Composition for maintaining hair health or growing hair

Abstract

The present invention relates to a composition for maintaining hair health or growing hair, the composition containing phycocyanin as an active ingredient.

Description

Composition for maintaining hair health or for hair growth

[0001] The present invention relates to a composition containing phycocyanin.

[0002] Cyanobacteria, particularly those of the genus Spirulina, are known to contain phycocyanin, which exhibits a blue color. Much research has been conducted on the biological activity of phycocyanin. For example, a lipase activity inhibitor containing phycocyanin as an active ingredient has been reported, stating that phycocyanin inhibits the activity of lipases such as pancreatic lipase (Patent Document 1). Furthermore, a serum lipid-lowering agent containing phycocyanin as an active ingredient has been reported, stating that phycocyanin has a higher serum lipid-improving effect than soy protein (Patent Document 2). Furthermore, a skin-beautifying food product containing phycocyanin as an active ingredient has been reported, stating that oral intake of phycocyanin exhibits skin-beautifying effects (Patent Document 3).

[0003] JP 2004-359638 A JP 2003-137805 A International Publication No. 2020 / 230347

[0004] The effect of phycocyanin on hair was not particularly known.

[0005] An object of the present invention is to provide a composition suitable for maintaining hair health or promoting hair growth.

[0006] As a result of intensive research to solve the above problems, the present inventors have discovered that phycocyanin has an effect suitable for maintaining hair health or promoting hair growth, and have completed the present invention.

[0007] That is, the present invention encompasses the following aspects. [1] A composition for maintaining hair health or growing hair, comprising phycocyanin as an active ingredient. [2] The composition according to [1], wherein the phycocyanin is derived from cyanobacteria. [3] The composition according to [2], wherein the cyanobacteria is a cyanobacteria of the genus Spirulina. [4] The composition according to any one of [1] to [3], wherein the phycocyanin is C-phycocyanin, allophycocyanin, or a combination thereof. [5] The composition according to any one of [1] to [4], wherein the maintenance of hair health is hair moisturizing, improving hair strength, improving hair thickness, suppressing or improving hair loss, improving the cuticle, maintaining a normal cuticle, improving frizz, improving hair firmness and body, improving hair suppleness, improving hair luster, improving hair volume, or maintaining normal hair color. [6] The composition according to any one of [1] to [5], which is ingested or administered to a subject in which expression of at least one gene selected from the group consisting of LIPH, TCHHL1, KRTAP6-1, KRT71, KRTAP6-2, KRTAP19-1, KRTAP19-3, KRTAP19-4, KRT36, SCD, KRT25, KRT33A, KRTAP13-1, and KRTAP13-2 in hair follicle cells is reduced compared to normal individuals. [7] The composition according to any one of [1] to [6], which is an oral composition. [8] The composition according to any one of [1] to [6], which is an external composition. [9] The composition according to any one of [1] to [6], which is a composition for scalp and / or hair.

[0008] The composition of the present invention has an effect suitable for maintaining hair health or for promoting hair growth, and therefore can be used for these purposes.

[0009] 1 is a graph showing the hair regeneration scores of mice administered 50% ethanol, a commercially available hair growth agent, and phycocyanin in Test Example 2. 2 is a photograph of the 50% ethanol and phycocyanin administration groups on days 9, 13, and 17 of administration in Test Example 2.

[0010] Preferred embodiments of the present invention will be described in detail below. However, the present invention is not limited to the following embodiments. In this specification, the upper and lower limits individually described can be combined in any combination.

[0011] As used herein, the term "cyanoalgae" is used synonymously with the division Cyanobacteria and encompasses the class Cyanophyceae, including the genus Spirulina. Furthermore, as used herein, the terms red algae and cryptophytes are used synonymously with the division Rhodophyta and Cryptophyta, respectively.

[0012] As used herein, "phycocyanin" (PC) is a general term for pigment proteins bound to at least phycocyanobilin, and includes allophycocyanin, C-phycocyanin, R-phycocyanin, and complexes thereof (e.g., the entire or partial phycobilisomes containing phycocyanobilin). Phycocyanin is known to be present in organisms of cyanobacteria, red algae, cryptophytes, Glaucophyta, and the Paulinella genera. As used herein, "C-phycocyanin" (CPC) is a type of phycocyanin found primarily in cyanobacteria, containing only phycocyanobilin as a chromophore. C-phycocyanin is typically composed of α and β subunits, with one chromophore in the α subunit and two in the β subunit, and is known to constitute the peripheral rod-shaped portion of phycobilisomes. In this specification, "R-phycocyanin" refers to a type of phycocyanin that contains phycoerythrobilin as well as phycocyanobilin as a chromophore. R-phycocyanin is also typically composed of α and β subunits, with one chromophore in the α subunit and two in the β subunit. In this specification, "allophycocyanin" (APC) refers to a type of phycocyanin and is the main protein component of the phycobilisome core structure. Allophycocyanin is a cylindrical trimer (αβ) in which one molecule of phycocyanobilin is bound to each of the α and β subunits. ３ The structure is as follows:

[0013] Gene names in this specification are represented by the human gene symbols (official symbols of the Human Genome Nomenclature Committee (HGNC)) used in the NCBI Gene database (https: / / www.ncbi.nlm.nih.gov / gene). When a specific gene name is represented by a human gene symbol, in animals other than humans, it refers to the ortholog of the gene for each animal (for example, one identified in the above-mentioned Gene database, etc.).

[0014] As used herein, "improvement" of a disease, symptom, health condition, or aesthetic condition refers to curing, improving, or alleviating the disease, symptom, health condition, or aesthetic condition; preventing or slowing the deterioration of the disease, symptom, health condition, or aesthetic condition; or reversing, preventing, or slowing the progression of the disease or symptom.

[0015] As used herein, "prevention" refers to preventing or delaying the onset of a disease or condition, or reducing the incidence or risk of a disease or condition.

[0016] As used herein, the term "subject" refers to a living organism to which a composition is administered. As used herein, the term "patient" is interpreted as being synonymous with "subject."

[0017] [Composition for maintaining healthy hair or for growing hair] The composition of the present invention for maintaining healthy hair or for growing hair contains phycocyanin as an active ingredient.

[0018] (Active ingredient: phycocyanin) Examples of organisms from which phycocyanin is derived include at least one selected from the group consisting of cyanobacteria, red algae, cryptophytes, glaucophytes, and Paulinella. Among these, at least one selected from cyanobacteria is preferred because they can be harvested industrially in large quantities. Examples of cyanobacteria include the genus Spirulina, Arthrospira, Aphanizomenon, Fisherella, Anabaena, Nostoc, Synechocystis, Synechococcus, Tolypothrix, Aphanothece, Mastigoclaus, and Pleurocapsa. Among these, cyanobacteria of the genus Spirulina and Arthrospira are preferred, as they are produced on an industrial scale and their safety has been confirmed, cyanobacteria of the genus Spirulina are more preferred, and Spirulina (Arhrospira platensis) is even more preferred.

[0019] As the raw material for preparing phycocyanin, a raw organism can be used, or a product that has been processed by drying or the like can be used. The dried product of the organism can be a raw organism that has been dried according to a conventional method, or a commercially available dried product can be used. A single organism can be used as the raw organism, or multiple organisms can be mixed and used. Alternatively, multiple biological raw materials can be processed separately, and the resulting phycocyanins can be used in combination.

[0020] From the viewpoints of quality, safety, availability, etc., the phycocyanin preferably contains at least C-phycocyanin, and more preferably contains C-phycocyanin as the main component. When the total content of phycocyanin in the composition of the present invention is taken as 100% by mass, the content of C-phycocyanin is preferably 50% by mass or more, and can be, for example, 60% by mass or more, 70% by mass or more, 80% by mass or more, 90% by mass or more, or 95% by mass or more.

[0021] In a more preferred embodiment, the phycocyanin is a composition containing C-phycocyanin and allophycocyanin. The C-phycocyanin and allophycocyanin are preferably derived from cyanobacteria of the genus Spirulina. The ratio of C-phycocyanin to allophycocyanin, by mass, is preferably 3-9.5:0.5-7, more preferably 6-9.5:0.5-4, and even more preferably 7-8:2-3.

[0022] In another embodiment, the composition of the present invention may be a composition containing only C-phycocyanin as the phycocyanin.

[0023] (Method for Producing Phycocyanin) Phycocyanin can be obtained, for example, by suspending a biological material containing phycocyanin in water or a buffer solution such as a phosphate buffer or a citrate buffer, and extracting the phycocyanin from the cyanobacteria. There are no particular limitations on the method for extracting phycocyanin, and any commonly known method can be used. By appropriately selecting the extraction conditions in the extraction method, the abundance ratio of each component (allophycocyanin, C-phycocyanin, R-phycocyanin, etc.) contained in the phycocyanin can be adjusted. Alternatively, the abundance ratio of each component can be adjusted as desired by mixing multiple phycocyanins with different abundance ratios of each component.

[0024] A preferred embodiment of the extraction method, for example, when blue-green algae are used as the raw material, is the extraction method described in JP 2006-230272 A. Specifically, the extraction method described in the following extraction method (i) can be mentioned. By this extraction method (i), phycocyanin with high purity and vivid color can be obtained.

[0025] <Extraction method (i)> Extraction method (i) comprises the following steps: a first step of extracting phycocyanin from cyanobacteria into an aqueous suspension to obtain an extract; a second step of reacting a calcium salt with a phosphate salt in the extract to produce calcium phosphate and adsorbing impurities of phycocyanin onto the calcium phosphate to obtain an adsorbate; and a third step of removing cyanobacterial residues and the adsorbate from the extract.

[0026] Furthermore, it is more preferable that the above extraction method (i) is the following extraction method (ii). <Extraction method (ii)> Extraction method (ii) comprises the following steps: a first step of extracting phycocyanin from cyanobacteria into an aqueous suspension to obtain an extract; a second step of reacting a calcium salt with a phosphate salt in the extract to produce calcium phosphate and adsorbing phycocyanin impurities onto the calcium phosphate to obtain an adsorbate; a third step of removing cyanobacteria residues and the adsorbate from the extract; and a step of adding a chelating agent to the extract prior to the third step.

[0027] By using the above extraction method (i) or (ii), high-quality phycocyanin can be extracted from cyanobacteria. In particular, by using the above extraction method (i) or (ii) for cyanobacteria of the genus Spirulina, high-quality phycocyanin with a good mixture ratio of C-phycocyanin and allophycocyanin can be extracted.

[0028] (Action, Use) As shown in the examples below, phycocyanin has the effect of promoting the expression of at least one gene selected from the group consisting of LIPH, TCHHL1, KRT71, KRTAP6-1, KRTAP6-2, KRTAP19-1, KRTAP19-3, KRTAP19-4, KRT36, SCD, KRT25, KRT33A, KRTAP13-1, and KRTAP13-2. Here, promotion of gene expression refers to an increase in the expression level (transcription level) of mRNA in the skin (e.g., hair follicles).

[0029] LIPH is a gene encoding a lipase that synthesizes 2-acylphosphatidic acid from phosphatidic acid (1,2-diacylglycerol 3-phosphate). It is known to be one of the causative genes for woolly hair, a human genetic disorder, and to be essential for normal hair follicle differentiation. LIPH expression is also known to be localized in the inner root sheath within hair follicles. TCHHL1 protein is also localized distal to the inner root sheath (close to the surface of the skin) and is thought to be involved in hair growth from the epidermal surface. Therefore, abnormal expression of LIPH and TCHHL1 can result in abnormal differentiation or activation of hair follicles (particularly the inner root sheath). Furthermore, within hair follicles, the inner root sheath keratinizes early in hair follicle differentiation and serves as a "mold" for unkeratinized hair. As can be seen from the above, decreased expression of LIPH and TCHHL1 can lead to abnormalities in the hair shaft and even hair morphology (abnormalities in hair strength or thickness, abnormalities in the cuticle, frizzy hair, abnormalities in hair firmness, body, luster or suppleness, abnormalities in hair volume, etc.). Furthermore, the innermost layer of the inner root sheath, the cuticle, "interlocks" with the hair cuticle (so-called "cuticle") to prevent the hair shaft from falling out of the hair follicle. Therefore, decreased expression of LIPH and TCHHL1 can lead to hair loss and hair loss.

[0030] Keratin is a protein produced specifically by epithelial cells and is a major component of hair. KRT71 is a gene encoding type II epithelial keratin, a causative gene for woolly hair that occurs in hypotrichosis, and is known to be expressed throughout the entire inner root sheath. KRT25 is a gene encoding type I epithelial keratin, a causative gene for woolly hair, and is known to be expressed in the inner root sheath. KRT36 is a gene encoding type I epithelial keratin. KRT33A is a gene encoding type I hair keratin. KRTAP6-1, KRTAP6-2, KRTAP19-1, KRTAP19-3, KRTAP19-4, KRTAP13-1, and KRTAP13-2 are genes encoding keratin-associated proteins, respectively. In other words, decreased expression of genes encoding these keratins and keratin-associated proteins can cause abnormalities in hair moisture retention, abnormalities in hair shape, abnormalities in hair color, hair loss, etc. through a decrease in the main components of hair.

[0031] SCD is a gene that encodes an enzyme that desaturates palmitoyl coenzyme A (palmitoyl CoA) by introducing a double bond between the 9th and 10th carbons. SCD has been reported to be expressed in the hair matrix, the lowest part of the hair follicle, and may be involved in hair matrix activation. Therefore, decreased SCD expression can cause hair thinning, hair loss, etc. through decreased hair matrix function.

[0032] Therefore, the composition of the present invention can be used for maintaining hair health (e.g., moisturizing hair, improving hair strength, improving hair thickness, suppressing or improving hair loss, improving the cuticle, maintaining a normal cuticle, improving frizzy hair, improving hair firmness and body, improving hair suppleness, improving hair gloss, improving hair volume, or maintaining normal hair color) or for hair growth (e.g., promoting hair growth, promoting hair growth, or preventing or improving thinning hair). It can be understood that the composition of the present application is suitable for these uses based on the action of each of the above-mentioned genes, but it is possible that actions suitable for these uses may be exhibited by combining multiple genes.

[0033] In one embodiment, maintaining hair health or promoting hair growth relates to the prevention or amelioration of diseases, conditions, health conditions or aesthetic conditions caused in part by ultraviolet radiation.

[0034] The composition of the present invention may be a composition that displays the above-mentioned use as an efficacy or function. Examples of such display include a direct display of the above-mentioned use or a display that substantially implies the above-mentioned use. Display that substantially displays the above-mentioned use include, for example, a figurative display of the above-mentioned use, a display of a target for which ingestion or administration is recommended, and a display indicating an effect on a specific disease, symptom, or health condition. Examples of such displays include, but are not limited to, "for healthy hair," "for those with lackluster hair," "for those concerned about hair loss," "for those concerned about hair quality (including, for example, hair shine, color, cuticle, etc.)," ​​"hair care," "scalp care," "healthy hair," "fluffy," "(hair) volume increase," "black hair," "approach to hair follicles," "stimulates hair follicles," etc.

[0035] Furthermore, the compositions of the present invention can also be used for activating hair follicles, for example activating the inner root sheath (cells) or activating the hair matrix (cells).

[0036] The composition of the present invention can be ingested or administered to a subject (e.g., a human or non-human animal) for each of the above uses. The non-human animal can be, for example, a mammal, more specifically, a mammal known as a companion animal or livestock, such as a dog, cat, rabbit, horse, cow, sheep, pig, rat, hamster, or mouse.

[0037] The compositions of the present invention can be used for therapeutic purposes (e.g., medical treatments) as well as non-therapeutic purposes. Non-therapeutic purposes refer to uses that do not involve medical procedures, such as surgical, therapeutic, or diagnostic procedures on a subject by a physician or a person under the supervision of a physician (e.g., uses related to maintaining or improving health, or cosmetics).

[0038] The compositions of the present invention can be ingested or administered to, for example, a subject in need of maintaining healthy hair or promoting hair growth (e.g., a subject experiencing or at risk of hair moisture retention, abnormalities in hair shape or color, or hair loss or alopecia). Such a subject is, for example, a subject in which expression of at least one gene selected from the group consisting of LIPH, TCHHL1, KRTAP6-1, KRT71, KRTAP6-2, KRTAP19-1, KRTAP19-3, KRTAP19-4, KRT36, SCD, KRT25, KRT33A, KRTAP13-1, and KRTAP13-2 in hair follicle cells is reduced relative to normal individuals. Here, the reduction in gene expression can be such that the expression level of the gene (e.g., mRNA expression level) relative to normal individuals is 70% or less, 50% or less, 30% or less, 20% or less, or 10% or less.

[0039] The age of the subject is not particularly limited, and for example, the subject is an adult, preferably in his / her 30s or older, more preferably in his / her 40s or older, and even more preferably in his / her 50s or older. The gender of the subject is not particularly limited, and for example, the subject may be male or female.

[0040] (Form of composition) The composition of the present invention is a composition for oral or external use. As can be seen from the examples, the composition of the present invention is surprisingly suitable for the use of the present invention even when taken orally. The composition of the present invention may be, for example, a pharmaceutical product, a quasi-drug, a cosmetic product, a food or drink, a feed or a feed, or a preparation for producing them (e.g., an additive, etc.).

[0041] Examples of pharmaceutical forms include oral preparations (liquids, tablets, powders, granules, capsules, powders, troches, syrups), topical preparations (liquids, creams, emulsions, gels, lotions, poultices, liniments, mists, other transdermal preparations, etc.), and injections.

[0042] Oral quasi-drugs include, for example, liquids, tablets, powders, granules, capsules, powders, lozenges, and syrups.

[0043] Examples of external quasi-drugs or cosmetics include shampoos, rinses, hair conditioners, hair tonics, lotions, hair liquids, hair sprays, waxes, gels, mousses, styling agents, coloring agents, perm solutions, ointments, lotions, emulsions, creams, and mascaras.

[0044] Examples of foods and beverages include confectioneries (caramel, candy, capsule candy, tablet candy, etc.), frozen desserts (ice cream, etc.), dairy products (yogurt, etc.), jellies, various processed foods, various seasonings, various beverages, functional foods, nutritional supplements, and supplements. The foods and beverages are preferably foods labeled with the use or efficacy or function related to the use of the present invention, and more preferably foods for specified health uses or foods with functional claims. Furthermore, examples of preparations for producing foods and beverages include preparations treated as food or food additives.

[0045] When the composition of the present invention is in the form of a beverage, the total amount of phycocyanin is preferably 0.01% by mass or more, more preferably 0.06% by mass or more, and preferably 10% by mass or less, more preferably 3% by mass or less, based on the total amount of the composition. When the composition of the present invention is in the form of a food, the total amount of phycocyanin is preferably 0.008% by mass or more, more preferably 0.06% by mass or more, and preferably 99% by mass or less, more preferably 70% by mass or less, based on the total amount of the composition. When the composition of the present invention is in the form of a tablet, the total amount of phycocyanin is preferably 2.5% by mass or more, more preferably 20% by mass or more, and preferably 98% by mass or less, more preferably 70% by mass or less, based on the total amount of the composition.

[0046] The composition of the present invention may further contain, depending on its form and intended use, at least one selected from the group consisting of hair growth active ingredients [minoxidil, finasteride, dutasteride, carpronium chloride, bimatoprost, trans-3,4'-dimethyl-3-hydroxyflavanone (t-flavanone), 6-benzylaminopurine (cytopurine), pentadecanoic acid glyceride (pentadecane), adenosine, panthenol (D-pantothenyl alcohol), pantothenyl ethyl ether, DL-α-tocopherol acetate, dipotassium glycyrrhizinate, ginseng extract, Swertia japonica extract, Scutellaria Root extract, and Ginkgo biloba extract] and vitamins (vitamins A, B, C, D, E, etc.).

[0047] The composition of the present invention may further contain ingredients other than phycocyanin, the above-mentioned hair growth active ingredient, and vitamins (referred to as "other ingredients"). When the composition of the present invention is in the form of a drug, quasi-drug, cosmetic, or a preparation for producing them, the other ingredients may include, but are not limited to, any one or a combination of two or more selected from the group consisting of pharmaceutically acceptable carriers (excipients, disintegrants, binders, lubricants, colorants, plasticizers, antioxidants, pH adjusters, thickeners, flavoring agents, coating agents, surfactants, stabilizers, preservatives, fragrances, fluidizing agents, liquid media, foaming agents, UV absorbers, antioxidants, moisturizers, etc.). When the oral composition of the present invention is in the form of a food or drink, feed or bait, or a preparation for producing them, examples of other ingredients include, but are not limited to, any one or a combination of two or more selected from the group consisting of fish meat, vegetables, grains, dairy products, fermented products, spices, proteins, amino acids, sugars, various seasonings, sweeteners, flavorings, fragrances, oils and fats, vitamins, thickeners, gelling agents, antioxidants, preservatives, chelating agents, pH adjusters, and colorants.

[0048] (Dosage and Administration) When the composition of the present invention is an oral composition or an injection, the dosage or intake amount can be selected according to the subject's body weight, sex, age, condition, or other factors. The daily dose, in terms of the total amount of phycocyanin, can be, for example, 0.1 mg / kg body weight or more, 0.2 mg / kg body weight or more, 0.5 mg / kg body weight or more, or 1 mg / kg body weight or more, and can be 5,000 mg / kg body weight or less, 1,000 mg / kg body weight or less, 500 mg / kg body weight or less, or 100 mg / kg body weight or less.

[0049] When the composition of the present invention is a topical composition, it can be administered to, for example, the scalp, eyebrows, eyelids (where eyelashes grow), chin, around the mouth, chest, etc., but is preferably administered to the scalp. The amount of a single dose can be selected depending on the subject's weight, sex, age, condition, administration site, or other factors. ２ A single dose per may be, for example, 0.1 mg or more, 0.5 mg or more, 1 mg or more, 2 mg or more, 5 mg or more, 10 mg or more, 20 mg or more, 50 mg or more, or 100 mg or more, and may be, for example, 5,000 mg or less, 2,000 mg or less, 1,000 mg or less, 500 mg or less, or 200 mg or less.

[0050] The compositions of the present invention are ingested or administered, for example, 1 to 5 times, 1 to 3 times, or 2 to 3 times per day. The period of ingestion or administration can be, for example, one month or more, three months or more, six months or more, or one year or more. The period of ingestion or administration may optionally include intervals of, for example, several days to several weeks, during which no ingestion or administration is performed.

[0051] The composition of the present invention may be in a form containing, as a single unit dose, the above-mentioned daily dose (e.g., the dose for an adult at 60 kg body weight) or an amount obtained by dividing that by the number of times of ingestion or administration per day (e.g., 1, 2, 3, 4, or 5 times).

[0052] The active ingredient phycocyanin or its contents can be used for the same purposes, in the same manner, and in the same dosage as the above-mentioned composition of the present invention. Phycocyanin can also be used to manufacture the above-mentioned composition of the present invention.

[0053] In addition to the above-mentioned aspects, the present invention can also provide the following aspects, for example. Use of the active ingredient for producing a composition for maintaining hair health or growing hair. Use of the active ingredient for maintaining hair health or growing hair. Use of a composition containing the active ingredient for maintaining hair health or growing hair. Use of the active ingredient by oral administration for maintaining hair health or growing hair. Use of a composition containing the active ingredient by oral administration for maintaining hair health or growing hair. Use of the active ingredient by dermal application for maintaining hair health or growing hair. Use of a composition containing the active ingredient by dermal application for maintaining hair health or growing hair. Phycocyanin for maintaining hair health or growing hair. A method for maintaining hair health or growing hair, comprising administering an effective amount of the active ingredient to a subject in need of maintaining hair health or growing hair. A method for maintaining hair health or growing hair, comprising orally administering an effective amount of the active ingredient to a subject in need of maintaining hair health or growing hair. A method for maintaining healthy hair or promoting hair growth, comprising applying an effective amount of the active ingredient to the skin (scalp) of a subject in need of such treatment. A method for preventing or treating a condition that can be prevented or treated by maintaining healthy hair or promoting hair growth, comprising administering a prophylactically effective or therapeutically effective amount of the active ingredient to a subject in need of such treatment. The method is preferably a non-therapeutic method, more preferably when the subject is healthy, and is also preferably used in cosmetics. A method for preventing or treating a condition that can be prevented or treated by maintaining healthy hair or promoting hair growth, comprising orally administering a prophylactically effective or therapeutically effective amount of the active ingredient to a subject in need of such treatment. The method is preferably a non-therapeutic method, more preferably when the subject is healthy, and is also preferably used in cosmetics. A method for preventing or treating a condition that can be prevented or treated by maintaining healthy hair or promoting hair growth, comprising applying a prophylactically effective or therapeutically effective amount of the active ingredient to the skin of a subject in need of such treatment. Preferably, the method is a non-therapeutic method, more preferably the subject is the skin of a healthy subject, and preferably is used in cosmetic applications.

[0054] The present invention will be described in more detail below using examples, but the present invention is not limited to these examples.

[0055] Test Example 1: Oral Administration of Phycocyanin to Mice In this test example, phycocyanin was repeatedly orally administered to mice as a test substance, and the effects on gene expression in hair follicles and skin under ultraviolet light irradiation were examined.

[0056] (Phycocyanin Production) <Phycocyanin Extraction> 65 kg of dried Spirulina (Arhrospira platensis) algae (spray-dried product) produced in an outdoor culture tank was added to 1300 L of 1% calcium chloride (anhydrous) solution, and the mixture was stirred for 15 minutes to form a uniform suspension. The suspension was then left to stand at 20°C for 15 hours to extract the phycocyanin from the cyanobacteria. 32 kg of sodium dihydrogen phosphate was added to the extract, which was then stirred for 0.5 hours and then allowed to stand at 20°C for 2.5 hours to produce calcium phosphate, which adsorbed phycocyanin impurities to the calcium phosphate, yielding an adsorbate. The extract was then centrifuged at a gravitational acceleration of 10,000 g for 15 minutes to remove the cyanobacteria residue and adsorbed material from the extract. The obtained phycocyanin extract was subjected to ultrafiltration using a separation membrane with a molecular weight cutoff of 10,000 to remove low-molecular-weight components and salts, and then freeze-dried to obtain 5 kg of dried phycocyanin pigment. This was designated Phycocyanin 1. The phycocyanin content of 100% by mass of Phycocyanin 1 pigment powder was approximately 70% by mass (C-phycocyanin: approximately 53% by mass, allophycocyanin: approximately 16% by mass).

[0057] <Phycocyanin Analysis Method> Phycocyanin was quantified according to the method described in Journal of AOAC International, 2008, Vol. 91, Issue 3, pp. 524-529. The sample was dissolved in 100 mM phosphate buffer (pH 6.0), and the absorbance at 620 nm and 650 nm (OD ) were measured. ６２０ , O.C. ６５０The amounts of C-phycocyanin (CPC) and allophycocyanin (APC) were calculated according to the following formula: CPC (mg / mL) = 0.162 × OD ６２０ -0.098 x OD ６５０ APC (mg / mL) = 0.180 x OD ６５０ -0.042 x OD ６２０

[0058] (Oral administration to mice) Four-week-old mice (DBA / 2CrS1c, SPF, male) were acclimated for 10 days and then divided into four experimental groups (n=6 per group) as shown in Table 1 below. The test substance was repeatedly administered once a day for 21 days using a disposable syringe and a disposable oral probe. For the UV-irradiated group in Table 1, UV irradiation treatment was carried out 1 hour after administration of the test substance on days 15 to 21 of administration. The mice were exposed to UV-B light at an average irradiance of 0.3 mW / cm on the ears. ２ The irradiation device used was a UV treatment device (Dermaray (registered trademark)-200 [equipped with UVA and broadband UVB], manufactured by Clinical Supply Co., Ltd.).

[0059]

[0060] (Collection of ear skin and RNA extraction) (1) On the day following the 21st day of administration, mice were euthanized by carbon dioxide inhalation. The skin of the left and right ears of each mouse was collected near the edge of the earlobe using a biopsy trephine (6 mm diameter, model number: BP-60F, Kai Industries, Inc.). The ear skin was treated with RNAlater (Thermo Fisher Scientific) and stored frozen. (2) Total RNA was extracted from the left ear skin using RNAiso plus (Takara Bio Inc.) by standard methods, followed by DNase treatment in liquid phase and purification using the RNeasy MinElute Cleanup Kit (QIAGEN). (3) The absorbance of the obtained total RNA was measured to confirm that the A260 / 230 ratio was 1.5 or higher, the A260 / 280 ratio was 1.8 or higher, and the RNA Integrity Number (RIN) measured by a microchip electrophoresis device (2100 Bioanalyzer, Agilent Technologies) was equal to or higher than the reference value (5.9). The RNA was stored frozen (below -70°C) until use.

[0061] (DNA Microarray Analysis) DNA microarray analysis was performed using the obtained ear skin total RNA according to the following procedure. (1) Preparation of labeled cRNA: Equal amounts of individual RNA samples were pooled for each group (n = 6), and labeled cRNA was prepared from the RNA samples using a Low Input Quick Amp Labeling Kit (Agilent). (2) Hybridization: Each labeled cRNA was fragmented using a Gene Expression Hybridization Kit (Agilent), applied to a Whole Human Genome Microarray Ver. 2.0 (Agilent), and hybridized at 65°C for 17 hours. (3) Array washing: The array slide was washed using Gene Expression Wash Buffer 1 and 2 (Agilent). (4) Scanning, fluorescence intensity digitization, and analysis: Array images scanned with a microarray scanner (GenePix 4000B, Molecular Devices) were digitized using the array analysis software GenePix Pro (Molecular Devices). Fluorescence intensity values ​​were normalized, and the intensity ratio (Ratio) of each sample group relative to the control was calculated.

[0062] (Evaluation of Increase in Gene Expression) The intensity ratio of group A3 (phycocyanin (PC) group) was divided by the intensity ratio of group A2 to obtain the value (PC relaxation rate). A PC relaxation rate of greater than 1 indicates that an increase in the expression level of the gene was observed due to the test substance. The increase in gene expression was ranked as follows: A > B > C indicates that the gene expression level was significantly increased due to the test substance. <Evaluation of Increase in Gene Expression> A: PC relaxation rate 2.0 or more B: PC relaxation rate 1.5 or more but less than 2.0 C: PC relaxation rate greater than 1.0 but less than 1.5

[0063] [Results] The results of the evaluation of the gene expression levels are shown in Table 2. For each group, genes not listed in Table 2 were not evaluated.

[0064]

[0065] The genes in Table 2 are all genes whose expression and functional involvement are observed in hair follicles, particularly in inner root sheath cells and hair matrix cells. As described above, it has become clear that administration of PC is suitable for maintaining normal hair health and promoting hair growth through hair follicle activation.

[0066] In this test, the expression levels of all of the genes in Table 2 were lower in Group A2 compared to Group A1. Therefore, it was found that ultraviolet rays are one of the causes of the decline in hair follicle function associated with the decline in these genes.

[0067] [Test Example 2. Test of topical administration of phycocyanin to mice] In this test example, phycocyanin was administered topically to mice as a test substance.

[0068] We investigated the hair growth promoting effect of topical administration of

[0069] Extraction and analysis of phycocyanin were carried out in the same manner as in Test Example 1.

[0070] <Preparation of Substance for Topical Administration> The test substance was prepared immediately before use. Phycocyanin 1 obtained by the preparation method exemplified in Test Example 1 was suspended in 50% ethanol to a concentration of 5% and used for administration.

[0071] (Topical Administration to Mice) Two to three days before the start of administration, the hair on the backs of 7-week-old mice (C3H / HeNJcl, SPF, male) was clipped using pet clippers (Panasonic Corporation ER803PP-A) to an area of ​​approximately 4 cm lengthwise from the base of the tail and approximately 3 cm wide, and then the hair was removed using a razor to prepare an observation (evaluation) site, after which the mice were divided into three experimental groups (n=6 per group) as shown in Table 3 below. The test substance was administered topically once daily for 24 days. 50% ethanol served as a negative control, and a commercially available hair growth agent (Taisho Pharmaceutical Co., Ltd.'s "Riaup X5" (registered trademark)) served as a positive control.

[0072]

[0073] (Observation of the administration site) Photographs showing the hair growth status were taken under certain conditions on the first day of administration (day 1 of administration), days 9, 13, 17, and 21, and the day after the final administration (day 25 of administration), and the evaluation site (approximately 3 × 4 cm) was evaluated according to the hair regeneration score evaluation criteria shown in Table 4.

[0074]

[0075] [Results] The results of the evaluation of hair regeneration scores on the first day of administration (day 1), days 9, 13, 17, and 21, and the day after the final administration (day 25) for Group B1 (50% ethanol application group), Group B2 (commercial hair growth agent application group), and Group B3 (phycocyanin application group) are shown in Table 5 and are also shown as a graph in Figure 1. For Group B1 and Group B3, the state of hair regeneration on the backs of the mice on days 9, 13, and 17 of administration is shown as a photograph in Figure 2.

[0076]

[0077] The hair regeneration score increase from 13 to 17 days after administration in the phycocyanin application group was 1.4, the hair regeneration score increase from 13 to 17 days after administration in the positive control group treated with a commercially available hair growth agent was 0.9, and the hair regeneration score increase from 13 to 17 days after administration in the negative control group treated with 50% ethanol was 0.6. These results show that the application of phycocyanin promoted hair regeneration in a short period of time in the early stages.

Claims

1. A hair growth composition containing phycocyanin as an active ingredient.

2. The composition according to claim 1, wherein the phycocyanin is derived from cyanobacteria of the genus Spirulina.

3. The composition according to claim 1, wherein the phycocyanin is C-phycocyanin, allophycocyanin, or a combination thereof.

4. The composition according to claim 1, which is ingested or administered to a subject in which the expression of at least one gene selected from the group consisting of LIPH, TCHHL1, KRTAP6-1, KRT71, KRTAP6-2, KRTAP19-1, KRTAP19-3, KRTAP19-4, KRT36, SCD, KRT25, KRT33A, KRTAP13-1, and KRTAP13-2 is reduced compared to a normal individual in hair follicle cells.

5. A composition for oral use, according to any one of claims 1 to 4.

6. A composition for external use, as described in any one of claims 1 to 4.