JP7460134B2 - Desmoglein reducer - Google Patents

Description

The present invention relates to desmoglein reducing agents.

It is known that an increase in desmosomal proteins is one of the causes of deterioration of skin conditions, such as rough skin and desquamation 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.

By the way, in Patent Documents 5 to 7, extracts of Wild Thyme (T. serpyllum) and extracts of Thymus plants of the Lamiaceae family have an effect of improving water retention function, an anti-inflammatory effect, It has been reported to have skin anti-aging effects and UV protection effects.

Special Publication No. 7-505383 Japanese Patent Application Publication No. 2016-37501 Patent No. 4002635 Patent No. 4197194 Japanese Patent Application Publication No. 2001-122729 JP 2006-16337 A JP2009-256270A

While there are the above-mentioned prior art techniques, it is an object of the present invention to provide a novel technique for reducing desmogleins.

As a result of intensive research efforts, the inventors discovered that an extract of Wild Thyme (T. serpyllum) 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 Wild Thyme (T. serpyllum) 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 present invention, the active ingredient is an extract of the aerial parts of Wild Thyme (T. serpyllum).

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 form of the present invention, the desmoglein reducing agent is used to prevent or improve sensitive skin.

In a preferred form of the invention, the desmoglein reducing agent is used to improve skin brightness.

In a preferred form of the present invention, the desmoglein reducing agent is used to improve skin transparency.

In a preferred form of the invention, the desmoglein reducing agent is used for skin softening.

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 improve and/or prevent stratum corneum desquamation.

In a preferred embodiment of the present invention, the active ingredients are a composition containing an extract of Wild Thyme (T. serpyllum) and 1,3-butylene glycol.

The present invention also provides an external skin composition for reducing desmoglein, which includes the desmoglein reducing agent described above.

In a preferred embodiment of the present invention, the composition for topical application to the skin for reducing desmoglein is a cosmetic.

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

The present invention provides a new technology for reducing desmogleins.

3 is a diagram showing test results of Example 1. FIG. 1 is a graph showing test results of Example 1. 3 is a graph showing measurement results of L ^* values in Example 2. 1 is a graph showing the measurement results of a ^* value in Example 2. 3 is a graph showing the measurement results of b ^* values in Example 2. 3 is a graph showing hardness measurement results in Example 2. 1 is a graph showing the measurement results of the amount of desmoglein 1 in Example 2. 3 is a diagram (representative) showing the measurement results of 1 amount of desmoglein in Example 2. FIG. 3 is a diagram (representative) showing the measurement results of the amount of desmoglein in Example 3. FIG. 3 is a graph (6 weeks) showing the measurement results of 1 amount of desmoglein in Example 3. 3 is a graph (12 weeks) showing the measurement results of 1 amount of desmoglein in Example 3.

The desmoglein reducing agent of the present invention contains an extract of Wild Thyme (T. serpyllum) as an active ingredient.
Among these, it is preferable that the desmoglein reducing agent of the present invention contains an extract of the aerial part of Wild Thyme (T. serpyllum) as an active ingredient.

For the extract of Wild Thyme, T. serpyllum, a commercially available extract of Wild Thyme, T. serpyllum, which is sold by a company that handles plant materials, is purchased and used. be able to.

Furthermore, an extract of Wild Thyme, T. serpyllum can be produced by extracting wild Thyme, T. serpyllum.
Here, it is desirable to prepare an extract of Wild Thyme, T. serpyllum, by extracting the aerial part of Wild Thyme, T. serpyllum.

When extracting Wild Thyme (T. serpyllum), it is preferable to process the above-ground parts of Wild Thyme (T. serpyllum) in advance by crushing or shredding them to improve the extraction efficiency.

The extract can be obtained, for example, by the following method. First, 1 to 30 parts by mass of a solvent is added to 1 part by mass of the above-ground part of Wild Thyme (T. serpyllum) or its dried material, and the mixture is incubated at room temperature for several days or at a temperature near the boiling point. Soak for several hours. After immersion, the solution is cooled to room temperature, and if desired, insoluble matter is removed, and then the solvent is removed by concentrating under reduced pressure or the like. Thereafter, the desired extract can be obtained by fractionation and purification using column chromatography packed with silica gel or ion exchange resin.

The extraction solvent is preferably a polar solvent, and suitable examples thereof include one or more selected from water, alcohols such as ethanol, 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.
Among these, 1,3-butylene glycol is preferred as the extraction solvent.

The content of the extract of Wild Thyme (T. serpyllum) in the desmoglein-reducing agent of the present invention is preferably 0.02% by mass or more, more preferably 0.05% by mass or more, even more preferably 0.08% by mass or more, and particularly preferably 0.1% by mass or more.
The content of the extract of Wild Thyme (T. serpyllum) is 2% by mass or less, preferably 1.5% by mass or less, and more preferably 1.2% by mass or less.
In another embodiment, the content of the extract of Wild Thyme (T. serpyllum) is preferably 1% by mass or less, more preferably 0.5% by mass or less, even more preferably 0.3% by mass or less, and particularly preferably 0.2% by mass or less.

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

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

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).

Furthermore, the desmoglein reducing agent of the present invention is preferably used to suppress intercellular adhesion of human epidermal keratinocytes.
In this specification, the term "cell-to-cell adhesion" refers to an abnormality in the stratum corneum adhesion function due to an increase in desmosomal proteins (a group of proteins including desmoglein) (progressing stratification of the stratum corneum due to increased cell adhesion). (If necessary, refer to Patent No. 4197194 and Patent No. 4002635).
Furthermore, in this specification, the concept of "inhibiting cell-cell adhesion" includes both prevention and improvement of cell-cell adhesion.

Here, by suppressing intercellular adhesion of human epidermal keratinocytes, stratification of the stratum corneum is suppressed, and an effect of suppressing delamination of the stratum corneum can be obtained.
That is, the desmoglein reducing agent of the present invention can be used to prevent or improve stratum corneum exfoliation.
In this specification, "stratum corneum layer exfoliation" refers to peeling off from the skin surface in a plurality of overlapping layers of the stratum corneum. Whether or not the skin is in a state where the stratum corneum layer is exfoliated can be confirmed by collecting the stratum corneum using a tape stripping method, staining the collected stratum corneum, and observing it.

Furthermore, as shown in the examples described later, a large amount of desmoglein 1 exists in areas where the skin brightness is low. Here, the decrease in skin brightness is considered to be due to stratification of the stratum corneum.
In view of the above findings, by reducing desmoglein with the active ingredient of the present invention, an effect of improving skin brightness can be obtained.
That is, the active ingredient of the present invention can also be used as a skin brightness improving agent.
Here, in this specification, "skin brightness improvement" refers to a reduction in the difference in brightness between skin in a normal state and skin whose brightness has decreased due to the excessive presence of desmoglein.

Further, as shown in the Examples described below, the active ingredients of the present invention can provide a more transparent skin condition. That is, the active ingredient of the present invention can also be used as an agent for improving skin clarity.

Furthermore, as shown in the Examples described later, areas of high skin hardness contain a large amount of desmoglein 1. Here, the cause of skin hardness is thought to be the stratification of the stratum corneum.
Based on the above findings, reducing desmoglein makes the skin softer.
In other words, the active ingredient of the present invention can reduce desmoglein, thereby softening the skin, and therefore 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, and therefore by reducing desmoglein with the active ingredient 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 can also be used as an agent for aiding the penetration of medicinal ingredients into the skin.

In particular, the active ingredient of the present invention is preferably used as a skin whitening component penetration aid for use in combination with a whitening agent.

In the present invention, it is preferable to use an extract of Wild Thyme (T. serpyllum) in combination with 1,3-butylene glycol.
Desmoglein can be reduced more efficiently by combining an extract of Wild Thyme (T. serpyllum) with 1,3-butylene glycol.

The content of 1,3-butylene glycol in the desmoglein-reducing agent of the present invention is preferably 20% by mass or more, and more preferably 50% by mass or more.
Furthermore, the content of 1,3-butylene glycol in the desmoglein reducing agent of the present invention is 70% by mass or less, preferably 50% by mass or less.

The ratio of the extract of Wild Thyme (T. serpyllum) to 1,3-butylene glycol in the desmoglein reducing agent of the present invention is preferably 1:20 to 1:80, more preferably 1:50 to 1:70.

Desmoglein can be reduced more efficiently by setting the contents of Wild Thyme (T. serpyllum) extract and 1,3-butylene glycol within the above ranges.

The desmoglein reducing agent of the present invention can be in the form of a composition for external use on the skin or a composition for oral use.
Among these, 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 external 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, serum, cream, gel, sun care product, etc.

The content of the extract of Wild Thyme (T. serpyllum) in the composition for external skin application 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 Wild Thyme (T. serpyllum) 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.
The content of the extract of Thymus vulgare is preferably 1 mass % or less, more preferably 0.5 mass % or less, even more preferably 0.3 mass % or less, and particularly preferably 0.2 mass % or less.

The content of 1,3-butylene glycol in the composition for external use on skin is preferably 20% by mass or more, and more preferably 50% by mass or more.
The content of 1,3-butylene glycol in the composition for external use on skin is 70% by mass or less, preferably 50% by mass or less.
In another embodiment, the content of 1,3-butylene glycol in the composition for external use on skin is preferably 0.5% by mass or more, more preferably 1% by mass or more, and even more preferably 3% by mass or more.
Furthermore, the content of 1,3-butylene glycol in the composition for external use on skin is preferably 20% by mass or less, more preferably 15% by mass or less, even more preferably 10% by mass or less, and particularly preferably 7% by mass or less.

Desmoglein can be more effectively reduced by setting the contents of Wild Thyme (T. serpyllum) extract and 1,3-butylene glycol within the above ranges.

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, and the like; higher fatty acids such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol, and the like; cetyl isooctanoate, isopropyl myristate, Synthetic ester oils such as hexyldecyl isostearate, 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 stearyl trimethylammonium chloride, benzalkonium chloride, and laurylamine oxide; imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy 2-sodium salts, etc.), betaine surfactants (alkyl betaines, amido betaines, sulfobetaines, 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 nonionic surfactants such as POE nonylphenyl ether, Pluronic 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, etc. 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 a dosage form such as a general food, tablet, granule, or drink.

Here, regarding the content and content ratio of the extract of Wild Thyme (T. serpyllum) in the oral composition, the description of the preferred embodiment of the skin external composition described above may be applied mutatis mutandis. Can be done.

In addition, when preparing an oral composition, optional ingredients may be added as appropriate within a range that does not impair the effects of the present invention.

The present invention may also be in the form of a kit comprising a composition containing an extract of Wild Thyme (T. serpyllum) and a composition containing 1,3-butylene glycol.
In a kit including a composition containing an extract of Wild Thyme (T. serpyllum) and a composition containing 1,3-butylene glycol, the contents of the active ingredients when the composition containing an extract of Wild Thyme (T. serpyllum) and the composition containing 1,3-butylene glycol are mixed can be determined in the above-mentioned manner.

The present invention can also be a method for adjusting desmoglein on the skin, which comprises applying an extract of Wild Thyme (T. serpyllum) to the skin.
According to the present invention, sensitive skin can be prevented or improved by applying an extract of Wild Thyme (T. serpyllum) to the skin.

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

In addition, by applying an extract of Wild Thyme (T. serpyllum) and 1,3-butylene glycol to the skin and adjusting desmoglein as described above, sensitive skin can be prevented or improved. Can be done.

The above-mentioned contents can be applied to the preferred embodiment of the method for adjusting desmoglein on the skin of the present invention.

[Example 1] Measurement of desmoglein reducing effect The desmoglein reducing ability of the above-ground extract of Wild Thyme (T. serpyllum) was investigated below.

<Preparation process of stratum corneum cell sample>
The stratum corneum collected from the outside of a human forearm with cellophane tape was transferred onto a glass slide ("MAS coat", manufactured by Matsunami Glass Industries Co., Ltd.), and the slide glass was immersed in xylene overnight.
Next, the slide glass was washed twice with xylene for 10 minutes, and then washed with PBS to prepare a stratum corneum cell sample.

<Preparation of test samples and control samples>
Wild thyme extract (Wild Thyme Extract, manufactured by Ichimaru Falcos Co., Ltd., aboveground extract of Wild Thyme, T. serpyllum) was prepared at a concentration of 0.1% by mass, 0.05% by mass, and 0% by mass, respectively. A test sample was obtained by diluting the sample with PBS to a concentration of 0.025% by mass and 0.0125% by mass.

<Preparation of measurement sample>
Corner layer cells were immersed in the above-mentioned test sample overnight to obtain measurement samples of Examples 1 to 4.
In addition, as a reference example (control), stratum corneum cells that were not immersed in the above-mentioned test sample were prepared.

<Immunostaining test>
After washing the measurement sample with PBS, it was incubated in 4% PFA/phosphate buffer at room temperature for 15 minutes. After the incubation, the measurement sample was washed with PBS and incubated in a 20% Block Ace (manufactured by DS Biopharma Medical) aqueous solution at room temperature for 1 hour.
Thereafter, a primary antibody (Anti-Desmoglein 1, Mouse-Mono) was added and incubated for 2 hours at room temperature in a humid chamber.

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, images for analysis were taken of each measurement sample using a confocal laser microscope, and fluorescence (staining) was confirmed. The results are shown in Figure 1.

In addition, the fluorescent (stained) areas of the measurement samples in Examples 1 to 4 were calculated by taking the area of the fluorescent (stained) areas of the control as 100%. The results are shown in Table 1 and Figure 2. In Figure 2, * means p<0.05, and *** means p<0.001.

As shown in Figures 1 and 2, it was found that the expression level of desmoglein 1 was reduced in the measurement samples (Examples 1 to 4) to which wild thyme extract was added, compared to the sample (Reference Example 1) to which wild thyme extract was not added.
These results demonstrate that wild thyme extract has the effect of reducing the amount of desmoglein 1.

[Example 2] Relationship between skin brightness and/or skin hardness and the amount of desmoglein 1 The relationship between the difference in skin brightness and the amount of desmoglein 1, and the amount of desmoglein 1 present Test results supporting the relationship between skin hardness are shown.

(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 (the dark areas, see area A in the figure) and an area in a normal state (see area B in the figure) 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 (the dark areas, see area A in the figure) showed significantly lower values than the areas in a normal state (see area B in the figure). In other words, the areas where a decrease in brightness was visually confirmed (the dark areas, see area A in the figure) were confirmed to be darker than the areas in a normal state (see area B in the figure).

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

Regarding the b ^* value, the areas where it was confirmed by visual inspection that the brightness had decreased (the dark areas, see area A in the figure) showed significantly higher values than the areas in the normal state (see area B in the figure). In other words, it was confirmed that the areas where it was confirmed by visual inspection that the brightness had decreased (the dark areas, see area A in the figure) were more yellowish than the areas in the normal state (see area B in the figure).

(2-2) Hardness Measurement Hardness was measured by pressing an indentation meter against each measurement site subjected to the test in (2-1) above and performing the measurement five times.
The results are shown in FIG.

Regarding hardness, the areas where it was confirmed that the brightness had decreased by visual inspection (darkish area, see area A in the figure) were significantly lower than the areas in a normal state (see area B in the figure). The value was shown. In other words, the area where the brightness was visually confirmed to have decreased (darkish area, see area A in the figure) was confirmed to be harder than the area in a normal state (see area B in the figure). did it.

(2-3) Desmoglein 1 (Dsg1) staining The stratum corneum cells from each measurement site used in the tests (2-1) and (2-2) above were collected by tape stripping and pasted on a glass slide.

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

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, dried, and then (photographed) observed using a confocal laser microscope (Nikon A1) at ×20.
The results are shown in FIGS. 7 and 8.

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

(3) Discussion As shown in FIGS. 3 to 8, it was found that desmoglein 1 was present in large amounts in areas with low skin brightness. Here, the decrease in skin brightness is thought to be due to the dense overlapping of the stratum corneum (stratification of the stratum corneum) due to the low skin pH and low activity of serine protease that degrades desmoglein 1. Conceivable.

As mentioned above, the active ingredient of the present invention exhibits a desmoglein reducing effect.
In view of the above findings, it was found that the active ingredient of the present invention has an effect of improving skin brightness by reducing desmoglein 1.

As shown in Figures 3 to 8, 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 the effect of softening the skin by reducing desmoglein 1.

Example 3: Verification of the desmoglein-reducing effect of a composition containing wild thyme extract The desmoglein-reducing ability of a composition containing a component having a desmoglein-reducing effect was examined below.

<Preparation of Composition>
A cosmetic preparation (lotion) containing wild thyme extract was prepared according to the recipe shown in Table 2 below. That is, the ingredients (a), (b), and (c) were each heated to 70° C., (c) was added to (b) to neutralize it, (a) was gradually added while stirring to emulsify, the mixture was homogenized using a homogenizer, and then cooled while stirring to obtain the lotion of Production Example 1.

(2) 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 (92 areas) on the subject's face where blemishes were present were selected as measurement areas.
The emulsion of Preparation Example 1 was applied twice a day to the areas of the skin where blemishes existed on each subject, and this was continued for 12 weeks.

(3) Measurement Before using the emulsion of Production Example 1, after 6 weeks of continued use, and after 12 weeks, the emulsion was measured according to the method described in "<Preparation step of stratum corneum cell sample>" of Example 1. Corner layer cells were collected from the subject's site where the stain was present, and a corneum cell sample was prepared. Next, desmoglein 1 in each corneal cell sample was stained according to the method described in "<Immunostaining test>" in Example 1, and the results were stained before and after use in Production Example 1 using a confocal laser microscope. Images for analysis were taken and fluorescence (staining) was confirmed.

The area of the entire image for analysis was taken as 100%, and the area of the fluorescent portion was selected. The results are shown in Tables 3 and 4, and Figures 9 (representative), 10 and 11. Note that data after 12 weeks of using lotion was compiled from data for 80 out of 93 spots on the skin, and compared with data for the same spots before using lotion. In Figure 10, ** means p<0.01, and in Figure 11, *** means p<0.001.

As shown in Tables 3 and 4 and Figures 9 to 11, it was confirmed that the amount of desmoglein 1 in the blemished area was significantly reduced by continuously using a lotion containing wild thyme extract.

The present invention can be applied to cosmetics.

Claims (14)

A desmoglein reducing agent containing an extract of Wild Thyme (T. serpyllum) as an active ingredient. The desmoglein reducing agent according to claim 1, wherein the active ingredient is an extract of the aerial parts of Wild Thyme (T. serpyllum). The desmoglein reducing agent according to claim 1 or 2, for suppressing 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. The 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. The desmoglein reducing agent according to any one of claims 1 to 4, which is used to assist the penetration of medicinal ingredients into the skin. The desmoglein reducing agent according to any one of claims 1 to 4 for improving and/or preventing stratum corneum desquamation. The desmoglein reducer according to any one of claims 1 to 10, comprising as an active ingredient a composition containing an extract of Wild Thyme (T. serpyllum) and 1,3-butylene glycol. A composition for external application to the skin for reducing desmoglein, comprising the desmoglein reducing agent according to any one of claims 1 to 11. The composition for external application to the skin for reducing desmoglein according to claim 12, which is a cosmetic product. The composition for topical application to the skin for reducing desmoglein according to claim 12, which is a pharmaceutical product.