Hair growth promoter, cosmetic, hair care agent, and shampoo soap

Abstract

To obtain a hair growth promoter, a cosmetic, a hair care agent and a soap for shampooing, having slight adverse effects and safely usable. To provide a hair growth promoter, a cosmetic, a hair care agent and a shampoo soap which can be used even by a consumer who could not use a conventional hair growing agent, hair tonic, hair tonic and cosmetic due to a problem of compatibility or the like.SOLUTION: The hair growth promoter, cosmetic, hair care agent and shampoo of the present invention contain whale oil as an active ingredient. The whale oil which is an active ingredient of the hair-growth promoter, cosmetic, hair-care agent and shampoo soap of the present invention is, for example, a whale oil derived from whale belonging to the family Mysticeti. The hair growth-promoting agent of the present invention promotes hair growth, for example. The dosage form of the hair growth promoter of the present invention is, for example, a transdermal preparation.SELECTED DRAWING: Figure 2

Description

[Technical Field]

[0001] The present invention relates to a hair growth promoter, a cosmetic, a hair care agent, and a hair washing soap. [Background technology]

[0002] Hair not only protects the head from external stimuli such as temperature, ultraviolet rays, and bruises, but also plays a role in excreting toxic substances such as heavy metals. Hair also has a significant impact on the visual effect of a person's appearance and the psychological effect of increasing self-confidence. Demand for hair care products to maintain beautiful hair is increasing, especially as economic activity recovers following the easing of restrictions imposed by the Covid-19 pandemic. In particular, the issue of hair loss due to aging and other factors is expected to contribute significantly to the growth of the hair care product market.

[0003] Currently, two drugs, minoxidil and finasteride, are approved by the U.S. Food and Drug Administration (FDA) to treat hair loss. Although the details of the mechanisms of action of these two drugs are largely unknown, minoxidil is presumed to prolong the duration of the hair growth phase (Non-Patent Documents 1 to 4).Finasteride is said to prevent male pattern baldness by inhibiting the activity of 5α-reductase, which affects the metabolism of male hormones (Non-Patent Document 5). However, minoxidil has been reported to have side effects such as scalp problems including rash and itching, while finasteride has been reported to have side effects such as liver dysfunction, erectile dysfunction, and rash (Non-Patent Documents 6-7).

[0004] For this reason, the hair care product market is in high demand for hair growth and hair care products that do not have these side effects. Natural ingredients have been attracting attention as ingredients in hair growth agents and hair restoration agents that meet this demand, and various hair growth agents and hair restoration agents that use natural ingredients as active ingredients have been proposed.

[0005] Patent Document 1 discloses a hair growth promoter containing an extract of Phellinus linteus (Mulberry yellow).

[0006] Patent Document 2 discloses a hair composition containing an extract of ginseng of the genus Panax in the family Araliaceae and an extract of tea plant of the genus Camellia in the family Theaceae. The active ingredient of the hair composition disclosed in Patent Document 2 is said to be a high concentration of saponin.

[0007] Patent Document 3 discloses a hair growth agent containing, as an active ingredient, an extract from one or more plants selected from the group consisting of Arisaema asiatica, marjoram, Actinidia polygama, and the fruit parts of plum.

[0008] Patent Document 4 discloses a hair agent having a hair growth effect, which contains, as an active ingredient, an extract extracted from the runners of a plant belonging to the genus Rubus in the family Rosaceae.

[0009] Patent Document 5 discloses a hair growth agent containing an extract of Snow Reishi. The Snow Reishi extract, which is the active ingredient of the hair growth agent disclosed in Patent Document 5, is said to have an excellent testosterone-5α-reductase inhibitory effect.

[0010] However, these known hair growth and hair restoration agents may not be able to exert sufficient hair growth and hair restoration effects, or may not be usable due to compatibility issues with the user (allergies, etc.). For this reason, there is still a great demand for the development of new hair growth and hair care products that use natural ingredients. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] Japanese Patent Application Laid-Open No. 2009-263250 [Patent Document 2] Japanese Patent Application Publication No. 2019-026634 [Patent Document 3] Japanese Patent Application Laid-Open No. 2014-185131 [Patent Document 4] Japanese Patent Application Laid-Open No. 2013-071908 [Patent Document 5] Special Announcement No. 2012-176921 [Non-licensed literature]

[0012] [Non-licensed Document 1] Meisheri, KD; Cipkus, LA; Taylor, CJ Mechanism of action of minoxidil sulfate-induced vasodilation: a role for increased 498 K+ permeability. J. Pharmacol. Exp. Ther. 1988, 245, 751-760. [Non-licensed Document 2] Buhl, AE; Waldon, DJ; Conrad, SJ; Mulholland, MJ; Shull, KL; Kubicek, MF, Johnson, GA, Brunden, MN, Stefanski, 500 KJ, Stehle, RG, et al. Potassium channel conductance: a mechanism affecting hair growth both in vitro and in vivo. J. Invest. 501 Dermatol. 1992, 98, 315-319. [Non-licensed Document 3] Lachgar, S.; Charveron, M.; Gall, Y.; Bonafe, JL Minoxidil upregulates the expression of vascular endothelial growth factor in 503 human hair dermal papilla cells. Br. J. Dermatol. 1988, 138, 407-411. [Non-licensed Document 4] Messenger, AG; Rundegren, J. Minoxidil: mechanisms of action on hair growth. Br. J. Dermatol. 2004, 150, 186–194.

Direct Environment 5

Outdoor Configuration6

Direct Environment 7

Outdoor Track 8

[0013] The present invention has been made in view of the above-mentioned background art. An object of the present invention is to provide a novel hair growth promoter, cosmetic, hair care agent, and hair washing soap that have few side effects and can be used safely. [Means for solving the problem]

[0014] The present inventors have conducted extensive research to solve the above problems and have discovered the following facts, which have led to the completion of the present invention.

[0015] In other words, whale oil, an oil extracted from cetaceans, is a natural ingredient that has traditionally been used for a variety of purposes, and the inventors' research has confirmed that whale oil has hair growth and hair care effects.

[0016] The present invention thus completed is as follows:

[0017] A hair growth promoter characterized by having whale oil as an active ingredient.

[0018] Cosmetics characterized by whale oil as an active ingredient.

[0019] A hair care agent characterized by containing whale oil as an active ingredient.

[0020] A hair washing soap characterized by having whale oil as an active ingredient. [Effects of the Invention]

[0021] According to the present invention, it is possible to provide novel hair growth promoters, cosmetics, hair care agents, and hair washing soaps that have few side effects and can be used safely.

[0022] The hair growth promoter of the present invention contains whale oil as an active ingredient, which has not previously been known as an ingredient in hair growth and hair restoration agents. Even consumers who have been unable to use conventional hair growth and hair restoration agents due to compatibility issues or other reasons may be able to use the hair growth promoter of the present invention, and the present invention can provide consumers with a new option for hair growth and hair restoration agents.

[0023] It has been reported that conventional hair growth agents such as minoxidil often cause telogen effluvium 4 to 10 weeks after use (Non-Patent Document 8). In this regard, the hair growth promoter of the present invention does not cause telogen effluvium, and therefore, according to the present invention, it is possible to provide a hair growth agent, hair restoration agent, etc. that consumers can use with confidence. [Brief explanation of the drawings]

[0024] [Figure 1] 1 is a photograph of the back of a mouse taken in Evaluation Example 1. [Figure 2] 1 is a graph showing the hair growth rate calculated in Evaluation Example 1 (**: p<0.01). [Figure 3] 1 shows cross-sectional images of mouse skin tissue taken in Evaluation Example 2. [Figure 4] 1 is a graph showing the average number of hair follicles in mice calculated in Evaluation Example 2 (*: p<0.05, **: p<0.01). [Figure 5] 1 is a graph showing the average thickness of the subcutaneous tissue of mice calculated in Evaluation Example 2 (*: p<0.05, **: p<0.01). [Figure 6] 1 is a graph showing the cell viability calculated in Evaluation Example 3 (*: p<0.05, **: p<0.01). [Figure 7] 1 shows details of primers specific to the FGF7 gene and the VEGF gene used in RT-PCR analysis in Evaluation Example 4. [Figure 8] 1 is a graph showing the expression level of the FGF7 gene calculated in Evaluation Example 4 (*: p<0.05). [Figure 9] 1 is a graph showing the expression levels of the VEGF gene calculated in Evaluation Example 4 (*: p<0.05, **: p<0.01). [Figure 10] 1 is a graph showing (a) hair moisture content, (b) hair glossiness, and (c) hair tensile strength calculated in Evaluation Example 8 (**: p<0.01 vs. whale oil application). [Figure 11] 1 is a graph showing (a) the surface bending rigidity and (b) the surface bending hysteresis calculated in Evaluation Example 8 (**: p<0.01 vs. whale oil application). [Figure 12] 1 is a graph showing (a) the average coefficient of friction and (b) the average deviation of the coefficient of friction calculated in Evaluation Example 8 (**: p<0.01 vs. whale oil application). [Figure 13] 10 shows photographs of the subject's forehead, parietal region, and occipital region taken in Evaluation Example 9. [Figure 14] 1 is a graph showing (a) the number of hairs and (b) the hair diameter calculated in Evaluation Example 9 (**: p<0.01 vs. whale oil application). [Figure 15] 10 is a graph showing the number of fallen hairs calculated in Evaluation Example 9. DETAILED DESCRIPTION OF THE INVENTION

[0025] The present invention will be described below, but the present invention is not limited to the following embodiments and can be practiced with any modifications.

[0026] The present invention relates to a hair growth promoter, a cosmetic, a hair care agent, and a hair washing soap containing whale oil as an active ingredient.

[0027] The "hair growth promoter" in the present invention is an agent that has the effect of promoting the growth of body hair in a subject to which it is administered. The mechanism of hair growth is thought to involve proliferation of hair papilla cells, an increase in the number of hair follicles, and an increase in the thickness of the subcutaneous tissue.

[0028] The hair growth promoter of the present invention may be a hair growth agent that has the effect of growing new hair from the hair root, a hair restoration agent that causes existing hair to grow thicker and longer, or a hair care agent that supports hair growth.

[0029] Whale oil, the active ingredient of the hair growth promoter of the present invention, is oil extracted from cetaceans. Whale oil has traditionally been used for purposes such as fuel oil for lighting, a candle material, machine lubricant, leather detergent, and a margarine material, and can be considered a highly safe, naturally derived ingredient.

[0030] Whale oil contains triglycerides of fatty acids such as linoleic acid, linoleic acid, octadecathotolaenoic acid, lauric acid, tridecanoic acid, myristoleic acid, myristic acid, pentadecenoic acid, pentadecylic acid, pentadecanoic acid, palmitoleic acid, palmitic acid, hexadecadienoic acid, heptadecenoic acid, heptadecanoic acid, margaric acid, linoleic acid, linoleic acid, oleic acid, cis-vaccenic acid, stearic acid, nonadecenoic acid, nonadecanoic acid, eicosapentaenoic acid, arachidonic acid, arachidic acid, eicosatetraenoic acid, eicosatrienoic acid, eicosadienoic acid, eicosenoic acid, heneicosapentaenoic acid, gadoleic acid, docosahexaenoic acid, docosapentaenoic acid, docosatetraenoic acid, behenic acid, and erucic acid; aliphatic aldehydes; acetic acid; ketones; hydrocarbons; and the like.

[0031] n-3 polyunsaturated fatty acids such as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA) are generally known to have high antioxidant properties, reduce inflammation throughout the body, and prevent atherosclerosis, and their intake is known to contribute to various health-promoting effects. Furthermore, stearic acid is an ingredient that helps improve barrier function and has traditionally been used in conditioners and treatments, making whale oil, which contains stearic acid, suitable as a cosmetic ingredient.

[0032] The whale oil of the present invention may be extracted from any type of cetacean. Examples of cetacean animals that are sources of whale oil include whales belonging to the suborder Mysticeti, such as fin whales, humpback whales, blue whales, minke whales, Antarctic minke whales, sei whales, Bryde's whales, Bryde's whales, Beretta whales, Siberian right whales, rice whales, bowhead whales, right whales, southern right whales, North Atlantic right whales, and pygmy right whales; and whales belonging to the suborder Odontoceti, such as sperm whales, beaked whales, short-finned pilot whales, Risso's whales, false killer whales, and short-finned pilot whales. In terms of the high hair growth promoting effect and ease of availability, the whale oil used in the present invention is preferably derived from whales belonging to the suborder Mysticeti.

[0033] The whale oil of the present invention may be extracted from any part of a cetacean. Examples of whale oil sources include skin, bones, internal organs, and muscles. Among these, whale oil extracted from skin is preferred as the whale oil of the present invention because it is easy to extract.

[0034] There is no particular limitation on the method for collecting whale oil in the present invention, and known methods such as boiling and decoction can be used.

[0035] In the present invention, whale oil collected by the above-mentioned method may be subjected to some kind of treatment, and the resulting treated whale oil may be used as a hair growth promoter. Specific examples of such processed whale oil products include concentrates; pastes; dried products such as spray-dried products, vacuum-dried products, and drum-dried products; liquefied products; diluted products; crushed products; and sterilized products.

[0036] There are no particular limitations on the subjects to which the hair growth promoter of the present invention can be administered, and examples include humans; monkeys; pets such as cats and dogs; livestock such as horses, cows, pigs, goats, and chickens; and laboratory animals such as mice and rats. In view of demand, humans are the most suitable recipients of the hair growth promoter of the present invention.

[0037] There are no particular limitations on the type of hair whose growth is promoted by the hair growth promoter of the present invention, and examples include scalp hair, beard, eyelashes, eyebrows, ear hair, nose hair, armpit hair, chest hair, abdominal hair, pubic hair, buttocks hair, leg hair, downy hair, and finger hair. In view of demand, it is desirable that the hair growth promoter of the present invention promotes hair growth.

[0038] The dosage form of the hair growth promoter of the present invention is not particularly limited, and examples thereof include transdermal agents, oral agents, injections, sprays, inhalants, and dusting agents. The hair growth promoting agent of the present invention is particularly preferably a transdermal agent, since it can be administered only to the necessary area and is easy to administer.

[0039] When the hair growth promoter of the present invention is a transdermal agent, it can be in the form of, for example, a lotion, ointment, cream, spray, tape, or cataplasm.

[0040] The hair growth promoter of the present invention may be a pharmaceutical product or a quasi-drug. In addition, if it is a quasi-drug, it may be a medicated cosmetic product such as a medicated shampoo or a medicated rinse.

[0041] In addition to the active ingredient whale oil, other ingredients can be added to the hair growth promoter of the present invention. Specific examples of such "other ingredients" include known ingredients other than whale oil that have hair growth and hair restoration effects, excipients, binders, disintegrants, lubricants, colorants, solvents, solubilizers, suspending agents, isotonicity agents, buffers, soothing agents, preservatives, antioxidants, solubilizers, adsorbents, pH adjusters, and fragrances. The hair growth promoter of the present invention may contain only one of these "other ingredients" or two or more of them.

[0042] The content of the whale oil-derived component in the hair growth promoter of the present invention is preferably 0.01% by mass or more, more preferably 0.1% by mass or more, and particularly preferably 1% by mass or more, and is preferably 99% by mass or less, more preferably 90% by mass or less, and particularly preferably 80% by mass or less. The above-mentioned preferred range of content is the range of content at the time of use of the hair growth promoter of the present invention. The hair growth promoter of the present invention may be stored in a form in which the active ingredient, etc. is concentrated.

[0043] There are no particular limitations on the dosage of the hair growth promoter of the present invention, and it can be appropriately selected depending on the age and weight of the individual to be administered, the degree of the desired effect, the method of administration, and the like. For example, in the case of a transdermal preparation, the daily dose for an adult is preferably 1 mg or more, more preferably 10 mg or more, and particularly preferably 100 mg or more, calculated as the amount of whale oil-derived components, and is preferably 100 g or less, more preferably 10 g or less, and particularly preferably 1 g or less.

[0044] There is no particular limitation on the administration frequency of the hair growth promoter of the present invention. The administration frequency is preferably at least once every three days, more preferably at least once a day, and particularly preferably at least three times a day. The administration frequency is preferably 30 times or less a day, more preferably 20 times or less a day, and particularly preferably 10 times or less a day.

[0045] As will be shown in the evaluation examples described below, the hair growth-promoting agent of the present invention promotes the growth of body hair in a subject by proliferating dermal papilla cells and increasing the number of hair follicles in the subject.

[0046] The hair growth-promoting agent of the present invention also increases the thickness of the subcutaneous tissue in which hair follicles are located. The thickness of the subcutaneous tissue is thought to be involved in defense against environmental factors and external stress, protection of hair follicles by strengthening the formation of the dermis and fatty tissue layer below the hair follicle, including moisture retention and heat retention, and promotion of the supply of necessary nutrients. It is believed that the hair growth promoter of the present invention increases the thickness of the subcutaneous tissue, thereby creating an environment that promotes healthy hair growth and further improving hair growth.

[0047] The present invention also relates to cosmetics containing whale oil as an active ingredient. The cosmetic product of the present invention can be in the form of, for example, shampoo, rinse, conditioner, hair tonic, hair cream, hair liquid, lotion, or emulsion.

[0048] The cosmetic product of the present invention contains whale oil as an active ingredient, and as described above, whale oil has the effect of promoting the growth of body hair in the subject to which it is administered. For this reason, the cosmetic product of the present invention is mainly used by applying it to the head.

[0049] The present invention also relates to a hair care agent characterized by containing whale oil as an active ingredient. The hair care agent of the present invention is used for the purpose of improving hair quality, caring for damaged hair, and the like. The hair care agent of the present invention is used mainly by applying it to the scalp.

[0050] The hair care agent of the present invention has effects such as increasing the moisture content of hair (improving moisturizing properties), improving hair luster, increasing hair tensile strength, improving hair flexibility (bending rigidity / bending hysteresis), and smoothing hair by reducing friction on the hair surface.

[0051] The present invention also relates to a hair washing soap characterized by containing whale oil as an active ingredient. The hair washing soap of the present invention may be a solid soap or a liquid soap. The hair washing soap of the present invention is used to wash hair, and since whale oil has the effect of promoting hair growth in the recipient, it is suitable as a soap to be used, for example, by people who wish to promote hair growth, hair care, and hair care. [Example]

[0052] The present invention will be explained in more detail below by way of evaluation examples, but the present invention is not limited to these evaluation examples as long as they do not depart from the gist of the invention.

[0053] Whale oil was obtained by collecting the skin of Bryde's whale, extracting it with heated steam at 200°C for 20 minutes, and then freezing it. The whale oil thus obtained was used as a whale oil sample in each evaluation example described below.

[0054] Evaluation example 1 (evaluation of hair growth) A sample was prepared by dissolving whale oil in a mixed solution of propylene glycol:ethanol:water = 5:3:2 (volume ratio) to a concentration of 20% by mass, and a sample was prepared by dissolving minoxidil in the same mixed solution to a concentration of 3% by mass. The mixed solution itself was used as the control sample.

[0055] C57BL / 6J mice were divided into three groups (6 mice per group) and housed at 22±2°C, 6:00 AM to 6:00 PM, with a 12-hour / 12-hour light / dark cycle. Each group of mice received daily application of 200 μL of one of the prepared samples to the back of the mice. The backs of the mice were shaved weekly.

[0056] The condition of the backs of the mice was observed and photographed at weeks 0, 1, and 3. The photographs were analyzed using Image J software (National Institutes of Health and the Laboratory for Optical and Computational Instrumentation, University of Wisconsin), and the ratio of the hair-grown area to the shaved area was calculated to calculate the hair growth rate (%). The results were expressed as mean ± standard error using Tukey's method, with p<0.05 considered to indicate a significant difference (the same applies to other evaluation examples).

[0057] The results are shown in Figures 1 and 2. In Figures 1 and 2, "WO" is the group to which whale oil was applied, "MXD" is the group to which minoxidil was applied, and "Vehicle" is the control group.

[0058] The group that received whale oil showed a significantly higher hair growth promotion effect in the third week. Furthermore, while there was individual variation in the hair growth promotion effect in the group that received minoxidil, there was almost no individual variation in the group that received whale oil, and a high hair growth promotion effect was confirmed in all individuals.

[0059] Evaluation example 2 (observation of hair follicles and subcutaneous tissue) In Evaluation Example 1, after the completion of observation and photography at the third week, the skin tissue from the back of the mice was fixed in 4% paraformaldehyde solution and replaced with phosphate-buffered saline (PBS). Paraffin sections were dehydrated and infiltrated with xylene for paraffin embedding. 5 μm paraffin sections were treated with hematoxylin / eosin staining, dehydrated, permeabilized, and sealed with a cover glass.

[0060] Cross-sectional images were taken at 40x magnification using a BZ-X710 Brightfield System (Keyence Corporation), and the images were analyzed using Image J software. The total number of hair follicles in the dermis and subcutaneous tissue and the thickness of the subcutaneous tissue were measured, and the average number of hair follicles and the average thickness of the subcutaneous tissue were calculated (n = 6).

[0061] The results are shown in Figures 3 to 5. In Figures 3 to 5, "WO" is the group to which whale oil was applied, "MXD" is the group to which minoxidil was applied, and "Vehicle" is the control group.

[0062] The number of hair follicles increased significantly in both the whale oil and minoxidil groups, but the increase in the number of hair follicles was particularly remarkable in the whale oil group (Figure 4). Furthermore, in the whale oil group, it was also possible to confirm in a skin microscope photograph (Figure 3) that the hair follicles were densely packed together. Furthermore, a significant increase in the thickness of the subcutaneous tissue was observed in the group to which whale oil was applied.

[0063] Evaluation Example 3 (Evaluation of the proliferation effect of human hair dermal papilla cells by MTT assay) 3 x 10 human hair dermal papilla cells (HFDPC) 4 The cells were seeded into a 96-well culture plate at 100 μL / well and pre-incubated for 24 hours at 37°C and 5% CO. The cultures were grown in DMEM medium (Dulbecco's Modified Eagle's Medium "Nissui" (2), Nissui Pharma) supplemented with 10% FBS (fetal bovine serum) and 1% antibiotic (Sigma A5955) at a ratio of 1%.

[0064] After pre-culture, the medium was replaced with fresh medium, and whale oil, docosapentaenoic acid (DPA), or eicosapentaenoic acid (EPA) adjusted to a concentration of 0.0-5.0% in D / E solution (a 1:1 volumetric mixture of dimethyl sulfoxide and ethanol), or minoxidil adjusted to a concentration of 5-30 μM in D / E solution was added.

[0065] After culturing for 24 to 120 hours under conditions of 37°C and 5% CO2, the cells were incubated with MTT (3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide) from Cell Proliferation Kit I (Roche Life Sciences). After culturing for 4 hours under conditions of 37°C and 5% CO2, 100 μL / well of solubilization solution (0.01 M HCl, 10% SDS) was added, and the cells were cultured for 24 to 120 hours under conditions of 37°C and 5% CO2. Then, 100 μL / well of solubilization solution (0.01 M HCl, 10% SDS) was added, and the cells were cultured at 37° C. and 5% CO 2 for 17 hours.

[0066] After incubation, absorbance at 570 nm was measured using an iMark microplate reader (Bio-Rad) with a reference wavelength of 650 nm. The number of viable cells without the addition of the evaluation sample was set at 100, and the number of viable cells with the addition of the evaluation sample was measured to calculate the cell viability (%). Each measurement was repeated twice, and the average value (average of the values ​​from each of the 12 wells) was calculated.

[0067] The results are shown in Figure 6. In Figure 6, "WO" is the group to which whale oil was added, "DPA" is the group to which docosapentaenoic acid was added, "EPA" is the group to which eicosapentaenoic acid was added, "MXD" is the group to which minoxidil was added, and "D / E" is the control group.

[0068] In the whale oil group and minoxidil group, a significant cell proliferation effect was confirmed compared to the control group at all concentrations of 10 μM, 15 μM, and 30 μM. The proliferation effect on human hair dermal papilla cells in the whale oil group was greater than that in the docosapentaenoic acid or eicosapentaenoic acid groups, suggesting that components other than docosapentaenoic acid and eicosapentaenoic acid are involved in this proliferation effect.

[0069] Evaluation example 4 (Confirmation of gene expression related to hair growth) Total RNA was extracted from the mixture of whale oil, docosapentaenoic acid, eicosapentaenoic acid, or minoxidil in D / E solution and human hair papilla cells prepared in Evaluation Example 3, and incubated for 30, 60, and 90 minutes using an RNeasy Micro Kit (Qiagen). Total RNA extraction was performed according to standard protocols.

[0070] RNA quality was confirmed by measuring the RNA concentration and the absorbance ratios at 260 nm / 280 nm and 260 nm / 230 nm using a spectrophotometer. cDNA was synthesized using 100–500 ng of RNA with the Affinity Script qPCR cDNA Synthesis Kit (Agilent), followed by RT-PCR analysis using specific primers for the FGF7 and VEGF genes (Figure 7) using the Emerald Amp PCR Master (Takara Bio). PCR reactions (initial denaturation at 97°C for 5 minutes, heat denaturation at 95°C for 45 seconds, annealing at 55°C for 45 seconds, and extension at 72°C for 1 minute) were performed for 27–45 cycles.

[0071] After PCR, the PCR products were separated on a 1.5% agarose gel and visualized with ethidium bromide and ultraviolet light. The visualized expression levels of the target genes were normalized with the expression level of the housekeeping gene, GAPDH, and plotted (n = 3).

[0072] The results of FGF7 gene expression are shown in Figure 8, and the results of VEGF gene expression are shown in Figure 9. In Figures 8 and 9, "WO" is the group to which whale oil was added, "DPA" is the group to which docosapentaenoic acid was added, "EPA" is the group to which eicosapentaenoic acid was added, "MXD" is the group to which minoxidil was added, and "D / E" is the control group.

[0073] Evaluation example 5 (refining and component analysis of whale oil) Whale oil contains many hydrophilic impurities and fats that remain solid at room temperature, which affect the uniformity of quality when it is made into a hair growth agent. For this reason, the whale oil was refined as follows, and only the components that remain liquid at room temperature were extracted.

[0074] First, whale oil at room temperature and hexane (Fujifilm Wako) were mixed in a volume ratio of 1:1 and centrifuged (20°C, 7830 rpm, 15 min) to precipitate and remove impurities other than the lipophilic components contained in the whale oil. Next, the solution from which the impurities had been removed was heated to 60°C, and the hexane was evaporated and removed. The resulting oil was then left to stand overnight at room temperature, and after standing, it was centrifuged (4°C, 307830 rpm, 30 min) to separate the liquid oil components. This procedure was repeated twice to extract the liquid oil components at room temperature.

[0075] Next, activated carbon (Fujifilm Wako Co., Ltd.) was added in an amount 10 times the amount of the liquid oil and fat components, and the mixture was deodorized by stirring overnight with a stirrer. After stirring, the activated carbon was removed by suction filtration, and the filtered liquid was allowed to stand overnight at 4°C. Finally, the liquid was centrifuged (5°C, 7830 rpm, 120 min), and only the supernatant liquid was collected and further sterilized by filtration using a membrane filter (IWAKI Co., Ltd.).

[0076] An appropriate amount of filter-sterilized sample was dissolved in 50 mL of a chloroform / methanol mixture (2:1). An aliquot of the dissolved material was added to an internal standard (heptadecanoic acid). A 0.5 mol / L solution of sodium hydroxide in methanol was then added for saponification. A solution of boron trifluoride-methanol complex in methanol was added to the saponified solution for methyl esterification. Hexane and saturated saline were then added, and the hexane layer was used as the sample.

[0077] The samples were analyzed by GC / FID (gas chromatography / flame ionization detector) using a capillary gas chromatograph (GC-2025, Shimadzu Corporation). Components were identified from the sample's retention time, and the content of each component was calculated from the peak area of ​​the gas chromatogram. The analytical results are shown in Table 1 (amount of fatty acids contained in refined whale oil) and Table 2 (quantitative analysis of 45 fatty acids contained in refined whale oil by GC and their content ratios).

[0078] [Table 1]

[0079] [Table 2]

[0080] Refined whale oil contained many unsaturated fatty acids (Table 1). In particular, n-6 unsaturated fatty acids and n-9 unsaturated fatty acids have antioxidant properties and inhibit arteriosclerosis, and are often found in hair growth supplements. Furthermore, n-3 unsaturated fatty acids may promote blood flow to hair matrix cells in hair follicles, which are important for hair growth and development. Additionally, omega-3 has been reported to promote hair growth. Furthermore, refined whale oil also retained large amounts of EPA, DHA, and DPA, which are abundant in whale oil (Table 2).

[0081] Evaluation Example 6 (Preparation of Hair Growth Agent Sample) Using the whale oil refined in Evaluation Example 5, a hair growth agent sample containing whale oil as an active ingredient was prepared according to the component composition shown in Table 3 (hereinafter referred to as "whale oil hair growth agent A").

[0082] [Table 3]

[0083] The solubilizing agent, cetanol (Fujifilm Wako Co., Ltd.), was added to the base vehicle, ethanol (Fujifilm Wako Co., Ltd.), and dissolved. To the solution was then added Tween 60 (Fujifilm Wako Co., Ltd., "TWEEN" is a registered trademark), a highly hydrophobic solubilizing agent, and further refined whale oil was added and stirred overnight. After confirming complete dissolution, phenoxyethanol (Fujifilm Wako) was added as a preservative, tocopherol (Fujifilm Wako) as an antioxidant, and peppermint essential oil (Pronalom) was added for fragrance.

[0084] A safety test was conducted by analyzing the ingredients of the prepared whale oil hair growth agent A. The safety test was conducted in accordance with the standards set out in the "Cosmetics Standards (Ministry of Health, Labour and Welfare Notification No. 331)" based on Article 42, Paragraph 2 of the Pharmaceutical and Medical Device Act (Act on Ensuring Quality, Efficacy and Safety of Pharmaceuticals, Medical Devices, etc.), and included an analysis of prohibited ingredients, upper limit ingredients, recommended testing items, and cosmetic testing. Table 4 shows the prohibited ingredients, upper limit of ingredients, and analytical methods.

[0085] [Table 4]

[0086] The results of the component analysis of whale oil hair growth agent A are shown in Table 5. As for prohibited ingredients and ingredients with upper limit of inclusion, almost no substances were detected other than phenoxyethanol, which was added as a preservative. The amount of phenoxyethanol detected was also within the upper limit of 1%. This confirms that whale oil hair growth agent A complies with cosmetic standards and is safe.

[0087] [Table 5]

[0088] Evaluation Example 7 (Safety test of hair growth agent samples in humans) A safety test of whale oil hair growth agent A was conducted using a closed patch test (24-hour closed method) on six Japanese men and 14 Japanese women aged 20 to 60 years (those who did not meet certain exclusion criteria) as subjects. The 24-hour occlusion method was performed by applying a skin test patch tape (Torii Pharmaceutical Co., Ltd.) filled with 30 μL of whale oil hair growth agent A to the subject. After 24 hours, the patch tape was removed, and a doctor assessed safety 60 minutes after removal. 24 hours later, a doctor again assessed safety.

[0089] All 20 subjects showed no reaction 60 minutes after peeling and 24 hours after peeling, and none of the subjects developed any skin abnormalities. The skin irritation index was also 0.0, and Whale Oil Hair Growth Agent A was classified as a safe product.

[0090] Evaluation Example 8 (Evaluation of hair care effects using human hair) The test product was made by applying whale oil hair growth agent A to human hair that had been damaged by ultraviolet light, and the following evaluations were carried out on the test product. In addition, a control test product in which camellia oil (Kurobara Honpo Co., Ltd.) was applied to the same hair, and an untreated test product in which nothing was applied to the same hair were also evaluated in the same way.

[0091] (8-1) Hair moisture content (measurement of gloss, elasticity, and body) Measurements were taken using an infrared moisture meter FD-660 (Kett Electric Laboratory Co., Ltd.). Approximately 1 g of test hair bundles were cut into small pieces of 5-10 mm, and each bundle was heated to 90°C five times to dry the moisture content before measurement. The average value was calculated from the measurement results. The results are shown in Figure 10(a).

[0092] (8-2) Hair gloss (measurement of luster) Measurements were taken using a gloss meter MPA-5 (Integral Corporation). A probe was pressed against the center of the hair and four adjacent locations (a total of five locations), and the reflection of the light emitted from the probe was automatically measured. Measurements were taken five times per hair bundle, and the average value was calculated. The results are shown in Figure 10(b).

[0093] (8-3) Hair tensile strength (measurement of flexibility and strength) Measurements were taken using a digital force gauge ZP-20N (IMADA Co., Ltd.). Five random hairs from the hair bundle were attached to both ends of the attachment, and the force (N) at which the hair broke was measured by pulling the hair at a speed of 5 mm / s with a motor. Measurements were taken five times per hair bundle. The average value was calculated. The results are shown in Figure 10(c).

[0094] (8-4) Multipurpose pure bending test (strength, stiffness) Measurements were performed using a multipurpose pure bending tester (KES-FB2-S / Kato Tech Co., Ltd.). Prior to measurement, the hair bundle was cut to a length of 10 cm from the root. Since a measurement sample of 100 hairs was used, the hairs cut from the root of the bundle were counted, and the ends were aligned and secured with cellophane tape. After securing the 100 hairs with cellophane tape, any excess hair extending beyond the tape was cut off. The sample was set so that the front of the pure bending tester was the root and the back was the tip, and measurements were performed after each measurement. After each measurement, the next measurement was performed without moving the sample, for a total of five measurements. The average of the five measurements was calculated. The results are shown in Figure 11. The surface bending stiffness (Figure 11(a)) is an index of the flexibility of hair, with the softer the hair, the smaller the value. The surface bending hysteresis (Figure 11(b)) indicates that the smaller the value, the higher the recovery.

[0095] (8-5) Hair analysis (smoothness and smoothness) Measurements were taken using a friction tester KES-SE (Kato Tech Co., Ltd.). After fixing the hair bundle to the sample stand, it was fixed with a square holding frame to maintain tension in the hair bundle. After setting the friction element at a point about 10 cm from the root, measurements were taken five times and the average value was calculated. The results are shown in Figure 12. The average friction coefficient (Figure 12(a)) indicates how slippery the hair surface is, with smaller values ​​indicating more slipperiness. The average deviation of the friction coefficient (Figure 12(b)) indicates how smooth the hair surface is, with smaller values ​​indicating smoother hair.

[0096] [Discussion of results] Test products in which whale oil hair growth agent A was applied to hair damaged by UV exposure had significantly greater moisture content, gloss, and tensile strength than control test products in which camellia oil was applied, or untreated test products in which nothing was applied (Figure 10). Furthermore, the test items to which whale oil hair growth agent A was applied had significantly smaller surface bending stiffness and surface bending hysteresis than the control test items and untreated test items (Figure 11). Furthermore, the test items to which whale oil hair growth agent A was applied had significantly smaller average friction coefficients and average deviations in the friction coefficients than the control test items and untreated test items (Figure 12).

[0097] These results demonstrate that whale oil hair growth agent A has superior hair care effects in all evaluation items, surpassing camellia oil. In particular, significant differences were observed in terms of improved hair moisture retention, smoothness, elasticity and strength. This suggests that whale oil hair growth agent A can be used not only as a hair growth agent, but also as a wide range of hair care products aimed at improving hair quality and caring for damage.

[0098] Evaluation Example 9 (Verification of hair growth effect on human scalp) The hair growth effect on the human scalp was examined using 16 Japanese men aged 40 to 69 years who were aware of thinning hair (those who did not meet certain exclusion criteria) as subjects. Five pumps (1 mL) of Whale Oil Hair Growth Tonic A were applied to the entire head once a day before going to bed for 24 weeks (168 days), and the following analyses were conducted based on the before and after evaluations.

[0099] (9-1) Digital photography of the head (front, top, and back of the head) The head was divided equally into left and right halves from the center, and the frontal, parietal, and occipital regions were photographed.

[0100] After a 24-week continuous application test, a tendency toward hair growth promotion was confirmed in all photographed areas. In particular, in the forehead and top of the head, where thinning hair was most prevalent, increases in hair density and thickness were relatively clear, giving the visual impression of increased hair volume. A similar tendency was observed in the back of the head, but the changes were relatively mild (Figure 13).

[0101] (9-2) Analysis using a digital microscope (number of hairs and hair diameter) After photographing the top of the head with a microscope, the number of hairs and hair diameter were analyzed using image analysis software (ImageJ) and compared over time using Dunnett's test. The results are shown in FIG.

[0102] [Table 6]

[0103] The number and diameter of hairs increased significantly after 12 and 24 weeks of use. Generally, hair thickness is considered one of the indicators of healthy hair, and it is said that thinning hair becomes more noticeable as the diameter decreases. It was shown that applying whale oil hair growth agent A can improve the health of hair.

[0104] (9-3) Photographing the shaved area The hair was shaved to an area of ​​approximately 1 cm x 1 cm and approximately 1 mm around the intersection of the midline of the head and the top of the left and right ear helices, and the shaved area was photographed.

[0105] (9-4) Hair loss analysis The experts analyzed the number of hairs washed and the number of hairs that fell out during the wash. The results are shown in FIG.

[0106] [Table 7]

[0107] No change in the number of hairs that fell out was observed after application of whale oil hair growth agent A. It has been reported that many hair growth agents, such as minoxidil, cause telogen effluvium 4 to 10 weeks after use. On the other hand, whale oil hair growth agent A did not show a significant increase in the number of hairs that fell out.

[0108] [Discussion of results] Whale oil hair growth agent A was found to have a hair growth promoting effect in humans. Furthermore, unlike existing hair growth agents, whale oil hair growth agent A showed almost no short-term hair loss after starting use. This suggests that whale oil may be used as an ingredient in new hair growth agents with fewer side effects such as initial hair loss.

Claims

1. A hair growth promoter characterized by having whale oil as an active ingredient.

2. 2. The hair growth promoter according to claim 1, wherein the whale oil is derived from a whale belonging to the suborder Mysticeti.

3. The hair growth promoter according to claim 1, which promotes scalp hair growth.

4. The hair growth promoter according to claim 1, which is a transdermal agent.

5. Cosmetics characterized by whale oil as an active ingredient.

6. A hair care agent characterized by containing whale oil as an active ingredient.

7. A hair washing soap characterized by having whale oil as an active ingredient.

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