Ceramide synthesis promoter

Abstract

To provide a novel agent for promoting ceramide synthesis.SOLUTION: The present invention relates to an agent for promoting ceramide synthesis, comprising ionized alkaline water obtained by electrolyzing an aqueous solution comprising a mineral salt containing silicon and phosphorus.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a ceramide synthesis promoter. [Background technology]

[0002] Skin ceramides are biosynthesized in epidermal keratinocytes by forming a sphingosine skeleton from the amino acids L-serine and palmitic acid, followed by various enzymatic reactions. Healthy ceramide metabolism can be impaired by aging, UV exposure, dry skin, rough skin, atopic dermatitis, senile xerosis, psoriasis, and other conditions. As a result, the amount of ceramide in each layer decreases, resulting in, for example, a decline in the skin's moisturizing and barrier functions. Recently, it has been reported that acylceramides, which have a structure in which linoleic acid is bound to the ω-hydroxyl group of ceramide and have a fatty acid chain length of C28 or longer, are highly hydrophobic and essential for skin barrier function (Non-Patent Document 1).

[0003] Thus, it is expected that promoting the production of ceramides, particularly acylceramides, will promote the improvement or normalization of skin barrier function.

[0004] Recently, research and development into substances that promote the production of ceramides, particularly acylceramides, that can be used in topical skin preparations, foods, beverages, etc. has been actively carried out. However, the results are not sufficient, and there is a need for the development of safe and effective components that promote ceramide synthesis. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] YAKUGAKU ZASSHI 137(10) 1201-1208 (2017) Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a novel ceramide synthesis promoter. [Means for solving the problem]

[0007] As a result of extensive research conducted by the present inventors to develop a novel ceramide synthesis promoter, they discovered that alkaline ionized water produced by a specific production method has the effect of promoting the expression of genes involved in ceramide synthesis, and that the use of this alkaline ionized water promotes ceramide synthesis. The present invention was completed based on this finding.

[0008] That is, the present invention is as follows. Section 1. A ceramide synthesis promoter comprising alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus. Section 2. Item 2. The ceramide synthesis promoter according to item 1, wherein the alkaline ionized water contains 0.4 to 400 mass ppm of silicon and 2 to 800 mass ppm of phosphorus. Section 3. 3. The ceramide synthesis promoter according to item 1 or 2, wherein the alkaline ionized water further contains at least one mineral selected from the group consisting of calcium, potassium, sodium, and magnesium. Section 4. 4. The ceramide synthesis promoter according to any one of items 1 to 3, wherein the alkaline ionized water further contains chlorine. Section 5. 5. The ceramide synthesis promoter according to any one of items 1 to 4, wherein the alkaline ionized water has a pH of 10 to 12.5. Section 6. 6. The ceramide synthesis promoter according to any one of items 1 to 5, wherein the alkaline ionized water has a surface tension of 55 to 68 mN / m. Section 7. 7. The ceramide synthesis promoter according to any one of items 1 to 6, wherein the alkaline ionized water is obtained by an electrolysis step of electrolyzing an aqueous solution containing water and a mineral salt containing silicon and phosphorus, and an electron supply step of mixing the alkaline water on the cathode side obtained in the electrolysis step with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and supplying electrons to the mixed solution. Section 8. 8. The ceramide synthesis promoter according to any one of items 1 to 7, wherein the ceramide synthesis promoter comprises a ceramide synthesis gene expression promoter. Section 9. Item 9. The ceramide synthesis promoter according to any one of items 1 to 8, wherein the ceramide synthesis gene expression promoter comprises at least one gene expression promoter selected from the group consisting of an ELOVL4 gene expression promoter, an ELOVL7 gene expression promoter, a CerS3 gene expression promoter, and an SPTLC2 gene expression promoter. Section 10. 10. The ceramide synthesis promoter according to any one of items 1 to 9, which comprises an ELOVL4 gene expression promoter. Section 11. 10. The ceramide synthesis promoter according to any one of items 1 to 9, which comprises an ELOVL7 gene expression promoter. Section 12. 10. The ceramide synthesis promoter according to any one of items 1 to 9, wherein the ceramide synthesis promoter comprises a CerS3 gene expression promoter. Section 13. 10. The ceramide synthesis promoter according to any one of items 1 to 9, comprising an SPTLC2 gene expression promoter. Section 14. 14. A pharmaceutical product comprising the ceramide synthesis promoter according to any one of items 1 to 13. Section 15. 14. An external skin preparation containing the ceramide synthesis promoter according to any one of items 1 to 13. Section 16. 14. A cosmetic preparation comprising the ceramide synthesis accelerator according to any one of items 1 to 13. Section 17. 14. A food or drink composition comprising the ceramide synthesis promoter according to any one of items 1 to 13. Section 18. 14. The food or drink composition for promoting ceramide synthesis according to any one of items 1 to 13.

[0009] Section 19. 1. Use of alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus for promoting ceramide synthesis. Section 20. 20. The use according to item 19, wherein the alkaline ionized water contains 0.4 to 400 mass ppm of silicon and 2 to 800 mass ppm of phosphorus. Section 21. 21. The use according to item 19 or 20, wherein the alkaline ionized water further contains at least one mineral selected from the group consisting of calcium, potassium, sodium, and magnesium. Section 22. 22. The use according to any one of items 19 to 21, wherein the alkaline ionized water further contains chlorine. Section 23. 23. The use according to any one of items 19 to 22, wherein the alkaline ionized water has a pH of 10 to 12.5. Section 24. 24. The use according to any one of items 19 to 23, wherein the alkaline ionized water has a surface tension of 55 to 68 mN / m. Section 25. 25. The use according to any one of items 19 to 24, wherein the alkaline ionized water is obtained by an electrolysis step of electrolyzing an aqueous solution containing water and a mineral salt containing silicon and phosphorus, and an electron supply step of mixing the alkaline water on the cathode side obtained in the electrolysis step with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and supplying electrons to the mixed solution.

[0010] Section 26. 1. A method for promoting ceramide synthesis, comprising: A method using alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus. Section 27. 27. The method according to item 26, wherein the alkaline ionized water contains 0.4 to 400 mass ppm of silicon and 2 to 800 mass ppm of phosphorus. Section 28. 28. The method according to item 26 or 27, wherein the alkaline ionized water further contains at least one mineral selected from the group consisting of calcium, potassium, sodium, and magnesium. Section 29. 29. The method according to any one of items 26 to 28, wherein the alkaline ionized water further contains chlorine. Section 30. 30. The method according to any one of items 26 to 29, wherein the alkaline ionized water has a pH of 10 to 12.5. Section 31. 31. The method according to any one of items 26 to 30, wherein the alkaline ionized water has a surface tension of 55 to 68 mN / m. Section 32. 32. The method according to any one of items 26 to 31, wherein the alkaline ionized water is obtained by an electrolysis step of electrolyzing an aqueous solution containing water and a mineral salt containing silicon and phosphorus, and an electron supply step of mixing the alkaline water on the cathode side obtained in the electrolysis step with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and supplying electrons to the mixed solution.

[0011] Section 33. A method for producing a ceramide synthesis promoter, comprising: A method for producing a ceramide synthesis promoter, comprising a step of electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus. Section 34. Item 34. The method for producing a ceramide synthesis promoter according to Item 33, wherein the alkaline ionized water contains 0.4 to 400 mass ppm of silicon and 2 to 800 mass ppm of phosphorus. Section 35. Item 35. The method for producing a ceramide synthesis promoter according to Item 33 or 34, wherein the alkaline ionized water further contains at least one mineral selected from the group consisting of calcium, potassium, sodium, and magnesium. Section 36. 36. The method for producing a ceramide synthesis promoter according to any one of items 33 to 35, wherein the alkaline ionized water further contains chlorine. Section 37. 37. The method for producing a ceramide synthesis promoter according to any one of items 33 to 36, wherein the alkaline ionized water has a pH of 10 to 12.5. Section 38. 38. The method for producing a ceramide synthesis promoter according to any one of items 33 to 37, wherein the alkaline ionized water has a surface tension of 55 to 68 mN / m. Section 39. 39. The method for producing a ceramide synthesis promoter according to any one of items 33 to 38, wherein the alkaline ionized water is obtained by an electrolysis step of electrolyzing an aqueous solution containing water and a mineral salt containing silicon and phosphorus, and an electron supply step of mixing the alkaline water on the cathode side obtained in the electrolysis step with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and supplying electrons to the mixed solution.

[0012] At present, it is impossible or impractically difficult to completely identify the structure of the above-mentioned ceramide synthesis promoter, gene expression promoter, pharmaceutical, topical skin preparation, cosmetic, and food and beverage composition, and therefore the invention of each product is described using a product-by-process claim. [Effects of the Invention]

[0013] According to the present invention, a ceramide synthesis promoter can be provided, which contains alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus. The alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus has the effect of promoting the expression of the ELOVL4 gene, the ELOVL7 gene, the CerS3 gene, and the SPTLC2 gene, which are involved in the synthesis of ceramide, particularly acylceramide, and therefore can promote ceramide synthesis. Therefore, the ceramide synthesis promoter of the present invention is effective in preventing or treating skin disorders induced by a decrease in ceramide. [Brief explanation of the drawings]

[0014] [Figure 1] FIG. 1 is a schematic diagram illustrating an apparatus for producing alkaline ionized water. [Figure 2] FIG. 2 shows the gene sequence of human ELOVL4 (SEQ ID NO: 1). [Figure 3] FIG. 3 shows the gene sequence of human ELOVL7 (SEQ ID NO: 2). [Figure 4] FIG. 4 shows the gene sequence of human CerS3 (SEQ ID NO: 3). [Figure 5A] FIG. 5A shows the gene sequence of human SPTLC2 (SEQ ID NO: 4). [Figure 5B] FIG. 5B is a diagram showing the gene sequence of human SPTLC2 (SEQ ID NO: 4), and is a continuation of FIG. 5A. [Figure 6] FIG. 6 is a graph showing the expression level of ELOVL4 mRNA in alkaline ionized water. [Figure 7] FIG. 7 is a graph showing the expression level of ELOVL7 mRNA in alkaline ionized water. [Figure 8] FIG. 8 is a graph showing the amount of CerS3 mRNA expressed in alkaline ionized water. [Figure 9] FIG. 9 is a graph showing the expression level of SPTLC2 mRNA in alkaline ionized water. [Figure 10]FIG. 10 shows the results of evaluating the changes in expression of ELOVL4 and CerS3 proteins by Western blotting. [Figure 11] FIG. 11 shows the results of evaluation by thin layer chromatography. [Figure 12] FIG. 12 is a graph showing the rate of increase in color intensity of the GlcCer region at each concentration in FIG. [Figure 13] FIG. 13 is a graph showing the rate of increase in color intensity of the Cer region at each concentration in FIG. DETAILED DESCRIPTION OF THE INVENTION

[0015] The ceramide synthesis promoter of the present invention is characterized by containing alkaline ionized water (hereinafter sometimes simply referred to as "alkaline ionized water") obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus. Alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus has the effect of promoting the expression of genes involved in ceramide synthesis and can promote the synthesis of ceramide, so the ceramide synthesis promoter of the present invention is effective in preventing or treating skin disorders induced by a decrease in ceramide.

[0016] Alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus contains silicon and phosphorus. Specifically, the alkaline ionized water contains silicon (Si) at about 0.4 to 400 mass ppm (mg / L), preferably about 0.8 to 390 mass ppm, and more preferably about 1.0 to 380 mass ppm, and phosphorus (P) at about 2 to 800 mass ppm, preferably about 4 to 780 mass ppm, and more preferably about 10 to 770 mass ppm. Elemental analysis was performed using inductively coupled plasma (ICP) optical emission spectroscopy (SPECTR ACROS II, manufactured by Hitachi High-Tech Science Corporation).

[0017] The alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus may contain elements other than phosphorus and silicon, and may contain, for example, calcium (Ca) at about 0.0001 to 900 mass ppb, preferably about 0.001 to 800 mass ppm, and more preferably about 0.01 to 700 mass ppm; potassium (K) at about 0.0005 to 3000 mass ppm, preferably about 1 to 2900 mass ppm, and more preferably about 5 to 2800 mass ppm; magnesium (Mg) at about 0.0001 to 300 mass ppb, preferably about 0.001 to 250 mass ppm, and more preferably about 0.01 to 200 mass ppm; and sodium (Na) at about 34 to 8000 mass ppm, preferably about 40 to 7900 mass ppm, and more preferably about 50 to 7800 mass ppm. Furthermore, alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus may contain chlorine (Cl), for example, about 1.5 to 450 mg / kg of chlorine (Cl), preferably about 1.5 to 440 mg / kg, and more preferably about 1.5 to 430 mg / kg.

[0018] The silicon contained in the alkaline ionized water is a silicate ion, for example, H3SiO4 - , H5Si2O7 - , H5Si3O9 - It can exist in such a state.

[0019] The phosphorus contained in the alkaline ionized water is a phosphate ion, for example, H2PO4 - , H3P2O7 - , H4P3O 10 - It can exist in such a state.

[0020] The pH of alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus is usually about 10 to 12.5. The surface tension is usually 70 mN / m or less, preferably about 50 to 69 mN / m, and more preferably about 55 to 68 mN / m. Furthermore, the alkaline ionized water contains a physical excess of electrons.

[0021] The alkaline ionized water used in the present invention can be obtained by an electrolysis step in which an aqueous solution containing water and a mineral salt containing silicon and phosphorus is electrolyzed, and an electron supply step in which the alkaline water on the cathode side obtained in the electrolysis step is mixed with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and electrons are supplied to the mixed solution. As the mineral salt containing silicon and phosphorus, for example, an electrolyte containing silicon and phosphorus obtained from seawater can be used. The seawater can be used without any particular limitation, and for example, seawater from Japan can be used. The mineral salt containing silicon and phosphorus can be added in an amount of approximately 0.001 to 3 mass% of the amount of water used.

[0022] In addition to the above steps, the method for producing alkaline ionized water further includes a deoxidation step of deoxidizing an aqueous solution containing water and a mineral salt containing silicon and phosphorus to reduce the dissolved oxygen concentration to 1 ppm or less, and a step of adding 1 kg / cm 3 of ionized water to the mixed solution of the alkaline water on the cathode side and the mineral salt containing silicon and phosphorus. 2 ~12kg / cm 2 Here, the mineral salt containing silicon and phosphorus that is mixed with the alkaline water on the cathode side can be the same as the mineral salt containing silicon and phosphorus that is used in the electrolysis step, and for example, mineral salt collected from seawater in Japan can be used.

[0023] The method for producing alkaline ionized water will be described below. The first step is an electrolysis step in which an aqueous solution containing water and a mineral salt containing silicon and phosphorus is electrolyzed.

[0024] Pure water, ion-exchanged water, etc. can be used as the raw material water. Mineral salts containing silicon and phosphorus obtained from seawater are dissolved in the water as a supporting electrolyte. It is preferable to use water that has been deoxidized to reduce the dissolved oxygen concentration to 1 ppm or less. Deoxidation methods include physical and chemical methods, and conventionally, chemical methods have been used alone or in combination. Physical methods include degassing using a thermal degasser or a membrane degasser. Chemical methods include adding hydrazine, sodium sulfite, or sugars (such as glucose) as a deoxidizer. For deoxidation, for example, the above-mentioned oxygen remover can be added to the water.

[0025] Electrolysis is preferably carried out in a sealed space such as an electrolytic cell. The electrolytic cell is preferably filled with an inert atmosphere such as nitrogen or carbon dioxide. Instead of using water that has been previously deoxidized to reduce the dissolved oxygen concentration to 1 ppm or less, or in addition to using water that has been previously deoxidized, it is also possible to add a deoxidizer to the electrolytic cell when electrolysis is carried out in the electrolytic cell. The electric energy applied in the electrolysis is preferably 1000 W to 3000 W.

[0026] The second step is an electron supply step in which the alkaline water on the cathode side obtained in the first step is mixed with a mineral salt containing silicon and phosphorus to obtain a mixed solution, and electrons are supplied to the mixed solution.

[0027] The alkaline water on the cathode side obtained by electrolysis is preferably extracted from the electrolytic cell and supplied to a mixing tank, which is preferably provided with a means for supplying mineral salts containing silicon and phosphorus obtained from seawater and an electron supply means for supplying electricity to the alkaline water.

[0028] Examples of raw materials containing silicon (or mineral salts containing silicon) include alkali metal silicates such as sodium silicate and potassium silicate, alkaline earth metal silicates such as calcium silicate, magnesium silicate, etc. The silicon content in the mixed liquid is preferably about 0.005 to 5% by mass, and more preferably 0.01 to 1% by mass, when the total amount of the mixed liquid is taken as 100% by mass.

[0029] Examples of raw materials containing phosphorus (or mineral salts containing phosphorus) include alkali metal phosphates such as sodium phosphate and potassium phosphate; alkaline earth metal phosphates such as calcium phosphate; magnesium phosphate, etc. The content of phosphorus in the mixed solution is preferably about 0.005 to 5% by mass, and more preferably 0.01 to 1% by mass, when the total amount of the mixed solution is taken as 100% by mass.

[0030] The mass ratio of the silicon-containing mineral salt to the phosphorus-containing mineral salt added to the alkaline water is preferably 1:0.5 to 1.5.

[0031] An example of the electron supply means is a cathode terminal. Electrons can be supplied by bringing the cathode terminal into contact with the alkaline water on the cathode side obtained by electrolysis. Electrons are discharged from the cathode terminal, and many electrons are supplied to the alkaline water.

[0032] It is preferable that a direct current be applied to the cathode terminal. The voltage of the direct current applied to the cathode terminal is, for example, 10 to 1000 V, preferably 50 to 300 V, and more preferably 100 to 300 V. It is believed that by supplying electrons to the mixed solution, the produced alkaline ionized water can effectively exert the effect of expressing ceramide synthesis genes.

[0033] A direct current with a voltage of 100 to 300 V is applied to the mixing tank, and the amount of electricity emitted from the cathode electrons is preferably 1000 to 3000 W.

[0034] The mixing tank contains 1 kg / cm 2 ~12kg / cm 2 It is preferable to apply a pressure of 2 kg / cm (98 to 1177 kPa). 2 ~6kg / cm 2 (196 to 588 kPa) is more preferable. Applying pressure improves the stability of alkaline ionized water.

[0035] The mixing tank is preferably filled with an inert atmosphere such as nitrogen or carbon dioxide. An oxygen scavenger may also be added to the mixing tank. The mixing tank is preferably insulated from the ambient temperature by a heat insulating means so as not to be affected by the ambient temperature, and the temperature inside the mixing tank is preferably adjusted to a range of -5°C to 25°C by a temperature adjusting means. The temperature of the mixed liquid is preferably set to 0°C to 10°C.

[0036] The mixing tank is preferably placed in an isolated tank that is isolated from the outside atmosphere, and the isolated tank is preferably filled with nitrogen or carbon dioxide. This makes it possible to prevent deterioration of the alkaline ionized water, and therefore allows performance to be maintained for a long period of time.

[0037] The alkaline ionized water is stabilized in the mixing tank for 24 to 36 hours to obtain the final product, alkaline ionized water. The obtained alkaline ionized water can be taken out directly from the alkaline ionized water outlet. Alternatively, an alcohol can be mixed with the alkaline ionized water and taken out from the mixed alkaline ionized water outlet. As the alcohol, it is preferable to use an alcohol that is highly compatible with water, such as methanol, ethanol, or isopropyl alcohol.

[0038] A manufacturing device that can be used to manufacture alkaline ionized water will be described below. FIG. 1 shows a schematic diagram illustrating an apparatus for producing alkaline ionized water. The alkaline ionized water production apparatus 101 is equipped with an electrolytic cell 1 and a mixing cell 11, and the electrolytic cell 1 and the mixing cell 11 are connected by an alkaline water outlet pipe 9. A water conduit 2 is connected to the electrolytic cell 1, and raw water (an aqueous solution containing water and mineral salts containing silicon and phosphorus) is supplied through this water conduit 2. The electrolytic cell 1 has an anode chamber 4 and a cathode chamber 5 separated by a diaphragm 3. An anode 6 is provided in the anode chamber 4, and a cathode 7 is provided in the cathode chamber 5. An electrolytic current is applied to both electrodes, and acidic water is extracted from an acidic water outlet pipe 8 connected to the anode chamber 4, and alkaline water is extracted from an alkaline water outlet pipe 9 connected to the cathode chamber 5. The alkaline water is cooled as it passes through the alkaline water outlet pipe 9 by a cooling device 10 equipped with a cooling pipe through which a refrigerant is circulated, and then introduced into the mixing cell 11.

[0039] The mixing tank 11 is connected to the alkaline water outlet pipe 9, the raw material liquid storage tank 12, and the outlet pipe 16. The raw material liquid is supplied to the mixing tank 11 from the raw material liquid storage tank 12 via an inflow rate adjuster 13. The supply rate of the raw material liquid is adjusted by the amount of electricity applied, etc. Mineral salts containing silicon and phosphorus collected from seawater in Japan are used as the raw material liquid. An agitator 14 is provided within the mixing tank 11 so that the alkaline ionized water and the raw material liquid are mixed uniformly.

[0040] A cathode terminal 100 is placed in the mixing vessel 11. A direct current of 200 V is applied to the cathode terminal 100. The amount of electricity emitted from the cathode terminal 100 is preferably 1000 W to 3000 W.

[0041] Mixing tank 11 is insulated by heat insulating means 15 so as not to be affected by the ambient temperature, and the internal temperature is adjusted to about 15°C to 25°C by temperature adjusting means (not shown). The temperature of mixing tank 11 is preferably set to about 0°C to 10°C.

[0042] The pH of the alkaline ionized water discharged from the mixing tank 11 is measured by a pH measuring means 17 provided in the outlet pipe 16. The pH of the alkaline ionized water taken out from the mixing tank 11 is preferably about 10 to 12.5.

[0043] An outlet pipe 16 connected to the mixing tank 11 branches into an alkaline ionized water outlet pipe 18 and a mixed alkaline ionized water outlet pipe 21. The alkaline ionized water in the mixing tank 11 is directly extracted from the alkaline ionized water outlet pipe 18. The alkaline ionized water can also be mixed with alcohol to produce mixed alkaline ionized water. In this case, the alkaline ionized water and alcohol can be mixed in an alcohol mixing tank 20 into which alcohol is injected from an alcohol storage tank 19, and the mixed alkaline ionized water can be extracted from the mixed alkaline ionized water outlet pipe 21. The alcohol to be mixed with the alkaline ionized water can be an alcohol that is highly miscible with water, such as methanol, ethanol, or isopropyl alcohol.

[0044] The alkaline ionized water production device 101 described above is isolated from the outside air by being housed in the isolated chamber 22. The isolated chamber 22 is connected to the outside atmosphere by an aeration device that is substituted with nitrogen or filled with carbon dioxide, an oxygen remover, or the like. By providing the alkaline ionized water production device 101 inside the isolated chamber 22, it is possible to prevent deterioration of the alkaline ionized water and maintain its performance for a long period of time.

[0046] The alkaline ionized water obtained by the above-described production method has an excellent effect of promoting the expression of ceramide synthesis genes, and can be used as a ceramide synthesis gene expression promoter.

[0047] Alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus promotes gene expression related to ceramide synthesis, particularly acylceramide synthesis. Specifically, it promotes the expression of the ELOVL (very long-chain fatty acid elongase) 4 gene (ELOVL4 gene expression promotion), which is essential for the synthesis of fatty acids of C28 or longer that are abundant in the epidermis; the ELOVL7 gene, which plays a central role in the synthesis of C16, C18, and C20 fatty acids (ELOVL7 gene expression promotion); and the CerS3 gene, a ceramide synthase enzyme that synthesizes ceramide by binding acyl-CoA to a long-chain base, and is active on very long-chain acyl-CoA (CerS3 gene expression promotion). It also promotes the expression of the SPTLC2 gene, a subunit of serine palmitoyltransferase (SPT), an enzyme involved in ceramide synthesis in epidermal keratinocytes (SPTLC2 gene expression promotion). Therefore, alkaline ionized water containing silicon and phosphorus can be used as an ELOVL4 gene expression promoter, an ELOVL7 gene expression promoter, a CerS3 gene expression promoter, and an SPTLC2 gene expression promoter.

[0048] The ceramide synthesis promoter of the present invention can be used alone or can contain additives. Examples of additives include sodium chloride, various vitamin preparations, glucose, etc. The ceramide synthesis promoter of the present invention can be incorporated into various compositions such as pharmaceuticals, quasi-drugs, foods and beverages, and cosmetics to obtain a composition having a ceramide synthesis promoting effect.

[0049] The form of the food or drink containing the ceramide synthesis promoter of the present invention is not particularly limited, and specific forms of the food or drink include general foods, general drinks, supplements, health foods, functional foods with functional claims, specific health foods and foods for specific uses, soft drinks, tea drinks, health supplements, alcoholic drinks such as wine, confectioneries, cooked rice dishes, breads, noodles, prepared dishes, seasonings, etc.

[0050] The form of a drug or quasi-drug containing the ceramide synthesis promoter of the present invention is not particularly limited. Specific forms include, for example, an internal preparation and an external preparation, and it is preferably used as an external preparation for skin.

[0051] The form of the cosmetic or quasi-drug containing the ceramide synthesis accelerator of the present invention is not particularly limited, and specific dosage forms include capsules, tablets, powders, granules, liquids, etc.

[0052] The dosage of the ceramide synthesis promoter of the present invention can be determined taking into consideration the age, sex, weight, method of use, dosage, etc. Examples of methods of use include topical application, oral administration, intravenous injection (intravenous drip), etc.

[0053] For example, in the case of topical application, alkaline ionized water containing 0.001 to 1.5% by mass of a silicon compound and a phosphorus compound can be applied to the skin. When the ceramide synthesis gene expression promoter of the present invention is orally administered, the dosage is preferably about 100 to 200 mL / day of alkaline ionized water containing 0.001 to 1.5% by mass of a silicon compound and a phosphorus compound. When the ceramide synthesis gene expression promoter of the present invention is intravenously injected, the dosage is preferably about 100 to 500 mL / day of alkaline ionized water containing 0.001 to 1.5% by mass of a silicon compound and a phosphorus compound.

[0054] Compositions such as pharmaceuticals containing the ceramide synthesis promoter of the present invention are highly effective in promoting ceramide synthesis by promoting the expression of ceramide synthesis genes, and are therefore effective in preventing or treating dermatitis and improving damaged skin sites. Pharmaceuticals or quasi-drugs containing the ceramide synthesis promoter of the present invention can be used as agents for preventing or treating, for example, atopic dermatitis, acne, burns, wounds, gangrene, bedsores, scabies, pyoderma, shingles, scalp eczema, epidermal abscess, herpes simplex, chickenpox, malignant skin tumors, athlete's foot, etc. Cosmetics containing the ceramide synthesis promoter of the present invention can be used as skin moisturizers, agents for preventing dry skin, agents for preventing rough skin, sunburn repair agents, whitening agents, etc.

[0055] The present invention also includes the use of alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus for promoting ceramide synthesis. The present invention also includes a method for promoting ceramide synthesis, which uses alkaline ionized water obtained by electrolyzing an aqueous solution containing mineral salts containing silicon and phosphorus. The present invention also includes a method for producing a ceramide synthesis promoter, the method comprising the step of electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus. [Example]

[0056] The present invention will be explained in more detail below with reference to examples, but the technical scope of the present invention is not limited to these examples.

[0057] Example 1 Using the alkaline ionized water production device 101 described above, alkaline ionized water was produced as follows. Ion-exchanged water was prepared as raw water. The ion-exchanged water was produced by removing impurities from tap water using an ion exchange resin (manufactured by Samsung). An aqueous solution of ion-exchanged water with 1% by mass of mineral salt containing silicon and phosphorus extracted from Japanese seawater added was introduced into electrolytic cell 1 through water pipe 2, and electricity was applied to anode 6 and cathode 7. The electrolysis conditions were a voltage of 200 V and electrical energy of 2500 W. The ion-exchanged water was electrolyzed, producing acidic water in anode chamber 4 and alkaline water in cathode chamber 5. The acidic water produced in anode chamber 4 was discharged through acidic water discharge pipe 8. The alkaline water produced in cathode chamber 5 was discharged into alkaline water discharge pipe 9, and the alkaline water passing through alkaline water discharge pipe 9 was cooled by cooling device 10 and introduced into mixing tank 11.

[0058] In mixing tank 11, cooled alkaline water and mineral salt containing silicon and phosphorus collected from Japanese seawater, supplied from raw material liquid storage tank 12, were mixed by agitator 14. The mineral salt containing silicon and phosphorus collected from Japanese seawater in raw material liquid storage tank 12 was supplied to mixing tank 11 after being adjusted by inflow adjuster 13 so that the content of mineral salt containing silicon and phosphorus collected from Japanese seawater in the mixed liquid was 0.3 mass%. The mixed liquid was stored for 24 hours at a pressure of 294 kPa and a temperature of 4±3°C. During storage, electrons were emitted from cathode terminal 100 into the mixed liquid. Thereafter, the alkaline ionized water was led out from outlet pipe 16, and after the pH of the alkaline ionized water was measured by pH measuring means 17, it was taken out from alkaline ionized water outlet pipe .

[0059] The alkaline ionized water of Example 1 taken out from the alkaline ionized water outlet pipe was subjected to elemental analysis using ICP optical emission spectroscopy (manufacturer: Hitachi High-Tech Science Corporation, product name: SPECTRO ARCOSII). As a result, it was found to contain 100 mass ppm of silicon and 760 mass ppm of phosphorus. In more detail, it was found to contain 7500 mass ppm of sodium, 2700 mass ppm of potassium, 0.2 mass ppm of calcium, and 0.1 mass ppm or less of magnesium.

[0060] Furthermore, because the above-mentioned ICP optical emission spectroscopy cannot detect gases such as chlorine, the alkaline ionized water of Example 1 (sample 1) taken out from the electrolyzed water outlet pipe was separately measured at the Japan Food Research Center General Incorporated Foundation using the combustion-coulometric titration method described below. As the combustion - coulometric titration method, using a chlorine and sulfur analyzer TSX - 10 type manufactured by Nitto Seiko Analytic Co., Ltd., about 10 mg of the above sample 1 was combusted, and the concentration of chloride ions was calculated by coulometric titration. By this measurement, the amount of chlorine in the sample can be determined regardless of organic chlorine or inorganic chlorine. The amount of organic chlorine was determined using the value obtained by subtracting the amount of inorganic chlorine from the total amount of chlorine. The amount of inorganic chlorine was determined by extracting 5 g of the sample with hot water and measuring the chloride ions in the extract by ion chromatography. As a result, it was found that the alkaline ion water of Example 1 contained 410 mg / kg of chlorine.

[0061] <Measurement of pH> When the pH of the alkaline ion water obtained in Example 1 was measured using a pH meter (manufactured by Horiba, Ltd., HORIBA F - 74SP), the pH value was 12.

[0062] <Measurement of surface tension> The surface tension of the alkaline ion water obtained in Example 1 was measured using the Wilhelmy method. As a result, the surface tension of the alkaline ion water obtained in Example 1 was about 65.41 mN / m (20 °C). When the surface tension of purified water was measured in the same way, it was about 72.96 mN / m (20 °C). Therefore, the surface tension of the alkaline ion water containing the mineral salt containing silicon and phosphorus used in the present invention was about 7.55 mN / m lower than that of purified water.

[0063] (Example 2) Alkaline ion water was produced in the same manner as in Example 1, except that the raw material of the mineral salt containing silicon and phosphorus was changed from seawater in Japan to seawater in Okinawa Prefecture. The obtained alkaline ionized water was subjected to elemental analysis using ICP emission spectroscopy in the same manner as in Example 1, and the results showed that it contained 370 mass ppm of silicon and 450 mass ppm of phosphorus. Furthermore, in more detail, it was found that it contained 6900 mass ppm of sodium, 1 mass ppm or less of potassium, 0.9 mass ppm of calcium, and 0.3 mass ppm or less of magnesium. Furthermore, the chlorine contained in the alkaline ionized water obtained in Example 2 was measured using the combustion-coulometric titration method at the Japan Food Analysis Center General Incorporated Foundation, in the same manner as in Example 1, and it was found that the alkaline ionized water of Example 2 contained 410 mg / kg of chlorine.

[0064] (Test Example 1) The alkaline ionized water produced in Example 1 was examined for its effect on the expression of genes for enzymes related to ceramide synthesis. Note that, on actual skin, alkaline ionized water is neutralized by sebum or sweat on the skin before reaching keratinocytes, or is captured in the stratum corneum, and therefore in Test Example 1, the alkaline ionized water of Example 1 was diluted 20 times, and evaluation was carried out using alkaline ionized water with this concentration as the reference (100%). The same was true for Test Examples 2 and 3.

[0065] Among the enzymes involved in ceramide synthesis, we evaluated the expression changes of very long-chain fatty acid elongase 4 (ELOVL4), very long-chain fatty acid elongase 7 (ELOVL7), ceramide synthase 3 (CerS3), and ceramide synthase (SPTLC2). Primers for the target genes were designed based on sequences reported in human RT-PCR (Shirakura et al.: Lipid in Health and Disease, 11,108 (2012).).

[0066] Forward and reverse primers were synthesized for human ELOVL4, ELOVL7, CerS3, and SPTLC2, respectively. Synthesis of PCR primers was outsourced to FASMAC Corporation. Figure 2 shows the human ELOVL4 gene sequence (SEQ ID NO: 1). Figure 3 shows the human ELOVL7 gene sequence (SEQ ID NO: 2). Figure 4 shows the human CerS3 gene sequence (SEQ ID NO: 3). Figures 5A and 5B show the human SPTLC2 gene sequence (SEQ ID NO: 4). Table 1 shows the forward primer sequence (SEQ ID NO: 5) and reverse primer sequence (SEQ ID NO: 6) for ELOVL4, the forward primer sequence (SEQ ID NO: 7) and reverse primer sequence (SEQ ID NO: 8) for ELOVL7, the forward primer sequence (SEQ ID NO: 9) and reverse primer sequence (SEQ ID NO: 10) for CerS3, the forward primer sequence (SEQ ID NO: 11) and reverse primer sequence (SEQ ID NO: 12) for SPTLC2, and the forward primer sequence (SEQ ID NO: 13) and reverse primer sequence (SEQ ID NO: 14) for β-actin (endogenous control). In PCR, primers bind to the complementary strands of the forward and reverse primers, and a DNA fragment is amplified between them. In Figures 2 to 5A and 5B, the bolded black boxed portion in each sequence indicates the sequence to which the primers bind (the complementary strands of the forward and reverse primers), and the underlined portion indicates the sequence amplified by PCR.

[0067] [Table 1]

[0068] <Reagents> Formazan (MTT): manufactured by Dojindo Laboratories Cell culture microtest plate: 96-well flat-bottom low-evaporation type, manufactured by FALCON Fetal Bovine Serum (FBS): Biosera · Dulbecco’s Modified Eagle Medium (DMEM): manufactured by WAKO · TriPure Isolation Reagent: manufactured by Takara Bio Inc. · Random Primer: manufactured by Takara Bio Inc. · Reverse Transcriptase: RevatraAce, manufactured by TOYOBO · THUNDERBIRD (registered trademark) SYBR qPCR Mix: manufactured by TOYOBO <Apparatus> · Microplate Reader: ARVO MX1420 MULTIABEL COUNTER manufactured by WALLAC was used. · Real Time PCR: Step One Real Time PCR (48 well) manufactured by Thermo Fisher Scientific was used.

[0069] <Preparation of DNA> Normal Human Epidermal Keratinocytes (NHEK) were seeded at a density of 1 × 10 cells per well in a 6-well plate and cultured for 24 hours. Then, DMEM containing 10%, 20%, or 100% (v / v%) alkaline ion water and 10% FBS was added to each well and cultured for 72 hours. As a control, DMEM without alkaline ion water (containing 10% FBS) was used. After culturing for 72 hours, total RNA was extracted using TriPur Reagent from Takara Bio Inc. according to the attached instructions. The mRNA of ELOVL4, ELOVL7, CerS3, and SPTLC2 was quantified by Real Time-PCR using the prepared cDNA. β-actin was used as an endogenous control. The Comparative ΔΔCt method was used for data analysis.<​​​​​​Real-time PCR was performed using Step One (Thermo Fisher Scientific) and TOYOBO THUNDERBIRD (registered trademark) SYBR qPCR Mix.

[0071] The results of ELOVL4 mRNA expression by alkaline ionized water are shown in Figure 6, the results of ELOVL7 mRNA expression are shown in Figure 7, the results of CerS3 mRNA expression are shown in Figure 8, and the results of SPTLC2 mRNA expression are shown in Figure 9. The mRNA expression levels are shown as relative values ​​when the expression level of β-actin is set to 1.

[0072] From Figure 6, it can be seen that alkaline ionized water promoted the expression of ELOVL4 mRNA. From Figure 7, it can be seen that the expression of ELOVL7 mRNA was also observed. From Figure 8, it can be seen that alkaline ionized water promoted the expression of CerS3 mRNA. From Figure 9, it can be seen that alkaline ionized water promoted the expression of SPTLC2 mRNA.

[0073] (Test Example 2) The changes in the expression of ELOVL4 and CerS3 proteins were evaluated by Western blotting using specific antibodies. Specifically, normal human epidermal keratinocytes (NHEK) were placed in a 6-well plate at 1 × 10 per well. 5Cells were seeded at a density of 1000 μg / well and cultured for 24 hours. Then, DMEM containing 10%, 20%, or 100% (v / v%) alkaline ionized water and 10% FBS was added to each well and cultured for 72 hours. DMEM without alkaline ionized water (containing 10% FBS) was used as a control. After 72 hours of culture, the cells were washed with phosphate buffer. Cells were then harvested using cell lysis solution (50 mM Tris-HCl, 150 mM NaCl, 1% NP-40, 1 mM EDTA, pH 7.4) and centrifuged (10,000 rpm, 10 minutes, 4°C). The supernatant was used as the protein extract. Proteins in the protein extract were separated by SDS-PAGE (SDS-polyacrylamide gel electrophoresis), transferred to a nitrocellulose membrane, and blocked with phosphate buffer containing 2% skim milk. CerS3 was detected using rabbit anti-CerS3 antibody (Cusabio, 1:2000 dilution), ELOVL4 was detected using rabbit anti-ELOVL4 antibody (Cusabio, 1:1000 dilution), and β-actin was detected using mouse anti-β-actin antibody (Wako Pure Chemical Industries, 1:3000 dilution). After thorough washing with phosphate buffer, the target proteins were labeled with peroxidase using peroxidase-conjugated goat anti-rabbit IgG antibody (KPL, 1:5000 dilution) or peroxidase-conjugated goat anti-mouse IgG antibody (Jackson, 1:3000 dilution). Detection was performed using a luminescence reagent (Immunostar LD, Wako Pure Chemical Industries, Ltd.), and images were captured using a luminograph (ATTO). The results are shown in Figure 10.

[0074] FIG. 10 shows that alkaline ionized water promoted the expression of CerS3 protein in a concentration-dependent manner.

[0075] (Test Example 3) Whether alkaline ionized water can actually promote the synthesis of ceramide (Cer) and glucosylceramide (GlcCer) was evaluated by thin-layer chromatography (TLC). Specifically, normal human epidermal keratinocytes (NHEK) were placed in a 6-well plate at 1 × 10 per well. 5 Cells were seeded at a density of 1000 μg / well and cultured for 24 hours. Then, DMEM containing 10%, 20%, or 100% (v / v%) alkaline ionized water and 10% FBS was added to each well and cultured for 72 hours. DMEM without alkaline ionized water (containing 10% FBS) was used as a control. After 48 hours of culture, the cells were washed with phosphate buffer, and the intracellular lipids were extracted with chloroform:methanol (1:2, v / v) by shaking for 1 hour, and the supernatant was collected. The remaining residue was then extracted with chloroform:methanol:water (1:2:0.5, v / v) by shaking, and the supernatant was collected. A 2.5% (w / v) aqueous potassium chloride solution was added to the extracted sample, and the mixture was centrifuged (5 minutes, 900 x g, room temperature). The lower layer was then collected in a new tube. The remaining lipids in the upper layer were collected by adding chloroform, stirring, and centrifuging (5 minutes, 900 x g, room temperature). The lipid fraction was dried using a centrifugal evaporator, dissolved in chloroform:methanol (2:1, v / v), and then stored at −20° C. until analysis. Thin-layer chromatography was performed in the following order: (1) chloroform:methanol:water (40:10:1, v / v) was used to develop the sample up to 2 cm from the point of application, and the plate was then dried. (2) chloroform:methanol:water (40:10:1, v / v) was used to develop the sample up to 5 cm from the point of application, and the plate was then dried. (3) chloroform:methanol:acetic acid (47:2:0.5, v / v) was used to develop the sample up to 1.5 cm from the edge of the plate, and the plate was then dried. (4) hexane:diethyl ether:acetic acid (65:35:1, v / v) was used to develop the sample up to 0.5 cm from the edge of the plate, and the plate was then dried. Lipids on the developed thin layer plate were detected by thoroughly spraying it with 8% (v / v) phosphoric acid aqueous solution containing 3% (w / v) copper sulfate, and then heating it on a hot plate at 180°C. The results are shown in Figure 11. In Figure 11, VLC indicates very long chain (generally a carbon chain longer than 20 carbons), and ULC indicates ultra long chain (generally a carbon chain of 26 carbons or more).

[0076] Next, Table 2 shows the Rf values ​​of the main spots in the GlcCer region and the Cer region in FIG.

[0077] [Table 2]

[0078] The color intensity of each region was measured using the image processing software ImageJ (NIH) after capturing images using a LumiCube camera (manufactured by Liponics Co., Ltd.). Table 3 and FIG. 12 show the rate of increase in color intensity of the GlcCer region when the color intensity of the GlcCer region at 0% alkaline ionized water (when cultured in a medium (DMEM) without added alkaline ionized water) is set to 1, and Table 3 and FIG. 13 show the rate of increase in color intensity of the Cer region when the color intensity of the Cer region at 0% alkaline ionized water is set to 1.

[0079] [Table 3]

[0080] It was confirmed from Figure 11 that the higher the concentration of alkaline ionized water, the larger the size of the spots in the GlcCer region and Cer region (especially the Cer(ULC) region) and the deeper the color. Furthermore, Table 3 shows that the higher the concentration of alkaline ionized water, the greater the rate of increase in color intensity in the GlcCer region and Cer region. These results indicate that the higher the concentration of alkaline ionized water, the greater the amount of ceramide synthesized, and it can be seen that alkaline ionized water concentration-dependently promoted the synthesis of GlcCer and Cer, and increased the intracellular content of GlcCer and Cer.

[0081] Furthermore, from Figure 11 and Table 2, it was confirmed that the higher the concentration of alkaline ionized water, the larger the Rf value (longer the development distance) of the spots in the GlcCer region and Cer region (especially the Cer(ULC) region). Table 3 shows that the higher the concentration of alkaline ionized water, the greater the rate of increase in color intensity in the GlcCer region and Cer region. These results indicate that a higher concentration of alkaline ionized water promotes the synthesis of fatty acids with longer carbon chains, and that as fatty acids become longer, their liposolubility increases (they become more hydrophobic), which increases the development distance and allows spots to be detected further up. These findings suggest that alkaline ionized water promotes the synthesis of ceramides (acylceramides) composed of very long-chain fatty acids in keratinocytes. [Explanation of symbols]

[0082] 1 electrolytic cell 2 Water pipes 3 Diaphragm 4 Anode chamber 5. Cathode Chamber 6 Anode 7 Cathode 8 Acidic water discharge pipe 9 Alkaline water outlet pipe 10 Cooling device 11 Mixing tank 12 Raw material liquid storage tank 13 Inflow adjustment device 14 Stirring device 15. Insulation measures 16 Outlet pipe 17 pH measuring means 18 Alkaline ionized water outlet pipe 19 Alcohol storage tank 20 Alcohol mixing tank 21 Mixed alkaline ionized water outlet pipe 22 Isolation room 100 cathode electrons 101 Alkaline ionized water production equipment

Claims

1. A ceramide synthesis promoter comprising, as an active ingredient, alkaline ionized water obtained by electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus, The ceramide synthesis promoter contains silicon, phosphorus, sodium, and chlorine in the alkaline ionized water.

2. 2. The ceramide synthesis promoter according to claim 1, wherein the alkaline ionized water contains 0.4 to 400 mass ppm of silicon, 400 to 800 mass ppm of phosphorus, and 34 to 8000 mass ppm of sodium.

3. 3. The ceramide synthesis promoter according to claim 1, wherein the alkaline ionized water further contains at least one mineral selected from the group consisting of calcium and magnesium.

4. The ceramide synthesis promoter according to any one of claims 1 to 3, wherein the alkaline ionized water has a pH of 10 to 12.

5.

5. The ceramide synthesis promoter according to any one of claims 1 to 4, wherein the alkaline ionized water has a surface tension of 55 to 68 mN / m.

6. an electrolysis step of electrolyzing an aqueous solution containing a mineral salt containing silicon and phosphorus and water to obtain the alkaline ionized water; A mineral salt mixing step of mixing the alkaline water on the cathode side obtained in the above step with a mineral salt containing silicon and phosphorus to obtain a mixed liquid; and The mixture obtained in the mineral salt mixing step is supplied with electrons emitted from a cathode terminal in an electron supply step. The ceramide synthesis promoter according to any one of claims 1 to 5, wherein the alkaline ionized water contains silicon, phosphorus, sodium, and chlorine.

7. The ceramide synthesis promoter according to any one of claims 1 to 6, which is a ceramide synthesis gene expression promoter.

8. The ceramide synthesis promoter according to any one of claims 1 to 7, wherein the ceramide synthesis gene expression promoter is at least one gene expression promoter selected from the group consisting of an ELOVL4 gene expression promoter, an ELOVL7 gene expression promoter, a CerS3 gene expression promoter, and an SPTLC2 gene expression promoter.

9. A pharmaceutical for promoting ceramide synthesis, comprising the ceramide synthesis promoter according to any one of claims 1 to 8.

10. A skin external preparation for promoting ceramide synthesis, comprising the ceramide synthesis promoter according to any one of claims 1 to 8.

11. A cosmetic preparation for promoting ceramide synthesis, comprising the ceramide synthesis promoter according to any one of claims 1 to 8.

12. A food or beverage composition for promoting ceramide synthesis, comprising the ceramide synthesis promoter according to any one of claims 1 to 8.

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