Composition for promoting ceramide production or aquaporin 3 production

Abstract

To provide a novel substance that promotes ceramide production or aquaporin 3 production. [Solution] A composition for promoting ceramide production or aquaporin 3 production, containing a residue obtained by extracting pigment from safflower.

Description

Technical Field

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[0001] The present invention relates to a composition for promoting ceramide production or aquaporin 3 production.

Background Art

[0002] Ceramide exists as a main component of the intercellular lipids of epidermal cells and is known as a substance that plays a barrier function to prevent water evaporation from the inside of the skin. The search for useful components that promote ceramide production has been underway (Patent Document 1, Patent Document 2).

[0003] Aquaporin (AQP) proteins are known as proteins that selectively permeate water on the cell membrane. In humans, the existence of 13 types of AQP is known, and in epidermal cells, mainly aquaporin 3 (AQP3) exists. AQP3 is important for improving the physiological barrier function of the skin by promoting the transport of water and glycerol. By promoting AQP3 production, an effective improvement in the barrier function can be expected (Patent Document 3, Patent Document 4).

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Patent Document 2

Patent Document 3

Patent Document 4

Summary of the Invention

Problems to be Solved by the Invention

[0005] While substances that promote ceramide production are known, there is a desire to expand the options in this technological field and broaden the possibilities for new research and applications by providing novel substances that can replace known substances. Similarly, while substances that promote AQP3 production are known, there is a desire to expand the options in this technological field and broaden the possibilities for new research and applications by providing novel substances that can replace known substances. [Means for solving the problem]

[0006] As a result of diligent research to solve the above problems, the present inventors discovered that a substance that promotes ceramide production or AQP3 production can be produced using the residue remaining after extracting pigment from safflower, leading to the present invention.

[0007] In other words, the present invention relates to at least the following: [Invention 1] A composition for promoting ceramide production or aquaporin 3 production, containing a residue obtained by extracting pigment from safflower. [Invention 2] The composition according to Invention 1, wherein the aforementioned residue is an extract obtained using an organic solvent from the residue obtained by extracting pigment from safflower. [Invention 3] The composition according to Invention 2, wherein the organic solvent is an aliphatic compound containing one or more hydroxyl groups. [Invention 4] A cosmetic, cosmetic additive, or humectant containing the composition described in any of Inventions 1 to 3. [Invention 5] A method for producing a composition that promotes ceramide production or aquaporin 3 production, comprising the step of extracting an extract from the residue after extracting pigment from safflower using an organic solvent. [Invention 6] The method according to Invention 5, wherein the organic solvent is an aliphatic compound containing one or more hydroxyl groups. [Invention 7] A method for producing a cosmetic, a cosmetic additive, or a humectant, comprising carrying out the method described in invention 5 or 6.

Advantages of the Invention

[0008] In the present application, the present invention provides a novel composition for promoting ceramide production or AQP3 production. In one aspect, the present invention is a cosmetic, a cosmetic additive, or a moisturizer containing the composition.

Brief Description of the Drawings

[0009] [Figure 1] It is a graph showing that the expression of AQP3 mRNA is significantly enhanced by the safflower flower extract of the present invention. [Figure 2] It is a graph showing that the expression of NQO1 mRNA is significantly enhanced by the safflower flower extract of the present invention. [Figure 3] It is a graph showing that the expression of GCLC mRNA is significantly enhanced by the safflower flower extract of the present invention. [Figure 4] It is a graph showing that the expression of SPTLC1 mRNA is significantly enhanced by the safflower flower extract of the present invention. [Figure 5] It is a graph showing that the safflower flower extract of the present invention has a high ceramide production promoting effect. [Figure 6] It is a graph showing that the safflower flower extract of the present invention significantly increases the amount of water in the stratum corneum.

Embodiments for Carrying Out the Invention

[0010] The composition of the present invention contains the residue obtained by extracting the pigment from safflower.

[0011] Safflower pigment extract In preparing the residue, the method for extracting the pigment from safflower is not particularly limited and can be carried out by a known method.

[0012] As the safflower used for the extraction of the pigment, safflower flowers may be used, crude drugs (Kouka) obtained by drying the flowers may be used, or manjyu may be used.

[0013] The safflower may be subjected to an extraction operation as it is, or extraction may be carried out after performing treatments such as cutting into small pieces, drying, and pulverizing. The extraction is carried out by immersing it in an extraction solvent. Stirring may be performed or homogenization may be carried out in the extraction solvent to increase the extraction efficiency. The extraction temperature and extraction time can be appropriately selected.

[0014] The solvent for extracting the pigment is not particularly limited. For example, water, physiological saline, phosphate buffer, or phosphate buffered saline can be used alone or in combination as a mixture of two or more types at any mixing ratio. Only water can be used, or a solvent such as ethanol that is easy to remove after extraction can be employed and used alone or as a mixture with water at any mixing ratio. Preferably, a solvent containing 50% by weight or more of water (that is, a solvent containing no organic solvent or a mixed solvent containing less than 50% by weight of an organic solvent) is used. The solvent may contain an acid or a base component.

[0015] Residue after pigment extraction The residue contained in the composition of the present invention may be the residue itself obtained by extracting the pigment from safflower, or may be obtained through further purification steps such as extraction and filtration from the residue.

[0016] When further extraction is carried out, it is preferable to use a solvent containing 50% by weight or more of an organic solvent (that is, only an organic solvent or a mixed solvent containing 50% by weight or more of an organic solvent). The solvent may contain an acid or a base component. As the organic solvent, an alcohol, that is, an aliphatic compound containing one or more hydroxy groups (for example, a monohydric alcohol such as methanol, ethanol, propyl alcohol, isopropyl alcohol, etc., or a polyhydric alcohol such as propylene glycol, dipropylene glycol, 1,3 - butylene glycol, 1,2 - pentylene glycol, glycerin, etc.) is preferable.

[0017] In one embodiment, the composition of the present invention can be used as an active ingredient in cosmetics, cosmetic additives, or moisturizers. Depending on the application, the formulation (the proportion of the composition and the types and proportions of other ingredients) can be appropriately selected from conventional formulation examples in accordance with the common technical knowledge of those skilled in the art. For example, the composition of the present invention can be used in the formulation of conventional topical skin preparations. The type or form of the cosmetic of the present invention can be applied as, for example, lotion, emulsion, cream, ointment, patch, pack, cleanser, lipstick, lip balm, foundation, blush, eyeshadow, eyebrow, etc. Furthermore, the composition of the present invention can be used in topical pharmaceuticals, quasi-drugs, etc. for the skin, etc.

[0018] Other ingredients The compositions, cosmetics, cosmetic additives, and humectants of the present invention may, as necessary and within the limits that do not impair the effects of the present invention, appropriately incorporate various other components, such as mineral oil, alcohols, UV absorbers, moisturizing components, aqueous components, antioxidants, cosmetic components, preservatives, water-soluble polymers, film-forming agents, color-fastening agents, defoaming agents, fragrances, etc., in order to impart various effects.

[0019] Other components may be added at any stage in the production of the composition of the present invention.

[0020] Examples of mineral oils include petrolatum, synthetic waxes, and microcrystalline waxes.

[0021] Examples of alcohols include natural monoalcohols such as ethanol and phenoxyethanol, synthetic monoalcohols such as 2-hexyldecanol, and polyhydric alcohols such as ethylene glycol and pentylene glycol.

[0022] Examples of UV absorbers include benzophenone-based, PABA-based, cinnamic acid-based, and salicylic acid-based UV absorbers, as well as 4-tert-butyl-4'-methoxydibenzoylmethane and oxybenzone.

[0023] Examples of moisturizing ingredients include proteins, mucopolysaccharides, collagen, elastin, and keratin.

[0024] The aqueous component can be any water or any water-soluble component. In addition to water, examples include glycerols such as glycerin, diglycerin, and polyglycerin, and plant extracts such as aloe vera, witch hazel, cucumber, lemon, lavender, and rose.

[0025] Examples of antioxidants include α-tocopherol and ascorbic acid.

[0026] Examples of beauty ingredients include vitamins, anti-inflammatory agents, and herbal medicines.

[0027] Examples of preservatives include parahydroxybenzoic acid esters and phenoxyethanol.

[0028] Ceramide production promotion The composition of the present invention promotes ceramide production. This can be confirmed by known methods, for example, by the methods described in the examples herein.

[0029] Promotion of AQP3 production gene expression The composition of the present invention promotes the expression of a gene that produces AQP3, thereby promoting the production of AQP3. The expression of the AQP3-producing gene can be confirmed by known methods, for example, by the method described in the examples herein.

[0030] NQO1 production promotion In one embodiment, the compositions of the present invention may also promote the expression of a gene that produces NAD(P)H quinone dehydrogenase 1 (NQO1), thereby promoting the production of NQO1. The expression of the NQO1-producing gene can be confirmed by known methods, for example, by the methods described in the examples herein.

[0031] Promoting GCLC production In one embodiment, the compositions of the present invention may also promote the expression of a gene that produces glutamate cysteine ​​ligase catalytic subunit (GCLC), thereby promoting GCLC production. The expression of the GCLC-producing gene can be confirmed by known methods, for example, by the methods described in the examples herein.

[0032] SPTLC1 gene expression promotion In one embodiment, the composition of the present invention may also promote the expression of a gene that produces serine palmitoyltransferase, long chain base subunit 1 (SPTLC1), thereby promoting the production of SPTLC1. The expression of the SPTLC1-producing gene can be confirmed by known methods, for example, by the method described in the examples herein.

[0033] Method for producing the composition of the present invention The compositions, cosmetics, etc. of the present invention can be manufactured by commonly used methods. For example, a solid cosmetic can be manufactured by heating and mixing all the ingredients, degassing, filling into a mold or container, and then cooling and solidifying it. [Examples]

[0034] The present invention will be described in more detail below with reference to examples, but the present invention is not limited to the following examples.

[0035] Pigment extraction from safflower Red safflower cakes obtained from the Yamagata Prefecture Safflower Producers' Association were soaked in water and kneaded to extract the yellow pigment. The process was repeated, changing the water as needed, until no more yellow liquid came out. The red safflower cakes, from which no more yellow liquid came out, were then soaked in a weak alkaline solution (wood ash water, straw ash water, 5% potassium carbonate solution) and kneaded to extract the red pigment. The process was repeated, changing the weak alkaline solution as needed, until the red safflower cakes turned a color similar to wet straw scraps, thus obtaining the safflower cake residue.

[0036] Processing of residue after pigment extraction 50 g of the above residue, obtained after extracting pigment from safflower-derived red rice cakes, was immersed in 1 kg of pentylene glycol for 3 hours. This was filtered and purified to obtain a safflower flower extract sample to be used in the tests described later. As can be seen from the above description, in this specification, "safflower flower extract" refers to the extract obtained from the residue after obtaining the pigment, not the extract obtained when obtaining the pigment from safflower flowers.

[0037] Aquaporin 3 gene expression promotion NHEK cells (Normal Human Epidermal Keratinocytes) were seeded in a 96-well plate. KG2 (Kurabo) was used as the culture medium during seeding. The above safflower extract sample was filtered and sterilized, then mixed with culture medium KB2 (Kurabo) at a concentration of 0.3%. Evaluation substance samples of different concentrations were prepared using pentylene glycol in a 3-fold dilution series (i.e., 0.3%, 0.1%, and 0.03%). In addition, a negative control sample was prepared by adding pentylene glycol to culture medium KB2 to a final concentration of 0.3%. After seeding NHEK cells in culture medium KG2 and culturing for 24 hours, the NHEK cells were treated with evaluation substance samples at various concentrations and a negative control sample. qPCR was performed 24 hours after the sample was added to check for genetic changes. For the preparation of reverse transcripts used in qPCR, we used the SuperPrep® II Cell Lysis & RT Kit for qPCR (Toyobo) and performed the reverse transcription reaction according to the protocol provided by the manufacturer. Using the resulting reverse transcript as a template and THUNDERBIRD® SYBR® qPCR Mix (Toyobo) as the qPCR reagent, we performed gene expression analysis of AQP3 and Peptidylprolyl Isomerase A (PPIA) as an internal standard. Figure 1 shows the measurement results of AQP3 mRNA levels. As can be seen from these results, it was found that the expression of AQP3 mRNA was significantly enhanced by the safflower extract of the present invention.

[0038] NQO1 gene expression promotion NQO1 mRNA expression was performed using the same method as described above for the promotion and confirmation of aquaporin 3 gene expression, and its expression level was measured. The measurement results are shown in Figure 2. As can be seen from these results, it was found that the expression of NQO1 mRNA was significantly enhanced by the safflower extract of the present invention.

[0039] GCLC gene expression promotion GCLC mRNA expression was performed using the same method as described above for the promotion and confirmation of aquaporin 3 gene expression, and its expression level was measured. The measurement results are shown in Figure 3. As can be seen from these results, GCLC mRNA expression was significantly enhanced with the safflower extract of the present invention.

[0040] SPTLC1 gene expression promotion SPTLC1 mRNA expression was performed using the same method as described above for the promotion and confirmation of aquaporin 3 gene expression, and its expression level was measured. The measurement results are shown in Figure 4. As can be seen from these results, it was found that the expression of SPTLC1 mRNA was significantly enhanced by the safflower extract of the present invention.

[0041] Ceramide production promotion Culture cups containing human epidermal tissue from LabCyte EPI-MODEL 24 (J-TEC) were placed in each well of an assay plate dispensed with 500 μL of assay medium (J-TEC), and the cultures were incubated in a CO2 incubator. The above safflower extract sample was diluted to 0.1% with HEPES buffer (Kurabo) to prepare the evaluation sample. A negative control sample was prepared by diluting pentylene glycol (Symrise) to 0.1% with HEPES buffer. After culturing for 24 hours as described above, the culture medium was replaced with fresh medium, and 50 μL of the prepared sample (evaluation substance sample or negative control sample) was exposed to each tissue. After 1 hour, the samples were removed, and the surface of each tissue was washed three times with 50 μL of HEPES buffer before incubation for 24 hours. This series of operations was performed for 4 days. After washing on the 4th day, the bottom of the culture cup was immersed in Trypsin / EDTA solution (Kurabo) for 1 hour, and the stratum corneum was collected from the cup. A ceramide sample was obtained through extraction, lipid fractionation, and concentration. Specifically, 3 mL of methanol and 100 μL of 1 ppm CERAMIDE III (Evonik) were added to the stratum corneum recovered above. After sonication for 3 minutes, the methanol was completely evaporated under reduced pressure, and 1 mL of chloroform was added and sonication was performed for 1 minute. This was the total lipid sample. Sep-Pak Silica 3 cc Vac Cartridge (Waters) was conditioned with 10 mL of chloroform / methanol (99.5 / 0.5 volume ratio) followed by 10 mL of chloroform, and then 1 mL of the total lipid sample was injected into the cartridge and held in the substrate. After cleaning by passing 10 mL of chloroform through it, 10 mL of methanol was passed through it to obtain the ceramide fraction. The ceramide fraction was completely dried under reduced pressure, and 1 mL of methanol was added and sonication was performed for 1 minute. This was collected in a vial, 100 μL of 1 ppm Ceramide Bio (Symrise) was added, and then it was dried to nitrogen. After drying, the ceramide sample was obtained by redissolving it in 100 μL of heptane / chloroform / methanol (in a volume ratio of 95 / 2.5 / 2.5). The total amount of ceramide in the obtained ceramide sample was quantified using LC-MS. The quantification results are shown in Figure 5. As can be seen from these results, it was confirmed that the safflower extract of the present invention has a high ceramide production promoting effect.

[0042] Increased moisture content in the stratum corneum of the lips Lip balms containing the safflower extract sample of the present invention and a negative control lip balm were prepared using the formulations shown in Table 1. Next, for six subjects with no bias in gender or age, the moisture content of the stratum corneum of the lips before lip balm application was measured at a total of 12 points on the lips (3 points on the upper left, 3 points on the lower left, 3 points on the upper right, and 3 points on the lower right) using a Corneometer CM825 (Courage+Khazaka). After measurement, a negative control lip balm was applied to the upper and lower left sides of the lips, and the lip balm of the present invention was applied to the upper and lower right sides. Thereafter, the lip balm was applied at times when the subjects felt that no lip balm was present, such as upon waking, after lunch, and before going to bed, and the application state was maintained for one week. During this time, the use of other lip cosmetics was avoided. After one week, the lip balm was wiped off, and the moisture content was measured again, and the percentage change in moisture content between the start and end of the test was compared. The results are shown in Table 1 and Figure 6. As these results show, the lip balm containing safflower extract of the present invention significantly increased the moisture content of the stratum corneum. [Table 1] [Industrial applicability]

[0043] According to the present invention, it is possible to provide a cosmetic composition having excellent ceramide production ability, and therefore it can be used in the cosmetics industry and the like.

Claims

1. The use of an extract obtained by extracting pigment from safflower using an organic solvent for the production of a ceramide production promoter, wherein the use does not include the steps of adding yeast to the residue and fermenting it, enzymatically decomposing the residue, adding yeast to the extract and fermenting it, or enzymatically decomposing the extract. The organic solvent is an aliphatic compound containing one or more hydroxyl groups. use.

2. The use according to claim 1, wherein the ceramide production promoter is used as an active ingredient in a cosmetic, a cosmetic additive, or a moisturizer.

3. A use for producing an aquaporin 3 production promoter, comprising using an extract obtained from the residue of safflower from which pigment has been extracted using an organic solvent, wherein the use does not include the steps of adding yeast to the residue and fermenting it, enzymatically decomposing the residue, adding yeast to the extract and fermenting it, or enzymatically decomposing the extract. The organic solvent is an aliphatic compound containing one or more hydroxyl groups. use.

4. The use according to claim 3, wherein the aquaporin 3 production promoter is used as an active ingredient in a cosmetic, a cosmetic additive, or a moisturizer.

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