Hair growth product manufacturing method

Abstract

To provide a hair growth agent that comprises a swiftlet nest extract decomposed by weak acid or solid acid, while demonstrating an enhanced hair growth effect.SOLUTION: A hair growth agent comprises: a sialic acid-containing extract, comprising a swiftlet nest extract decomposed by weak acid or solid acid; and a human hair root stem cell culture supernatant.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present invention relates to a hair growth agent used for growing human hair and the like.

Background Art

[0002] There are not a few people who are troubled by thinning hair and baldness of the head caused by aging, illness, etc., and the development of a more effective hair growth agent is expected. Here, Patent Document 1 discloses a hair growth agent obtained by decomposing a material derived from a natural product of a nest extract of an Asian swiftlet and / or animal saliva and / or royal jelly with a weak acid such as acetic acid, citric acid, malic acid, or lactic acid, or a solid acid such as silica alumina, zeolite, or acid clay, and containing the obtained monomolecular and low molecular weight sialic acid and epithelial growth factor and / or fibroblast growth factor, or a mixture of all of these. Further, Patent Document 2 discloses a hair growth agent containing a pulp stem cell culture supernatant and a dermal papilla cell culture supernatant. Further, Patent Document 3 discloses a hair growth agent containing a stem cell culture supernatant, and preferred stem cells include mesenchymal stem cells derived from umbilical cord (paragraph 0020 of the specification).

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0004] As described in Patent Document 1, by including hydrolyzed swiftlet nest extract decomposed with a weak acid or solid acid, it is possible to create a hair growth agent that contains a large amount of low-molecular-weight sialic acid, EGF (epidermal growth factor), and FGF (fibroblast growth factor), and that is well absorbed by the scalp. However, if it is possible to further enhance the hair growth effect while including low-molecular-weight sialic acid, it would contribute even more to resolving concerns such as thinning hair. Note that the hair growth agents described in Patent Documents 2 and 3 do not contain swiftlet nest extract decomposed with a weak acid or solid acid. Therefore, the present invention aims to provide a hair growth agent that contains swiftlet nest extract decomposed with a weak acid or solid acid, while having superior hair growth effects compared to conventional products. [Means for solving the problem]

[0005] Hair growth agent of the present invention as described in claim 1 Manufacturing method It is decomposed by a weak acid or solid acid. , and also contains sialic acid Sialic acid-containing extract including swiftlet nest extract and human hair follicle stem cell culture supernatant A hair growth product is manufactured by mixing these ingredients and ensuring that the amount of human hair follicle stem cell culture supernatant is greater than the amount of sialic acid-containing extract. It is characterized by the following: Claim 2 The present invention described herein is the hair growth agent described in claim 1. Manufacturing method In, The aforementioned hair growth agent, It is characterized by containing placenta extract. Claim 3 The present invention described herein is as follows: 2 Hair growth products listed in any one of the items below Manufacturing method teeth, The aforementioned hair growth agent, It is characterized by being a shampoo, conditioner, or tonic. [Effects of the Invention]

[0006] According to the present invention, it is possible to provide a hair growth agent that contains swiftlet nest extract decomposed with a weak acid or solid acid and has a superior hair growth effect compared to conventional products. [Brief explanation of the drawing]

[0007] [Figure 1]Figure showing the results of the efficacy verification test of a hair growth agent containing the culture supernatant of human hair follicle stem cells according to an embodiment of the present invention

Mode for Carrying Out the Invention

[0008] The hair growth agent according to the first embodiment of the present invention contains a sialic acid-containing extract containing an edible bird's nest extract decomposed by a weak acid or a solid acid, and a culture supernatant of human hair follicle stem cells. According to this embodiment, a hair growth agent with a higher effect than conventional ones can be provided.

[0009] The second embodiment of the present invention is a hair growth agent according to the first embodiment, in which the content of the culture supernatant of human hair follicle stem cells is made higher than the content of the sialic acid-containing extract. According to this embodiment, while enhancing the hair growth effect, precious edible bird's nest extract can be saved.

[0010] The third embodiment of the present invention is a hair growth agent according to the first embodiment, which contains placenta extract. According to this embodiment, the hair growth effect can be further enhanced.

[0011] The fourth embodiment of the present invention is a hair growth agent according to any one of the first to third embodiments, made into a shampoo, conditioner, or tonic. According to this embodiment, a hair growth shampoo, hair growth conditioner, or hair growth tonic with a higher hair growth effect than conventional ones can be provided.

Examples

[0012] Hereinafter, the hair growth agent according to the embodiment of the present invention will be described. The hair growth agent contains a sialic acid-containing extract containing an edible bird's nest extract (edible bird's nest extract) decomposed by a weak acid or a solid acid, and a culture supernatant of human hair follicle stem cells (culture supernatant of human hair follicle stem cells, conditioned culture solution of human hair follicle stem cells). The nest of the edible-nest swiftlet is made by solidifying the saliva secreted from the developed salivary glands during the spawning period of the edible-nest swiftlet over about one month. Regarding the consumption of the nest of the edible-nest swiftlet, it is known to have nourishing, beauty, strengthening effects, as well as infection prevention and the effect of producing testosterone (male hormone). The nest of the edible-nest swiftlet contains extremely high amounts of sialic acid at over 10%, and contains 200 to 1500 times more sialic acid compared to humans and honeybees. The edible-nest swiftlet nest extract of this example is obtained by decomposing the nest of the edible-nest swiftlet with a weak acid such as acetic acid, citric acid or lactic acid, or a solid acid such as silica alumina, zeolite or acid clay. Thereby, in addition to sialic acid with a reduced molecular weight, it contains a large amount of EGF and FGF, and an edible-nest swiftlet nest extract excellent in absorbability to the scalp can be obtained. The edible-nest swiftlet nest extract promotes the production of IGF-1 (insulin-like growth factor) and promotes the division of hair matrix cells.

[0013] Regarding the culture supernatant of human hair root stem cells, stem cells are defined as unspecialized cells having self-renewal ability and differentiation ability. Specifically, they are cells that have "differentiation ability" to differentiate into various tissues and "self-renewal ability" to maintain an undifferentiated state even after going through multiple cell division cycles as their characteristics. In mammals, there are various types of stem cells depending on the location of stem cells in the body, including pancreatic stem cells, hepatic stem cells, hematopoietic stem cells, vascular stem cells, mesenchymal stem cells, neural stem cells, retinal stem cells, etc. Among them, examples of mesenchymal stem cells include epithelial stem cells, hair follicle stem cells, adipose stem cells, bone marrow stem cells, umbilical cord stem cells, placental stem cells, dental pulp stem cells, skeletal muscle stem cells, etc. In the medical field, regenerative medicine using stem cells has attracted attention as an alternative therapy for diseases that are difficult to treat with pharmaceuticals. In stem cell transplantation medicine, it has been found that not only the stem cells themselves bring about a therapeutic effect, but various bioactive substances such as cytokines and exosomes secreted by the stem cells also greatly contribute to the therapeutic effect. Cytokines are a general term for physiologically active substances secreted by cells that are involved in intercellular interactions. When they reach a cell, they trigger responses such as cell proliferation, differentiation, and functional expression. Exosomes are vesicles secreted by cells that contain various proteins and nucleic acids, and their potential role in intercellular communication is becoming increasingly clear. When stem cells are artificially cultured, these bioactive substances are released from the cells into the culture medium. Therefore, the culture medium from which the cells have been removed can be recovered as the culture supernatant and effectively utilized.

[0014] Hair follicle stem cells are located in the bulge region and hair bulb of the hair follicle. They are deeply involved not only in the formation of the hair follicle, which is the foundation for hair growth, but also in hair growth by differentiating into hair matrix cells, dermal papilla cells, and pigment cells. Hair roots secrete large amounts of KGF (keratinocyte growth factor) and IGF-1, and secrete abundant cytokines necessary for hair formation. Human hair follicle stem cell culture supernatant is a supernatant obtained by culturing hair follicle stem cells obtained from human hair roots, and is rich in secretions (nutrients) necessary for hair formation, such as EGF, KGF, and IGF-1. Human hair follicle stem cell culture supernatant also contains exosomes. Human hair follicle stem cell culture supernatant exerts an effect that supports hair follicle formation not only in the neonatal and anagen phases, but also in the catagen and telogen phases of the hair cycle (the cycle from when a single hair begins to grow until it falls out), which consists of the neonatal, anagen, catagen, and telogen phases, and can activate the entire hair root throughout the entire hair cycle.

[0015] In the production of human hair follicle stem cell culture supernatant, the usual processes and conditions can be applied without any particular limitations. Furthermore, during manufacturing, sufficient attention will be paid to safety, including: collecting tissue from donors in facilities with adequate hygiene management; conducting prescribed tests on donors to confirm their health; confirming the safety of the collected hair follicle stem cells through prescribed tests; and collecting only the supernatant fluid that does not contain cells.

[0016] In the production of human hair follicle stem cell culture supernatant, hair follicles are first collected from donors, stored at around 5°C until the next processing step, and cell separation is performed within 24 hours. Next, hair follicle stem cells are isolated. The hair follicles are finely chopped and attached to culture plates, then cultured in Dulbecco's modified Eagle medium-low glucose (DMEM-LG) medium supplemented with antibiotics as needed. After 12 to 96 hours, remove non-adherent cells, and culture the resulting adherent cells in Dulbecco's Modified Eagle Medium - Low Glucose (DMEM-LG) containing 10% MSCGS at 37°C in a humidified atmosphere with 5% CO2. Change the medium every other day. Collect cells at subconfluence using trypsin and perform the necessary subculturing. 1.0 × 10 7 After growing the cells to a volume of / mL, they are harvested to obtain human hair follicle stem cells. Subsequently, the culture supernatant of human hair follicle stem cells is prepared, and if necessary, the cells are stored in liquid nitrogen and thawed as needed before use.

[0017] Next, human hair follicle stem cell culture supernatant is prepared. "Culture supernatant (supernatant)" refers to the supernatant obtained by culturing cells in a suitable cell culture medium under conditions that allow cell proliferation, and then removing the cells from the culture medium after cultivation. Specifically, "culture medium" refers to a liquid for cell culture that has been prepared to contain the necessary components for cell culture, but is not in contact with the cells. The liquid obtained as a result of culturing with the culture medium in contact with the cells is called "culture solution," regardless of whether cells are present or not. "Culture supernatant" is obtained by clearly removing cells from the "culture solution," and the removal of cells from the culture solution can be achieved by appropriately performing methods such as centrifugation, dialysis, or membrane separation. The culture medium used to produce the conditioned medium of human hair follicle stem cells can be a commercially available product. For example, DMEM, Ham F-12, MEMα, EMEM, IMDM, RPMI-1640, etc. can be used as a base medium, and various vitamins and minerals can be added to prepare the medium as appropriate. Furthermore, it is preferable that human hair follicle stem cell culture supernatant be free of animal serum to enhance safety. To achieve this, multiple subculturing cycles are performed during the culture of hair follicle stem cells, and serum-free medium is used for all or the last few subculturing cycles, thereby obtaining hair follicle stem cell culture supernatant free of animal serum. Alternatively, serum can be removed from human hair follicle stem cell culture supernatant using methods such as dialysis or solvent replacement. In this example, human hair follicle stem cells are cultured for 120 to 240 hours in a humidified atmosphere containing 5% CO2 and at 37°C, with serum added as needed, while subculturing as appropriate. The resulting culture medium is collected, and human hair follicle stem cell culture supernatant is obtained using a separation membrane that does not allow cells to pass through.

[0018] The culture supernatant obtained using stem cells is called "stem cell culture supernatant." When a specific origin name is listed, it indicates that stem cells isolated from that tissue were used. "Human hair follicle stem cell culture supernatant" refers to the culture supernatant obtained by culturing human hair follicle-derived stem cells. It is known that stem cells produce and secrete various cytokines, such as EGF and KGF, into the culture medium during culture. The concentrations of these cytokines can be easily measured using commercially available ELISA kits for each cytokine.

[0019] The recovered human hair follicle stem cell culture supernatant may be used as is as a raw material for hair growth products such as hair growth shampoos, hair growth conditioners, or hair growth tonics, or it may be used as a raw material for hair growth products after appropriate adjustments such as concentration, solvent substitution, freezing, drying, freeze-drying, dilution, and desalting. In addition, lecithin, cholesterol, etc. may be added to the human hair follicle stem cell culture supernatant and then liposomalized. Culture supernatant is sometimes called "conditioned medium" or "conditioned medium" as a cosmetic ingredient, but it is synonymous with "culture supernatant" as used here. Also, even when referred to as "culture medium," if it is clearly confirmed that cells have been removed, it is synonymous with "culture supernatant." Since the culture supernatant does not contain cells themselves or cellular components, there is no concern about immune rejection, and it can be used without problems even if it is prepared from hair follicle stem cells of a donor other than the user. In addition, because it is not the cells themselves, it is easy to manage and store, and the culture supernatant can be prepared in advance and stored and used immediately when needed.

[0020] This section describes the efficacy verification test of a hair growth agent containing the human hair follicle stem cell culture supernatant of this embodiment. Table 1 shows the components of the liquid in Comparative Example 1, Table 2 shows the components of the hair growth agent in Example 1, and Table 3 shows the components of the hair growth agent in Example 2. The preparation method involved adding B to A and mixing, then adding C and mixing, and finally adding D and mixing. The blending ratios are in weight percent. The hair growth agents of Example 1 and Example 2 contain human hair follicle stem cell culture supernatant in addition to the sialic acid-containing extract contained in the liquid of Comparative Example 1. In Example 1, the hair growth agent had the same concentration of sialic acid-containing extract and human hair follicle stem cell culture supernatant. In Example 2, the concentration of sialic acid-containing extract was 2% less than the concentration of human hair follicle stem cell culture supernatant. [Table 1] [Table 2] [Table 3]

[0021] As shown in Tables 1 to 3, the liquid of Comparative Example 1, and the hair growth agents of Examples 1 and 2, contain BG (1,3-butylene glycol), phenoxyethanol, polysorbate 80 (polyoxyethylene sorbitan oleate), and ethylhexylglycerin. BG is a moisturizer with low irritation and toxicity to the skin, possessing antibacterial properties, and is used in creams, lotions, and other products. Polysorbate 80 is a relatively safe nonionic surfactant that is used not only for emulsification but also for solubilizing fragrances and other substances. Ethylhexylglycerin is structurally stable, less susceptible to changes in pH and temperature, exhibits high compatibility with various oily components, and also possesses bacteriostatic properties, making it suitable for use as a preservative aid. By including BG, phenoxyethanol, polysorbate 80, and ethylhexylglycerin in the hair growth tonic, the feel and other properties when applied to the scalp can be improved.

[0022] The liquid in Comparative Example 1, as well as the sialic acid-containing extract contained in the hair growth agents of Example 1 and Example 2, contains swiftlet nest extract. Table 4 below shows the components of the sialic acid-containing extract. The proportions are given in weight percent. [Table 4]

[0023] As shown in Table 4, the sialic acid-containing extract also contains placenta extract. Placenta extract, extracted from the placenta, an organ unique to mammals, contains various beneficial components such as amino acids and enzymes. Therefore, including placenta extract in hair growth products can enhance their hair growth effects. Placenta extract is also used as an ingredient in health foods and cosmetics, and among these, placenta extracts extracted from pigs, sheep, and horses are particularly commonly used due to their safety. In this example, placenta extract extracted from horse placenta is used.

[0024] In the efficacy verification test, several male subjects were selected, and each subject's scalp was divided into multiple regions. The liquid from Comparative Example 1, the hair growth agent from Example 1, or the hair growth agent from Example 2 was applied to each region twice daily, morning and evening, to evaluate the hair growth effect. The regions were divided so that the crown and temples of the head were included in every region. Measurements in the efficacy verification test were taken before use of the liquid or hair growth agent (hereinafter referred to as "hair growth agent, etc.") (first measurement), and at 2 weeks (second measurement), 4 weeks (third measurement), 6 weeks (fourth measurement), and 8 weeks (fifth measurement) after starting use of the hair growth agent, etc. The measurement procedure at each measurement timing was as follows. 1. Approximately 1 cm in each area of ​​the subject 2 Trim the hair in the area using scissors and a T-shaped razor, and take a photograph of the area immediately after trimming. 2. Take another photograph of the area two days after the hair was trimmed. 3. Compare a photograph taken immediately after hair cutting with a photograph taken two days later to analyze the length of hair growth over two days and calculate the amount of growth over those two days. The reason for trimming the hair before each measurement is that measuring with long hair could reduce measurement accuracy due to the hair's natural curl and expansion / contraction.

[0025] Figure 1 shows the results of the efficacy verification test, comparing the hair elongation rate 8 weeks after the start of use (5th measurement) for the liquid of Comparative Example 1 and the hair growth agents of Examples 1 and 2. The elongation rate (growth rate) is based on the amount of hair growth before the start of use of the hair growth agent (1st measurement), and was calculated using the formula "amount of hair growth at 5th measurement ÷ amount of hair growth at 1st measurement × 100". The elongation rate of Comparative Example 1 is shown as the baseline (100%). As shown in Figure 1, the elongation rate of Example 1 was approximately 160% higher than that of Comparative Example 1. This indicates that the elongation rate of Example 1, which contains human hair follicle stem cell culture supernatant, is significantly higher than that of Comparative Example 1, which does not contain human hair follicle stem cell culture supernatant. Furthermore, the elongation rate of Example 2 was approximately 140% of that of Comparative Example 1. Although it did not reach the level of Example 1, a significant increase in elongation was still observed compared to Comparative Example 1. These results confirm that a high hair growth effect can be obtained by including a sialic acid-containing extract, which contains swiftlet nest extract decomposed with a weak acid or solid acid, and human hair follicle stem cell culture supernatant. Furthermore, since a significant hair growth effect can be obtained even with a lower amount of sialic acid-containing extract, including swiftlet nest extract, than with human hair follicle stem cell culture supernatant, it is possible to conserve valuable swiftlet nest extract. [Industrial applicability]

[0026] The present invention can be used as a hair growth agent or hair regrowth agent in the form of a hair growth shampoo, hair growth conditioner, or hair growth tonic.

Claims

1. A sialic acid-containing extract, which contains swiftlet nest extract that has been decomposed with a weak acid or solid acid and contains sialic acid, is mixed with human hair follicle stem cell culture supernatant. A method for producing a hair growth agent, characterized by producing the hair growth agent by increasing the amount of human hair follicle stem cell culture supernatant to a greater amount than the amount of sialic acid-containing extract.

2. The method for producing the hair growth agent according to Claim 1, characterized in that the hair growth agent contains placenta extract.

3. The method for producing a hair growth agent according to any one of claims 1 to 2, characterized in that the hair growth agent is a shampoo, conditioner, or tonic.

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