Agent for activating hair dermal papilla cells

Abstract

The purpose of the present invention is to provide a pharmaceutical preparation that effectively activates hair dermal papilla cells. Provided is a composition for activating hair dermal papilla cells, a composition for promoting hair growth or for hair restoration, a composition for improving hair quality, and a composition for promoting production of FGF-2, FGF-7, or VEGF, the compositions containing microalgae culture supernatant and / or microalgae extracellular vesicle.

Description

Dermal papilla cell activator

[0001] The present invention relates to a hair papilla cell activator.

[0002] The hair cycle consists of three phases: growth, regression, and resting, during which hair repeatedly grows and falls out. Many types of cells are involved in the hair cycle, but the one that plays a role that could be called the control tower is the dermal papilla cell. Dermal papilla cells are cells located at the center bottom of the hair tissue, and play an important role in hair differentiation, such as promoting the proliferation of surrounding hair matrix cells.

[0003] When the function of hair papilla cells declines due to various factors, such as aging, stress in daily life, changes in hormone balance, and side effects of medication, the activation and differentiation of hair matrix cells are suppressed, which can shorten the hair cycle and cause hair loss.

[0004] In recent years, due to the various factors mentioned above, the number of people, both men and women, suffering from thinning hair and hair loss has been increasing. In response to this social situation, various hair growth promoters and hair restorers have been developed.

[0005] In the pharmaceutical field, for example, preparations containing minoxidil (6-(1-piperidinyl)-2,4-pyrimidinediamine-3-oxide) have been marketed as a treatment for AGA (androgenetic alopecia).

[0006] Minoxidil is known to promote blood circulation by dilating peripheral blood vessels and to stimulate hair papilla cells, thereby promoting hair growth.

[0007] In addition to pharmaceutical ingredients, various ingredients have been studied for their effects on hair growth and hair development. For example, Patent Document 1 discloses that taurine has an excellent hair growth promoting effect because it extends the hair growth phase.

[0008] Patent Document 2 discloses that rice extract promotes the production of hepatocyte growth factor and therefore has an excellent effect of promoting hair growth.

[0009] Patent Document 3 discloses that bamboo shoot extracts activate hair papilla cells and promote the production of vascular endothelial growth factor and fibroblast growth factor, and therefore have excellent hair growth promoting effects.

[0010] JP 2002-097116 A JP 2004-099503 A JP 2013-180999 A

[0011] As described above, it is presumed that any component that can activate hair papilla cells will be effective in promoting hair growth and hair development, but there have not yet been sufficient reports on what components are suitable for inclusion in a hair papilla cell activator.

[0012] Therefore, an object of the present invention is to provide a preparation that is effective in activating dermal papilla cells.

[0013] In order to solve the above problems, the present inventors conducted extensive research and found that culture supernatant and / or extracellular vesicles of microalgae have an effect on activating hair papilla cells, leading to the completion of the present invention.

[0014] That is, the present invention provides the compositions listed below.

[0015] [1] A composition for activating hair papilla cells, comprising a culture supernatant and / or extracellular vesicles of microalgae.

[0016] [2] A composition for promoting hair growth or hair care, comprising a culture supernatant and / or extracellular vesicles of microalgae.

[0017] [3] A hair quality improving composition containing a culture supernatant and / or extracellular vesicles of microalgae.

[0018] [4] A composition for promoting FGF-2, FGF-7, or VEGF production, comprising a culture supernatant and / or extracellular vesicles of microalgae.

[0019] [5] The composition according to any one of [1] to [4] above, wherein the microalgae are microalgae belonging to the genus Pavlova, Euglena, Spirulina, or Chlorella.

[0020] [6] The composition according to any one of [1] to [5] above, which is a food or drink, a cosmetic, a pharmaceutical, or a quasi-drug.

[0021] [7] Use of a culture supernatant and / or extracellular vesicles of a microalga in the manufacture of a composition used for activating hair follicle papilla cells.

[0022] [8] Use of a culture supernatant and / or extracellular vesicles of a microalga in the manufacture of a composition for promoting hair growth or hair development.

[0023] [9] Use of a culture supernatant and / or extracellular vesicles of a microalga in the production of a composition for improving hair quality.

[0024]

[10] Use of a culture supernatant and / or extracellular vesicles of a microalga in the manufacture of a composition used to promote the production of FGF-2, FGF-7, or VEGF.

[0025]

[11] The use according to any one of [7] to

[10] above, wherein the microalgae are microalgae belonging to the genus Pavlova, Euglena, Spirulina, or Chlorella.

[0026]

[12] The use according to any one of the above [7] to

[11] , wherein the composition is a food or drink, a cosmetic, a pharmaceutical, or a quasi-drug.

[0027] According to the present invention, hair growth and hair development can be promoted by using a culture supernatant and / or extracellular vesicles of microalgae.

[0028] Fig. 1 is a graph showing the results of a test in Test Example 1 in which Pavlova-derived extracellular vesicles were added to dermal papilla cells. Fig. 2 is a graph showing the results of a test in Test Example 1 in which Spirulina-derived extracellular vesicles were added to dermal papilla cells. Fig. 3 is a graph showing the results of a test in Test Example 1 in which Chlorella-derived extracellular vesicles were added to dermal papilla cells.

[0029] [Composition for Activating Hair Papilla Cells] In one embodiment, the composition for activating hair papilla cells of the present invention contains a culture supernatant and / or extracellular vesicles of microalgae.

[0030] (Culture Supernatant and / or Extracellular Vesicles of Microalgae) In this specification, microalgae refer to organisms that perform oxygen-producing photosynthesis, excluding mosses, ferns, and spermatophytes, and have a cell size of 0.1 μm to 1000 μm in diameter. Labyrinthula, which are protists closely related to microalgae, are also included.

[0031] Microalgae include, for example, prokaryotic organisms of the phylum Cyanobacteria, as well as eukaryotic organisms of the phylum Glaucophyta (Glaucophyta), phylum Rhodophyta (red algae), phylum Chlorophyta (Chlorophyta), phylum Cryptophyta (Cryptophyta), phylum Haptophyta (Haptophyta), phylum Heterokontophyta (Heterokontophyta), phylum Dinophyta (Dinoflagellates), phylum Euglenida, and phylum Chlorarachniophyta.

[0032] Examples of microalgae include microalgae belonging to the genus Pavlova, the genus Euglena (Japanese name: Midorimushi, protozoa / green algae), the genus Spirulina (cyanophyceae), the genus Chlorella (green algae), the genus Dunaliella (green algae), the genus Nannochloropsis (true eyespot algae), the genus Licoricerum (green algae), the genus Botryococcus (green algae), and the Labyrinthulomycete Thraustochytrids. The microalgae exemplified above may be used singly or in combination of two or more.

[0033] Among microalgae, from the viewpoint of significantly achieving the effects of the present invention, microalgae belonging to at least one species selected from the group consisting of the genus Pavlova, the genus Euglena, the genus Spirulina, and the genus Chlorella are preferred, microalgae belonging to at least one species selected from the group consisting of the genus Pavlova, the genus Euglena, and the genus Spirulina are more preferred, microalgae belonging to at least one species selected from the group consisting of the genus Pavlova and the genus Euglena are even more preferred, and microalgae belonging to the genus Pavlova are particularly preferred.

[0034] Pavlova is included in the haptophyte division, and examples of microalgae of the genus Pavlova include P. calceolate, P. granifera, P. gyrans, P. lutheri, P. pinguis, and P. salina, with P. granifera and / or P. gyrans being preferred.

[0035] Examples of microalgae belonging to the genus Euglena include E. gracilis, E. longa, E. caudata, E. oxyuris, E. tripteris, E. proxima, E. Viridis, E. socialis, E. ehrenbergii, E. Deses, E. pisciformis, E. spirogyra, E. acus, E. geneculata, E. intermedia, E. mutabilis, E. sanguinea, E. stellata, E. terricola, E. Klebsi, E. rubra, E. Examples of suitable strains include E. cyclopicola, and E. gracilis and / or E. longa are preferred.

[0036] Examples of microalgae of the genus Spirulina include the genus Altrospira, which has a scientific name change from Spirulina, and include S. platensis (A. platensis) and S. maxima (A. maxima), with S. platensis being preferred.

[0037] Examples of microalgae of the genus Chlorella include C. vulgaris, C. saccharophila, C. ellipsoidea, C. pyrenoidosa, C. sorokiniana, and C. lobophora, with C. vulgaris being preferred.

[0038] The microalgae exemplified above are widely distributed in seawater, freshwater such as ponds and marshes, and brackish water, and may be separated from these and used, or any microalgae that has already been isolated may be used.

[0039] The microalgae exemplified above include closely related species and mutant strains thereof, as long as they exhibit the effects of the present invention. Mutant strains also include those obtained by genetic methods such as gene recombination, transduction, and transformation.

[0040] As used herein, the term "microalgae culture supernatant" refers to the supernatant of a culture solution obtained after culturing microalgae. For example, the term "microalgae culture supernatant" refers to the supernatant of a culture solution obtained by culturing microalgae using a known culture method and removing the microalgae using a known separation means such as centrifugation.

[0041] The culture conditions for preparing the culture supernatant of microalgae are not particularly limited as long as the effects of the present invention are achieved, and known methods can be used, including the following examples.

[0042] (Cultivation Conditions) Microalgae can be cultured using a culture medium. A carbon source is added to the culture medium as a nutrient source. The carbon source is an inorganic carbon source (CO ２ , NaHCO ３ , Na ２ CO ３ The culture medium is not limited, but it is preferable to use an autotrophic medium that does not contain an organic carbon source such as glucose as a nutrient source. Examples of the culture medium include a culture medium supplemented with nutrient salts such as a nitrogen source, a phosphorus source, and minerals, such as Cramer-Myers medium and modified Cramer-Myers medium.

[0043] Depending on the type of microalgae, when seawater is used, it is possible to use a culture medium for marine microalgae or artificial seawater, or it is also possible to use commercially available products such as IMK medium.

[0044] The pH of the culture medium is not particularly limited as long as the effects of the present invention are achieved, and examples include 2 to 8, 3 to 8, 4 to 8, 5 to 8, 2 to 7.5, 3 to 7.5, 4 to 7.5, and 5 to 7.5.

[0045] The culture temperature is not particularly limited as long as the effects of the present invention are achieved, but examples include 15 to 40°C, 20 to 34°C, and 23 to 28°C.

[0046] The culture period is not particularly limited as long as the effects of the present invention are achieved, but examples include 4 to 30 days, 4 to 20 days, and 5 to 15 days.

[0047] Depending on the type of microalgae, the type of light source, the presence or absence of light and dark, aerobic conditions, anaerobic conditions, etc. can be appropriately selected.

[0048] In one embodiment, when culturing marine microalgae such as pavlova, any known method can be used as long as the effects of the present invention are achieved. For example, the culture can be carried out under conditions that satisfy at least one of the following: Seawater concentration: 50% seawater Culture medium concentration: 2x IMK medium Culture solution volume: 800 mL Light source: Side: fluorescent light (100 to 150 μmol) Light / dark cycle: 12 hours of light and 12 hours of dark pH: pH of about 7.4 at the start of each culture Aeration: performed Mechanical stirring: none Culture period: approximately 10 days Culture temperature: 25°C to 28°C

[0049] (Recovery of culture supernatant) After culturing microalgae under the culture conditions exemplified above, the culture supernatant is recovered using a known separation means such as centrifugation, but there are no particular limitations on the method as long as it can appropriately recover the culture supernatant of microalgae.

[0050] In the present invention, extracellular vesicles of microalgae can also be used. As used herein, extracellular vesicles refer to vesicles secreted from microalgae that typically have an average diameter of 10 nm to 1 μm. Although not limited thereto, when extracellular vesicles of microalgae are used in the composition of the present invention, it is preferable that the composition does not contain the microalgae cells themselves.

[0051] The method for separating extracellular vesicles can be any known separation means and is not particularly limited. For example, the method for separating extracellular vesicles can be, but is not limited to, a technique such as centrifugation.

[0052] In one embodiment, the method for producing extracellular vesicles includes, but is not limited to, (1) centrifuging a culture supernatant of microalgae to obtain a supernatant; and (2) further centrifuging the obtained product, if necessary, to obtain a precipitate. This method can produce microalgae-derived extracellular vesicles that exhibit effective activity.

[0053] In exemplary embodiments, the centrifugation in steps (1) and (2) may be carried out for 30 to 60 minutes at a centrifugal force of, for example, 1,000 to 20,000 x g, 1,500 to 20,000 x g, 1,500 to 15,000 x g, or 1,500 to 10,000 x g. In this case, the centrifugation may be carried out stepwise by varying the speed or time. For example, the culture supernatant may be separated by centrifugation at a low speed of 1,500 to 2,000 x g, and then further centrifuged at a high speed of 10,000 to 20,000 x g to remove cells or cell-related debris and residues.

[0054] [Uses] In one embodiment, the composition of the present invention is suitable for use in activating dermal papilla cells. Dermal papilla cells are present at the center base of hair tissue and, upon activation, secrete various growth factors, angiogenesis factors, and the like, promoting the proliferation of surrounding hair matrix cells and the supply of nutrients to hair matrix cells. Preparations containing minoxidil have been marketed as treatments for androgenetic alopecia (AGA), and the hair growth effect of minoxidil is thought to be due in part to the activation of dermal papilla cells. For example, it is known that the expression of VEGF (vascular endothelial growth factor) is reduced in hair tissue in patients with androgenetic alopecia, but minoxidil can promote the production of VEGF in dermal papilla cells. Therefore, it is speculated that any component capable of promoting VEGF production in dermal papilla cells will promote the supply of nutrients to surrounding hair matrix cells and contribute to hair growth and development. In the examples (test examples) described below, the effect on VEGF-A was confirmed. It is known that VEGF-A develops a capillary network around hair follicles, enlarges hair follicle tissue, and is involved in promoting hair growth. In addition, the composition of the present invention is preferably used to activate hair papilla cells, leading to an improvement in the environment around hair follicles, and is therefore expected to have an effect of improving not only male pattern baldness, but also female pattern baldness (female thinning hair) such as androgenetic alopecia, alopecia areata, stress-induced alopecia, and alopecia in animals such as pets.

[0055] It is also known that in dermal papilla cells of androgenetic alopecia, the expression level of the FGF-7 (fibroblast growth factor) gene is reduced to about half. Adenosine, which is used in quasi-drugs and the like, is known to have the effect of promoting FGF-7 production and VEGF production, and it is believed that these actions contribute to hair growth and development. Therefore, it is speculated that any component that can promote FGF-7 production in dermal papilla cells will promote the proliferation and differentiation of surrounding hair matrix cells, contributing to hair growth and development.

[0056] Furthermore, FGF-2 is known to have a strong angiogenic effect. Its angiogenic effect can be more effective than VEGF and FGF-7, which are also known to have such effects, and its involvement in hair growth and development has attracted attention. Therefore, it is presumed that a component that can promote FGF-2 production in dermal papilla cells, or further a component that can promote the production of both FGF-2 and VEGF in dermal papilla cells, will promote the supply of nutrients to surrounding hair matrix cells and contribute to hair growth and development.

[0057] It is known that IGF-1 (insulin-like growth factor-1) produced by dermal papilla cells can promote the proliferation of epithelial cells in hair follicles and contribute to the extension of the anagen phase of the hair cycle. This leads to longer hair growth during the anagen phase of the hair cycle and the growth and maintenance of the hair bulb morphology. Therefore, it is speculated that any component that can promote IGF-1 production in dermal papilla cells will contribute to the growth of long, healthy hair.

[0058] Activation of dermal papilla cells not only contributes to hair growth, but is also effective in improving the quantity and quality of hair. As described above, promoting the production of FGF-2 and VEGF improves blood supply to hair tissue through angiogenesis, leading to growth of the hair bulb due to proliferation and differentiation of hair matrix cells, which leads to thicker hair growth, and by acting on melanocytes adjacent to the hair matrix cells, leads to thicker, lustrous hair growth. Therefore, it is presumed that any component that can promote the production of at least one selected from the group consisting of FGF-2, FGF-7, IGF-1, and VEGF in dermal papilla cells will contribute to improving hair quality.

[0059] The improvement in hair quality is not limited to, but may be, for example, at least one selected from the group consisting of hair thickness, hair color depth, hair texture, and hair hardness.

[0060] In the Examples (Test Examples) described below, it has been confirmed that microalgae culture supernatant and / or extracellular vesicles can promote the production of at least one selected from the group consisting of FGF-2, FGF-7, IGF-1, and VEGF in hair papilla cells. Thus, microalgae culture supernatant and / or extracellular vesicles are suitable for use in activating hair papilla cells, promoting hair growth and hair care by promoting the production of the above components, and further, improving hair quality.

[0061] The composition of the present invention can be added to or mixed with pharmaceuticals, quasi-drugs, cosmetics, foods, beverages, feed, or pet foods, or can be used as is as pharmaceuticals, quasi-drugs, cosmetics, foods, beverages, feed, or pet foods.

[0062] When the composition of the present invention is used by adding to or mixing with so-called external preparations such as pharmaceuticals, quasi-drugs, and cosmetics, the composition of the present invention can be used as pharmaceuticals, quasi-drugs, and cosmetics that explicitly or implicitly have the efficacy, effect, or function of activating hair papilla cells, promoting hair growth, hair restoration, and promoting or improving hair quality.

[0063] When the above functionality is not explicitly stated, the composition of the present invention can be used by, for example, labeling it as suitable for those who want firmness from the roots, those who want strength from the roots, those who are concerned about their hairline, those who want volume, those who want hair roots that are less likely to fall out, those who want early hair care, those who want to grow their hair from the scalp, those who want to guide their hair into the correct hair cycle, those who want to nourish their hair, those who want to improve their hair environment, those who want to stimulate scalp cells, those who want to support the roots of their hair, those who want a voluminous look, those who want to improve weak hair, or similar labeling.

[0064] When used in so-called external preparations such as pharmaceuticals, quasi-drugs, cosmetics, etc., the composition of the present invention is preferably prepared as a product for hair regeneration, hair regeneration, hair growth, hair care, or hair loss prevention.

[0065] When used in so-called external preparations such as pharmaceuticals, quasi-drugs, and cosmetics, the daily dose of microalgal extracellular vesicles for an adult can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, ingestion or administration route, ingestion or administration schedule, formulation form, and other factors. Examples of the daily dose of microalgal extracellular vesicles for an adult include 1 μg or more, 5 μg or more, 10 μg or more, 15 μg or more, 20 μg or more, 30 μg or more, 40 μg or more, 50 μg or more, 70 μg or more, 100 μg or more, 150 μg or more, and 200 μg or more. In addition, the daily dosage of microalgal extracellular vesicles for an adult can be, for example, 5 mg or less, 3 mg or less, 1 mg or less, 900 μg or less, 800 μg or less, 700 μg or less, 600 μg or less, 500 μg or less, 400 μg or less, 300 μg or less, 200 μg or less, or 100 μg or less. The daily dose of microalgal extracellular vesicles for adults is, for example, 1 to 1000 μg, 1 to 900 μg, 1 to 800 μg, 1 to 700 μg, 1 to 600 μg, 1 to 500 μg, 1 to 400 μg, 1 to 300 μg, 1 to 200 μg, 1 to 100 μg, 5 to 1000 μg, 5 to 900 μg, 5 to 800 μg, 5 to 700 μg, 5 to 600 μg, 5 to 500 μg, 5 to 400 μg, 5 to 300 μg, 5 to 200 μg, 5 to 100 μg. g, 10 to 1000 μg, 10 to 900 μg, 10 to 800 μg, 10 to 700 μg, 10 to 600 μg, 10 to 500 μg, 10 to 400 μg, 10 to 300 μg, 10 to 200 μg, 10 to 100 μg, 20 to 1000 μg, 20 to 900 μg, 20 to 800 μg, 20 to 700 μg, 20 to 600 μg, 20 to 500 μg, 20 to 400 μg, 20 to 300 μg, 20 to 200 μg, and 20 to 100 μg. The dosage of microalgal extracellular vesicles varies depending on various factors, such as the degree of progression of symptoms, the time of onset of the disease, the age, health condition, and complications of the subject. However, based on an adult, the dosage is generally 1 μg / kg to 200 mg / kg of the composition, and in another aspect, 50 μg / kg to 50 mg / kg may be administered in divided doses one to three times a day. The dosage does not limit the scope of the present specification in any way.

[0066] When used in so-called external preparations such as pharmaceuticals, quasi-drugs, and cosmetics, the daily adult dosage of microalgal extracellular vesicles can be appropriately determined depending on the individual's condition, body weight, sex, age, activity of the material, ingestion or administration route, ingestion or administration schedule, formulation form, and other factors. Examples of the daily adult dosage of microalgal extracellular vesicles in terms of protein amount include 1 μg or more, 5 μg or more, 10 μg or more, 15 μg or more, 20 μg or more, 30 μg or more, 40 μg or more, 50 μg or more, 70 μg or more, 100 μg or more, 150 μg or more, and 200 μg or more. In addition, the daily dosage of microalgal extracellular vesicles for an adult can be, for example, 5 mg or less, 3 mg or less, 1 mg or less, 900 μg or less, 800 μg or less, 700 μg or less, 600 μg or less, 500 μg or less, 400 μg or less, 300 μg or less, 200 μg or less, or 100 μg or less. The daily dose of microalgal extracellular vesicles for adults is, for example, 1 to 1000 μg, 1 to 900 μg, 1 to 800 μg, 1 to 700 μg, 1 to 600 μg, 1 to 500 μg, 1 to 400 μg, 1 to 300 μg, 1 to 200 μg, 1 to 100 μg, 5 to 1000 μg, 5 to 900 μg, 5 to 800 μg, 5 to 700 μg, 5 to 600 μg, 5 to 500 μg, 5 to 400 μg, 5 to 300 μg, 5 to 200 μg, 5 to 100 μg. g, 10 to 1000 μg, 10 to 900 μg, 10 to 800 μg, 10 to 700 μg, 10 to 600 μg, 10 to 500 μg, 10 to 400 μg, 10 to 300 μg, 10 to 200 μg, 10 to 100 μg, 20 to 1000 μg, 20 to 900 μg, 20 to 800 μg, 20 to 700 μg, 20 to 600 μg, 20 to 500 μg, 20 to 400 μg, 20 to 300 μg, 20 to 200 μg, and 20 to 100 μg. The dosage of microalgal extracellular vesicles varies depending on various factors, such as the degree of progression of symptoms, the time of onset of the disease, the age, health condition, and complications of the subject. However, based on an adult, the dosage may be 1 μg / kg to 200 mg / kg of the composition, or in another aspect, 50 μg / kg to 50 mg / kg, administered 1 to 3 times a day. The dosage does not limit the scope of the present specification in any way.

[0067] In another embodiment, the daily dose of microalgal extracellular vesicles for an adult is, for example, 1 x 10 ７ pcs or more, 1×10 ８ pcs or more, 1×10 ９ The daily dose of extracellular vesicles of microalgae for an adult is, for example, 1 x 10 １４ Less than or equal to 1 x 10 １３ Less than or equal to 1 x 10 １２ Less than or equal to 1 x 10 １１ Less than or equal to 1 x 10 １０ The daily dose of extracellular vesicles of microalgae for an adult is, for example, 1 x 10 ７ ~1 x 10 １４ pieces, 1×10 ７ ~1 x 10 １３ pieces, 1×10 ７ ~1 x 10 １２ pieces, 1×10 ７ ~1 x 10 １１ pieces, 1×10 ８ ~1 x 10 １４ pieces, 1×10 ８ ~1 x 10 １３ pieces, 1×10 ８ ~1 x 10 １２ pieces, 1×10 ８ ~1 x 10 １１ pieces, 1×10 ９ ~1 x 10 １４ pieces, 1×10 ９ ~1 x 10 １３ pieces, 1×10 ９ ~1 x 10 １２ pieces, 1×10 ９ ~1 x 10 １１ Examples include:

[0068] The daily dose of the microalgae culture supernatant for an adult can be determined appropriately depending on the individual's condition, body weight, sex, age, activity of the material, administration schedule, formulation form, or other factors. The daily dose of the microalgae culture supernatant for an adult can be, for example, the amount of microalgae extracellular vesicles contained in the above-mentioned intake or administration amount.

[0069] The composition of the present invention can be ingested or administered once to several times a day, typically 1 to 6 times a day, 1 to 3 times a day, 1 to 2 times a day, or at any period and interval, but once a day is preferred.

[0070] When the composition of the present invention is added to or mixed with food, beverage, feed, or pet food, the composition of the present invention can be used as a food or beverage that explicitly or implicitly claims to have the functionality of activating hair papilla cells, promoting hair growth, hair restoration, promoting or improving hair quality, etc., i.e., a health food, a functional food, a food for patients, or a food for specified health uses. Furthermore, even if the above functionality is not explicitly or implicitly claimed, the composition can be used as a so-called doctor's supplement recommended or presented by doctors in internal medicine, orthopedic surgery, veterinary clinics, etc. in hospitals and / or clinics.

[0071] When the above functionality is not explicitly stated, the composition of the present invention can be used by, for example, labeling it as suitable for those who want firmness from the roots, those who want strength from the roots, those who are concerned about their hairline, those who want volume, those who want hair roots that are less likely to fall out, those who want early hair care, those who want to grow their hair from the scalp, those who want to guide their hair into the correct hair cycle, those who want to nourish their hair, those who want to improve their hair environment, those who want to stimulate scalp cells, those who want to support the roots of their hair, those who want a voluminous look, those who want to improve weak hair, or similar labeling.

[0072] Health foods, functional foods, foods for the sick, and foods for specified health uses can be produced by conventional means in the same way as known pharmaceutical preparations.

[0073] The composition of the present invention can be applied in the form of oral administration, internal administration, etc. When used as a pharmaceutical composition, it may be used therapeutically or non-therapeutically.

[0074] The daily oral intake or administration amount of microalgal extracellular vesicles for an adult can be determined appropriately depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. Examples of the daily oral intake or administration amount of microalgal extracellular vesicles for an adult include 1 μg or more, 5 μg or more, 10 μg or more, 15 μg or more, 20 μg or more, 30 μg or more, 40 μg or more, 50 μg or more, 70 μg or more, 100 μg or more, 150 μg or more, and 200 μg or more. Furthermore, the daily oral intake or administration amount of microalgal extracellular vesicles for an adult can be, for example, 5 mg or less, 3 mg or less, 1 mg or less, 900 μg or less, 800 μg or less, 700 μg or less, 600 μg or less, 500 μg or less, 400 μg or less, 300 μg or less, 200 μg or less, or 100 μg or less. The daily oral intake or administration amount of microalgal extracellular vesicles for adults is, for example, 1 to 1000 μg, 1 to 900 μg, 1 to 800 μg, 1 to 700 μg, 1 to 600 μg, 1 to 500 μg, 1 to 400 μg, 1 to 300 μg, 1 to 200 μg, 1 to 100 μg, 5 to 1000 μg, 5 to 900 μg, 5 to 800 μg, 5 to 700 μg, 5 to 600 μg, 5 to 500 μg, 5 to 400 μg, 5 to 300 μg, 5 to 200 μg, 5 to 500 μg, 5 to 600 μg, 5 to 200 μg, 5 to 500 μg, 5 to 500 μg, 5 to 400 μg, 5 to 300 μg, 5 to 200 μg, 5 to 500 μg, 5 to 500 μg, 5 to 500 μg, 5 to 500 μg, 5 to 500 μg, 5 to 500 μg, 5 to 500 μg, 5 to 6 ... Examples include up to 100 μg, 10 to 1000 μg, 10 to 900 μg, 10 to 800 μg, 10 to 700 μg, 10 to 600 μg, 10 to 500 μg, 10 to 400 μg, 10 to 300 μg, 10 to 200 μg, 10 to 100 μg, 20 to 1000 μg, 20 to 900 μg, 20 to 800 μg, 20 to 700 μg, 20 to 600 μg, 20 to 500 μg, 20 to 400 μg, 20 to 300 μg, 20 to 200 μg, and 20 to 100 μg.

[0075] In another embodiment, the daily oral intake or administration amount of microalgal extracellular vesicles for an adult is, for example, 1 × 10 ７ pcs or more, 1×10 ８ pcs or more, 1×10 ９ The daily oral intake or administration amount of extracellular vesicles of microalgae for an adult is, in terms of the number of particles, for example, 1 × 10 １４ Less than or equal to 1 x 10１３ Less than or equal to 1 x 10 １２ Less than or equal to 1 x 10 １１ Less than or equal to 1 x 10 １０ The daily oral intake or administration amount of extracellular vesicles of microalgae for an adult is, in terms of the number of particles, for example, 1 x 10 ７ ~1 x 10 １４ pieces, 1×10 ７ ~1 x 10 １３ pieces, 1×10 ７ ~1 x 10 １２ pieces, 1×10 ７ ~1 x 10 １１ pieces, 1×10 ８ ~1 x 10 １４ pieces, 1×10 ８ ~1 x 10 １３ pieces, 1×10 ８ ~1 x 10 １２ pieces, 1×10 ８ ~1 x 10 １１ pieces, 1×10 ９ ~1 x 10 １４ pieces, 1×10 ９ ~1 x 10 １３ pieces, 1×10 ９ ~1 x 10 １２ pieces, 1×10 ９ ~1 x 10 １１ Examples include:

[0076] The daily oral intake or administration amount of a microalgae culture supernatant for an adult can be determined appropriately depending on the individual's condition, body weight, sex, age, activity of the material, intake or administration route, intake or administration schedule, formulation form, or other factors. The daily oral intake or administration amount of a microalgae culture supernatant for an adult can be, for example, the amount of microalgae extracellular vesicles contained in the above-mentioned intake or administration amount.

[0077] The oral intake or dosage per day for an adult may be divided into 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules depending on the dosage form.

[0078] The composition of the present invention can be ingested or administered once to several times a day, typically 1 to 6 times a day, 1 to 3 times a day, 1 to 2 times a day, or at any period and interval, but once a day is preferred.

[0079] When the present invention is used as feed, pet food, or the like, the target organisms are not particularly limited, but are preferably mammals, reptiles, amphibians, birds, or fish, and more preferably mammals other than humans, such as platypus, echidna, opossum, quoll, kangaroo, aardvark, rock badger, elephant, armadillo, sloth, anteater, tree shrew, flying lemur, chimpanzee, rabbit, degu, dormouse, squirrel, raccoon, mouse, hedgehog, chinchilla, ferret, camel, wild boar, giraffe, deer, cow, goat, hippopotamus, whale, dolphin, horse, rhinoceros, tapir, bat, monkey, tiger, wolf, weasel, bear, seal, dog, cat, parakeet, parrot, finch, owl, and horned owl, and more preferably dog ​​or cat.

[0080] When the present invention is used as a feed or pet food, it may be given to a pet in divided doses several times a day by adding it to the staple food, or it may be given to a pet as a snack at any time.

[0081] Next, the present invention will be specifically explained by way of examples, but the present invention is not limited to the following examples.

[0082] Test Example 1: Test of adding culture supernatant and / or extracellular vesicles of microalgae to dermal papilla cells. 6 x 10 dermal papilla cells were added to a 24-well plate. ３ The cells were seeded at 1000 cells / well and washed twice with 5 mL / dish of HBSS (hereinafter simply referred to as medium). Accutase (Nacalai Tesque) was added at 1 mL / dish. The incubation temperature was 37°C, 5% CO ２ The cells were cultured in an incubator, and 3 mL of medium was added per dish. The cells were transferred to a 15 mL tube and centrifuged at 220 x g for 5 minutes at 4°C. The supernatant was removed, and the pellet was suspended in medium and the cell number was counted. The cell number was adjusted and the cells were seeded at 1 mL per well. The cells were incubated at 37°C, 5% CO ２ The cells were cultured in an incubator.

[0083] Pavlova (Pavlova sp.), Spirulina (Arthrospira platensis), or Chlorella (Chlorella vulgaris) was cultured in a commercially available algae medium by conventional methods and subjected to ultracentrifugation as described below. Each culture supernatant was transferred to a clear ultracentrifuge tube at 11 mL per tube. Ultracentrifugation was performed using an Optima XE-90 Ultracentrifuge (Beckman Coulter) at 35,000 rpm for 70 minutes at 4°C. The supernatant was removed and the precipitate was suspended in PBS(-). Ultracentrifugation was performed again in the same manner at 35,000 rpm for 70 minutes at 4°C, the supernatant was removed, and the precipitate was suspended in PBS(-). The resulting samples were stored at 4°C.

[0084] As a result of the above, the following samples were obtained: Pavlova culture supernatant ultracentrifuged sample (protein concentration 2500 μg / mL) Spirulina culture supernatant ultracentrifuged sample (protein concentration 226 μg / mL) Chlorella culture supernatant ultracentrifuged sample (protein concentration 84.4 μg / mL)

[0085] The above samples or PBS(-) were mixed with the above medium. The conditions for adding the above samples were as follows: Pavlova 40 μg / mL, Spirulina 40 μg / mL, Chlorella 10 μg / mL

[0086] Human dermal papilla cells (HFDPC cells, Cell Applications) seeded the day before were washed once. The sample-containing medium mixed above was added at 250 μL / well. The incubation was continued at 37°C, 5% CO ２ The cells were cultured in an incubator for 48 hours.

[0087] After treating each sample for 48 hours, the culture medium was removed from the human dermal papilla cells, which were then washed. QIAzol (QIAGEN) was added at 700 mL / well. The cells were collected into 1.5 mL tubes using a cell scraper and stored at -80°C. RNA was then extracted using the miRNeasy mini Kit (QIAGEN).

[0088] The RNA concentration of the RNA sample was measured using a NanoDrop ND-1000 (Thermo Fisher Scientific), followed by DNase treatment and reverse transcription to produce cDNA. After reverse transcription, the cDNA sample was diluted 5-fold with RNase-free water and stored at -30°C.

[0089] Gene expression analysis of FGF-2, FGF-7, IGF-1, and VEGF was performed by real-time PCR using a TaqMan probe on a QuantStudio® 3 real-time PCR system (Thermo Fisher Scientific). A sample without each sample (a sample containing only PBS) was used as a control, GAPDH was used as an internal standard, and data analysis was performed using the ΔΔCT method. The results of the test in which Pavlova-derived extracellular vesicles were added to dermal papilla cells are shown in Figure 1, the results of the test in which Spirulina-derived extracellular vesicles were added to dermal papilla cells are shown in Figure 2, and the results of the test in which Chlorella-derived extracellular vesicles were added to dermal papilla cells are shown in Figure 3. Figures 1 to 3 show the relative results when each sample was added, with the control (a sample containing only PBS) set to 1.

[0090] 1 to 3, it was confirmed that adding microalgal culture supernatant and / or extracellular vesicles to dermal papilla cells can promote the production of at least one selected from the group consisting of FGF-2, FGF-7, IGF-1, and VEGF in the dermal papilla cells, thereby activating the dermal papilla cells. Thus, microalgal culture supernatant and / or extracellular vesicles are suitable for use in activating dermal papilla cells, and are suitable for promoting hair growth and hair care by promoting the production of the above components, and can also be suitable for use in improving hair quality.

Claims

1. A composition for activating hair papilla cells, comprising culture supernatant and / or extracellular vesicles of microalgae.

2. A composition for promoting hair growth or hair development, comprising culture supernatant and / or extracellular vesicles of microalgae.

3. A composition for improving hair quality, comprising culture supernatant and / or extracellular vesicles of microalgae.

4. A composition for promoting FGF-2, FGF-7, or VEGF production, comprising culture supernatant and / or extracellular vesicles of microalgae.

5. The composition according to any one of claims 1 to 4, wherein the microalgae is microalgae belonging to the genus Pavlova, Euglena, Spirulina, or Chlorella.

6. The composition according to any one of claims 1 to 4, which is a food or drink, a cosmetic, a medicine, or a quasi-drug.

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