JP7589413B2 - Desmoglein reducer - Google Patents

Description

The present invention relates to a desmoglein reducer.

Increased levels of desmosomal proteins are known to be one of the causes of deterioration of skin conditions, such as rough skin and peeling of the stratum corneum caused by sunburn or dryness.
As a method for controlling the amount of desmosomal proteins, a method has been reported in which decomposition of desmosomal proteins accumulated in the stratum corneum with proteases is used to improve acne, dandruff, and desquamation (Patent Document 1).

Among desmosomal proteins, desmoglein in particular is known to be involved in adhesion between epidermal cells and keratinocytes (Patent Documents 2-4).

As a prior art, Patent Document 2 reports an agent for reducing desmoglein in stratum corneum cells, which contains astaxanthin.
Furthermore, Patent Document 4 reports an orally administered agent for inhibiting intercellular adhesion in the skin, which contains licorice extract as an active ingredient.

Incidentally, Patent Documents 5 and 6 report that extracts of marsh mallow (Athaeea officinalis) have the effect of preventing and improving gray hair, and that tiliroside, a component of marsh mallow, has the effect of promoting fat metabolism.

Special Publication No. 7-505383 JP 2016-37501 A Patent No. 4002635 Patent No. 4197194 JP 2016-210755 A JP 2007-176858 A

Given the above prior art, the objective of the present invention is to provide a new technology for reducing desmogleins.

As a result of intensive research, the inventors discovered that an extract of marsh mallow (Athaea officinalis) has the effect of reducing desmoglein, and thus completed the present invention.

The present invention, which solves the above problems, provides a desmoglein reducer containing an extract of marsh mallow (Athaea officinalis) as an active ingredient.
In this specification, the concept of "reduction of desmoglein" includes both a reduction in desmoglein and inhibition of an increase in desmoglein.

In a preferred embodiment of the invention, the active ingredient is an extract of the roots of Marsh mallow (Athaeea officinalis).

In a preferred embodiment of the present invention, the desmoglein-reducing agent is used to inhibit intercellular adhesion of human epidermal keratinocytes.

In a preferred embodiment of the present invention, the desmoglein reducing agent of the present invention is used for reducing desmoglein 1.
Desmoglein 1 is mainly present in the skin. Therefore, the desmoglein-reducing agent of the present invention, which has the effect of reducing desmoglein 1, can more efficiently prevent or improve deterioration of skin conditions.

In a preferred embodiment of the present invention, the desmoglein reducer is used to improve and/or prevent sensitive skin.

In a preferred embodiment of the present invention, the desmoglein reducer is used to improve skin brightness.

In a preferred embodiment of the present invention, the desmoglein reducer is used to improve skin clarity.

In a preferred embodiment of the present invention, the desmoglein reducing agent is used to soften the skin.

In a preferred embodiment of the present invention, the desmoglein reducer is used to aid in the penetration of medicinal ingredients into the skin.

In a preferred embodiment of the present invention, the desmoglein reducing agent is used to prevent or improve stratum corneum desquamation.

The present invention, which solves the above-mentioned problems, is a composition for external application to the skin for suppressing desmoglein, which contains the above-mentioned desmoglein reducer.

In a preferred embodiment of the present invention, the above-mentioned composition for external application to the skin for inhibiting desmoglein is a cosmetic.

In a preferred embodiment of the present invention, the above-mentioned composition for topical application to the skin for inhibiting desmoglein is a pharmaceutical product.

The present invention provides a new technology for reducing desmogleins.

FIG. 1 is a diagram showing test results of Example 1. 1 is a graph showing test results of Example 1. 1 is a graph showing the measurement results of L ^* value in Example 2. 1 is a graph showing the measurement results of a ^* value in Example 2. 1 is a graph showing the measurement results of b ^* value in Example 2. 13 is a graph showing the results of hardness measurements in Example 2. 1 is a graph showing the measurement results of the amount of desmoglein 1 in Example 2. FIG. 1 is a representative diagram showing the measurement results of the amount of desmoglein 1 in Example 2.

The desmoglein reducing agent of the present invention contains an extract of marsh mallow (Athaea officinalis, hereinafter simply referred to as marsh mallow) as an active ingredient.
In particular, the desmoglein reducing agent of the present invention preferably contains an extract of the roots of Marshmallow as an active ingredient.

Marshmallow extract can be purchased and used from a commercially available marshmallow extract sold by a company that handles plant raw materials.

Also, a marshmallow extract can be produced by extracting wild or cultivated marshmallow.
It is also preferred to prepare an extract of Marshmallow by extracting the roots of Marshmallow.

When extracting marshmallow, it is preferable to process the marshmallow roots in advance by crushing or shredding them to improve the extraction efficiency.

The extract can be obtained, for example, by the following method. 1 to 30 parts by mass of a solvent is added to 1 part by mass of marshmallow root or its dried matter, and the mixture is immersed for several days at room temperature or for several hours at a temperature close to the boiling point. After immersion, the mixture is cooled to room temperature, and insoluble matter is removed if desired, and then the solvent is removed by concentrating under reduced pressure or the like. The desired extract can then be obtained by fractionation and purification using column chromatography filled with silica gel or ion exchange resin.

Suitable examples of the extraction solvent include one or more selected from the group consisting of water, alcohols such as ethanol, propanediol, glycerin, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butylene glycol and polypropylene glycol, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran.
As the extraction solvent, alcohols are particularly preferred, and among these, propanediol and glycerin are preferred.

The content of the marshmallow extract in the desmoglein-reducing agent of the present invention is preferably 0.02 mass % or more, more preferably 0.05 mass % or more, even more preferably 0.08 mass % or more, and particularly preferably 0.1 mass % or more.
The content of the extract of Marshmallow is 2% by mass or less, preferably 1.5% by mass or less, and more preferably 1.2% by mass or less.

The desmoglein reducer of the present invention preferably targets desmoglein 1.

The desmoglein-reducing agent of the present invention is preferably used for improving and/or preventing sensitive skin.
Here, in this specification, "sensitive skin" refers to a skin type with reduced resistance (reduced skin irritation value) to external stimuli (dryness, ultraviolet rays, application of chemical compounds).

More specifically, "sensitive skin" in this specification refers to the skin types listed in (1) and/or (2).

(1) "Skin that reacts to substances such as topical medicines, cosmetics, plants, ultraviolet rays, and metals and is prone to skin problems. Also, skin that is hypersensitive to allergens (pollen, fragrances, etc.) and irritants (alcohol, etc.)."
(2) "Skin that is temporarily more susceptible to skin problems due to lack of sleep, overwork, menstruation, seasonal changes, mental stress, etc."

In addition, in this specification, "for the improvement and/or prevention of sensitive skin" refers to the improvement of skin quality with reduced resistance to external stimuli (reduced skin irritation threshold) and/or the prevention of reduced resistance to external stimuli in skin quality (reduced skin irritation threshold).

In addition, the desmoglein-reducing agent of the present invention is preferably used for inhibiting intercellular adhesion of human epidermal keratinocytes.
In this specification, the term "intercellular adhesion" refers to abnormalities in the adhesive function of the stratum corneum (progression of stratification of the stratum corneum due to increased cell adhesiveness) caused by an increase in desmosomal proteins (a group of proteins including desmoglein) (see Patent Nos. 4,197,194 and 4,002,635 for details).
In addition, in this specification, the concept of "inhibiting intercellular adhesion" includes both prevention and improvement of intercellular adhesion.

Here, by inhibiting intercellular adhesion of human epidermal keratinocytes, stratification of the stratum corneum is inhibited, and an effect of inhibiting stratum corneum desquamation can be obtained.
In other words, the desmoglein-reducing agent of the present invention can be used to prevent or improve stratum corneum desquamation.
In this specification, the term "stratum corneum delamination" refers to the peeling off of the stratum corneum from the skin surface in a multi-layered state. Whether or not the skin is in a state where stratum corneum delamination can be confirmed by collecting the stratum corneum by tape stripping and staining and observing the collected stratum corneum.

Furthermore, as shown in the Examples described below, desmoglein 1 is present in large amounts in areas of the skin with low brightness, and it is believed that the decrease in skin brightness is due to stratification of the stratum corneum.
In view of the above findings, by reducing desmoglein with the active ingredient of the present invention, it is possible to obtain an effect of improving skin brightness.
That is, the active ingredient of the present invention can also be used as a skin lightening agent.
In this specification, "improving skin brightness" refers to reducing the difference in brightness between skin in a normal state and skin whose brightness has been reduced due to the excess of desmoglein.

In addition, as shown in the examples described below, the desmoglein-reducing agent of the present invention can provide a more transparent skin condition. In other words, the active ingredient of the present invention can also be used as an agent for improving skin transparency.

Furthermore, as shown in the Examples described below, desmoglein 1 is present in large amounts in areas of high skin hardness, and it is believed that the cause of skin hardness is the stratification of the stratum corneum.
Based on the above findings, the effect of softening the skin can be obtained by reducing desmoglein, that is, the active ingredient of the present invention can also be used as a skin softener.

Here, by softening the skin, other medicinal ingredients can penetrate more efficiently, so by reducing desmoglein with the desmoglein reducer of the present invention, it is possible to obtain the effect of allowing medicinal ingredients such as whitening agents to penetrate the skin more efficiently.
That is, the active ingredient of the present invention is preferably used as a whitening ingredient penetration aid for use in combination with a whitening agent.

In particular, the desmoglein reducing agent of the present invention is preferably used as a whitening ingredient penetration aid in combination with a whitening agent.

The desmoglein-reducing agent of the present invention can be a composition for external application to the skin or a composition for oral administration.
In particular, the desmoglein-reducing agent of the present invention is preferably in the form of a composition for external use on the skin.

Suitable examples of compositions for topical use on the skin include cosmetics and pharmaceuticals.

In particular, it is preferable for the product to be in the form of a cosmetic product that can be used continuously, such as a lotion, milky lotion, beauty essence, cream, gel, sun care product, etc.

The content of the extract of Marshmallow in the composition for external use on skin is 0.02% by mass or more, preferably 0.05% by mass or more, and more preferably 0.1% by mass or more.
The content of the extract of marsh mallow (Athaea officinalis) in the composition for external skin application is 2% by mass or less, preferably 1.5% by mass or less, and more preferably 1.2% by mass or less.

When the composition is provided in the form of a skin topical composition, the composition is not particularly limited, and it may contain any optional ingredient that is normally used, provided that the effect of the present invention is not impaired. Examples of such optional ingredients include oils and waxes such as macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin, hydrogenated coconut oil, hydrogenated oil, Japan wax, hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, Japanese laurel wax, lanolin, reduced lanolin, hard lanolin, and jojoba wax; liquid paraffin, squalane, pristane, ozokerite, and paraffin. Hydrocarbons such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid, etc.; higher fatty acids such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol, etc.; cetyl isooctanoate, isopropyl myristate, isopropyl alcohol, Synthetic ester oils such as hexyldecyl sostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, trimethylolpropane triisostearate, and pentane erythritol tetra-2-ethylhexanoate; anionic surfactants such as fatty acid soaps (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, and alkyl sulfate triethanolamine ether; cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, and laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt, etc. ), betaine surfactants (alkyl betaine, amido betaine, sulfobetaine, etc.), amphoteric surfactants such as acyl methyl taurine; sorbitan fatty acid esters (sorbitan monostearate, sorbitan sesquioleate, etc.), glycerin fatty acids (glycerin monostearate, etc.), propylene glycol fatty acid esters (propylene glycol monostearate, etc.), hydrogenated castor oil derivatives, glycerin alkyl ethers, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbitan fatty acid esters (POE-sorbitan monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoisostearate, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE 2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonionic surfactants such as POE/POP alkyl ethers (POE/POP 2-decyltetradecyl ether, etc.), Pluronic (registered trademark) types, POE/POP alkyl ethers (POE/POP 2-decyltetradecyl ether, etc.), Tetronics, POE castor oil/hydrogenated castor oil derivatives (POE castor oil, POE hydrogenated castor oil, etc.), sucrose fatty acid esters, and alkyl glucosides; polyethylene glycol, glycerin, erythritol, sorbitol, maltitol, propylene glycol, 2,4-hexanediol, Preferred examples include polyhydric alcohols such as ol; moisturizing ingredients such as sodium pyrrolidone carboxylate, lactic acid, and sodium lactate; para-aminobenzoic acid-based UV absorbers; anthranilic acid-based UV absorbers; salicylic acid-based UV absorbers; cinnamic acid-based UV absorbers; benzophenone-based UV absorbers; sugar-based UV absorbers; and UV absorbers such as 2-(2'-hydroxy-5'-t-octylphenyl)benzotriazole and 4-methoxy-4'-t-butyldibenzoylmethane.

When the composition is prepared as an oral composition, it is preferable to prepare a food composition containing the active ingredient of the present invention.
Specifically, the composition may be in the form of a supplement having the dosage form of a general food, tablet, granule, drink, or the like.

Here, the content and content ratio of the marshmallow extract in the oral composition may be the same as described above in the preferred embodiment of the topical skin composition.

When preparing an oral composition, optional ingredients may be added as appropriate within the scope that does not impair the effects of the present invention.

The present invention also relates to a method for preparing Desmoglace on the skin, which comprises applying an extract of Marshmallow to the skin.
According to the present invention, sensitive skin can be prevented or improved by applying an extract of Marshmallow to the skin.

The method for preparing desmoglein on the skin of the present invention is a non-therapeutic method, preferably a cosmetic method, more preferably a cosmetic method for the skin.

Example 1: Measurement of desmoglein-reducing effect The desmoglein-reducing ability of an extract of Marshmallow root was examined below.

<Step of preparing stratum corneum cell sample>
The stratum corneum was collected from the outer side of a human forearm using cellophane tape and transferred to a slide glass ("MAS Coat", Matsunami Glass Industry Co., Ltd.), and the slide glass was immersed in xylene overnight.
Next, the slide glass was washed twice with xylene for 10 minutes each, and then washed with PBS to prepare a keratinocyte sample.

<Preparation of test samples and target samples>
A test sample was obtained by diluting an extract of Marshmallow root (manufactured by Jurlique) with PBS to a concentration of 0.25% by mass.

<Preparation of measurement samples>
The stratum corneum cells were immersed in the above-mentioned test sample overnight to obtain a measurement sample for Example 1.
As a reference example (control), stratum corneum cells that were not immersed in the above-mentioned test samples were prepared.

<Immunostaining test>
The measurement sample was washed with PBS, and then incubated in 4% PFA-phosphate buffer at room temperature for 15 minutes. After incubation, the measurement sample was washed with PBS and incubated in a 20% aqueous solution of Block Ace (DS BioPharma Medical) at room temperature for 1 hour.
Thereafter, a primary antibody (Anti-Desmoglein 1, Mouse-Mono) was added, and the mixture was incubated in a humidified box at room temperature for 2 hours.

Each measurement sample was washed three times with PBS for 5 minutes each, and then a secondary antibody (Allexa Fluor@488 Goat Anti-mouse IgG) was added and incubated in a humidified box at room temperature for 1 hour.
Each measurement sample was washed three times with PBS for 5 minutes each, washed twice with distilled water, and then sealed using Fluoromout-G (manufactured by Southern Biotech).
Thereafter, analytical images of the measurement samples were taken using a confocal laser microscope, and the fluorescence (staining) was confirmed. The results are shown in FIG.

The area of the fluorescent (stained) portion of the measurement sample in Example 1 was calculated by taking the area of the fluorescent (stained) portion of the control as 100%. The results are shown in Table 1 and Figure 2.

As shown in Figures 1 and 2, it was found that the expression level of desmoglein 1 was suppressed in the measurement sample (Example 1) to which marshmallow extract was added, compared to the sample (Reference Example) to which marshmallow extract was not added.
These results demonstrate that the marshmallow extract has the effect of suppressing the amount of desmoglein 1.

[Example 2] Relationship between skin brightness and/or skin hardness and the amount of desmoglein 1 The following test results support the relationship between differences in skin brightness and the amount of desmoglein 1 present, and the relationship between the amount of desmoglein 1 present and skin hardness.

(1) Subjects and Measurement Sites In this example, 20 women aged between 40 and 52 years old were used as subjects.
In this embodiment, the areas on the subject's face where a decrease in brightness was visually confirmed (blackish areas, see area A in FIG. 8 ) and an area in a normal state (see area B in FIG. 8 ) were selected as measurement areas.

(2) Measurement (2-1) Measurement of L ^* value, a ^* value, and b ^* value A spectrophotometer was placed on the above measurement sites, and the L ^* value, a ^* value, and b ^* value were measured by measuring each site five times.
The results are shown in Figures 3 to 5.

Regarding the L ^* value, the areas where a decrease in brightness was visually confirmed (blackish areas, see area A in Figure 8) showed significantly lower values than the areas in a normal state (see area B in Figure 8). In other words, the areas where a decrease in brightness was visually confirmed (blackish areas, see area A in Figure 8) were confirmed to be darker than the areas in a normal state (see area B in Figure 8).

Regarding the a ^* value, the areas where the brightness was visually confirmed to have decreased (blackish areas, see area A in Figure 8) showed significantly higher values than the areas in the normal state (see area B in Figure 8). In other words, the areas where the brightness was visually confirmed to have decreased (blackish areas, see area A in Figure 8) were confirmed to have a stronger reddish hue than the areas in the normal state (see area B in Figure 8).

Regarding the b ^* value, the areas where the brightness was visually confirmed to have decreased (blackish areas, see area A in Figure 8) showed significantly higher values than the areas in the normal state (see area B in Figure 8). In other words, the areas where the brightness was visually confirmed to have decreased (blackish areas, see area A in Figure 8) were confirmed to be more yellowish than the areas in the normal state (see area B in Figure 8).

(2-2) Hardness Measurement The hardness was measured by pressing an indenter against each measurement site used in the test (2-1) above and measuring five times each.
The results are shown in Figure 6.

Regarding hardness, the areas where reduced brightness was visually confirmed (blackish areas, see area A in Figure 8) showed significantly lower values than the areas in a normal state (see area B in Figure 8). In other words, the areas where reduced brightness was visually confirmed (blackish areas, see area A in Figure 8) were confirmed to be harder than the areas in a normal state (see area B in Figure 8).

(2-3) Desmoglein 1 (Dsg1) Staining Corneal cells were collected from each measurement site used in the above tests (2-1) and (2-2) by tape stripping and attached to a slide glass.

The slide glass to which the keratinocytes were attached was immersed in xylene overnight.
After immersion, the sample was washed and air-dried, and then allowed to stand in 4% paraformaldehyde-phosphate buffer at room temperature for 15 minutes.

After 15 minutes, the stratum corneum cells were washed with PBS and immersed in a 0.5% TritonX (Nacalai Tesque)/PBS solution at room temperature for 5 minutes.

After immersion, the sample was washed and then left to stand in an aqueous solution of Block Ace (DS Biopharma Medical UK-B80) at room temperature for 1 hour.

After standing, the plate was framed with Liquid Blocker (manufactured by Daido Sangyo Co., Ltd.), a primary antibody was added, and the plate was left standing in a humid box at room temperature for 2 hours.
As the primary antibody, Anti Desmoglein 1, Mouse Mono (Dsg1 P23) Progen, 651110 stock solution (IgG concentration: 10-15 g/mL) was used.

After standing, the plate was washed with PBS, a secondary antibody was added, and the plate was left standing in a humidified box at room temperature for 1 hour.
As the secondary antibody, Allexa Fluor@488 Goat Anti-mouse IgG Invitrogen, A11001, 200-fold diluted solution in PBS was used.

After leaving it to stand, it was washed, sealed with Fluoromout-G (Southern Biotech), and dried at room temperature for 30 minutes.

After drying, the cover glass was fixed with nail polish and allowed to dry, and then (photographed) observed with a confocal laser microscope (Nikon A1) at ×20.
The results are shown in Figure 7.

Regarding the amount of desmoglein 1 (Dsg1), the areas where a decrease in brightness was confirmed by visual inspection (blackish areas, see area A in Figure 8) showed significantly higher values than the areas in a normal state (see area B in Figure 8). In other words, it was confirmed that the areas where a decrease in brightness was confirmed by visual inspection (blackish areas, see area A in Figure 8) had a higher amount of desmoglein 1 (Dsg1) than the areas in a normal state (see area B in Figure 8).

(3) Discussion As shown in Figures 3 to 5 and 7, it was found that desmoglein 1 was present in large amounts in areas of the skin with low brightness. Here, the decrease in skin brightness is thought to be due to the dense layering of the stratum corneum (stratification of the stratum corneum) caused by the low pH of the skin and low activity of serine protease that breaks down desmoglein 1.

As described above, the active ingredient of the present invention has the effect of reducing desmoglein.
In view of the above findings, it has been found that the active ingredient of the present invention has the effect of improving skin brightness by reducing desmoglein 1.

As shown in Figures 6 and 7, it was found that desmoglein 1 is present in large amounts in areas of high skin hardness. Here, the cause of skin hardness is thought to be the dense layering of the stratum corneum (stratification of the stratum corneum) due to the low pH of the skin and low activity of serine protease, which breaks down desmoglein 1.

Based on the above findings, it has been found that reducing desmoglein 1 can soften the skin.
As described above, the active ingredient of the present invention has the effect of reducing desmoglein.
In other words, it was found that the active ingredient of the present invention has a skin softening effect by reducing desmoglein 1.

The present invention is applicable to cosmetics.

Claims (13)

A skin preparation for external application containing an extract of marsh mallow (Athaeea officinalis) as an active ingredient, for application to dark areas of the skin;
In the darker areas of the skin,
To soften the firmness of said skin, or
To allow the medicinal ingredients to penetrate the skin efficiently
A desmoglein reducer used in . The desmoglein reducing agent according to claim 1, wherein the extract is an extract of the roots of marsh mallow (Athaea officinalis). The desmoglein-reducing agent according to claim 1 or 2 for inhibiting intercellular adhesion of human epidermal keratinocytes. The desmoglein reducer according to any one of claims 1 to 3, wherein the target to be reduced is desmoglein 1. A desmoglein-reducing agent according to any one of claims 1 to 4 for improving and/or preventing sensitive skin. A desmoglein reducer according to any one of claims 1 to 4 for improving skin brightness. A desmoglein reducer according to any one of claims 1 to 4 for improving skin clarity. A desmoglein reducing agent according to any one of claims 1 to 4 for softening the skin. A desmoglein reducer according to any one of claims 1 to 4 for aiding the penetration of medicinal ingredients into the skin. A desmoglein reducing agent according to any one of claims 1 to 4 for improving and/or preventing stratum corneum desquamation. A composition for external application to the skin for reducing desmoglein, comprising the desmoglein reducing agent according to any one of claims 1 to 4 . The composition for external application to the skin for reducing desmoglein according to claim 11, which is a cosmetic product. The composition for external application to the skin for reducing desmoglein according to claim 11, which is a pharmaceutical product.