Desmoglein reducer

Abstract

To provide a novel desmoglein reducing agent.SOLUTION: Provided is a desmoglein reducing agent comprising salvianophosphate A as an active ingredient.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] The present invention relates to a desmoglein-reducing agent and an external skin preparation containing the desmoglein-reducing agent. [Background technology]

[0002] Increased desmosomal proteins are known to be 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).

[0003] Among desmosomal proteins, desmoglein in particular is known to be involved in adhesion between epidermal cells and keratinocytes (Patent Documents 2-4).

[0004] As a prior art, Patent Document 2 reports an agent for reducing desmoglein in stratum corneum cells that 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.

[0005] Incidentally, Patent Documents 5-7 report that extracts of Wild Thyme (Thymus serpyllum, hereinafter referred to as Wild Thyme) and extracts of Thymus plants of the Lamiaceae family have effects of improving moisture retention, anti-inflammatory effects, preventing skin aging, and protecting against UV rays. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Special Publication No. 7-505383 [Patent Document 2] Japanese Patent Application Laid-Open No. 2016-37501 [Patent Document 3] Patent No. 4002635 [Patent Document 4] Patent No. 4197194 [Patent Document 5] Japanese Patent Application Laid-Open No. 2001-122729 [Patent Document 6] Japanese Patent Application Laid-Open No. 2006-16337 [Patent Document 7] Japanese Patent Application Laid-Open No. 2009-256270 Summary of the Invention [Problem to be solved by the invention]

[0007] In view of the above-mentioned prior art, an object of the present invention is to provide a novel technique for reducing desmogleins.

[0008] As a result of extensive research, the present inventors have discovered that salvianolic acid A has a desmoglein-reducing effect, and have completed the present invention. [Means for solving the problem]

[0009] The present invention, which solves the above problems, is a desmoglein reducer containing salvianolic acid A as an active ingredient. In this specification, the concept of "decreasing desmoglein" includes both a decrease in desmoglein and suppression of an increase in desmoglein.

[0010] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used to inhibit intercellular adhesion of human epidermal keratinocytes.

[0011] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used to reduce desmoglein 1.

[0012] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used for preventing or improving sensitive skin.

[0013] In a preferred embodiment of the present invention, the desmoglein reducing agent is used to improve skin brightness.

[0014] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used to improve skin clarity.

[0015] In a preferred embodiment of the present invention, the desmoglein reducing agent is used for softening the skin.

[0016] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used to aid the penetration of medicinal ingredients into the skin.

[0017] In a preferred embodiment of the present invention, the desmoglein-reducing agent is used for improving and / or preventing stratum corneum desquamation.

[0018] The present invention, which solves the above problem, provides a composition containing the desmoglein reducer.

[0019] In a preferred embodiment of the invention, the composition is a cosmetic or pharmaceutical product. [Effects of the Invention]

[0020] According to the present invention, it is possible to reduce the amount of desmoglein present on human skin. [Brief explanation of the drawings]

[0021] [Figure 1] FIG. 1 shows the experimental results of a test to confirm the desmoglein-reducing ability (control, 80% ethanol extract of wild thyme, and 60% methanol eluate). [Figure 2] FIG. 1 shows the experimental results (isolated components 1 to 3) of a test to confirm the desmoglein-reducing ability. [Figure 3] FIG. 1 shows the experimental results of a test to confirm the desmoglein-reducing ability (isolated components 4 to 6, component 7). DETAILED DESCRIPTION OF THE INVENTION

[0022] (1) Salvianolic Acid A The desmoglein-reducing agent of the present invention contains salvianolic acid A as an active ingredient.

[0023] The structure of salvianolic acid A is shown in Chemical Formula 2.

[0024] [ka]

[0025] Salvianolic acid A may be isolated from a plant extract or synthesized, or commercially available salvianolic acid A may be used.

[0026] Salvianolic acid A can be isolated from extracts of Salvia miltiorrhiza or wild thyme. To 1 part by weight of the plant body or its dried product, 1 to 250 parts by weight of solvent is added, and the plant body or its dried product is immersed for several days at room temperature or for several hours at temperatures near the boiling point. After immersion, salvianolic acid A can be obtained by fractionation and purification using column chromatography packed with silica gel, synthetic adsorbents, octadecylsilylated silica gel, ion exchange resins, etc.

[0027] The extraction solvent is preferably a polar solvent, and suitable examples 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, preferred extraction solvents include ethanol and a mixed solvent of water and ethanol.

[0028] (2) Desmoglein reducer The desmoglein-reducing agent of the present invention preferably targets desmoglein 1 for reduction.

[0029] 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 type with reduced resistance to external stimuli (dryness, ultraviolet rays, application of chemical compounds) (reduced skin irritation threshold).

[0030] More specifically, "sensitive skin" in this specification refers to the skin types listed in (I) and / or (II).

[0031] (I) "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.)" (II) "Skin that is temporarily more susceptible to skin problems due to lack of sleep, overwork, menstruation, seasonal changes, mental stress, etc."

[0032] 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).

[0033] Furthermore, the desmoglein-reducing agent of the present invention is preferably used to inhibit intercellular adhesion of human epidermal keratinocytes. In this specification, the term "intercellular adhesion" refers to abnormalities in stratum corneum adhesion (progression of stratum corneum stratification due to increased cell adhesion) caused by an increase in desmosomal proteins (a group of proteins including desmoglein) (see Patent Nos. 4197194 and 4002635 for details). In addition, in this specification, the concept of "inhibiting intercellular adhesion" includes both prevention and improvement of intercellular adhesion.

[0034] Here, by inhibiting intercellular adhesion of human epidermal keratinocytes, stratification of the stratum corneum is inhibited, and the effect of inhibiting stratum corneum desquamation can be obtained. That is, the desmoglein-reducing agent of the present invention can be used to prevent or improve stratum corneum desquamation. As used herein, "stratum corneum delamination" refers to the peeling off of multiple layers of the stratum corneum from the skin surface. Whether or not the skin is in a state where stratum corneum delamination is possible can be confirmed by collecting a sample of the stratum corneum by tape stripping, staining the collected sample, and observing it.

[0035] Furthermore, areas of the skin with low brightness have a high concentration of desmoglein 1. Here, the decrease in skin brightness is thought to be due to stratification of the stratum corneum. In view of the above findings, the active ingredient of the present invention can reduce desmoglein, thereby 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 decreased due to the excessive presence of desmoglein.

[0036] Furthermore, the desmoglein-reducing agent of the present invention can provide a skin condition with a more transparent appearance. That is, the active ingredient of the present invention can also be used as an agent for improving skin transparency.

[0037] Furthermore, areas with high skin hardness have a high concentration 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 providing a skin softening effect, and can also be used as a skin softener.

[0038] Here, softening the skin allows other medicinal ingredients to penetrate more efficiently, so 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 aid for the penetration of medicinal ingredients into the skin.

[0039] In particular, the active ingredient of the present invention is preferably used as a whitening ingredient penetration aid for use in combination with a whitening agent.

[0040] (3) Composition The present invention also relates to a composition containing the aforementioned desmoglein-reducing agent. Among these, it is preferable to use the composition in the form of a composition for external use on the skin or an oral composition.

[0041] Suitable examples of external skin compositions include cosmetics and pharmaceuticals.

[0042] Among these, it is preferable to use the cosmetic in the form of a cosmetic that can be used continuously, such as a lotion, emulsion, serum, cream, gel, sun care product, etc.

[0043] The concentration of salvianolic acid A in the total volume of the topical skin composition is preferably 3 μg / μL or more, more preferably 5 μg / μL or more, even more preferably 8 μg / μL or more, and particularly preferably 10 μg / μL or more.

[0044] When an oral composition is prepared, it is preferable to prepare a food composition containing the active ingredient of the present invention. Specifically, it can be in the form of a supplement having the dosage form of a general food, tablet, granule, drink, or the like.

[0045] The concentration of salvianolic acid A in the entire oral composition is preferably 3 μg / μL or more, more preferably 5 μg / μL or more, even more preferably 8 μg / μL or more, and particularly preferably 10 μg / μL or more. [Example]

[0046] [Test Example 1] Purification of components in 80% ethanol extract of wild thyme The wild thyme raw material (1.0 kg) was soaked in 80% ethanol to prepare an 80% ethanol extract of wild thyme. The prepared 80% ethanol extract of wild thyme was passed through a Diaion HP20 column to obtain an unadsorbed / water eluted fraction, a 60% methanol eluted fraction, a methanol eluted fraction, and an acetone eluted fraction. From the 60% eluate, various chromatographic procedures were used to isolate luteolin 7-O-glucuronide (isolated component 1, 87 mg), apigenin 7-O-glucuronide (isolated component 2, 278 mg), erythrichin (isolated component 3, 44 mg), salvianolic acid A (isolated component 4, 13 mg), luteolin 5-O-glucoside (isolated component 5, 607 mg), and diosmetin 5-O-glucoside (isolated component 6, 124 mg).

[0047] [Test Example 2] Test to confirm the desmoglein-reducing ability of wild thyme fractions <Preparation of test samples and control samples> The test samples were an 80% ethanol extract prepared from wild thyme raw material, a 60% methanol eluate, six isolated components, and rosmarinic acid (Wako Pure Chemical Industries, Ltd., component 7).

[0048] The 80% ethanol extract and 60% methanol eluate of wild thyme were adjusted to a concentration of 6 μg / μL with DMSO. 100 μL of the prepared solution (final concentration: 30 μg / mL) was added to 20 mL of PBS dispensed into a 50 mL tube and then vortexed to obtain the test samples. Isolated components 1 to 6 and component 7 were prepared in DMSO to a concentration of 2 μg / μL. 10 μL (final concentration 1 μg / mL, low-concentration sample) or 100 μL (final concentration 10 μg / mL, high-concentration sample) of the prepared solution was added to 20 mL of PBS dispensed into a 50 mL tube and stirred with a vortex mixer. For the low concentration sample, 90 μL of DMSO was added, and this was used as the test sample. In addition, 100 μL of DMSO (final concentration: 0.5%) was prepared as a control sample.

[0049] <Preparation process of stratum corneum cell samples> The fourth layer of the stratum corneum was collected from the outer surface of a human forearm using tape stripping. The cellophane tape containing the stratum corneum was cut into approximately 4mm squares and attached to a glass slide. The slide was immersed in xylene overnight, then immersed in xylene for 10 minutes twice, and air-dried until all water droplets had disappeared.

[0050] <Immunostaining test> After air drying, the slides were immersed in PBS containing a control sample or PBS containing a test sample for 24 hours, after which they were rinsed with PBS and immersed in 4% PFA for 15 minutes to fix the keratinocytes. The slides were then rinsed with PBS and permeabilized by immersion in 0.5% Triton X-100 / PBS for 5 minutes at room temperature. After rinsing with PBS, they were blocked with 20% Block Ace (KAC) for 1 hour at room temperature. Subsequently, they were incubated with a primary antibody (anti-Desmoglein 1 mouse monoclonal, DSG1-P23; PROGEN Biotechnik GmbH) for 2 hours at room temperature.

[0051] After immersion in PBS for 5 minutes three times, the sections were incubated with a secondary antibody (Goat anti-Mouse IgG (H+L) Cross-Adsorbed Secondary Antibody, Alexa Fluor 488; Invitrogen) for 1 hour at room temperature in the dark. After immersion in PBS for 5 minutes three times, the sections were rinsed twice with distilled water and mounted.

[0052] Then, four analytical images were taken of each sample using an all-in-one fluorescence microscope BZ-X810 (KEYENCE). The area of ​​the fluorescent portion (stained desmoglein 1) was calculated for each of the four analytical images, with the area of ​​the entire analysis region taken as 100%, and the average area of ​​the fluorescent portion was calculated. The results are shown in Figures 1, 2, and 3.

[0053] The rate of decrease in desmoglein 1 (Dsg-1 decrease rate) for each test sample relative to the control sample was calculated using the following formula. The results are shown in Table 1.

[0054] (Calculation formula for the rate of decrease in desmoglein 1) Desmoglein 1 reduction rate (%) = (1-A / B) x 100 A: Amount of Dsg-1 (%) per area of ​​the entire image for analysis when the test sample was added B: Dsg-1 amount (%) per area of ​​the entire image for analysis of the control sample

[0055] [Table 1]

[0056] The 80% ethanol extract of wild thyme, the 60% methanol extract of wild thyme, and isolated component 3 (erythrixine) and isolated component 4 (salvianolic acid A) were found to have a significant Dsg-1 reducing effect. [Industrial Applicability]

[0057] The present invention can be applied to cosmetics and pharmaceuticals that improve various skin conditions.

Claims

1. A desmoglein reducer containing salvianolic acid A as the active ingredient.

2. The desmoglein-reducing agent according to claim 1, for inhibiting intercellular adhesion of human epidermal keratinocytes.

3. The desmoglein-reducing agent according to claim 1 or 2, wherein the target to be reduced is desmoglein 1.

4. A desmoglein-reducing agent according to any one of claims 1 to 3 for improving and / or preventing sensitive skin.

5. The desmoglein-reducing agent according to any one of claims 1 to 3 for improving skin brightness.

6. A desmoglein-reducing agent according to any one of claims 1 to 3 for improving skin clarity.

7. A desmoglein-reducing agent according to any one of claims 1 to 3 for softening the skin.

8. Desmoglein according to any one of claims 1 to 3 for aiding penetration of medicinal ingredients into the skin. Reducer.

9. A desmoglein-reducing agent according to any one of claims 1 to 3 for improving and / or preventing stratum corneum desquamation.

10. A composition for reducing desmoglein, which comprises the desmoglein reducer according to any one of claims 1 to 3 and contains salvianolic acid A as an active ingredient, and which is intended to improve stratum corneum stratification, loss of skin brightness, and / or increased skin hardness caused by an increase in desmoglein.

11. The desmoglein-reducing composition according to claim 10, which is a cosmetic or pharmaceutical product.

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