Anti-inflammatory agents

Abstract

The object of the present invention is to provide an inflammation inhibitor that can reduce oxidative damage and allergic symptoms of the scalp by using a substance that does not interfere with the progress of oxidation reactions during hair dyeing and that preferentially removes more damaging peroxides. [Solution] This is an inflammation inhibitor that reduces inflammatory damage to the scalp and allergic inflammatory reactions that occur when dyeing hair with an oxidative hair dye, and contains an antioxidant that removes peroxides produced by the reaction between the oxidative dye contained in the hair dye and hydrogen peroxide without inhibiting the oxidation reaction during hair dyeing.

Description

[Technical Field]

[0001] The present invention relates to an inflammation inhibitor that reduces inflammatory damage and allergic inflammatory reactions of the scalp that occur during hair dyeing using an oxidation reaction. [Background technology]

[0002] Traditionally, most hair dye products use hair dyes containing oxidative dyes, such as the aromatic amine compound paraphenylenediamine (PPD). Some consumers are seeking natural and hypoallergenic hair dyes, and this demand is steadily increasing. As a result, the market is seeing an increase in oxidative-free hair dye options. However, hair dyes containing oxidative dyes are generally inexpensive and widely available, and offer more vibrant and durable color than other compounds, making them the preferred choice for many beauty professionals and consumers. Currently, they dominate the hair dye market. According to a 2010 International Agency for Research on Cancer (IARC) survey, an estimated 50–80% of women worldwide have used hair dye at least once in their lives. Furthermore, a previous survey found that 50.9% of Europeans had dyed their hair at least once in their lifetime, with 81.4% of these being women. A previous survey conducted in South Korea reported that more than 50% of people experienced gray hair in their early 40s, and 60% of these people continued to dye their hair regularly despite side effects. Hair dyes containing PPD are said to account for more than 70% of the market in Europe, the United States, and East Asia.

[0003] Hair dyes containing oxidative dyes are oxidized with hydrogen peroxide (H2O2) to produce pigments and dye hair. However, side reactions produce peroxides and Bandorovsky bases, which can cause allergic symptoms and skin inflammation. When diamine-based oxidative dyes such as PPD cause allergic contact dermatitis, mild cases can result in irritant contact dermatitis, such as burning and prickling, upon dyeing hair. Moderate cases can result in allergic contact dermatitis, such as persistent itching, redness, and bumps that persist the next day or later. Once the allergic reaction appears, it worsens with each use of the hair dye, and in severe cases, systemic symptoms can develop. Although there are individual differences, even in people without allergies, the oxides produced by the oxidation reaction of oxidative dyes can induce scalp inflammation, resulting in symptoms such as itching and redness. It is believed that these allergic symptoms and inflammatory reactions could be improved by inhibiting the oxidation reaction and the production of peroxides. However, reducing agents, even though they inhibit oxidative damage, cannot be used because they also inhibit the dyeing reaction, which is also an oxidative reaction. In addition, applying a protective cream or the like to the scalp can suppress inflammation and oxidative damage to the skin, but if the protective cream or the like gets on the hair, it will inhibit the dyeing of the hair with the dyeing agent, resulting in uneven dyeing, etc., so it is necessary to apply the protective cream or the like only to the scalp, avoiding the hair, which is time-consuming and difficult to handle. On the other hand, for example, Patent Document 1 discloses a dyeing composition that contains PPD and orthoaminophenol in a predetermined ratio to suppress inflammation of the skin. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Japanese Patent Application Publication No. 56-15205 Summary of the Invention [Problem to be solved by the invention]

[0005] According to Patent Document 1, the inclusion of ortho-aminophenol is said to prevent the formation of Bandorovsky base and suppress skin inflammation, but at present, no inflammation inhibitor has been developed that can effectively suppress skin inflammation without interfering with hair dyeing.

[0006] The present invention has been made in view of the above circumstances, and aims to provide an inflammation inhibitor that can reduce oxidative damage and allergic symptoms of the scalp by using a substance that does not interfere with the progression of the oxidation reaction during hair dyeing and that preferentially removes more damaging peroxides. [Means for solving the problem]

[0007] The inflammation inhibitor of the present invention, which meets the above-mentioned objectives, is an inflammation inhibitor that reduces inflammatory damage to the scalp and allergic inflammatory reactions that occur when dyeing hair using a hair dye that involves oxidation, and contains an antioxidant that removes peroxides produced by the reaction between the oxidative dye contained in the hair dye and hydrogen peroxide without inhibiting the oxidation reaction during hair dyeing. The antioxidant may be any substance capable of removing peroxides generated by the reaction between the oxidative dye and hydrogen peroxide, provided that it eliminates radicals without itself becoming a radical and does not inhibit the reaction between hydrogen peroxide and the oxidative dye. Because hair dyes that use oxidation dyes and oxidizing agents generally generate radicals as intermediates, the anti-inflammatory agent of the present invention can reduce damage and decomposition caused by radicals, making it effective when dyeing hair with oxidative dyes. Specific examples of oxidation dyes include dyes that oxidize amine groups, such as paraphenylenediamine, toluene-2,5-diamine, paraaminophenol, aminophenol, 2,4-diaminophenol, and bisaminophenol; dyes that oxidize carbonyl groups, such as 2,2'-dichloroindigo, alizarin, carminic acid, chrome orange, naphthol, and hematoxylinic acid; dyes that oxidize imine groups, such as rhodamine B, chlorineol, metallergic, alizarin red, and liglon; and dyes that oxidize azo groups, such as azobenzene, azo dye red, azo blue, azo purple, azo orange, and azo green. Since the oxidation reaction during hair dyeing with these dyes (hair dyes) is an oxidation reaction that generates radicals similar to that described above, the agent can be used.

[0008] In the inflammation suppressant of the present invention, the antioxidant is preferably contained in a soil-derived extract extracted from soil collected from a stratum formed by the deposition of marine plants and marine animals under anaerobic conditions more than 300,000 years ago. Here, soil collected from a stratum formed by the deposition of marine plants and marine animals under anaerobic conditions refers to, for example, a stratum formed when a portion of the ocean was isolated by land uplift or other factors more than 300,000 years ago and then covered by debris flows and / or volcanic ash, resulting in repeated fermentation and decomposition of the marine plants and marine animals in this covered ocean. Specifically, soil collected from a stratum 10 to 80 meters underground in the Isahaya region of Nagasaki Prefecture (commonly known as siamarin) is preferred, but is not limited thereto. Furthermore, soil collected from a stratum that was deposited under anaerobic conditions and has not yet been incinerated under such conditions may also be used, for example, from a stratum that is 100 million years old.

[0009] In the inflammation suppressant according to the present invention, the soil-derived extract is preferably obtained by extracting a water-soluble substance contained in the soil. Here, an aqueous solvent such as water, hydrous ethanol, or ethanol can be used as the extraction solvent. The obtained soil-derived extract may be in liquid form, or may be a dry powder obtained by freeze-drying or the like. One type of soil-derived extract (soil-derived extract = sialumarin extract (sialumarin is a registered trademark)) is a glyconutrient extract, and a specific example of this is Noguchi Catalyzer 21 (Catalyzer and Noguchi Catalyzer 21 are registered trademarks) manufactured and sold by the present applicant. The reaction rate constant between hydroxyl radicals and antioxidants is at least 10 9 M -1 ·S -1 In contrast, hydrogen peroxide is 10 1 ~10 4 M -1 ·S -1 and is about 10 5 ~10 8 There is a difference in the order of the reaction rate. Therefore, radical scavenging is possible within a concentration that does not inhibit the dye oxidation reaction caused by hydrogen peroxide. The concentration of hydrogen peroxide in the dye used is 10 -1 ~10 1 Since the concentration is about 10 M, it will hardly inhibit the dyeing at all, as long as it is a concentration that does not inhibit the oxidation reaction between hydrogen peroxide and the dye, i.e., a concentration of 1 / 100 or more of the hydrogen peroxide concentration used. Theoretically, the lower the reaction rate constant between the antioxidant and hydrogen peroxide, the more the concentration can be increased, but in reality, the addition of antioxidants has an effect on pH and has an effect on things other than the oxidation reaction, so it is desirable to add as little as possible. Specifically, antioxidants with a high reaction rate constant with hydrogen peroxide are recommended to be added at a concentration of 10 1 mM or less, as low as 10 2 At concentrations below this level, the dye reaction can be prevented without inhibiting the oxidative damage to the scalp. 1 M -1 ·S -1 The reaction with hydrogen peroxide is relatively mild, and because it contains antioxidants at a concentration of about 1 mM, it can be added at the maximum concentration that does not affect the viscosity of the hair dye. Therefore, there is no problem with adding soil containing such antioxidants to hair dye.

[0010] The inflammation inhibitor according to the present invention may be added to the hair dye. The anti-inflammatory agent can be added to the hair dye during hair dyeing to the extent that it does not affect the viscosity of the hair dye, but the addition ratio is preferably 50% or less. The anti-inflammatory agent may be added to the hair dye in the form of a soil-derived extract (soil-derived extract liquid) or in the form of soil (soil itself).

[0011] The inflammation suppressant according to the present invention may be applied to the scalp before dyeing hair with the hair dye. Here, the anti-inflammatory agent may be applied to the scalp in the form of a soil-derived extract (soil-derived extract liquid) or in the form of soil (soil itself). [Effects of the Invention]

[0012] The inflammation inhibitor of the present invention contains an antioxidant that removes peroxides produced by the reaction of the oxidative dye contained in the hair dye with hydrogen peroxide without inhibiting the oxidation reaction during hair dyeing, thereby effectively suppressing only oxidative damage to the scalp while maintaining a good dyeing reaction with the hair dye.

[0013] In the inflammation suppressant of the present invention, when the antioxidant is contained in a soil-derived extract extracted from soil collected from a stratum formed by the deposition of marine plants and marine animals under anaerobic conditions more than 300,000 years ago, the agent is highly safe.

[0014] In the inflammation suppressing agent according to the present invention, when the soil-derived extract is obtained by extracting water-soluble substances contained in soil, it is excellent in safety and mass productivity.

[0015] When the inflammation inhibitor according to the present invention is added to a hair dye, there is no need to apply the inflammation inhibitor separately from the hair dyeing process, which provides excellent workability.

[0016] When the inflammation inhibitor according to the present invention is applied to the scalp before dyeing hair with a hair dye, it can be highly effective in inhibiting inflammation of the scalp. [Brief explanation of the drawings]

[0017] [Figure 1] FIG. 1 is an explanatory diagram showing the reaction pathway between PPD and hydrogen peroxide during hair dyeing. [Figure 2] This is a diagram explaining the damage and inflammatory reaction that occurs in the body when hair dye containing PPD reacts with an oxidizing agent. [Figure 3] 1(A) to 1(E) are images showing the results of an evaluation test of the inflammation suppression effect of the inflammation suppressing agent according to the present invention. [Figure 4] 4 is a graph showing the cell viability measured from the images of FIGS. 3(A) to (E). [Figure 5] 2(A) to 2(C) are graphs showing the results of an evaluation test of the hydroxyl radical scavenging ability of the inflammation inhibitor according to the present invention. [Figure 6] 6 is a graph showing the peaks of hydroxy radicals in FIGS. 5(A) to 5(C). [Figure 7] 1 shows images confirming the effect of the inflammation inhibitor according to the present invention on the staining ability of PPD. [Figure 8] 1 is a graph showing the influence of the inflammation inhibitor according to the present invention on the staining ability of PPD as measured by absorbance. [Figure 9] 1 is an image confirming the effect of the inflammation inhibitor according to the present invention on hair dyeing with PPD. [Figure 10] (A) to (C) are images confirming the effect of the inflammation inhibitor according to the present invention on the hair cuticle. [Figure 11] (A) to (C) are images confirming the effect of the concentration of the inflammation inhibitor of the present invention on hair dyeing, and (D) is a graph showing the results of calculating the dyeing rate from the images (A) to (C). [Figure 12] These are images confirming the effects of various antioxidants on hair dyeing. [Figure 13] 13 is a graph showing the results of calculating the staining rate from the image of FIG. 12. [Figure 14] 1 is a graph showing the results of a test evaluating the inhibition of the production and release of inflammatory substances by the inflammation inhibitor according to the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0018] Next, embodiments of the present invention will be described with reference to the accompanying drawings to facilitate understanding of the present invention. As shown in Figures 1 and 2, hair dyes containing paraphenylenediamine (PPD), a representative example of an oxidative dye, are oxidized by hydrogen peroxide (HO). The reaction of PPD with HO produces QDI (quinone diimine), which then undergoes further oxidation, polymerization, and bonding with other dyes to form the final dye compound used to color hair. However, PPD itself has been shown to induce allergic reactions associated with increased intracellular ROS. Furthermore, it has been shown that PPD oxidized by hydrogen peroxide generates a mutagenic compound called Bandorovsky base and hydroxyl radicals through oxidation. In other words, as shown in Figure 2, the generation of (2) peroxides, (3) Bandorovsky base, and peroxides resulting from its oxidation leads to scalp damage and inflammation through processes (4), (5), and (6). Conversely, if the generation of (2) and (3) can be suppressed without inhibiting the generation of QDI, the resulting damage and inflammation can be suppressed or prevented.

[0019] The inflammation suppressant according to one embodiment of the present invention reduces inflammatory damage and allergic inflammatory reactions to the scalp that occur when dyeing hair with a hair dye containing an oxidative dye such as PPD. The anti-inflammatory agent may be any agent containing an antioxidant capable of removing peroxides generated by the reaction of the oxidative dye with hydrogen peroxide. Specifically, it is preferable to use an agent having a high reaction rate constant with hydrogen peroxide, such as 10 1 mM or less, as low as 10 2Products containing submillimeter levels of antioxidants can suppress oxidative damage to the scalp without inhibiting the reaction of hair dyes. Such antioxidants are contained, for example, in soil-derived extracts extracted from soil collected from a stratum (3 to 80 meters underground) formed by the deposition of marine plants and animals (fish, shellfish, seaweed, moss, etc.) under anaerobic conditions more than 300,000 years ago.

[0020] This soil-derived extract may be a dry powder obtained by freeze-drying the soil, for example, but is preferably in liquid form. A liquid soil-derived extract (soil-derived extract) is obtained by extracting water-soluble substances contained in the soil. Specifically, it is a solution obtained by extracting water-soluble substances from the soil that has been pulverized and microparticulated, and removing impurities. This soil-derived extract contains natural glyconutrients in addition to humic acid (humic acid and fulvic acid). It is preferable that the soil-derived extract be sterilized. Suitable extraction solvents include aqueous solvents such as water, hydrous ethanol, and ethanol, but are not limited to these. By adding this soil-derived extract (soil-derived extract) to hair dye as an anti-inflammatory agent, it is possible to suppress oxidative damage to the scalp without inhibiting the reaction of the hair dye. Instead of adding this soil-derived extract (soil-derived extract) to hair dye, this soil-derived extract (soil-derived extract) can also be applied to the scalp as an anti-inflammatory agent before dyeing hair with a standard (pre-made) hair dye containing PPD. Furthermore, soil itself can be used as an anti-inflammatory agent instead of the soil-derived extract (soil-derived extract). [Example]

[0021] Next, experiments conducted to confirm the effects of the present invention will be described, but the following examples are not intended to limit the technical scope of the present invention in any way.

[0022] <Inflammation (cytotoxicity) suppression evaluation test using keratinocyte cells> Using keratinocyte cells, the inflammation-suppressing effect of the inflammation-suppressing agent of the present invention on hair dye containing PPD as an oxidative dye was examined.

[0023] (Comparative Example 1) Human epidermal keratinocyte cell line HaCaT was subcultured in Dulbecco's modified Eagle's medium (DMEM, Gibco) containing 10% fetal bovine serum (FBS, West Bio). 4 The cells were seeded at 1000 cells / ml, and after 24 hours, MQ water (Millipore) was added. After 24 hours of incubation, the cells were photographed using a microscope (TE2000-E, Nikon Instech). The HaCaT cells used (provided by the Department of Cellular Engineering, Graduate School of Agriculture, Kyushu University) are a type of human skin keratinocyte cell line that is non-cancerous and widely used in skin research and cosmetic safety testing.

[0024] (Comparative Example 2) Evaluation was carried out in the same manner as in Comparative Example 1, except that 80 μg / ml of PPD and 2 mM of hydrogen peroxide (H 2 O 2 ) were added instead of MQ water (Millipore).

[0025] Example 1 The evaluation was carried out in the same manner as in Comparative Example 2, except that 1% of Noguchi Catalyzer 21 (a glyconutrient extract, manufactured by Noguchi Research Institute Co., Ltd.), a type of soil-derived extract, was added to PPD and H2O2 as an inflammation inhibitor according to the present invention.

[0026] Example 2 Evaluation was carried out in the same manner as in Example 1, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 5%.

[0027] Example 3 Evaluation was carried out in the same manner as in Example 1, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 10%.

[0028] When keratinocyte cells from Comparative Example 1 (untreated) were photographed, the presence of cells was confirmed across the entire image, as shown in Figure 3(A). In contrast, in Comparative Example 2, in which keratinocyte cells were treated only with PPD and H2O2, the cells were found to have detached and become patchy, as shown in Figure 3(B). Furthermore, in Examples 1 to 3, in which the anti-inflammatory agent of the present invention was added to PPD and H2O2, cell detachment decreased as the concentration of the added anti-inflammatory agent increased, as shown in Figures 3(C) to (E). Furthermore, the graph in Figure 4, which shows the cell counts measured in Figures 3(A) to (E), indicates that cells survived in a concentration-dependent manner. These results suggest that the anti-inflammatory agent inhibited cell damage and inflammation caused by PPD and H2O2. All tests were performed independently three times, and the graphs show the average values. Unless otherwise indicated, error bars indicate standard deviation (as in the following tests).

[0029] <Evaluation test of hydroxyl radical scavenging ability using magnetic resonance ESR> Using ESR magnetic resonance, we verified the hydroxyl radical scavenging ability of the anti-inflammatory agent of the present invention in response to hair dye containing PPD as an oxidative dye. Hydroxyl radicals generated by PPD and H2O2 can be trapped using DMPO, a radical spin trap, and detected by ESR.

[0030] (Comparative Example 3) The strength of the DMPO-OH adduct formed in a reaction solution (200 μL) consisting of 2 mg / mL PPD, 178 mM DMPO, and 2% HO was measured. The reaction mixture was prepared in a tube containing all reactants except the HO solution. ESR measurements were performed using a free radical monitor (JES-FR30, JEOL Ltd.). Data acquisition parameters were as follows: magnetic field: 336 ± 10 mT, microwave power: 10 mW, modulation frequency: 100 kHz, modulation amplitude: 0.1 mT, microwave frequency: 9.4271 GHz, sweep time: 1 min, time constant: 0.03 s.

[0031] Example 4 Evaluation was carried out in the same manner as in Comparative Example 3, except that 1% of Noguchi Catalyzer 21 (a glyconutrient extract, manufactured by Noguchi Research Institute Co., Ltd.), a type of soil-derived extract, was added to the reaction solution as an inflammation inhibitor according to the present invention.

[0032] Example 5 Evaluation was carried out in the same manner as in Example 4, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 10%.

[0033] The evaluation results of Comparative Example 3 and Examples 4 and 5 are shown in FIGS. As shown in Figures 5(A) and 6, in Comparative Example 3, in which only PPD and 2% HO were added without the anti-inflammatory agent of the present invention, radicals were trapped by DMPO and detected as a typical hydroxyl radical peak (DMPO-OH adduct). In Examples 4 and 5, in which the anti-inflammatory agent of the present invention was added to PPD and HO, the DMPO-OH adduct decreased as the concentration of the added anti-inflammatory agent increased, as shown in Figures 5(B), (C), and 6. These results suggest that the radicals generated by PPD and HO are reduced by the anti-inflammatory agent, and that the anti-inflammatory agent of the present invention functions effectively as a radical scavenger, suppressing various damages caused by radical (peroxide) generation.

[0034] <Confirming the effect of anti-inflammatory agents on pigment production> The effect of the inflammation inhibitor according to the present invention on the staining ability of PPD, an example of an oxidative dye, was examined.

[0035] Comparative Example 4 The colored state of the liquid to which only 2% H2O2 was added was photographed.

[0036] (Comparative Example 5) The coloring process of a reaction solution in which 2% PPD was added with 2% H2O2 was photographed. The absorbance of the reaction solution was also measured after 30 minutes to confirm the amount of pigment produced.

[0037] Example 6 Evaluation was performed in the same manner as in Comparative Example 5, except that 0.1% of Noguchi Catalyzer 21 (a glyconutrient extract, manufactured by Noguchi Research Institute Co., Ltd.), a type of soil-derived extract, was added to the reaction solution as an inflammation inhibitor according to the present invention.

[0038] Example 7 Evaluation was carried out in the same manner as in Example 6, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 1%.

[0039] Example 8 Evaluation was carried out in the same manner as in Example 6, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 2%.

[0040] Example 9 Evaluation was carried out in the same manner as in Example 6, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 5%.

[0041] Example 10 Evaluation was carried out in the same manner as in Example 6, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 10%.

[0042] (Comparative Example 6) Evaluation was carried out in the same manner as in Comparative Example 5, except that the PPD concentration was set to 0.2%.

[0043] Example 11 Evaluation was carried out in the same manner as in Example 6, except that the PPD concentration was 0.2%.

[0044] Example 12 Evaluation was carried out in the same manner as in Example 7, except that the PPD concentration was set to 0.2%.

[0045] Example 13 Evaluation was carried out in the same manner as in Example 8, except that the PPD concentration was set to 0.2%.

[0046] Example 14 Evaluation was carried out in the same manner as in Example 9, except that the PPD concentration was set to 0.2%.

[0047] Example 15 Evaluation was carried out in the same manner as in Example 10, except that the PPD concentration was 0.2%.

[0048] As shown in Figure 7, when H2O2 is added to PPD, coloring progresses over time, but a comparison of Examples 6 to 10 and Examples 11 to 15 confirms that coloring is promoted as the concentration of the added anti-inflammatory agent increases. This is also evident from the absorbance measurement results shown in Figure 8, and it can be said that the anti-inflammatory agent of the present invention does not inhibit hair dyeing with a hair dye containing PPD, but rather promotes it.

[0049] <Confirming the effect of anti-inflammatory agents on hair dyeing> The effect of the inflammation inhibitor according to the present invention on hair dyeing with PPD, an example of an oxidative dye, was examined.

[0050] 9 shows images of the dyed state of human gray hair 30 minutes after it was placed in the same reaction solutions as those used in Comparative Example 5, Examples 6 to 10, and Comparative Example 6, and Examples 11 to 15. Purchased human gray hair was used in the tests, and the gray hair was placed in the reaction solutions of Examples 6 to 10 and Examples 11 to 15 immediately after the addition of the inflammation inhibitor.

[0051] From the images in FIG. 9, no visible changes in the staining state were observed under any of the conditions (regardless of the concentration of the anti-inflammatory agent (including 0%)). Furthermore, as shown in Figures 10(A) to (C), the surfaces of hair (gray hair) that had not been subjected to a dyeing treatment, hair dyed under the same conditions as in Comparative Example 5, and hair dyed under the same conditions as in Example 10 were photographed to confirm the effect on the cuticle. No change was observed with or without the addition of an inflammation inhibitor. From the above results, it can be said that even in actual hair dyeing with a hair dye containing PPD, the inflammation inhibitor of the present invention does not have any visually observable effect on the dyeing reaction and does not damage the hair.

[0052] Next, the effect of the inflammation inhibitor according to the present invention on hair dyeing was verified using an actual commercially available dye. (method) When commercial dye solutions 1 and 2 were mixed, each containing 10% of the total amount of an anti-inflammatory agent at various concentrations (0, 10, or 30%) prepared with MQ water, was applied to human gray hair for dyeing. Figures 11(A) to 11(C) show photographs of gray hair in which the dyeing reaction was stopped by washing after various times (2.5, 5, 7.5, and 10 minutes) following application. Figure 11(D) is a graph showing the dyeing rates calculated using image analysis software (ImageJ) for Figures 11(A) to 11(C). As the dyeing time increased (5, 7.5, and 10 minutes), the dyeing promotion effect of the anti-inflammatory agent was not confirmed. However, at the initial stage (2.5 minutes), the dyeing promotion effect was confirmed depending on the concentration of the anti-inflammatory agent. Based on these results, it can be said that the anti-inflammatory agent of the present invention does not actually inhibit dyeing, but rather promotes it, or at least does not inhibit dyeing.

[0053] Next, the effects of various antioxidants on hair dyeing were examined using actual commercially available dyes. Figure 12 shows, from left to right, images (photographs) of hair dyed for 10 minutes with no additive, and with a mixture of the anti-inflammatory agent of the present invention, 10 mM ascorbic acid, 10 mM reduced glutathione, 10 mM cysteine, 10 mM gallic acid, and 10 mM hydroquinone. Figure 13 is a graph showing the dyeing rate of hair dyed under each condition, obtained by image analysis of each image in Figure 12.

[0054] When each antioxidant was mixed at a concentration of 10 mM, ascorbic acid was almost the same as the control without additives, but reduced glutathione was slightly affected compared to the control without additives, resulting in a lower staining rate. Cysteine, gallic acid, and hydroquinone were affected by the staining. Ascorbic acid has a low reaction rate constant with hydrogen peroxide (approximately 10 2 M -1 ·S -1), and it is thought that this did not inhibit staining. Alternatively, it may have been oxidized to form monodehydroascorbic acid radicals, which ultimately promoted the oxidation reaction. The higher inhibitory rate of cysteine ​​compared to glutathione is thought to be due to its reaction rate constant with hydrogen peroxide being approximately 10 times higher than that of reduced glutathione. Furthermore, the high inhibitory rates of gallic acid and hydroquinone are presumably due to the fact that they have three reducing groups, resulting in a quantitatively higher reduction reaction rate. Although ascorbic acid appears to be suitable as an anti-inflammatory agent, its own oxidation causes a chain reaction, making it a radical generator. Therefore, it is considered unsuitable as an anti-inflammatory agent for the present invention. Reduced glutathione and cysteine ​​both have thiols as reducing groups, and upon oxidation, they become oxidized glutathione and cystine, respectively, and rarely undergo radical formation themselves. Such antioxidants that do not generate radicals are suitable as anti-inflammatory agents, but considering the reaction rate with hydrogen peroxide, they must be added at a concentration below that does not affect staining. These results suggest that antioxidants suitable as anti-inflammatory agents are those that do not inhibit the oxidation reaction during dyeing and do not generate radicals after the reaction. In this case, it is important to use them at a concentration below that which does not inhibit the reaction between hydrogen peroxide and PPD. Alternatively, a method may be used in which a plurality of antioxidants are used to suppress radical generation by the antioxidants.

[0055] <Test to evaluate the inhibition of the production and release of inflammatory substances by anti-inflammatory agents> In an allergic reaction, mast cells scattered throughout the skin come into contact with an allergen and release inflammatory substances such as histamine, resulting in an inflammatory response (allergic symptoms). More specifically, when the high-affinity IgE receptor (FcεRI) on mast cells is stimulated by IgE antibodies that react with the allergen, the mast cells are activated, releasing inflammatory mediators such as histamine contained in their granules, causing inflammation. Using the rat basophil-like cell line RBL-2H3 cells, which are widely used as this model, the inhibitory effect of the inflammation inhibitor of the present invention on the production and release of inflammatory substances was examined.

[0056] (Comparative Example 7) RBL-2H3 cells were seeded in 96-well culture plates and sensitized with anti-dinitrophenyl (DNP) IgE antibodies for 2 hours. After removing the medium, the cells were washed with Tyrode's buffer (MT buffer) and then incubated in MT buffer for 10 minutes. The cells were then stimulated with DNP-conjugated human serum albumin (DNP-HSA) for 3 hours, after which the supernatant was collected on ice. The cells were then lysed in Tyrode's buffer containing 0.1% Triton X-100 and collected. Protein quantification was performed using the cell lysate. The collected culture supernatant and cell lysate were added with 4-nitrophenyl-2-acetamido-2-deoxy-β-D-glucopyranoside, a chromogenic substrate for β-hexosaminidase. After the enzyme reaction was stopped, the absorbance at 405 nm was measured using an absorbance microplate reader to determine the enzyme activity of β-hexosaminidase. Based on the measurement results, the cellular β-hexosaminidase release rate (amount of inflammatory substance released) per protein was calculated.

[0057] (Comparative Example 8) Evaluation was carried out in the same manner as in Comparative Example 7, except that 100 nM wortmannin was added to the MT buffer as a degranulation inhibitor when the cells were cultured for 10 minutes.

[0058] Example 16 The evaluation was performed in the same manner as in Comparative Example 7, except that when the cells were cultured for 10 minutes, Noguchi Catalyzer 21 (a glyconutrient extract, manufactured by Noguchi Research Institute Co., Ltd.), a type of soil-derived extract, was added to the MT buffer to a final concentration of 0.1% as an inflammation inhibitor according to the present invention.

[0059] Example 17 Evaluation was carried out in the same manner as in Example 16, except that the amount of the inflammation inhibitor (Noguchi Catalyzer 21) added was 1%.

[0060] Example 18 Evaluation was carried out in the same manner as in Example 16, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 5%.

[0061] Example 19 Evaluation was carried out in the same manner as in Example 16, except that the amount of the inflammation suppressant (Noguchi Catalyzer 21) added was 10%.

[0062] As shown in FIG. 14 , the amount of inflammatory substance release was greatest in Comparative Example 7, in which neither a degranulation inhibitor nor an inflammation inhibitor was added. In Comparative Example 8, in which wortmannin was added as a degranulation inhibitor, the amount of inflammatory substance release was suppressed compared to Comparative Example 7. Furthermore, in Examples 16 to 19, in which an inflammation inhibitor was added, the amount of inflammatory substance release was suppressed more than in Comparative Example 8. In particular, in Example 16, in which an inflammation inhibitor was added at 0.1%, and in Example 17, in which an inflammation inhibitor was added at 1%, concentration-dependent suppression of the amount of inflammatory substance release was confirmed, with Example 17 showing the lowest amount of inflammatory substance release. Furthermore, in Example 19, in which an inflammation inhibitor was added at 10%, significant suppression of the amount of inflammatory substance release was confirmed. From these results, it is speculated that the inflammation inhibitor of the present invention has the effect of suppressing degranulation by mast cells in allergic patients, and can be expected to similarly suppress allergic symptoms in people with PPD allergies.

[0063] The anti-inflammatory agent of the present invention (Noguchi Catalyzer 21) is a solution containing stable antioxidant properties. If the anti-inflammatory agent inhibited the oxidation reaction, the dyeing process itself would not proceed. However, the above test results showed that the dyeing process was not actually inhibited. This means that the oxidation reaction of hydrogen peroxide (H2O2), an essential component of hair dye, has little or no effect on the dyeing process. Therefore, if the target of the anti-inflammatory agent is a peroxide such as the hydroxyl radical, a scheme for suppressing cell damage without inhibiting the dyeing process is feasible. Because the hydroxyl radical is highly reactive and has a reaction rate constant approximately 10^6 larger than that of hydrogen peroxide, it is entirely possible that the anti-inflammatory agent selectively eliminates the hydroxyl radical without eliminating hydrogen peroxide.

[0064] Furthermore, even in people without allergies, the inflammation inhibitor can be expected to have a preventive effect on the onset of inflammation. This is because, when damage to the skin barrier occurs due to the generation of hydroxyl radicals and peroxides, these harmful substances become more easily penetrated, resulting in inflammation. However, if the inflammation inhibitor of the present invention is present, the skin barrier is protected by removing peroxides, thereby preventing inflammation by inhibiting the penetration of harmful substances and the penetration of allergens. In addition, inhibition of the inflammatory response by inhibiting the degranulation reaction of mast cells upon allergen invasion can be expected. In the above examples, PPD was used as the oxidative dye. However, as explained above, when dyeing hair with hair dyes containing oxidative dyes that involve the oxidation of amine groups, carbonyl groups, imine groups, or azo groups, an oxidation reaction accompanied by the generation of radicals occurs. Therefore, the inflammation inhibitor of the present invention can be widely applied to hair dyeing using hair dyes that involve oxidation, and can eliminate (remove) radicals (peroxides) to reduce inflammatory damage and allergic inflammatory reactions to the scalp.

[0065] Although the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, but also includes other embodiments and modifications that are possible within the scope of the claims, and all changes in conditions that do not deviate from the gist of the present invention are within the scope of application of the present invention.

Claims

1. An inflammation inhibitor that reduces scalp inflammation and allergic inflammatory reactions that occur when dyeing hair with an oxidative hair dye, The inflammation inhibitor contains a soil-derived water-soluble extract containing an antioxidant that removes radicals generated by the reaction of the oxidative dye contained in the hair dye with hydrogen peroxide without inhibiting the oxidation reaction during hair dyeing, and is characterized in that the soil-derived water-soluble extract is obtained by extracting with an aqueous solvent soil collected from a stratum 3 to 80 meters underground in the Isahaya region of Nagasaki Prefecture that was formed by the deposition of marine plants and marine animals under anaerobic conditions more than 300,000 years ago.

2. 2. The anti-inflammatory agent according to claim 1, wherein the agent is added to the hair dye.

3. 2. The anti-inflammatory agent according to claim 1, which is applied to the scalp before dyeing hair with the hair dye.

Images