WO2014208093A1

WO2014208093A1 - Moisturizing-related gene expression promoting agent related to improving skin moisturizing function

Info

Publication number: WO2014208093A1
Application number: PCT/JP2014/003420
Authority: WO
Inventors: Shoichi Yoda; Shuta FURUKI
Original assignee: Kewpie Corporation
Priority date: 2013-06-28
Filing date: 2014-06-26
Publication date: 2014-12-31
Also published as: JP2015063510A

Abstract

[Object] To provide a moisturizing-related gene expression promoting agent related to skin moisture. [Solving Means] A moisturizing-related gene expression promoting agent includes hyaluronic acid or its pharmaceutically acceptable salt as an effective component.

Description

MOISTURIZING-RELATED GENE EXPRESSION PROMOTING AGENT RELATED TO IMPROVING SKIN MOISTURIZING FUNCTION

The present invention relates to a moisturizing-related gene expression promoting agent, and in more detail, to a moisturizing-related gene expression promoting agent that promotes expression of a gene of a protein related to skin moisture, which controls movement of liquid between the inside and the outside of a cell.

Maintaining moisture in the skin leads to improvement in skin tone and firmness. Therefore, moisture is essential for beautiful skin. The human skin, i.e., skin, is the outermost organ of the human body and has a function of preventing water from evaporating from the inside of the body excessively as an important role. A decrease in the amount of water in the skin cell due to excess evaporation of water from the body results in deterioration of skin conditions, e.g., decrease in skin tone and firmness. Therefore, preventing water from excessively evaporating from the body and maintaining water uptake by a normal cell lead to improvement in skin conditions, e.g., maintained moisture in the skin and improvement in skin tone and firmness.

The skin has a three-layered structure of epidermis, dermis, and subcutaneous tissue, from the surface. In particular, the epidermis adjusts the amount of water by preventing water from evaporating from the body, i.e., plays a role of maintaining moisture in the skin. The epidermis is divided into stratum corneum, stratum granulosum, stratum spinosum, and stratum basale. The cycle in which keratinocytes generated in the stratum basale of the epidermis are pushed up while differentiating and become corneocytes through the stratum spinosum, stratum granulosum, and stratum corneum, and the corneocytes fall off, is repeated.

There are two main pathways for movement of liquid between the inside and the outside of a cell in the epidermis, which is important for the skin moisturizing function. One is a known pathway in which aquaporin being a transport protein on a cell membrane, which is known as water channel, is expressed on a cell membrane of keratinocytes of the stratum basale from the stratum granulosum of the epidermis, and plays a role of taking, in the cell, a low molecular substance such as water in intercellular space. The other is a known pathway in which tight junction being a cell adhesion structure controls water permeation between cells in the stratum granulosum due to adhesion of cells.

Thirteen types of aquaporin (AQP0 to AQP12) being a transport protein on a cell membrane, which is known as water channel, have been known in humans. In the epidermis, mainly aquaporin 3 exists. The aquaporin 3 is considered to play a role of taking in also a low molecular compound such as glycerol and urea, which are involved in water or moisture retention action.

Proteins called tight junction exist in a belt form around cells of the stratum granulosum of the epidermis, and control permeation of liquid such as water or substance between cells in the stratum corneum by bringing adjacent cells into close contact with each other. Examples of the protein forming tight junction includes occludin, claudin, and JAM (Junctional Adhesion Molecule).

Therefore, in order to maintain moisture in the skin and to achieve beautiful skin, it is very important to facilitate production of the protein that controls movement of liquid between the inside and the outside of a cell as described above.

Therefore, from the past, a skin barrier function improving agent including a cactus fruit extract that has tight junction formation promoting effects (PTL 1), and a stable chemical having excellent tight junction formation promoting effects (PTL 2)have been disclosed, for example.
In addition, it is disclosed that tocopheryl retinoate facilitates expression of aquaporin in a skin cell (PTL 3).

However, a safer material having a higher effect has been searched for in order to use the material for a pharmaceutical drug, a quasi-drug, cosmetics, and a food product.

Japanese Patent Application Laid-open No. 2012-121871 Japanese Patent Application Laid-open No. 2007-210948 Japanese Patent Application Laid-open No. 2006-290873

Summary

The present invention provides a moisturizing-related gene expression promoting agent that is effective for promoting expression of a moisturizing-related gene.

The present inventors have found that hyaluronic acid or its pharmaceutically acceptable salt promotes expression of a moisturizing-related gene being a gene of a protein that controls movement of liquid between the inside and the outside of a cell, and have completed the present invention.

Specifically, the present invention provides
(1) a moisturizing-related gene expression promoting agent that promotes expression of a moisturizing-related gene being a gene of a protein related to skin moisture, the agent including
hyaluronic acid or its pharmaceutically acceptable salt as an effective component, the moisturizing-related gene being a gene of a protein that controls movement of liquid between inside and outside of a cell,
(2) the moisturizing-related gene expression promoting agent according to (1), in which
the moisturizing-related gene is a gene of aquaporin, and
(3) the moisturizing-related gene expression promoting agent according to (1), in which
the moisturizing-related gene is at least one gene selected from the group consisting of occludin, claudin, and JAM.

According to the above-mentioned moisturizing-related gene expression promoting agent, production of a protein related to skin moisture, which controls movement of liquid between the inside and the outside of a cell, is promoted, thereby providing a new preparation that is effective for improving the ability to retain moisture from the inside of the skin.

Moreover, hyaluronic acid or its pharmaceutically acceptable salt can permeate from the epidermis of the skin to the stratum corneum, and exert its effects. Even when being taken via the oral route, it is absorbed from the intestinal tract, reaches the skin, and can exert its effects.

Hereinafter, the present invention will be described in detail with reference to the drawings. It should be noted that in the present invention, the term "parts" refers to "parts by mass," and the term "%" refers to "% by mass," unless otherwise noted.

<Moisturizing-related gene expression promoting agent>
The moisturizing-related gene expression promoting agent according the present invention is characterized by containing hyaluronic acid or its pharmaceutically acceptable salt as an effective component. In the present invention, the moisturizing-related gene refers to a gene of a protein related to improvement in skin moisturizing function. Furthermore, the moisturizing-related gene in the present disclosure is a gene of a protein that controls movement of liquid between the inside and the outside of a cell. The moisturizing-related gene expression promoting agent according to the present disclosure promotes expression of a moisturizing-related gene, thereby promoting production of the protein related to improvement in skin moisturizing function.
There are two main pathways for movement of liquid between the inside and the outside of a cell in the epidermis, which is important for the skin moisturizing function. One is a known pathway of a transport protein on a cell membrane, which is known as water channel, and the other is a known pathway in which a cell adhesion structure protein controls permeation between cells due to adhesion of cells.
Examples of the moisturizing-related protein that controls movement of liquid between the inside and the outside of a cell include aquaporin 3, aquaporin 5, and aquaporin 9 that control water transport on a cell membrane, and occludin, claudin, and JAM that form tight junction controlling liquid permeation between cells.

<Evaluation for gene expression promoting effects>
Regarding the moisturizing-related gene expression promoting effects according to the present invention, the gene expression promoting effects can be confirmed by extracting RNA from keratinocytes of the human epidermis after adding the moisturizing-related gene expression promoting agent, synthesizing cDNA, amplifying the cDNA by a real-time PCR method, and measuring expression of the mRNA, for example.

<Hyaluronic acid or its pharmaceutically acceptable salt>
In the present invention, the term "hyaluronic acid" refers to polysaccharide having one or more repeating units formed of disaccharide of Beta-D-glucuronic acid and Beta-D-N-acetylglucosamine. Specifically, the hyaluronic acid is di- or higher saccharide including at least one disaccharide unit in which the first position of Beta-D-glucuronic acid is bound to the third position of Beta-D-N-acetyl-glucosamine. On the other hand, examples of the "hyaluronic acid's pharmaceutically acceptable salt" include, but not limited to, sodium salt, potassium salt, calcium salt, zinc salt, magnesium salt, and ammonium salt.
Hereinafter, in the present invention, the hyaluronic acid or its pharmaceutically acceptable salt is also referred to simply as "hyaluronic acid."

<Derivation of hyaluronic acid>
The hyaluronic acid may be one extracted from a biological tissue (e.g., crista galli, umbilical cord, skin, and joint fluid) of an animal, for example. Alternatively, those obtained by culturing a microbe, an animal cell, or a plant cell (e.g., fermentation method using a bacterium of genus Streptococcus or the like), or those synthesized chemically or enzymatically, or the like can be used.

<Purity of hyaluronic acid>
It should be noted that the purity of hyaluronic acid used in the present invention only needs to be at a level that can be used for a pharmaceutical drug, to be favorably 90% or more, and to be more favorably 95% or more. This purity represents a value calculated from a glucuronic acid quantitative value that is measured by a carbazole-sulfuric acid method (e.g., The Japanese Pharmacopoeia).

<Method of measuring purity>
The carbazole-sulfuric acid method is a method in which a hyaluronic acid aqueous solution is added to a sodium borate and sulfuric acid solution, the obtained solution is mixed, the hyaluronic acid is thermally decomposed before it is cooled, a carbazole and ethanol solution is added thereto, the obtained solution is mixed, the absorbance (530nm) of a sample soloution obtained by allowing the mixed solution to cool after heating is measured. D-glucuronolactone subjected to the same process is used to generate a calibration curve, D-glucuronolactone coversion value is calculated before multiplying it by 1.102, and thus a glucuronic acid quantitative value is obtained. The glucuronic acid quantitative value thus obtained is multiplied by (molecular weight of hyaluronic acid/molecular weight of glucuronic acid) to calculate the content of hyaluronic acid.

<Molecular weight of hyaluronic acid>
Moreover, the average molecular weight of hyaluronic acid used for the moisturizing-related gene expression promoting agent according to the present invention is 100,000 or less, favorably 70,000 or less, more favorably 400 or more to 60,000 or less, more favorably 1,000 or more to 10,000 or less, from a viewpoint of obtaining sufficient gene expression promoting effects. If the average molecular weight of hyaluronic acid is more than 100,000, it cannot permeate through the horn when it is applied as an external preparation. Therefore, it may be hard to obtain the moisturizing-related gene expression promoting effects according to the present invention.

It should be noted that the average molecular weight of hyaluronic acid defined in the present disclosure is measured by the following method.

<Method of measuring molecular weight>
Specifically, about 0.05 g of purified hyaluronic acid is accurately weighed and is dissolved in a sodium chloride solution having a concentration of 0.2 mol/L to obtain accurately 100 mL of the solution. Accurately 8mL, 12mL, and 16mL of the solution are weighed, and a sodium chloride solution having a concentration of 0.2 mol/L is added thereto, thereby obtaining accurately 20 mL of the solutions, which are sample solutions. The specific viscosity of the sample solutions and the sodium chloride solution having a concentration of 0.2 mol/L is measured by The Japanese Pharmacopoeia (Fourteenth Edition) General Tests, Processes and Apparatus (Method I Viscosity measurement by capillary tube viscometer) at 30.0+/-0.1deg C (formula (1)), and the reduction viscosity at each concentration is calculated (formula (2)). A graph having the reduction viscosity on the vertical axis and the converted concentration (g/100mL) with respect to the dried object of this product on the horizontal axis is drawn, and the limiting viscosity is obtained based on an intersection point between a straight line connecting points and the vertical axis. The limiting viscosity obtained here is substituted into the Laurent's formula (formula (3)) to calculate the average molecular weight (T.C. Laurent, M. Ryan, A. Pietruszkiewicz,:B.B.A., 42, 476-485(1960)).
(formula 1)
Specific viscosity = {(necessary number of seconds for falling of sample solution)/(necessary number of seconds for falling of 0.2 mol/L of sodium chloride solution)}-1
(formula 2)
Reduction viscosity = specific viscosity/(converted concentration (g/100mL) with respect to the dried object of this product)
(formula 3)
Limiting viscosity = 3.6x10^-4M^0.78
M: average molecular weight

<Content of hyaluronic acid>
The content of hyaluronic acid in the moisturizing-related gene expression promoting agent according to the present invention may be any amount as long as the hyaluronic acid can function as an effective component, and is normally 1% by mass or more, and favorably 5 to 95% by mass.

<Action mechanism of gene expression promoting effects>
The action mechanism of promoting the expression of a moisturizing-related gene in keratinocytes by taking or applying the moisturizing-related gene expression promoting agent according to the present invention is not necessarily clear. However, it is speculated that hyaluronic acid or its pharmaceutically acceptable salt, which is the effective component according to the present invention, acts on keratinocytes of the skin and increases the expression of the moisturizing-related gene.
In the case where hyaluronic acid having a molecular weight of 100,000 or more is applied as an external preparation, it has a difficulty of permeating through the horn. Therefore, it may be hard to obtain the moisturizing-related gene expression promoting effects according to the present invention of the subject patent application.

<Another material used for moisturizing-related gene expression promoting agent>
The moisturizing-related gene expression promoting agent according to the present invention may include, in addition to hyaluronic acid being an effective component, another raw material in the range where the effect of the invention is not lost. Examples of such a raw material include water, an excipient, an antioxidant agent, a preservative, a wetting agent, a thickener, a buffer, an adsorbent, a solvent, an emulsifier, a stabilizer, a surfactant, a lubricant, a water-soluble polymer, a sweetener, a flavoring substance, an acidulant, and alcohols.
Furthermore, the moisturizing-related gene expression promoting agent according to the present invention may include another effective component in the range where the effect of the invention is not lost. Specific examples of the effective component include an antioxidant ingredient, an anti-aging ingredient, an anti-inflammatory component, a whitening ingredient, a cell activation component, vitamins, a blood circulation promotion component, a moisturizing ingredient, a component having a prevention and/or repair action of DNA damage, an anti-glycation ingredient, a peptide or its derivative, an amino acid or its derivative, and a hydroquinone glycoside and its esters. Furthermore, it may include a component that is expected to have the same moisturizing-related gene expression promoting effects as those of the moisturizing-related gene expression promoting agent according to the present invention. Examples of such a component include a ginger extract, a watercress extract, a schizonepeta herb extract, an ajuga turkestanica extract, a phosphatidic acid, a dipotassium glycyrrhizin, a hydrangea vine extract, a lavender extract, an isodon japonicus extract, a cactus fruit extract, a kinginka extract, a saffron extract, a retinoic acid, and a cryptoxanthin.

<Form of moisturizing-related gene expression promoting agent>
The form of the moisturizing-related gene expression promoting agent according to the present invention is not particularly limited. However, in the case where the moisturizing-related gene expression promoting agent according to the present invention is taken via the oral route, examples of the form include an orally-administered agent such as a powder, a fine granule, a granule, a capsule, a solid preparation such as a pill, a water agent, a suspension, a syrup, and a solution such as an emulsion.
On the other hand, in the case where it is applied as an external preparation, examples of the form include a solution, a suspension, an emulsion, a cream, an ointment, a gel, a liniment, a lotion, and a poultice.

<Amounts of taking and applying moisturizing-related gene expression promoting agent>
It is considered that even if a large amount of hyaluronic acid is taken or applied, no or very few side effects are caused because it is a biological substance. However, the amount(s) of hyaluronic acid taken as the moisturizing-related gene expression promoting agent according to the present invention and/or its salt can be usually 10 mg to 1000 mg, favorably, 100 to 500 mg. The number of times of administration can be selected from once or a plurality of times a day.
Moreover, in the case where it is applied as an external preparation, 0.01 mg to 1000 mg, favorably, 0.5 to 500 mg, i.e., a proper amount of it may be applied to the skin several times a day although it differs depending on the state of the skin serving as a usage target, age, gender, and the like.

Hereinafter, the present invention will be described in more detail with the examples. However, the present invention is not limited to the examples.

<Examples 1 and 2>
The following hyaluronic acid was dissolved in sterilized water to obtain each moisturizing-related gene expression promoting agent.
Example 1: hyaluronic acid (average molecular weight of 8,000, white powder, manufactured by Kewpie Corporation)
Example 2: hyaluronic acid (average molecular weight of 2,000, white powder, manufactured by Kewpie Corporation)

<Test example 1>
In a test example 1, after a culture medium containing low molecular hyaluronic acid was added to epidermis keratinocytes and incubated, the expression of various moisturizing-related gene was measured in order to confirm the moisturizing-related gene expression promoting effects of low molecular hyaluronic acid.

<Cell culturing and RNA extraction>
In a SUMILON 24well Multi Well Plate, 1x10⁵ normal human adult epidermis keratinocyte NHEK (KURABO INDUSTRIES LTD.)is dispensed, and is cultured under the conditions of CO₂ concentration of 5% and 37deg C, using normal human epidermis keratinocyte basal medium (HuMedia-KG2, KURABO INDUSTRIES LTD.). It is cultured until it becomes confluent, and the supernatant is removed. After that, the medium is replaced with a normal human epidermis keratinocyte basal medium containing 1.8 mM of calcium ion, and culturing is performed under the conditions of CO₂ concentration of 5% and 37deg C for 2 to 4 days. After the medium is removed, a normal human epidermis keratinocyte basal medium containing 1.8 mM of calcium ion, which contains the moisturizing-related gene expression promoting agent according to the example 1 or the example 2, is added so that the concentration of hyaluronic acid is 0.01 mcg/mL finally, and is incubated for 3 hours or 6 hours under the conditions of CO₂ concentration of 5% and 37deg C. After that, the supernatant is removed, cells are collected, and total RNA is adjusted.

<Real-time PCR>
Based on total RNA, Affinity Script cDNA Synthesis Kit (Agilent Technologies Japan, Ltd.) was used to synthesize cDNA. Then, Brilliant III Ultra Fast SYBR Green QPCR Master Mix (Agilent Technologies Japan, Ltd.) and various primers described in Table 1 (100 pmoL/mcL, Hokkaido System Science Co.,Ltd.) were used to quantify the amount of expression of mRNA of the moisturizing-related gene using a real-time PCR device (Mx3005, Agilent Technologies Japan, Ltd.). Specifically, amplifying reaction in which thermal denaturation (95deg C, 10 seconds) and annealing (60deg C, 20 seconds) were repeated 40 times was performed, and the amplification of the amount of expression of mRNA was measured by monitoring the fluorescence of SYBR Green bound to DNA at the same time. The amount of expression of mRNA of aquaporin 3, occluding, claudin 1, and JAM-1 as a moisturizing-related gene was obtained as a proportion to the amount of expression of mRNA of GAPDH being an internal standard, and represented by the ratio of expression with a control being 1. It should be noted that in a comparative example (control), only a culture medium is added. The results of measuring the amount of expression of each gene are shown in Table 2.

<Primer sequence>
The primer sequences used for the real-time PCR were shown in Table 1.

From Table 2, in the example1 and the example2 in which low molecular hyaluronic acid having an average molecular weight of 100,000 or less was added, it had been shown that the expression of the moisturizing-related gene in human epidermis keratinocytes was promoted.
Moreover, because the gene expression was more promoted in a short time of 3 hours of incubation than 6 hours of incubation, it was considered that low molecular hyaluronic acid had high effects for promoting the expression of a moisturizing-related gene in a short time.

Hereinafter, prescription examples of the moisturizing-related gene expression promoting agent according to the present invention will be shown. However, the present invention is not limited by the prescription examples.

<Prescription example 1>
The hyaluronic acid used in the example 1 (hyaluronic acid having an average molecular weight of 8,000) was used as a moisturizing-related gene expression promoting agent to produce a powder medicine (granule) having the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1)_10%
Lactose_60%
Corn starch_25%
Hypromellose_5%
-------------------------------------------------------
100%

<Prescription example 2>
The hyaluronic acid used in the example 1 (hyaluronic acid having an average molecular weight of 8,000) was used as a moisturizing-related gene expression promoting agent to produce a tablet having the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1)_25%
Lactose_24%
Crystalline cellulose_20%
Corn starch_15%
Dextrin_10%
Emulsifier_5%
Silicon dioxide_1%
-------------------------------------------------------
100%

<Prescription example 3>
The hyaluronic acid used in the example 2 (hyaluronic acid having an average molecular weight of 2,000) was used as a moisturizing-related gene expression promoting agent to produce a soft capsule with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 2)_20%
Olive oil_50%
Beeswax_10%
Medium-chain triglyceride_10%
Emulsifier_10%
-------------------------------------------------------
100%

<Prescription example 4>
The hyaluronic acid of the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce an emulsion with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.20%
Propylene glycol_7.71%
Trehalose_0.03%
Ceramide 3_0.05%
Hinokitiol_0.01%
Citrate_0.05%
Glycolic acid_0.05%
Mineral oil_3.00%
Trioctanoin_1.50%
Squalane_1.00%
Stearic acid_0.50%
Cetearyl alcohol_0.50%
Lanolin_0.30%
Paraffin_0.20%
Sorbitan stearate_1.40%
Tetra Oleic acid solbes-30_1.00%
Polysorbate 60_0.80%
Methylparaben_0.20%
Propylparaben_0.10%
Ethanol_0.01%
Phenoxyethanol_Proper amount
Carbomer_0.10%
Potassium hydroxide_0.10%
Arginine_0.05%
BHT_0.02%
Tocopherol_Proper amount
EDTA-2 sodium_0.02%
Purified water_Remaining amount
-------------------------------------------------------
100%

<Prescription example 5>
In this example, the hyaluronic acid in the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce a thermal sense stimulating cataplasm with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.20%
Sodium polyacrylate_4.00%
Sodium carboxymethyl cellulose_2.00%
Kaolin_5.00%
Gelatin_3.00%
Glycerin_20.00%
Red pepper extract_0.15%
hyaluronic acid_5.00%
Purified water_remaining amount
-------------------------------------------------------
100.00%

<Prescription example 6>
In this example, the hyaluronic acid according to the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce a salve with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.5%
White petrolatum_25.0%
Stearyl alcohol_20.0%
Propylene glycol_12.0%
Polyoxyethylene hydrogenated castor oil 60_4.0%
Glyceryl monostearate_1.0%
Methyl parahydroxybenzoate_0.1%
Propyl parahydroxybenzoate_0.1%
Purified water_Remaining amount
-------------------------------------------------------
100%

<Prescription example 7>
In this example, the hyaluronic acid according to the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce face lotion with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_ 0.2%
Acetylated hyaluronic acid_0.1%
Chondroitin sulfate_0.1%
Collagen peptide_0.1%
1,3 - butylene glycol_5.0%
Glycerin_3.0%
Isostearyl alcohol_0.1%
Tocopherol acetate_0.1%
POE (20) sorbitan monolaurate acid ester_0.5%
POE (15) lauryl alcohol ether_0.5%
Pyrrolidone carboxylic acid zinc_0.1%
Retinoic acid_0.5%
Lavender extract_0.3%
Ginger extract_0.3%
Watercress extract_0.3%
Schizonepeta herb extract_0.3%
Saffron extract_0.3%
Ethyl paraben_0.1%
Methylparaben_0.15%
Ethanol_5.0%
Fragrance_Proper amount
Purified water_Remaining amount
-------------------------------------------------------
100%

<Prescription example 8>
In this example, the hyaluronic acid of the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce milky lotion with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.3%
Pentylene glycol_5.0%
Glycerin_3.0%
Squalane_5.0%
Stearic acid_0.5%
Stearyl alcohol_2.0%
Vaseline_4.0%
Sorbitan stearate_1.0%
POE (10) monostearate_1.0%
Carboxy vinyl polymer_0.5%
Polyquaternium -51_0.1%
Glycyrrhizin dipotassium_0.1%
Phosphatidic acid_0.5%
Cryptoxanthin_0.5%
Ajuga turkestanica extract_0.3%
Armature crane extract_0.3%
Isodon japonicus extract_0.3%
Cactus fruit extract_0.3%
Kinginka extract_0.3%
Methylparaben_0.15%
Propylparaben_0.1%
Potassium hydroxide_0.1%
BHT_0.02%
EDTA-2 sodium_0.02%
Fragrance_Proper amount
Purified water_Remaining amount
-------------------------------------------------------
100%

<Prescription example 9>
In this example, the hyaluronic acid of the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce cream (emollient cream) with the content of the following composition.

(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.5%
Polyethylene glycol_4.0%
1,3 - propanediol_6.0%
Squalane_11.0%
Dimethicone_1.0%
Cetanol_6.0%
Stearic acid_2.0%
Hydrogenated coco-glycerides_4.0%
Tricaprylin_8.0%
Glyceryl monostearate_3.0%
POE (20) cetyl alcohol ether_2.0%
Coenzyme Q10_0.03%
Ceramide_0.1%
Dilauroyl glutamic acid lysine sodium_0.1%
EDTA-2 sodium_0.02%
Propylparaben_0.1%
Methylparaben_0.15%
Fragrance_Proper amount
Purified water_Remaining amount
-------------------------------------------------------
100%

<Prescription example 10>
In this example, the hyaluronic acid of the example 1 or 2 was used as a moisturizing-related gene expression promoting agent to produce a serum (whitening moisturizing essence) with the content of the following composition.
(Composition proportion)
Moisturizing-related gene expression promoting agent (hyaluronic acid of the example 1 or 2)_0.8%
1,3 - butylene glycol_5.0%
Glycerin_1.5%
POE sorbitan monostearate_1.0%
Sorbitan monostearate_0.5%
Xanthane gum_0.2%
Sodium alginate_0.2%
Carboxy vinyl polymer_0.2%
Potassium hydroxide_0.1%
Olive oil_0.2%
Tocopherol_0.1%
EDTA-2 sodium_0.02%
Arginine_0.15%
Glycyrrhizin dipotassium_0.05%
Arbutin_0.2%
Retinol palmitate_0.2%
Ginseng extract_0.2%
Seaweed extract_0.2%
Tranexamic acid_0.1%
Elastin_0.1%
Collagen_0.1%
Phosphate magnesium ascorbate_0.1%
Sodium citrate_1.0%
Citrate_0.1%
Propylparaben_0.1%
Methylparaben_0.15%
Fragrance_Proper amount
Purified water_Remaining amount
-------------------------------------------------------
100%

Claims

A moisturizing-related gene expression promoting agent that promotes expression of a moisturizing-related gene being a gene of a protein related to skin moisture, the agent comprising
hyaluronic acid or its pharmaceutically acceptable salt as an effective component, the moisturizing-related gene being a gene of a protein that controls movement of liquid between inside and outside of a cell.
The moisturizing-related gene expression promoting agent according to claim 1, wherein
the moisturizing-related gene is a gene of aquaporin.
The moisturizing-related gene expression promoting agent according to claim 1, wherein
the moisturizing-related gene is at least one gene selected from the group consisting of occludin, claudin, and JAM.