embodiment in the same manner as in Experimental Example 2-2. The culture was then washed with PBS and treated with 300 μL of Easy Blue (Intron, Korea) to isolate RNA. The isolated RNA was quantified using Nanodrop, and cDNA synthesis was performed using the RNA template with a cDNA synthesis kit (Enzynomics, Korea). PCR was performed using the primers and PCR premix (Enzynomics, Korea) listed in Table 2 below. Gene expression at the RNA level was confirmed by electrophoresis on a 1.5% agarose gel using a Bio-Rad gel image system.

[0077] [Table 2]

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

[0079] Experimental Example 3: Confirmation of the effect of suppressing the expression of hair loss-related factors In this experiment, the effect of a peptide according to one embodiment on the expression of DKK-1 (Dickkopf-1), a hair loss inducer, was confirmed. Specifically, human hair follicle dermal papilla cells were cultured in the same manner as in Experimental Example 2-2, and the peptide according to one embodiment was dispensed at 500 mM, 5 μM, or 50 μM, while dihydrotestosterone (DHT), a DKK-1 expression inducer, was added. Thereafter, DKK-1 (cell signaling, USA) antibody was used to confirm the protein expression level in the same manner as in Experimental Example 2-2. Meanwhile, an untreated group was used as a control group, and a group treated with 5 μM finasteride was used as a 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 factor that induces hair loss.

[0081] Experimental Example 4. Confirmation of the effect of promoting activation of outer root sheath cells In this experiment, the effect of the peptide according to one embodiment on the expression of Ha3-II (type I cuticular Ha3-II), Keratin 5, Keratin 14, and Keratin 19, which are cytokines associated with the activation of human hair outer root sheath cells (HHORSCs), was confirmed. Specifically, 4×10 human outer root sheath cells were cultured in a 200-well plate. 5 After seeding into a 6-well plate at a density of 100 cells / well, the cells were cultured in mesenchymal stem cell complete media for 24 hours. The cell culture medium was then replaced with serum-free mesenchymal stem cell complete media and cultured for another 24 hours. A peptide according to one embodiment was then added at 500 mM, 5 μM, or 50 μM and cultured at 37°C for 24 hours. Gene expression at the RNA level was then confirmed using the primers listed in Table 3 below in the same manner as in Experimental Example 2-3. An untreated group served as a control, and a group treated with 50 nM EGF served as a positive control.

[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 hair follicle cell activation In this experiment, the effects of the peptide according to one embodiment on the expression of HOXC13 (Homeobox protein Hox-C13), MSX2 (Msh homeobox 2), and FOXN1 (Forkhead box protein N1), which are transcription factors associated with the activation of human hair germinal matrix cells (HHGMC), were confirmed. Specifically, 4×10 human hair germinal matrix cells (HHGMC) were cultured in a 200-well plate. 5 After seeding into a 6-well plate at a density of 100 cells / well, the cells were cultured in mesenchymal stem cell complete media for 24 hours. The cell culture medium was then replaced with serum-free mesenchymal stem cell complete media and cultured again for 24 hours. A peptide according to one embodiment was then added at 500 mM, 5 μM, or 50 μM and cultured at 37°C for 24 hours. Gene expression at the RNA level was then confirmed using the primers listed in Table 4 below in the same manner as in Experimental Example 2-3. An untreated group served as a control, and a group treated with 50 nM EGF served as a positive control.

[0085] [Table 4]

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

[0087] Experimental Example 6: Preparation of a composition for improving damaged hair In this experimental example, a composition for improving damaged hair containing a peptide (Peptide-1) according to one embodiment as an active ingredient was prepared. Specifically, a mist-type composition of Example 1 was prepared using a method known in the art with the ingredients and amounts listed in Table 5 below. Also, an emulsion-type composition of Example 2 was prepared using a method known in the art with the ingredients and amounts listed in Table 6 below.

[0088] [Table 5]

[0089] [Table 6]

[0090] Experimental Example 7. Confirmation of the effect of improving tensile strength of damaged hair In this experiment, the effect of a peptide according to one embodiment on the tensile strength (elasticity) of damaged hair was examined. Specifically, 23 subjects were tested. After shampooing and drying their hair once a day for three days, the composition of Example 1 or the composition of Example 2 was evenly applied to the hair, massaged, and brushed for five minutes. Then, hair was collected from the back of the head of each subject, and the break load (the strength of the force at which the hair breaks, gmf) was measured over time 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 of Examples 1 and 2 for one of the subjects 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, it was confirmed that when treated with the composition of Example 1, the average Break Load value increased by 15.81% immediately after one use and 33.86% after three days of use, and when treated with the composition of Example 2, the average Break Load value increased by 18.22% immediately after one use and 38.96% after three days of use. Compared to before use in Examples 1 and 2, statistically significant differences were observed both immediately after one use and after three days of use (p<0.001).

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

[0096] Experimental Example 8: Confirmation of hair gloss improvement effect In this experiment, the effect of the peptide according to one embodiment on the shine (angel ring) of damaged hair was examined. Specifically, 20 human hair wigs were used. After shampooing and drying, the composition of Example 1 or the composition of Example 2 was evenly applied to the human hair wigs, massaged, brushed, and left on for 5 minutes. Then, the gloss unit value at 60° was measured using a gloss meter (Multi Gloss 268 PLUS, Konica Minolta, Japan).

[0097] The results of Examples 1 and 2 are shown in Tables 9 and 10 below, respectively. The higher the Gloss Unit value, the more improved the hair gloss. In addition, the results of digital camera photography after applying Examples 1 and 2 to one of the human hair wigs 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 the average Gloss Unit value increased by 78.61% immediately after one use when treated with the composition of Example 1, and increased by 82.15% immediately after one use when treated with the composition of Example 2. A statistically significant difference was observed immediately after one use compared to before use of Examples 1 and 2 (p<0.001).

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

[0102] Experimental Example 9: Confirmation of the effect of improving roughness of damaged hair In this experiment, the effect of a peptide according to one embodiment on the roughness (cuticle) of damaged hair was confirmed. Specifically, the composition of Example 1 or the composition of Example 2 was applied to 23 subjects in the same manner as in Experimental Example 7. Then, the hair on the back of the subjects' heads was photographed using a scanning electron microscope (S-4700, Hitachi, Japan), and the parameter 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. The lower the Ra value, the more the hair cuticle and hair roughness are improved. In addition, the results of electron microscope images of one of the subjects treated with 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, it was confirmed that when treated with the composition of Example 1, the average Ra value decreased by 20.04% immediately after one use and by 35.64% after three days of use, and when treated with the composition of Example 2, the average Ra value decreased by 23.59% immediately after one use and by 37.94% after three days of use. Compared to before use in Examples 1 and 2, statistically significant differences were observed both immediately after one use and after three days of use (p<0.001).

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

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

[0109] The results of Examples 1 and 2 are shown in Tables 13 and 14 below, respectively. The smaller the Mean Horizontal Force Out value, the more improved the friction force of the hair.

[0110] [Table 13]

[0111] [Table 14]

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

[0113] From the above results, it was confirmed that the peptide according to one embodiment is effective in improving the friction 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 any abnormal skin reactions after application of the composition. The results confirmed that no abnormal reactions such as allergic contact dermatitis or irritant contact dermatitis were observed. The subjects were also asked to report any erythema (redness), edema (swelling), scaling (dead skin cells), itching, stinging (pain), burning sensation, stiffness, or tingling sensation, and it was confirmed that no abnormal skin reactions were reported.

[0115] From the above results, it was confirmed that the composition containing the peptide according to one embodiment 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 with thorough stirring. The resulting solution was mixed with 5 g of lecithin, 0.3 ml of sodium oleate, 50 ml of ethanol, and a small amount of oil, and then the mixture was adjusted to 1 L with distilled water. The mixture was then emulsified under high pressure in a microfluidizer 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 prepared using a method known in the art.

[0118] [Table 15]

[0119] Dosage form example 3: Nutritious cream A nourishing cream containing a peptide according to one embodiment and having the following composition was prepared using a method known in the art.

[0120] [Table 16]

[0121] The above description of the present invention is for illustrative purposes only, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and are not limiting.

Claims

1. A peptide consisting of the amino acid sequence Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

2. 2. The peptide of claim 1, wherein the N-terminus of the peptide is bound to any 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 (-NH 2 ), tertiary alkyl groups and azides (-NHNH 2 2. The peptide of claim 1, wherein the peptide is bound to any one of the protecting groups selected from the group consisting of:

4. The peptide of claim 1, wherein the peptide exhibits one or more of the following properties: (a) promoting the proliferation or activation of hair follicle dermal papilla cells; (b) promoting the activation of outer root sheath cells; (c) promoting hair follicle cell activation; and (d) Suppression of DKK-1 (Dickkopf-related protein 1) expression.

5. A cosmetic composition for improving damaged hair, comprising as an active ingredient a peptide having an amino acid sequence represented by Arg(R)-Cys(C)-Cys(C)-Gly(G) or Glu(E)-Glu(E).

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

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

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