JP2012188399A - Skin humectant - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a material capable of improving skin humectant properties at a low cost and with safety.SOLUTION: The skin humectant includes Mentha arvensis or an extract thereof as an active ingredient.

Description

  The present invention relates to a skin moisturizer.

  Natural moisturizing factors (NMF) are amino acid-based molecules produced from filaggrin, a protein derived from keratohyaline granules in the stratum corneum. The keratohyalin granule in the granule cell contains a large amount of profilagrin, which is a precursor of filaggrin, and profilagrin is decomposed into filaggrin through dephosphorylation and the action of protease. Filaggrin aggregates keratin fibers in the cytoplasm of the corneocytes and then degrades into NMF in the upper stratum corneum. NMF is known to play an important role in the moisture retention of the skin stratum corneum and contribute to the moisturizing function of the skin.

  It is known that filaggrin plays an important role in skin moisture retention. It has been reported that filaggrin expression is reduced and amino acids in the stratum corneum are reduced in diseases associated with dry skin, such as atopic dermatitis, ichthyosis, and senile xeroderma. In mice bred in a dry environment, it has also been reported that the amount of water in the stratum corneum decreases and the expression level of filaggrin and the amount of amino acids in the stratum corneum decrease.

Filaggrin is produced by proteagulin, which is a precursor, is degraded by protease. Prostasin / CAP1 / Prss8 is known as one of proteases involved in the degradation of profilagrin. It has been reported that mice knocked out of CAP1 / Prss8 fall into severe dehydration, and that the production of filaggrin monomer from profilagrin is impaired in the skin of the knockout mice (Non-patent Document 1).
By promoting the expression of filaggrin or prostasin, the production amount of the natural moisturizing factor can be increased, and the moisturizing function of the skin can be improved.

  Conventionally, as a filaggrin production promoter, for example, licorice extract (Patent Document 1), flavanone glycoside liquiritin (Patent Document 2) contained in natural plants, wild thyme extract, clove extract, salvia extract, Royal jelly extract, shiitake extract, dio extract, chamomile extract, arnica extract, aloe extract, ougon extract or agaric extract (patent document 3), betaine compound, polyol, ascorbic acid, or derivatives thereof, And herbal medicines derived from sawtooth, marjoram, Ibukikaku, Ichiyakusou, lavender, hamamelis, ogon liquid, cherry leaf, Izayoi rose fruit, and palo azul, carqueja, macelamista (patent document 4), Citrus genus plant extract or Yeast extract (Patent Document 5) and glycolipid compound (Patent Document 6) are known.

  It is desirable to develop further substances that can promote the production of filaggrin, prostasin or NMF cheaply and safely and improve the skin moisturizing ability.

JP 2002-363054 A JP 2003-146886 A JP 2006-16337 A International Publication WO2002 / 053127 JP 2001-261568 A JP 2006-241095 A

Leyvraz et al, J Cell Biol, 2005, 170 (3): 487-496

  The present invention relates to the provision of a skin moisturizer. The present invention also relates to the provision of an NMF production promoter, filaggrin production promoter, and prostasin production promoter.

  The present inventors examined a material capable of improving the skin moisturizing ability, and mint or an extract thereof promotes filaggrin production and prostasin production, thereby producing natural moisturizing factor (NMF) production in the skin. It was found that the skin moisturizing ability can be improved.

That is, the present invention provides the following.
(1) A skin moisturizer comprising mint or an extract thereof as an active ingredient.
(2) The skin moisturizer according to (1), wherein the extract is a water extract or an ethanol aqueous solution extract.
(3) The skin moisturizer according to (2) or (3), which is an external preparation.
(4) A natural moisturizing factor production promoter comprising mint or an extract thereof as an active ingredient.
(5) The natural moisturizing factor production promoter according to (4), wherein the extract is a water extract or an aqueous ethanol extract.
(6) A filaggrin production promoter comprising mint or an extract thereof as an active ingredient.
(7) The filaggrin production promoter according to (6), wherein the extract is a water extract or an aqueous ethanol solution extract.
(8) A prostasin production promoter containing mint or an extract thereof as an active ingredient.
(9) The prostasin production promoter according to (8), wherein the extract is a water extract or an aqueous ethanol extract.

  According to the present invention, by promoting filaggrin production and prostasin production, it is possible to increase the amount of NMF in the skin and improve the skin moisturizing ability. INDUSTRIAL APPLICABILITY The present invention is useful for moisturizing skin and improving diseases or conditions associated with dry skin.

  In the present specification, “non-therapeutic” is a concept that does not include a medical act, that is, a treatment act on the human body by therapy.

  In the present specification, “improvement” refers to improvement of a disease, symptom or condition, prevention or delay of deterioration of the disease, symptom or condition, or reversal, prevention or delay of progression of the disease, symptom or condition.

In the present specification, “mint” refers to Mentha arvensis var. Piperascens (Japanese name mint) of the genus Labiatae.

  Unless specifically limited, the plant extracts listed above are any part of the plant, such as whole grass, leaves, stems, buds, flowers, buds, wood parts, bark, roots, rhizomes, seeds, fruits, or resins. Or any combination thereof. A preferred site for obtaining the extract is the leaf. The parts listed above may be subjected to the extraction step as they are, or may be subjected to the extraction step after being pulverized, cut or dried.

  As the above extract, commercially available ones may be used, or various solvent extracts obtained by a conventional method, or diluted solutions thereof, concentrated solutions thereof, dried powders thereof, pastes or activated carbon treatments thereof. There may be. As an example, the extract in the present invention can be obtained by extracting the plant at room temperature or under heating, or by extracting it using an extraction device such as a Soxhlet extractor.

  As the solvent for extraction, either a polar solvent or a nonpolar solvent can be used. Specific examples of the solvent include water; alcohols such as methanol, ethanol, propanol and butanol; polyhydric alcohols such as propylene glycol and butylene glycol; ketones such as acetone and methyl ethyl ketone; methyl acetate and ethyl acetate Examples include esters; linear and cyclic ethers such as tetrahydrofuran and diethyl ether; polyethers such as polyethylene glycol; and mixtures thereof. Among these, it is preferable to use water, alcohols, a water-alcohol mixed solution, propylene glycol, and butylene glycol, and it is more preferable to use an ethanol aqueous solution.

When the extraction solvent is a water-alcohol mixture, the alcohol concentration in the liquid may be in the range of less than 95% by weight, preferably 90% by weight or less, more preferably 80% by weight or less. More preferably, it is 70% by mass or less.
The extraction conditions for obtaining the extract used in the present invention are not particularly limited, depending on the solvent used.
Specific examples of the extraction means for obtaining the extract include solid-liquid extraction, liquid-liquid extraction, immersion, decoction, leaching, reflux extraction, ultrasonic extraction, microwave extraction, and stirring.
Examples of extraction conditions include 15 to 25 ° C. for 7 to 14 days, about 70 ° C. for 5 hours, and the like.
Moreover, when shortening extraction time, solid-liquid extraction with stirring is desirable. An example of suitable conditions for this solid-liquid extraction is stirring at 40 to 100 ° C. (preferably 50 to 70 ° C.) at 100 to 1000 rpm for 30 to 300 minutes.
In order to prevent the oxidation of the extract, a means for extracting under a so-called non-oxidizing atmosphere while removing dissolved oxygen by bubbling degassing or inert gas such as nitrogen gas may be used in combination.

  As shown in Examples described later, mint or an extract thereof has an action of significantly promoting cell filaggrin gene expression and prostasin gene expression. Therefore, mint or an extract thereof is useful as a filaggrin production promoter and a prostasin production promoter.

  Profilagrin, a precursor of filaggrin, is degraded by enzymes such as prostasin to become filaggrin (Leyvraz et al, J Cell Biol, 2005, 170 (3): 487-496). It is further degraded to become an amino acid that is one of the natural moisturizing factors (NMF) components that play an important role in skin moisture retention (Dale BA et al., Nature, 1978, 276: 729- 731). Therefore, mint or its extract can promote the production of natural moisturizing factors (NMF) through the action of promoting filaggrin production and prostasin production, thereby exerting a skin moisturizing effect. be able to. The peppermint or its extract is also useful for improving diseases or conditions associated with dry skin, such as rough skin, dry skin wrinkles, and diseases such as atopic dermatitis, ichthyosis, and senile xeroderma.

That is, mint or its extract can be used for skin moisturizing. Also, mint or its extract can be used to promote NMF production. Also, mint or its extract can be used to promote filaggrin production. Also, mint or its extract can be used to promote prostasin production. Alternatively, mint or its extract should be used to improve diseases or conditions associated with dry skin, such as rough skin, dry skin wrinkles, and diseases such as atopic dermatitis, ichthyosis, and senile xeroderma. Can do.
The mint or the extract thereof may be used alone or in combination. The use can be in humans or non-human animals, or tissues, organs, cells derived therefrom, and can be therapeutic or non-therapeutic.

Therefore, as one aspect, the present invention provides a skin moisturizer containing mint or an extract thereof as an active ingredient.
As another aspect, the present invention provides a natural moisturizing factor (NMF) production promoter containing mint or an extract thereof as an active ingredient.
As another aspect, the present invention provides a filaggrin production promoter containing mint or an extract thereof as an active ingredient.
As another aspect, the present invention provides a prostasin production promoter containing mint or an extract thereof as an active ingredient.
In one aspect, the agent consists essentially of mint or an extract thereof.

  The peppermint or its extract is used to moisturize the skin, promote NMF production, promote filaggrin production, promote prostasin production, or diseases or conditions associated with dry skin, such as rough skin, dry skin wrinkles, and It can be used for the production of a composition for improving diseases such as atopic dermatitis, ichthyosis, senile xerosis, pharmaceuticals, quasi drugs, cosmetics and the like. Such compositions, pharmaceuticals, quasi drugs, cosmetics and the like are also within the scope of the present invention.

  The above composition, medicine, quasi-drug, cosmetic and the like can be produced or used for human or non-human animals. The mint or its extract is blended in the composition, medicine, quasi-drug, cosmetics, etc., for moisturizing the skin, for promoting NMF production, for promoting filaggrin production, for promoting prostasin production, or It may be an active ingredient for the improvement of diseases or conditions associated with dry skin, such as rough skin, dry skin wrinkles, and diseases such as atopic dermatitis, ichthyosis, senile xeroderma.

The medicine or quasi-drug contains mint or an extract thereof as an active ingredient. The pharmaceutical or quasi drug can be administered in any dosage form. Administration may be oral or parenteral. Dosage forms for oral administration include, for example, solid dosage forms such as tablets, coated tablets, granules, powders, capsules, and liquid dosage forms such as elixirol, syrups and suspensions, Examples of the dosage form for parenteral administration include injection, infusion, topical, external preparation, transdermal, transmucosal, nasal, enteral, inhalation, suppository, bolus, patch and the like.
Preferably, the medicine or quasi drug may be in the form of a skin external preparation.

  The medicament or quasi-drug may contain mint or an extract thereof alone or in combination, or may further contain a combination with a pharmaceutically acceptable carrier. Examples of such carriers include excipients, coating agents, binders, extenders, disintegrating agents, lubricants, diluents, osmotic pressure adjusting agents, pH adjusting agents, dispersing agents, emulsifiers, preservatives, and stabilizers. , Antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, lubricants, activity enhancers, anti-inflammatory agents, bactericides, fragrances, flavoring agents, flavoring agents and the like. Moreover, the said pharmaceutical and quasi-drug may contain another active ingredient and a pharmacological component, as long as the action which promotes filaggrin production and prostasin production of mint or its extract is not lost.

  The pharmaceutical or quasi-drug can be produced by a conventional method from mint or an extract thereof or, if necessary, in combination with the carrier and / or other active ingredients and pharmacological ingredients. The content of mint or the extract thereof in the medicine or quasi-drug is preferably 0.00001 to 20% by mass as the dry weight of the extract, for example, when used as a skin external preparation. More preferably, it is contained in an amount of 10 to 10% by mass.

The cosmetic contains mint or an extract thereof as an active ingredient. The cosmetic may contain mint or an extract thereof alone or in combination, or may further contain in combination with a cosmetically acceptable carrier. Examples of such carriers include excipients, coating agents, binders, extenders, disintegrating agents, lubricants, diluents, osmotic pressure adjusting agents, pH adjusting agents, dispersing agents, emulsifiers, preservatives, and stabilizers. , Antioxidants, colorants, ultraviolet absorbers, humectants, thickeners, lubricants, activity enhancers, anti-inflammatory agents, bactericides, fragrances, flavoring agents, flavoring agents and the like. In addition, as long as the action of promoting the production of filaggrin and prostasin by mint or its extract is not lost, the cosmetics may contain other active ingredients and cosmetic ingredients such as moisturizers, whitening agents, UV protection agents, cells An activator, a detergent, a keratolytic agent, a makeup component (for example, makeup base, foundation, funny, powder, teak, lipstick, eye makeup, eyebrow, mascara, etc.) may be contained. Examples of the form of the cosmetic include creams, emulsions, lotions, suspensions, gels, powders, packs, sheets, patches, sticks, cakes, and the like, which can be used for cosmetics.
Preferably, the cosmetic may be a skin moisturizing cosmetic.

  The cosmetic can be produced by a conventional method from mint or an extract thereof, or in combination with the carrier and / or other active ingredients and cosmetic ingredients as necessary. The content of mint or the extract thereof in the cosmetic is preferably 0.00001 to 20% by mass, and more preferably 0.0001 to 10% by mass, as the dry weight of the extract.

The present invention also provides a method of promoting cellular filaggrin production. The method includes a step of adding mint or an extract thereof to cells that have filaggrin-producing ability and that want to promote filaggrin production.
The present invention also provides a method of promoting cellular prostasin production. The method includes a step of adding mint or an extract thereof to cells that have prostasin-producing ability and that want to promote prostasin production.

In the present invention, the “cell” that promotes filaggrin production or prostasin production is not particularly limited as long as it has natural or genetically engineered filaggrin expression ability or prostasin expression ability. Preferably, the cells include epithelial cells, and more preferably epidermal keratinocytes.
Alternatively, the “cell” may be a cell fragment or cell fraction of the cells listed above, or a tissue containing the cells listed above or a culture derived from the cells listed above. Also good. When the cell is a cell culture, the cell is preferably cultured in the presence of mint or an extract thereof.
The concentration of added mint or its extract is 0.0001-2% (w / v) in terms of the dry weight of the extract as the final concentration in the culture when the cells are a cell culture. Yes, preferably 0.0003 to 0.5% (w / v), more preferably 0.0005 to 0.1% (w / v).

The present invention also provides a method for promoting natural moisturizing factor production in cells. The method includes a step of adding mint or an extract thereof to cells that have the ability to produce natural moisturizing factors and that want to promote natural moisturizing factor production. Preferably, the cells are cultured in the presence of mint or an extract thereof.
The cells having the ability to produce natural moisturizing factor may be cells having the ability to express filaggrin and prostasin. Examples of the cells and the concentration of mint or the extract thereof added to the cells are the same as those in the above-described method for promoting filaggrin production or prostasin production.

In the present invention, mint or an extract thereof is used for moisturizing the skin, for promoting NMF production, for promoting filaggrin production, for promoting prostasin production, or for diseases or conditions associated with dry skin, such as rough skin, For the improvement of dry wrinkles and diseases such as atopic dermatitis, ichthyosis, senile xeroderma, it can be administered in an effective amount to a subject in need thereof.
In one embodiment, the administration is performed non-therapeutically for health promotion or cosmetic purposes.
Examples of the administration target include animals desiring to improve skin moisturizing ability. Alternatively, examples of the administration target include animals having diseases or conditions associated with dry skin, such as rough skin, dry skin wrinkles, and diseases such as atopic dermatitis, ichthyosis, and senile xeroderma. The animal is preferably a human or non-human mammal, more preferably a human.

  The preferred dosage may vary according to the subject's species, weight, sex, age, condition or other factors. The dose, route, interval, and amount and interval of intake can be determined appropriately by those skilled in the art. For example, in the case of topical administration to human skin, the dose is preferably 0.001 to 1000 mg / day per adult (60 kg) in terms of dry weight of mint extract, 0.01 to 100 mg / day Day is more preferred.

  EXAMPLES Hereinafter, an Example is shown and this invention is demonstrated more concretely.

Manufacture example Preparation of mint extract (Manufacture example 1) Preparation of mint water extract 40 grams of mint leaves (manufactured by Shinwa Bussan Co., Ltd.), add 400 mL of water, and extract under conditions of 70 degrees and 5 hours. went. Thereafter, filtration was performed to obtain 279 mL of a water extract of mint. About the obtained extract, when the evaporation residue was computed by the following method, the evaporation residue was 2.09% (w / v). This was concentrated to dryness and then diluted with a 10% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.
<Calculation of evaporation residue>
When 1 ml of mint extract was dried at 105 ° C. for 6 hours (using a dryer: DRY Thermo Unit DTU-1C (TAITEC CORPORATION)), 20.9 mg of a dried product was obtained. The evaporation residue of this extract was calculated as 20.9 / 1000 × 100 = 2.09% (w / v). In the following production examples, the evaporation residue of each extract was calculated in the same manner.

(Manufacture example 2) Preparation of mint 10% ethanol aqueous solution 10 g of mint leaves (manufactured by Shinwa Bussan Co., Ltd.), add 100 mL of 10% ethanol aqueous solution, and extract for 7 days at room temperature and standing went. Thereafter, filtration was performed to obtain 67 mL of a 10% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 2.30% (w / v). This was diluted 2.30 times with a 10% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 3) Preparation of mint 20% ethanol aqueous solution Extraction was performed using a 20% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 65 mL of 20% ethanol aqueous extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 2.12% (w / v). This was diluted 2.12 times with a 20% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 4) Preparation of peppermint 30% ethanol aqueous solution Extraction was performed using a 30% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 63 mL of 30% ethanol aqueous extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 2.13% (w / v). This was diluted 2.13 times with a 30% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 5) Preparation of mint 40% ethanol aqueous solution Extraction was performed using a 40% ethanol aqueous solution under the same conditions as in Production Example 2. Then, filtration was performed to obtain 62 mL of 40% ethanol aqueous extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 2.01% (w / v). This was diluted 2.01 times with a 40% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 6) Preparation of mint 50% ethanol aqueous solution 40 g of mint leaves (manufactured by Shinwa Bussan Co., Ltd.), chopped with 400 mL of 50% ethanol aqueous solution and extracted for 21 days at room temperature and standing went. Thereafter, filtration was performed to obtain 277 mL of a 50% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 1.65% (w / v). This was diluted 1.65 times with a 50% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 7) Preparation of mint 60% ethanol aqueous solution Extraction was performed using a 60% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 60 mL of a 60% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 1.77% (w / v). This was diluted 1.77 times with a 60% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 8) Preparation of mint 70% ethanol aqueous solution Extraction was performed using a 70% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 59 mL of 70% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 1.91% (w / v). This was diluted 1.91 times with a 70% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 9) Preparation of mint 80% ethanol aqueous solution Extraction was performed using an 80% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 55 mL of an 80% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 1.47% (w / v). This was diluted 1.47 times with an 80% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

(Production Example 10) Preparation of mint 90% ethanol aqueous solution Extraction was performed using a 90% ethanol aqueous solution under the same conditions as in Production Example 2. Thereafter, filtration was performed to obtain 56 mL of a 90% ethanol aqueous solution extract of mint. When the evaporation residue was calculated for the obtained extract, the evaporation residue was 1.02% (w / v). This was diluted 1.02 times with a 90% aqueous ethanol solution to prepare a 1.0% (w / v) (extract solid content) extract.

Example 1 Effect of mint extract on filaggrin production Human normal epidermal cells (NHEK) were cultured using EpiLife (registered trademark) (purchased from Kurabo) at 37 ° C. and 5% CO ₂ . Cells were seeded in a 6-well plate at 1 × 10 ⁴ cells / cm ² , cultured for 24 hours, the medium was replaced with a serum-free medium, and the mint extract described in Table 1 prepared according to the production example was It added to a culture medium so that it might become a density | concentration 0.1% (v / v) (final extract concentration 0.001 w / v%). After culturing for 48 hours, total RNA was extracted from the cells using RNeasy (registered trademark) Mini Kit (QIAGEN). The total RNA concentration was measured, and a reverse transcription reaction was performed using a certain amount of total RNA. High capacity RNA-to-cDNA Kit (Applied Biosystems) was used for the reverse transcription reaction.
The filaggrin gene expression was quantified from the obtained cDNA by Real-time RT-PCR. Quantification was carried out using Taqman (registered trademark) Probe (Applied Biosystems) with PRISM 7500 (Applied Biosystems). In a 50 μl reaction system, amplification conditions were 95 ° C., 15 seconds denaturation reaction, 60 ° C., 1 minute annealing, and extension reaction. Each gene expression level was corrected by the expression level of RPLP0 and shown as a relative value when the expression level of filaggrin in the control with no extract added (solvent only) was 1. The results are shown in Table 1.

Example 2 Effect of mint aqueous ethanol extract on filaggrin production Using the mint extract prepared according to Production Examples 2 to 10, filaggrin gene expression was quantified in the same procedure as in Example 1. Table 2 shows the mint extract used and the expression level of filaggrin.

Example 3 Effect of mint extract on prostasin production Prostasin gene expression was quantified in the same procedure as in Example 1. The expression level of prostasin is shown in Table 3.

Claims (9)

  A skin moisturizer comprising mint or its extract as an active ingredient.   The skin moisturizer according to claim 1, wherein the extract is a water extract or an aqueous ethanol extract.   The skin moisturizer according to claim 2 or 3, which is an external preparation.   A natural moisturizing factor production promoter comprising mint or its extract as an active ingredient.   The natural moisturizing factor production promoter according to claim 4, wherein the extract is a water extract or an aqueous ethanol extract.   A filaggrin production promoter comprising mint or an extract thereof as an active ingredient.   The filaggrin production promoter according to claim 6, wherein the extract is a water extract or an aqueous ethanol extract.   A prostasin production promoter containing mint or its extract as an active ingredient.   The prostasin production promoter according to claim 8, wherein the extract is a water extract or an aqueous ethanol extract.