Hair-growing or hair-growing agent based on keratin microparticles

Abstract

Provided is a hair-growing and hair-growing agent which exhibits a high hair-growing and hair-growing effect and at the same time has no or little side effects. Specifically provided is a hair-growing or hair-growing agent containing keratin microparticles as a hair-growing or hair-growing active ingredient.

Description

Technical Field

[0001] The present invention relates to a hair growth or growth agent containing keratin microparticles and also to a cosmetic or pharmaceutical composition containing the hair growth or growth agent. Background Art

[0002] Minoxidil, finasteride, and dutasteride are known representative chemicals that promote hair growth and hair growth. Minoxidil has a vasodilating effect, so by applying it, it can promote blood circulation in the capillary blood under the scalp and provide more nutrients to the hair root cells, so it can show a hair growth and hair growth effect. However, the dilation of blood vessels can reduce blood pressure, which sometimes has an adverse effect on the heart and other organs, and even worse, it can cause a critical state and lead to fatal consequences. Finasteride and dutasteride can inhibit the hair loss effect of dihydrotestosterone (DHT), so the effect of preventing hair loss and increasing hair growth has been observed, but there are reports of serious side effects, namely liver dysfunction.

[0003] On the other hand, keratin is the main component protein of the hair, claws, feathers, scales, hoofs, wool, and horns of animals such as birds, mammals, and reptiles. Keratin shows non-toxicity, biodegradability, and excellent cell survival rate, and has good biocompatibility to living cells, so it is usually relevant to the fields of pharmacy, medicine, cosmetics, food, agriculture, and biotechnology to utilize relevant research and application with keratin. The purposes of keratin not only have drug delivery, but also have the purposes such as cell adhesion, cell proliferation, tissue scaffold, tissue regeneration, cell proliferation, wound healing, trauma, bioactive surface, and medical implantation equipment (such as non-patent literature 1, 2). In this way, keratin can form various shapes (such as non-patent literature 2) in the form of porous foam, sponge, cushion, film, sheet, gel, microfiber, particulate, nanoparticle, and bulk material.

[0004] Prior art literature

[0005] Non-patent literature

[0006] Non-patent document 1: A. Aluigi, et al., Int. J. Biol. Macromol., 2007, 41, 266-273

[0007] Non-patent document 2: K. Katoh, et al., Biomaterials, 2004, 25, 2265–2272

[0008] Non-patent document 3: Seong Yeong An, et al., Research Square, archive, https: / / www.researchsquare.com / article / rs-101358 / v1

[0009] Non-patent literature 4: S.Hu et al., Sci Adv. 2020, 6(30):eaba1685 Summary of the invention

[0010] Problems to be solved by the invention

[0011] Regarding the various effects of keratin, there are reports that subcutaneous injection of keratin itself has the effect of hair growth and development (Non-Patent Document 3). There are also reports that 3D spheroids of hair papilla cells containing keratin can show a hair growth effect (Non-Patent Document 4). However, any effect is accompanied by pain caused by subcutaneous injection, which is highly invasive, so a technology that brings about the hair growth and development effect of keratin by non-invasive application without pain is needed.

[0012] In addition, although various existing hair growth and growth agents can show high hair growth and growth effects, there is still a need to develop hair growth and growth agents with no toxicity and side effects or with low toxicity and side effects.

[0013] Technical solutions to the problem

[0014] The inventors found that keratin particles, when applied, exhibited higher hair growth and development effects than ordinary keratin aqueous dispersions, and subsequently came to the conclusion that keratin particles that are non-toxic to the human body can be used for hair growth and development, skin care, and medical care, leading to the completion of the present invention.

[0015] For example, the present invention includes the following embodiments:

[0016] [1] A hair growth or hair restoration agent comprising keratin fine particles as a hair growth or hair restoration effective ingredient.

[0017] [2] The hair growth or growth agent according to [1], wherein the particle size of the keratin fine particles is 0.01 to 100 μm.

[0018] [3] The hair growth or growth agent according to [1] or [2], wherein the keratin microparticles are in the form of an aqueous solution, an aqueous dispersion or a hydrogel.

[0019] [4] The hair growth or growth agent according to any one of [1] to [3], wherein the keratin comprises human keratin, wool-derived keratin, or partially sulfonated or ionized water-soluble keratin.

[0020] [5] A cosmetic composition comprising the hair growth or growth agent according to any one of [1] to [4] and a cosmetically acceptable excipient.

[0021] [6] A pharmaceutical composition comprising the hair growth or growth agent according to any one of [1] to [4] and a pharmaceutically acceptable excipient.

[0022] [7] The pharmaceutical composition according to [6], which is a skin external preparation.

[0023] [8] A method for growing or restoring hair in a subject, comprising applying the hair growing or restoring agent or keratin microparticles described in any one of [1] to [4] to the subject.

[0024] [9] A method for treating or preventing hair loss in a subject, comprising applying the hair growth or growth agent or keratin microparticles described in any one of [1] to [4] to the subject.

[0025]

[10] The method of [8] or [9], wherein applying to the subject comprises applying to the subject's skin.

[0026]

[11] The method according to [9], wherein the hair loss includes alopecia areata, male pattern hair loss and female pattern hair loss, alopecia caused by systemic diseases (chronic thyroiditis, iron deficiency anemia, connective tissue disease, etc.), drug-induced alopecia (alopecia caused by anticancer drugs, etc.), etc.

[0027]

[12] Keratin microparticles for use in a hair growth or hair restoration agent.

[0028]

[13] Keratin microparticles for use in the growth or restoration of hair in a subject.

[0029] Effects of the Invention

[0030] The present invention provides a hair growth or growth agent that can achieve high hair growth and growth effects. Keratin particles as an active ingredient are non-toxic to animals including humans, and therefore can be particularly effective as cosmetics and medicines. In addition, keratin particles are biodegradable, and therefore have little environmental burden. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 This is a photo showing the hair growth effect of keratin microparticles on mice.

[0032] Figure 2 The principal component analysis (PCA) graph and Venn diagram show changes in gene expression (up-regulation, down-regulation) caused by treatment with 1% minoxidil, 1% keratin aqueous dispersion (1% keratin), or 1% keratin microparticle suspension (1% keratin microparticle).

[0033] Figure 3 It is a scattergram showing changes in gene expression (up-regulation, down-regulation) caused by treatment with 1% keratin aqueous dispersion (1% keratin) or 1% keratin microparticle suspension (1% keratin microparticles).

[0034] Figure 4 It is a heat map of hierarchical clustering of changes in gene expression (up-regulation, down-regulation) caused by treatment with 1% keratin aqueous dispersion (1% keratin) or 1% keratin microparticle suspension (1% keratin microparticle).

[0035] Figure 5 This is a diagram showing the gene ontology (GO) up-regulated by the treatment with the aqueous dispersion of keratin fine particles or keratin powder.

[0036] Fig. 6A This is a diagram showing gene pathways that are upregulated by treatment with an aqueous dispersion of keratin fine particles or keratin powder.

[0037] Figure 6B This is a diagram showing gene pathways that are downregulated by treatment with an aqueous dispersion of keratin fine particles or keratin powder.

[0038] Figure 7 It is a Venn diagram and a heat map of hierarchical clustering of genes that changed when treated with 1% keratin aqueous dispersion and 1% keratin microparticle suspension.

[0039] Figure 8 This is a graph showing the fold change in gene expression in various biological processes caused by keratin microparticles.

[0040] Fig.9A The graph shows the results of microarray verification (Ctnnb1, Notch 1) by RT-PCR.

[0041] Fig. 9B The graph shows the results of microarray verification (DUX, Cdh11) by RT-PCR.

[0042] Fig. 9C It is a figure which shows the result of microarray verification (Lnk, K14) by RT-PCR.

[0043] Fig.10 This is a photograph showing that a keratin microparticle suspension is applied to a leather surface and then adsorbed onto the leather surface.

[0044] Fig.11 This is a photograph showing that a keratin microparticle suspension is applied to a leather surface and then adsorbed onto the leather surface.

[0045] Fig.12The photographs show the state before and after drying after dropping a keratin particle suspension on a glass. A: shows keratin spherical particles (dry spheres); B: shows the state of keratin spherical particles dispersed in water (swollen spherical gel); C: shows the state of keratin spherical particles dispersed in water dropped on a glass and dried (dried disc-like structure). DETAILED DESCRIPTION

[0046] The present invention is described in detail below.

[0047] The present invention relates to a hair growth or growth agent containing keratin microparticles as an effective ingredient for hair growth or growth. In the present invention, "hair growth" and "hair growth" refer to promoting the growth or growth of hair (such as hair, eyebrows, eyelashes, beards, etc.), increasing the amount of hair (number, thickness, etc.), reducing or inhibiting hair loss, improving the state of the hair growth site (such as inflammatory state, etc.), etc.

[0048] In the present invention, "keratin" is the main component protein of animal hair, claws, feathers, scales, hooves, wool, and horns. If it is keratin that can be used as cosmetics or medicines, it can be either natural or synthetic. If it is keratin originally from natural organisms or the environment, it can be keratin that has been pre-treated by purification and extraction, or keratin recombinantly produced according to its sequence information. In addition, as long as it has the same function as keratin from natural sources, it can be a variant or homolog thereof, or a chemically modified product. The average molecular weight (number average molecular weight) of keratin is, for example, 100 to 100,000, preferably about 200 to 40,000. In a preferred embodiment, keratin includes human keratin, keratin from wool or feathers, or water-soluble keratin to which these keratins are partially sulfonated or ionized. The keratin used can be one type, or two or more types of keratins can be used in combination.

[0049] Keratin can be conventionally prepared by those skilled in the art, or a commercially available product can be purchased.

[0050] The present invention uses keratin as microparticles with a particle size of nanometer to micrometer level. In the present invention, "microparticles" refer to spherical or substantially spherical particles with a particle size of about 0.01 to 500 μm, but are not limited to spherical shapes as long as they are small. In one embodiment, the particle size of the keratin microparticles is preferably 0.1 to 100 μm, more preferably 1 to 10 μm. In another embodiment, at least 90% of the keratin microparticles have a particle size of 0.05 to 100 μm, preferably 0.1 to 20 μm, more preferably 1 to 10 μm.

[0051] Keratin microparticles can be prepared by any method known in the art. For example, the following methods can be used: a method for preparing microparticles using keratin regenerated from wool (F. Cilurzo, et al., Polym. Adv. Technol., 2013, 24, 1025-1028), a method for preparing keratin microparticles using keratin extracted from human hair (Y. Srisuwan and P. Srihanam, Orient. J. Chem., 2019, 35, 1112-1116), a method for preparing microcapsules based on wool keratin (H. Rajabinejad, et al., Ultrason. Sonochem., 2018, 40, 527-532), etc.

[0052] In a preferred embodiment, the keratin solution is provided to a sprayer, an air gun or a spray pen using adjusted pressurized gas, and sprayed directly into a poor solvent (e.g., ethyl acetate) to allow the keratin aerosol to precipitate in the poor solvent. The suspension is then allowed to stand for 6 to 24 hours to allow the suspension to precipitate, and the precipitated keratin particles are recovered.

[0053] This will form keratin microparticles in the solution. The formed keratin microparticles are recovered by centrifugation and then washed with an appropriate solvent to remove unnecessary reagents.

[0054] The hair growth or growth agent of the present invention contains keratin particles as a hair growth or growth active ingredient. The keratin particles can be used directly, preferably dissolved, dispersed or suspended in a solvent. The solvent available is not particularly limited, and preferably a solvent acceptable to cosmetics or medicines is used. For example, the keratin particles are dissolved, dispersed or suspended in a solvent such as water, a salt solution (such as physiological saline, etc.). Therefore, the keratin particles can exist in the form of, for example, an aqueous solution, an aqueous dispersion, a hydrogel, etc.

[0055] Regarding the hair growth and / or hair growth effect, the hair growth or hair growth agent of the present invention can achieve rapid and / or large hair growth and / or hair growth effect, for example, compared with the case where no treatment is performed. Preferably, the hair growth or hair growth agent of the present invention has no side effects or has low side effects. Such side effects include, for example, inflammation (itching, rash, redness, etc.), initial hair loss, sweating, chills, palpitations, dizziness, headache, abdominal pain, liver dysfunction (nausea, loss of appetite, etc.), jaundice, loss of libido, hirsutism, etc.

[0056] The hair growth or growth agent of the present invention is not particularly limited to the object (subject) to be used. For example, it can be applied to mammals, such as humans, primates (monkeys, chimpanzees, etc.), livestock animals (cows, horses, pigs, etc.), pets (dogs, cats, etc.), and experimental animals (mice, rats, monkeys, etc.). For example, the animal subject having a desired part for hair growth or growth, particularly preferably a human subject.

[0057] The hair growth or hair growth agent of the present invention can be used as a composition. For example, keratin particles can be mixed with a suitable carrier or diluted or suspended in a carrier to form a composition. Examples of suitable carriers include: saline solution (e.g., saline solution, etc.), lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil, etc. In addition, the composition can contain conventional excipients, surfactants, dispersants, buffers, preservatives, cosolvents, stabilizers, isotonic agents, etc. In addition, other components or compositions in the keratin particles can be mixed or coordinated. There is no limitation to other ingredients, and examples thereof include other known hair growth or hair development active ingredients (ascorbic acid, ginkgo leaf extract, steinmosin, minoxidil, finasteride and dutasteride, etc.), and ingredients with other effects (hair active ingredients, moisturizers, softeners, anti-inflammatory drugs, minerals, amino acids, etc.).

[0058] The content of the hair growth or growth agent (keratin microparticles) in the composition varies depending on the form of the composition, the target of use, the purpose of use, etc., and can be appropriately determined by a person skilled in the art in consideration of the above factors. For example, the composition may contain 0.01% to 50% by weight of keratin microparticles, preferably 0.05% to 20% by weight of keratin microparticles.

[0059] In one embodiment, the hair growth or hair growth agent (keratin particles) of the present invention can be combined with cosmetics and used as a cosmetic composition. The combined cosmetics are not particularly limited as long as they are suitable for parts (such as hair, beard, etc.) that need hair growth or hair growth. For example, they can include: hair care products (shampoo, conditioner (Treatment), hair gel, hair mask, hair styling lotion, hair growth oil (Heatonic), etc.), lotion, emulsion, cream, etc. The cosmetic composition can include cosmetically acceptable excipients or carriers, such as extenders, adhesives, wetting agents, lubricants, surfactants, dispersants, buffers, pH adjusters, preservatives, solubilizers, preservatives, absorption promoters, stabilizers, isotonic agents, fragrances, etc. The cosmetic composition may also include other known hair growth or hair growth active ingredients (ascorbic acid, ginkgo leaf extract, steinmosin, minoxidil, finasteride and dutasteride, etc.), and other ingredients (hair active ingredients, moisturizers, softeners, anti-inflammatory drugs, minerals, amino acids, etc.). The cosmetic composition can be manufactured according to its form by conventional methods, such as mixing and the like.

[0060] In another embodiment, the hair growth or hair growth agent (keratin particles) of the present invention is combined with a drug and used as a pharmaceutical composition (e.g., a skin topical agent). It should be noted that in this specification, "drug" also includes quasi-drugs. The dosage form of the pharmaceutical composition can be set to an appropriate dosage form according to the applicable object or purpose, preferably a skin topical agent, such as an ointment, cream, lotion (liquid), gel, aerosol, transdermal plaster, etc., and can also be set to a dosage form suitable for other non-oral (e.g., intradermal administration, subcutaneous administration). The pharmaceutical composition may include a pharmaceutically acceptable excipient or carrier, such as an extender, adhesive, wetting agent, lubricant, surfactant, dispersant, buffer, pH adjuster, preservative, solubilizer, preservative, absorption promoter, stabilizer, isotonic agent, etc. The pharmaceutical composition may also include other known hair growth or hair growth active ingredients (minoxidil, finasteride and dutasteride, etc.), and other ingredients (moisturizer, softener, anti-inflammatory drug, mineral, amino acid, etc.). The pharmaceutical composition can be manufactured by conventional methods such as mixing, applying, spraying and the like according to its form.

[0061] The pharmaceutical composition can be used as an external skin preparation, for example, to treat or prevent diseases or symptoms in areas where hair growth or development is required, such as alopecia areata, male-pattern and female-pattern hair loss, alopecia caused by systemic diseases (chronic thyroiditis, iron deficiency anemia, connective tissue disease, etc.), and drug-induced alopecia (alopecia caused by anticancer drugs, etc.).

[0062] The amount of the composition to be used, the number of times of use, and the duration of use can be appropriately set according to the amount of keratin particles contained, the amount of hair on the target, the condition, etc. When applied to the skin, generally 1 μg to 50 mg / cm can be used at a time. 2 It can be used at a frequency of 1 to 3 times a day, once every 2 days, once every 3 days, once a week for 1 week to 3 months, further for 1 month to 1 year or more, and can be used for several years to several decades when necessary.

[0063] The present invention also relates to a method for growing or restoring hair in a subject, comprising applying (e.g., applying to the skin) the hair growing or restoring agent (keratin microparticles) of the present invention to the subject. In addition, the present invention relates to a method for treating or preventing alopecia in a subject, comprising applying (e.g., applying topically to the skin, subcutaneously or intradermally, etc.) the hair growing or restoring agent (keratin microparticles) of the present invention to the subject.

[0064] In addition, by combining keratin particles with cells, for example, they can be used as materials for regenerative medicine. For example, by applying a keratin particle suspension on a 3D sphere and then administering or transplanting the 3D sphere to a subject, the effect of keratin can be exerted in the subject. Alternatively, by administering cells (such as hair matrix cells) together with keratin particles, the hair growth and hair growth effects of keratin can be exerted.

[0065] Example

[0066] The present invention is described in more detail below through examples and drawings. It should be noted that the following examples do not limit the present invention.

[0067] [Example 1] Preparation of keratin microparticles

[0068] Keratin microparticles (also called keratin spherical microparticles) are prepared as follows. 1 to 2 mL of a water-soluble keratin aqueous solution (concentration 0.1 wt% to 2 wt%, partially sulfonated, TCI) is provided to an ultrasonic aerosol generator, and the generated keratin aerosol is added dropwise to 100 mL of a poor solvent (e.g., EtOAc) filled in a 200 mL beaker. The resulting suspension is allowed to stand for 6 to 24 hours to allow the generated keratin microparticles to precipitate. The precipitated keratin microparticles are filtered and recovered, and washed several times with a poor solvent. Finally, the keratin microparticles are dried at 25°C, and then dried in a vacuum oven (40°C, -0.1 MPa) to remove the residual solvent.

[0069] [Example 2] Verification of the hair growth effect of keratin microparticles on mice

[0070] This example investigates the effect of keratin microparticles on hair growth in mice. The experiment was approved by the Animal Experiment Committee of the University of Tsukuba, Japan (No. 17-060), and the experimental procedures were carried out in accordance with the "Basic Guidelines on Animal Experiments in Physiology" approved by the Board of Directors of the Japanese Physiological Society.

[0071] Six-week-old male C57BL / 6 mice (Charles River Laboratories Japan, Inc.) were housed in cages and acclimated for 1 week, and then randomly divided into experimental groups (n=5 per group), namely, a 1% minoxidil treatment group, a 1% keratin treatment group, a 1% keratin microparticle treatment group, and a milli-Q water treatment group (control group). After the mice were anesthetized with isoflurane (Wako Pure Chemical Industries, Ltd.), the resting hair on the back of the mice was shaved with a shaver to induce the growth phase. Then, 1% minoxidil (Tokyo Chemical Industry Co., Ltd.), 1% keratin aqueous dispersion (Tokyo Chemical Industry Co., Ltd.), 1% keratin microparticle suspension (obtained by suspending the keratin microparticles prepared in Example 1 in water), or milli-Q water was topically applied to each mouse every day for 4 weeks. The shaved part of the back was observed, and photos were taken every day from the 14th day after treatment.

[0072] At the end of the treatment period, mice were killed by cervical dislocation, and the skin collected from the treated area was divided into four parts, washed with phosphate-buffered saline (PBS), and directly immersed in liquid nitrogen and kept at -80°C. Hair regrowth in the treated area was measured using the ImageJ processing program (National Institutes of Health, Bethesda, USA).

[0073] The results are shown in Figure 1 . In mice treated with the keratin aqueous dispersion, hair growth was observed to be the same as in mice treated with nothing (control). On the other hand, mice treated with keratin microparticles showed the same degree of hair regrowth phenotype as mice treated with minoxidil at approximately the same time (10 days after application), and the hair cycle was accelerated compared to the control and keratin aqueous dispersion, thus demonstrating a hair growth promoting effect.

[0074] [Example 3] Verification of the hair growth effect of keratin microparticles on mice

[0075] In this example, DNA microarray was used to perform gene analysis on the skin cells of the mice treated in Example 2.

[0076] RNA used as template was extracted from skin tissue collected from the treated area of ​​the mouse back in Example 2 using ISOGEN solution (Nippon Gene) according to the manufacturer's instructions. The extracted total RNA was amplified and labeled. The labeled fragmented RNA was then hybridized with Affymetrix mouse array strips (Affymetrix). The gene chip (mouse genome 4302.0 array) was washed and stained, and scanned using the Affymetrix GeneAtlas imaging workstation to obtain mRNA expression of various genes from the mouse genome.

[0077] Then, the gene ontology, biological process, and fold change of gene expression (2-fold change, control vs. 1% minoxidil, 1% keratin aqueous dispersion, 1% keratin microparticles) were analyzed using the Transcriptome Analysis Console (TAC) software (version 4.0.1) and the database for annotation, visualization, and integrated discovery (DAVID) bioinformatics resource 6.8. Hierarchical clustering of the mean signal log2 (control vs. 1% minoxidil, 1% keratin aqueous dispersion, 1% keratin microparticles) was performed using the Euclidean distance and average linkage algorithm of TIGR Mev version 3.0.3 software (American Genome Institute, MD, USA).

[0078] The results are shown in Figure 2 ~9. It should be noted that the results are expressed as mean ± standard deviation (SD) (n = 2 for each). When comparing two value groups (control 1% minoxidil, 1% keratin aqueous dispersion, 1% keratin microparticles), Student's t-test was used for statistical analysis. P value ≤ 0.05 was considered significant. In order to evaluate the significance level between treatment groups in vivo, ANOVA [p value (condition pair) < 0.05] was performed. It should be noted that in Figure 2 In 9, the 1% keratin aqueous dispersion is simply referred to as keratin.

[0079] Figure 2 The genes up-regulated (>2) or down-regulated (<-2) by each treatment (1% minoxidil, 1% keratin aqueous dispersion, 1% keratin microparticles) compared to the control were represented by principal component analysis (PCA), Venn Diagram and graph representing differentially expressed genes. Figure 2 The results showed that any treatment observed different gene expression from the control. The total number of differentially expressed genes, the number of up-regulated genes, and the number of down-regulated genes are summarized in Table 1 below. The table gives the number of expressed genes sufficient for analysis.

[0080] [Table 1]

[0081]

[0082] Then, through the scatter plot ( Figure 3 ) and Hierarchical clustering ( Figure 4 )Analyze the changes in gene expression (up-regulated, down-regulated) when treated with 1% keratin aqueous dispersion and 1% keratin microparticle suspension compared with the control. Of the total 45,077 genes examined, 11,018 (89.87%) of the 12,260 genes (27.2%) that met the criteria were up-regulated, and 1,242 (10.13%) were down-regulated when treated with 1% keratin microparticle suspension. Of the 9,387 (20.82%) genes that met the criteria, 8,403 (89.52%) were up-regulated, and 984 (10.48%) were down-regulated when treated with 1% keratin microparticle suspension. Figure 3 As shown in the scatter plot, the changes in gene expression caused by 1% keratin aqueous dispersion and 1% keratin microparticles are different from those of the control. Figure 4 The hierarchical clustering heatmap shown shows that different gene expression patterns were produced by keratin aqueous dispersion and keratin microparticles when compared to the control.

[0083] Figure 5 Indicates the gene ontology (GO) that is upregulated by the treatment of keratin microparticles or keratin aqueous dispersion compared to the control. For example, compared to the control, genes related to epithelial development, cellular response to stress, metabolic process, cell cycle, homeostatic process, tissue development, mitochondrial activity, and oxidative phosphorylation are upregulated, and keratin microparticles contribute to the upregulation of genes related to the cell cycle. In other words, keratin aqueous dispersion and keratin microparticles upregulate the gene ontology of hair development, epidermal development, and cell adhesion, and this effect is more significant in the keratin microparticle treatment group.

[0084] Fig. 6A and Figure 6BThe diagram shows gene pathways that are upregulated (A) or downregulated (B) by the treatment with keratin microparticles or keratin aqueous dispersions compared to the control. For example, compared to the control, genes in the Hallmark of Myc Target, VEGF Signaling Pathway, and Innate Immune System pathways are upregulated, and genes in the Pathways of Neurodegeneration are downregulated.

[0085] Then, the gene expression induced by the treatment with keratin microparticles and keratin aqueous dispersion was compared. Of the 1,148 genes that met the criteria out of a total of 45,077 genes examined, 616 genes were upregulated and 532 genes were downregulated by the application of keratin microparticles. Figure 7 The Venn diagram and the heat map of hierarchical clustering show the genes that change when treated with 1% keratin aqueous dispersion (keratin) and 1% keratin microparticle suspension (keratin microparticle). Figure 8 The expression of genes in each biological process is shown as a multiple of change. For example, the expression of genes involved in the processes of cell adhesion, hair follicle morphogenesis, and muscle contraction regulation increased significantly. These results indicate that the treatment with keratin microparticles increases genes related to hair growth, such as the cell cycle and hair cycle, and decreases genes related to inflammation and stress, indicating a mechanism that brings about hair growth and development without showing toxicity and side effects.

[0086] The results of microarray validation by RT-PCR are shown in Figures 9A to 9C In addition, the function and expression fold change of each gene are summarized in the following Table 2.

[0087] [Table 2]

[0088]

[0089] From the above results, the mechanism of the keratin microparticle treatment group is to activate cell adhesion molecules as targets, which is mainly associated with the activation of hair growth-related pathways. The results showed that the hair growth-related pathways Notch, shh, JAK / STAT, and BMP were activated. The expression of genes was significantly increased compared with the keratin water dispersion treatment group.

[0090] In addition, it was shown that the expression of genes related to the migration of epidermal cells and mesenchymal stem cells increased, the communication between keratinocytes, melanocytes, and hair papillae was promoted, and the hair growth cycle was further strengthened. Since the differentiation ability of epidermal stem cells and keratinocytes, which are the origin of hair matrix cells, is improved, the effect on epidermal stem cells can be expected to be greater.

[0091] Mesenchymal stem cells such as those in the hair papilla are located in the deepest part of the skin, the dermis, and the keratin in the keratin aqueous dispersion was not completely dissolved, so it may not have reached there. This is because the genes for growth factors and keratin located in the epidermis were targeted, and these molecules activated genes related to hair growth in the hair papilla. In addition, Notch pathway-related genes such as Notch 1, which are known to inhibit the proliferation of keratinocytes and promote differentiation related to hair follicle development, were activated, which further supports the above explanation.

[0092] [Example 4] Application characteristics of keratin microparticle suspension on leather surface

[0093] This example tests the adsorption and drying state of keratin microparticle suspensions on leather surfaces after applying them. Specifically, fluorescent pigment (Acid Red 52) was added to the keratin microparticles (1% suspension) and keratin aqueous solution (1% suspension) prepared in Example 2, and then each suspension was applied to the surface of pig leather to confirm its adsorption on the leather surface. In addition, a 1% aqueous dispersion of keratin powder was also applied to the surface of pig leather to confirm its adsorption on the leather surface.

[0094] The results are shown in Fig.10 , 11 .like Fig.10 As shown, when a keratin aqueous solution or a 1% aqueous dispersion of keratin powder was applied, keratin aggregated after drying; whereas, when a 1% aqueous dispersion of keratin microparticles was applied, keratin microparticles were uniformly dispersed over the entire leather surface. Fig.11 The fluorescence microscopy images shown also clearly show that keratin microparticles are dispersed throughout the surface.

[0095] In addition, after applying a suspension of keratin particles on glass and letting it dry naturally, it was observed that disc-shaped (disk-shaped) structures covered the surface ( Fig.12 ).like Fig.12As shown, the particle size of keratin spherical particles (spheres in a dry state) is 1 to 10 μm (A); the particle size of the state in which keratin spherical particles are dispersed in water (swollen spherical gel) is 10 to 100 μm (B); the diameter of the state after the keratin spherical particles dispersed in water are dropped on glass and dried (dried disc-like structure) is 10 to 100 μm (C). This shows that the keratin particles form a swollen spherical gel in water and are not completely dissolved. From this, it can be inferred that keratin forms a three-dimensional mesh structure in water, which will induce the same characteristics as the hair growth effect of keratin in 3D cells.

[0096] [Example 5] Application of Keratin Microparticle Suspension to Human Subjects

[0097] The effect of applying keratin microparticles to human subjects was investigated according to the following steps: The 1% keratin microparticle aqueous solution prepared in Example 2 was used as the test solution.

[0098] (1) Use an electric razor and hair clipper to shave the sideburns on both sides.

[0099] (2) Use a cotton swab to apply a 1% aqueous solution of keratin microparticles to an area of ​​about 3 x 6 cm, including the right temple. Apply about 0.4 mL each time, twice a day, in the morning (after waking up) and at night (after bathing).

[0100] The results of the above test showed that no side effects such as allergies and rashes occurred after 14 days. In addition, it was confirmed that the hair growth area of ​​the applied part of the right temple extended further downward compared to the unapplied left temple.

[0101] All publications, patents and patent applications cited in this specification are incorporated herein by direct reference.

[0102] Industrial Applicability

[0103] The present invention provides a hair growth or growth agent that can achieve high hair growth and growth effects. The present invention can be used in the fields of cosmetic raw materials, cosmetics, medicines, etc.

Claims

1. A hair growth or growth agent comprising keratin microparticles as a hair growth or growth effective ingredient.

2. The hair growth or growth agent according to claim 1, wherein The particle size of the keratin particles is 0.01-100 μm.

3. The hair growth or growth agent according to claim 1, wherein The keratin microparticles are in the form of an aqueous solution, an aqueous dispersion or a hydrogel.

4. The hair growth or growth agent according to claim 1, wherein The keratin includes human keratin, wool-derived keratin, or partially sulfonated or ionized water-soluble keratin.

5. A cosmetic composition comprising the hair growth or growth agent according to claim 1 and a cosmetically acceptable excipient.

6. A pharmaceutical composition comprising the hair growth or growth agent according to claim 1 and a pharmaceutically acceptable excipient. The pharmaceutical composition according to claim 6 , which is an external preparation for skin.

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