KR102385250B1 - Novel amide compounds and composition for strengthening skin barrier comprising the same - Google Patents

Abstract

The present invention relates to a novel amide compound and a composition for strengthening the skin barrier comprising the same. The amide compound provided in one aspect of the present invention excellently enhances the expression of TJ-associated proteins which play an important role in strengthening the skin barrier and thus can be usefully used as a composition for strengthening the skin barrier for alleviating wrinkles, preventing skin aging, alleviating atopy, moisturizing skin, and the like and as an external preparation for the skin.

Description

A novel amide compound and a composition for strengthening skin barrier comprising the same {Novel amide compounds and composition for strengthening skin barrier comprising the same}

The present invention relates to a novel amide compound and a composition for strengthening skin barrier comprising the same.

The skin performs various functions essential for the survival of the human body. The barrier function to maintain homeostasis inside the human body in response to environmental changes, the sensory function to recognize external changes, and the body temperature control function are among the most representative functions of the skin. It plays a role in maintaining homeostasis. Among various functions of the skin, in particular, the barrier function of the skin is mainly expressed by the epidermis located at the outermost layer of the skin. The epidermis maintains epidermal homeostasis by constantly repeating the processes of epidermal cell formation, differentiation, and exfoliation.

It is the tight junction (Tj) of cells that plays an important function for strengthening the barrier function of the epidermis. Close junction protects internal organs from the external environment and plays a beneficial role in maintaining homeostasis of the human body. In such tight junctions, TJ-associated proteins such as occludin (OC), claudin, and junctional adhesion molecules play an important role. They use zona occludens-1 and cingulin, which are plaque proteins present in the cytoplasm, as mediators to form the actin-cytoskeleton and cytosolic regulatory proteins. proteins), and has a biological barrier function that regulates adhesion between cells and the movement of water-containing solute and water through the pericellular space.

Because the skin is always exposed to the external environment, this skin barrier function cannot be free from physical and chemical damage such as chronic ultraviolet rays, heat, and irritating coating agents. Even long-term intake of foods deficient in essential fatty acids can weaken or impair the barrier function (Kim Do-won, The Journal of Skin Barrier Research, 2006). In addition, even when inflammation or an autoimmune reaction occurs in the skin, the skin barrier is damaged and the homeostasis of the epidermis cannot be maintained. As described above, the causes of damage to the skin barrier are very diverse in addition to inflammation or autoimmunity. Accordingly, the present inventors have developed the present invention by identifying compounds that are effective in wrinkle improvement, skin aging prevention, atopy improvement, skin moisturizing, etc. by excellently increasing the expression of close junction-associated proteins while developing a skin barrier strengthening agent.

One object of the present invention is to provide a novel amide compound.

Another object of the present invention is to provide a composition for strengthening the skin barrier comprising an amide compound having an effect of increasing the expression of a cell adhesion-related protein.

Another object of the present invention is to provide an external preparation for skin barrier strengthening comprising the composition for skin barrier strengthening.

In order to achieve the above object,

One aspect of the present invention provides a compound represented by the following Chemical Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _{1-10 alkyl} , C _{2-10 alkenyl} , or C _2-10 alkynyl;

R ³ is substituted or unsubstituted C _1-10 alkoxy;

L is substituted or unsubstituted C _1-5 alkylene; and

_{The substituted} C _{1-10 alkyl} , C _2-10 alkenyl, C _2-10 alkynyl, C _1-10 alkoxy, _{and C 1-5 alkylene} are each independently the group consisting of C _{1-5 alkyl and} halogen. One or more substituents selected from is substituted.

Another aspect of the present invention provides a composition for strengthening the skin barrier comprising a compound represented by Formula 1 below, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In Formula 1,

R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _1-10 alkyl, C _2-10 alkenyl, or C _2-10 alkynyl;

R ³ is substituted or unsubstituted C _1-10 alkoxy;

L is substituted or unsubstituted C _1-5 alkylene; and

The substituted C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl, C _1-10 alkoxy, and C _1-5 alkylene are each independently the group consisting of C _1-5 alkyl and halogen. One or more substituents selected from is substituted.

In addition, another aspect of the present invention provides an external preparation for skin barrier strengthening comprising the composition for strengthening the skin barrier.

The amide compound provided in one aspect of the present invention excellently improves the expression of TJ-associated proteins, which play an important role in strengthening the skin barrier, thus improving wrinkles, preventing skin aging, improving atopy, improving skin It can be usefully used as a composition for strengthening the skin barrier for moisturizing, etc. and as an external preparation for skin.

1 is a claudin-1 mRNA expression when UV-irradiated Human Epidermal Keratinocytes were treated with Example 1, Comparative Example 1, and Comparative Examples 3 to 6 according to the present invention; It is a diagram showing the result of analyzing the effect.
2 is a result of analyzing the effect of claudin-4 mRNA expression when UV-irradiated human keratinocytes were treated with Example 1, Comparative Example 1, and Comparative Examples 3 to 6 according to the present invention; is a diagram showing
3 is a view showing the results of analyzing the occludin mRNA expression effect when UV-irradiated human keratinocytes were treated with Example 1, Comparative Example 1, and Comparative Examples 3 to 6 according to the present invention; am.
4 and 5 show the effect of claudin-1 (claudin-1) expression when UV-irradiated artificial skin is treated with Comparative Examples 2, 3 and 1 according to the present invention with a fluorescence microscope image ( 4), a view showing the result of quantitative analysis (FIG. 5).
6 and 7 show the effect of claudin-4 expression when treated with UV-irradiated artificial skin according to the present invention in Comparative Examples 2, 3, and 1 with a fluorescence microscope image ( 6) is a diagram showing the result of quantitative analysis (FIG. 7).
8 and 9 show the effect of occludin expression when UV-irradiated artificial skin was treated with Comparative Examples 2, 3, and 1 according to the present invention by a fluorescence microscope image (FIG. 8). , a diagram showing the result of quantitative analysis (FIG. 9).
10 and 11 is a fluorescence microscope image of the filaggrin expression effect when Comparative Example 2, Comparative Example 3, and Example 1 according to the present invention were treated on artificial skin irradiated with ultraviolet light (FIG. 10) , a diagram showing the result of quantitative analysis (FIG. 11).

Hereinafter, the present invention will be described in detail.

On the other hand, the embodiment of the present invention can be modified in various other forms, the scope of the present invention is not limited to the embodiments described below. In addition, the embodiments of the present invention are provided in order to more completely explain the present invention to those of ordinary skill in the art. Furthermore, in the entire specification, "including" a certain element means that other elements may be further included, rather than excluding other elements, unless otherwise stated.

One aspect of the present invention provides a compound represented by the following Chemical Formula 1, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.

[Formula 1]

In Formula 1,

R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _{1-10 alkyl} , C _{2-10 alkenyl} , or C _2-10 alkynyl;

R ³ is substituted or unsubstituted C _1-10 alkoxy;

L is substituted or unsubstituted C _1-5 alkylene; and

_{The substituted} C _{1-10 alkyl} , C _2-10 alkenyl, C _2-10 alkynyl, C _1-10 alkoxy, _{and C 1-5 alkylene} are each independently the group consisting of C _{1-5 alkyl and} halogen. One or more substituents selected from is substituted.

In this case, R ¹ and R ² are each independently substituted or unsubstituted C _{1-8 alkyl} , C _{2-8 alkenyl} , or C _2-8 alkynyl;

wherein R ³ is substituted or unsubstituted C _1-8 alkoxy;

wherein R ⁴ is substituted or unsubstituted C _1-8 alkyl;

L is substituted or unsubstituted C _1-3 alkylene; and

_{The substituted C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 1-8 alkoxy , and C 1-3 alkylene are each} independently the group consisting of _{C 1-3 alkyl and} halogen. One or more substituents selected from may be substituted.

In addition, R ¹ and R ² are each independently _substituted or unsubstituted C _{1-6 alkyl} , C _{2-6 alkenyl} , or C _2-6 alkynyl;

wherein R ³ is substituted or unsubstituted _{C 1-3 alkoxy} ;

wherein R ⁴ is substituted or unsubstituted C _{1-5 alkyl} ;

L is substituted or unsubstituted _{C 1-2} alkylene; and

The _substituted C _{1-6 alkyl} , C _{2-6 alkenyl} , C _2-6 alkynyl, _{C 1-3 alkoxy} , C _{1-5 alkyl} , _and C _1-2 alkylene are each independently _{C 1-3} One or more substituents selected from the group consisting of alkyl and halogen may be substituted.

_In addition, R ¹ and R ² are each independently substituted or unsubstituted C _3-8 alkyl, C _{3-8 alkenyl, or C 3-8 alkynyl ;}

wherein R ³ is substituted or unsubstituted _{C 1-3 alkoxy} ;

wherein R ⁴ is substituted or unsubstituted _{C 1-3 alkyl} ;

L is substituted or unsubstituted _{C 1-2} alkylene; and

wherein the substituted C _{3-8 alkyl} , C _{3-8 alkenyl} , C _{3-8 alkynyl} , _{C 1-3 alkoxy} , C _{1-3 alkyl, and C 1-2 alkylene are} each independently _{C 1-3} One or more substituents selected from the group consisting of alkyl and halogen may be substituted.

Alternatively, R ¹ and R ² may be C ₆ alkyl or —(CH ₂ ) ₅ CH ₃ , R ³ may be —OCH ₃ , R ⁴ may be —CH ₃ , and L is — CH ₂ - may be.

Specifically, the compound represented by Formula 1 may be a compound represented by Formula 1-1 below.

[Formula 1-1]

The compound represented by Formula 1 of the present invention may be used in the form of a pharmaceutically acceptable salt, and as the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid, phosphorous acid, etc., aliphatic mono and dicarboxylates, phenyl-substituted alkanoates, hydroxy alkanoates and alkanes. Non-toxic organic acids such as dioates, aromatic acids, aliphatic and aromatic sulfonic acids, etc., organic acids such as trifluoroacetic acid, acetate, benzoic acid, citric acid, lactic acid, maleic acid, gluconic acid, methanesulfonic acid, 4-toluenesulfonic acid, tartaric acid, fumaric acid, etc. get it from Examples of such pharmaceutically non-toxic salts include sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate chloride, bromide, and Odide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, sube Late, sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycolate, maleate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, mandelate and the like.

The acid addition salt according to the present invention can be prepared by a conventional method, for example, a precipitate formed by dissolving the derivative of Formula 1 in an organic solvent such as methanol, ethanol, acetone, methylene chloride, acetonitrile, etc. and adding an organic or inorganic acid It can be prepared by filtration and drying, or by distilling the solvent and excess acid under reduced pressure, followed by drying and crystallization in an organic solvent.

In addition, a pharmaceutically acceptable metal salt can be prepared using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, it is pharmaceutically suitable to prepare a sodium, potassium or calcium salt as the metal salt. The corresponding salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable negative salt (eg silver nitrate).

Furthermore, the present invention includes not only the compound represented by Formula 1 and pharmaceutically acceptable salts thereof, but also solvates, optical isomers, hydrates, and the like, which can be prepared therefrom.

Another aspect of the present invention provides a composition for strengthening the skin barrier comprising a compound represented by Formula 1 below, a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient.

[Formula 1]

In Formula 1,

R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _1-10 alkyl, C _2-10 alkenyl, or C _2-10 alkynyl;

R ³ is substituted or unsubstituted C _{1-10 alkoxy} ;

L is substituted or unsubstituted C _{1-5 alkylene} ; and

_{The substituted} C _{1-10 alkyl} , C _2-10 alkenyl, C _2-10 alkynyl, C _1-10 alkoxy, _{and C 1-5 alkylene} are each independently the group consisting of C _{1-5 alkyl and} halogen. One or more substituents selected from is substituted.

In this case, R ¹ and R ² are each independently _substituted or unsubstituted C _{1-8 alkyl} , C _{2-8 alkenyl} , or C _2-8 alkynyl;

wherein R ³ is substituted or unsubstituted C _{1-8 alkoxy} ;

wherein R ⁴ is substituted or unsubstituted C _{1-8 alkyl} ;

L is substituted or unsubstituted _{C 1-3 alkylene} ; and

_{The substituted C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, C 1-8 alkoxy , and C 1-3 alkylene are each} independently the group consisting of _{C 1-3 alkyl and} halogen. One or more substituents selected from may be substituted.

In addition, R ¹ and R ² are each independently _substituted or unsubstituted C _{1-6 alkyl} , C _{2-6 alkenyl} , or C _2-6 alkynyl;

wherein R ³ is substituted or unsubstituted _{C 1-3 alkoxy} ;

wherein R ⁴ is substituted or unsubstituted C _{1-5 alkyl} ;

L is substituted or unsubstituted _{C 1-2} alkylene; and

The _substituted C _{1-6 alkyl} , C _{2-6 alkenyl} , C _2-6 alkynyl, _{C 1-3 alkoxy} , C _{1-5 alkyl} , _and C _1-2 alkylene are each independently _{C 1-3} One or more substituents selected from the group consisting of alkyl and halogen may be substituted.

_In addition, R ¹ and R ² are each independently substituted or unsubstituted C _3-8 alkyl, C _{3-8 alkenyl, or C 3-8 alkynyl ;}

wherein R ³ is substituted or unsubstituted _{C 1-3 alkoxy} ;

wherein R ⁴ is substituted or unsubstituted _{C 1-3 alkyl} ;

L is substituted or unsubstituted _{C 1-2} alkylene; and

wherein the substituted C _{3-8 alkyl} , C _{3-8 alkenyl} , C _{3-8 alkynyl} , _{C 1-3 alkoxy} , C _{1-3 alkyl, and C 1-2 alkylene are} each independently _{C 1-3} One or more substituents selected from the group consisting of alkyl and halogen may be substituted.

Alternatively, R ¹ and R ² may be C ₆ alkyl or —(CH ₂ ) ₅ CH ₃ , R ³ may be —OCH ₃ , R ⁴ may be —CH ₃ , and L is — CH ₂ - may be.

Specifically, the compound represented by Formula 1 may be a compound represented by Formula 1-1 below.

[Formula 1-1]

In addition, the compound can strengthen the skin barrier by strengthening the tight junction between cells, which is important for strengthening the skin barrier. The compound may serve to protect internal organs from the external environment and maintain homeostasis of the human body by strengthening the close junction. The compound has an effect of enhancing the expression of TJ-associated proteins that play an important role in tight junctions, and examples of the tight junction-associated proteins include claudin-1, claudin- It may be to improve the expression of 4 (claudin-4), occludin, or filaggrin. By strengthening the skin barrier through this effect, the compound may be usefully applied as a composition for strengthening the skin barrier for the purpose of improving wrinkles, preventing skin aging, improving atopy, and moisturizing the skin.

The compound may be included as an active ingredient in the composition for strengthening the skin barrier, and based on the total weight of the composition, 0.001 wt% to 70 wt%, 0.001 wt% to 50 wt%, 0.1 wt% to 30 wt%, or 0.01 wt% to It may be included in an amount of 20% by weight, etc., and is not necessarily limited to the above weight range.

Another aspect of the present invention provides an external preparation for skin barrier strengthening comprising the composition for skin barrier strengthening.

In this case, the external preparation may be formulated to further include an appropriate additive commonly used. In this case, non-limiting examples of the additive include a stabilizer, an emulsifier, a thickener, a moisturizer, a liquid crystal film strengthening agent, a pH adjuster, an antibacterial agent, a water-soluble polymer, a film agent, a metal ion sequestrant, an amino acid, an organic amine, a polymer emulsion, a pH adjuster , one or more aqueous additives selected from skin nutrients, antioxidants, antioxidants, preservatives, fragrances, and the like; and at least one oily additive selected from oils and fats, waxes, higher fatty acids and higher alcohols.

At this time, the aqueous additive is not limited as long as it is a raw material generally used in the art, and non-limiting examples thereof include glycerin, dipropylene glycol, butylene glycol, pentylene glycol, methylpropanediol, sorbitol, Diglycerin, erythritol, pentaerythritol, polybutylene glycol-10, polyglycerin-3, polyglycerin-4, polyglycerol-6, polyglycerin-10, polyglycerol-20, polyglycerin-40, sorbet-5 , Sorbeth-6, Sorbeth-20, Sorbeth-30, Sorbeth-40, Inositol, Maltitol, Maltose, Mannan, Mannitol, Mannose, Lactitol, Lactose, Dihydroxypropyl PG-Glucoside, Dithioctane Diol, fructose, glucamine, methylglucamine, glucose, 1,2,6-hexanethiol, methylgluceth-10, methylglucose-20, ozonized glycerin, phytantriol, thioglycerin, threitol , trimethylolpropane, xylitol, EDTA, guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxy Methyl Cellulose, Methyl Hydroxypropyl Cellulose, Chondroitin Sulfate, Dermatan Sulfate, Glycogen, Heparan Sulfate, Hyaluronic Acid, Sodium Hyaluronate, Gum Tragant, Keratan Sulfate, Chondroitin, Mucoitin Sulfate, Hydroxyethyl Guar Gum, Carboxylic Acid Methyl guar gum, dextran, kerat sulfate, locust bean gum, succinoglucan, caronic acid, chitin, chitosan, carboxymethyl chitin, agar, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, sodium polyacrylate, It may be at least one selected from polyethylene glycol, bentonite, methylparaben, propylparaben, phenoxyethanol, 1,2-hexanediol, ethylhexylglycerin, and the like. In addition, the oily additive is not limited as long as it is a raw material generally used in the art, and non-limiting examples thereof include liquid fats and oils such as triglyceride, glycerol trioctanoate, glycerin triisopalmitate, and palm oil, hydrogenated palm oil, and palm oil. , hydrogenated oil, solid oils such as hydrogenated castor oil, beeswax, candelilla wax, carnauba wax, lanolin, jojoba wax, and the like. Examples of higher fatty acids include waxes such as lauric acid, myristic acid, palmitic acid, stearic acid, and behenic acid, and higher alcohols such as cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, and cetostearyl alcohol. and the like can be exemplified. In this case, the additive may be included in an amount of 0.001 wt% to 20 wt%, specifically 0.01 wt% to 10 wt%, or 0.05 wt% to 10 wt%, based on the total weight of the external preparation, but is not limited thereto.

The external preparation is a cosmetic, and may be formulated in a formulation such as softening lotion, astringent lotion, nutrient lotion, eye cream, nutrient cream, massage cream, cleansing cream, cleansing foam, cleansing water, powder, essence, or pack.

Hereinafter, Examples and Experimental Examples of the present invention will be specifically illustrated and described below. However, the Examples and Experimental Examples to be described below are merely illustrative of a part of the present invention, and the present invention is not limited thereto.

< Example 1> Preparation of the compound of Formula 1-1

[Scheme 1]

Step 1: Preparation of 4-heptylidene-3-hexyloxetan-2-one (octylketenedimer)

In a 250 mL reactor under a nitrogen atmosphere, 90 mL of ethyl acetate was added, and 13 mL (92.89 mmol) of TEA was added. After cooling the reactor temperature to 0° C. or lower using an ice bath, 15 g (92.22 mmol) of octanoyl chloride was slowly added dropwise. After completion of the dropwise addition, the ice bath was removed, the temperature of the reactor was slowly raised to room temperature, and the mixture was stirred overnight. After adding purified water to the reaction mixture, c-HCl was added to adjust the pH to 3, purified water was further added, stirred, and then the organic layer was separated. The organic layer was washed once more with brine, dried over MgSO ₄ , filtered, and the filtrate was concentrated under reduced pressure to form a pale yellow liquid, 4-heptylidene-3-hexyloxetan-2-one (4-heptylidene- 3-hexyloxetan-2-one) (Octyketendimer) was obtained.

Step 2: Preparation of dimethyl (2-hexyl-3-oxodecanoyl) aspartate

Into a 250 mL reactor, 50 mL of methanol was added, 9.1 g (46.11 mmol) of dimethyl aspartate hydrochloride was added, and then 5.1 g (50.72 mmol) of TEA was added. After the reaction was stirred for 10 minutes, 11.6 g (46.11 mmol) of 4-heptylidene-3-hexyloxetan-2-one was slowly added and stirred overnight. The reaction was confirmed by TLC, and the solvent was removed by concentration under reduced pressure. Ethyl acetate and purified water were added to the concentrate, and the pH was adjusted to 3 by adding c-HCl. After stirring and standing, the organic layer was separated. The organic layer was dried over MgSO ₄ , and the filtered filtrate was concentrated under reduced pressure to completely remove the solvent. The residue was separated by column chromatography (dichloromethane:methanol = 50:1) and dimethyl (2-hexyl-3-oxodecanoyl) as a white solid as dimethyl (2-hexyl-3-oxodecanoyl) aspartate) (7.2 g, 17.44 mmol, yield 38%).

MS (ESI pos. ion) m/z: 414 (MH+). Calc'd exact mass for C ₂₂ H ₃₉ NO ₆ : 413.56

¹ H NMR (400 MHz, CDCl ₃ ): 7.18 - 7.06 (m, 1H), 4.87 - 4.83 (m, 1H), 3.75 (s, 3H), 3.70 (s, 3H), 3.40 - 3.36 (m, 1H) ), 3.04 - 2.98 (m, 1H), 2.87 - 2.81 (m, 1H), 2.54 - 2.50 (m, 2H), 1.83 - 1.80 (m, 2H), 1.57 - 1.55 (m, 2H), 1.32 - 1.18 (m, 16H), 0.89 - 0.85 (m, 6H).

< comparative example 1>

Ascorbic acid, which is known to have a skin barrier strengthening effect, was prepared.

< comparative example 2>

Retinoic acid, which is known to have a skin barrier strengthening effect, was prepared.

< comparative example 3 to comparative example 6>

For the analysis of the effect of strengthening the skin barrier with Example Compound 1, amide compounds of Comparative Examples 3 to 6 were prepared as shown in Table 1 below.

comparative example
No. chemical formula comparative example
No. chemical formula 3

4

5

6

< Experimental example 1> Close connection- Associated protein mRNA Expression analysis

In order to analyze the mRNA expression effect of the tight junction-associated protein, which plays an important role in the tight junction of the compound according to Example 1, an experiment was conducted as follows.

First, human Epidermal Keratinocytes (purchased from Thermo Fisher Scientific) were cultured in 150 mm cell culture dishes every other day using EpiLife culture medium containing Human Keratinocyte Growth Supplement (HKGS). After culturing in a 37 °C 5% CO ₂ incubator while seeding at a seed density of 1 x 10 ⁶ cells/well, the number of human keratinocytes was 1.0 x 10 ⁵ cells/well in a 6-well plate. Passage as much as 37 ° C. 5% CO ₂ Incubated overnight in an incubator. After harvesting the culture solution of cultured human keratinocytes, in order to stimulate the cells, 1 mL of DPBS per well was treated with UV-A 20 mJ/cm ² , and after UV-A irradiation was completed, DPBS was removed. Next, Example 1, Comparative Example 1, and Comparative Examples 3 to 6 at a concentration of 50 μM were treated for 24 hours using EpiLife culture medium not containing HKGS, at this time, only the culture medium without HKGS was treated as a control. A control 1 and a control 2 treated with a culture medium without HKGS after UV-A 20 mJ/cm ² irradiation were prepared. Then, cells were harvested, total RNA was isolated, qPCR was performed, and gene amplification values were confirmed using claudin-1, claudin-4, and ocludin primers, which are proteins constituting the tight junction. The results are shown in FIGS. 1 to 3 .

As a result, as can be seen in FIGS. 1 to 3 , by irradiating the medium treated with only the culture medium with ultraviolet light, the mRNA expression of claudin-1, claudin-4, and ocludin, which are closely junction-associated proteins, was decreased, but in Example 1 It was possible to very well increase gene expression by treating the compound according to Specifically, in the case of claudin-1 mRNA, when the compound of Example 1 was treated, the expression was significantly increased by about 5-fold or more, and in the case of claudin-4 and ocludin mRNA, Comparative Examples 3 to 6 were treated Edo was able to increase the gene expression level compared to ascorbic acid, but when Example 1 was treated, the expression level was most significantly increased by about 2.5 times or more and about 7 times, respectively.

< Experimental example 2> Close junction in artificial skin- Associated protein Expression analysis

In order to analyze the mRNA expression effect of the tight junction-associated protein, which plays an important role in the close junction of the compound according to Example 1, in artificial skin, an experiment was conducted as follows.

First, artificial skin (place of purchase: EpiDermFT from MatTek, Ashland, USA) was stabilized for 18 hours in a dedicated medium (EFT-400), and then UV-A 500 mJ/cm ² was irradiated to stimulate the skin. At this time, as controls, Control 1, which is an untreated medium, and Control 2 treated only with UV-A 500 mJ/cm ² were prepared. Then, 30 μL of Example 1, Comparative Example 2, and Comparative Example 3 were applied and cultured for 48 hours. Then, artificial skin tissue was fixed with 10% formaldehyde and embedded with paraffin, cut into 5 μm, and then prepared, Control 1 And after staining the skin tissues treated with Control 2 and Example 1, Comparative Example 2 and Comparative Example 3 using IHC (ImmunoHistoChemistry) staining method, it was observed with a fluorescent microscopic image, and the microscopic image was obtained using the image J program was quantified and quantitatively analyzed, and changes in the activity of claudin-1, claudin-4, and ocludin, which are closely junction-associated proteins, and filaggrin, a protein constituting the skin barrier, were observed. The results are shown in FIGS. 4 to 11 .

As a result, as can be seen in FIGS. 4 to 11, the close junction-associated proteins claudin-1, claudin-4, and occludin, and filaggrin, a protein constituting the skin barrier, are expressed by irradiating the untreated medium with ultraviolet rays. Although this was significantly reduced, the expression of the protein was excellently increased by treating the compound according to Example 1. Specifically, even when Comparative Example 2 was treated, the expression of each protein could be increased, but only slightly increased, whereas in the case of Example 1, Claudine-1 was restored to a level of about 50% of the expression level. , Claudin-4 and Ocludin recovered to about 80% and 60% or more, respectively, and, in particular, filaggrin showed more protein expression effect than the amount before UV irradiation.

Therefore, by using the compound according to Example 1 of the present invention to increase the expression of a closely junction-associated protein and increase the expression of a protein constituting the skin barrier, damage to the skin barrier can be improved or the skin barrier can be strengthened, By using this, it can be usefully applied for various purposes related to skin barrier strengthening.

As mentioned above, although the present invention has been described in detail through preferred examples and experimental examples, the scope of the present invention is not limited to specific examples, and should be construed according to the appended claims. In addition, those skilled in the art should understand that many modifications and variations are possible without departing from the scope of the present invention.

Claims (11)

A compound represented by the following formula (1), a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1]

In Formula 1,
R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _1-10 alkyl;
R ³ is substituted or unsubstituted C _1-10 alkoxy;
L is substituted or unsubstituted C _1-5 alkylene; and
The substituted C _1-10 alkyl, C _1-10 alkoxy, and C _1-5 alkylene are each independently substituted with one or more substituents selected from the group consisting of C _1-5 alkyl and halogen.
According to claim 1,
wherein R ¹ and R ² are each independently substituted or unsubstituted C _1-8 alkyl;
wherein R ³ is substituted or unsubstituted C _1-8 alkoxy;
wherein R ⁴ is substituted or unsubstituted C _1-8 alkyl;
L is substituted or unsubstituted C _1-3 alkylene; and
The substituted C _1-8 alkyl, C _1-8 alkoxy, and C _1-3 alkylene are each independently substituted with one or more substituents selected from the group consisting of C _1-3 alkyl and halogen. , a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof.
The method of claim 1,
wherein R ¹ and R ² are each independently substituted or unsubstituted C _1-6 alkyl;
wherein R ³ is substituted or unsubstituted C _1-3 alkoxy;
wherein R ⁴ is substituted or unsubstituted C _1-5 alkyl; and
Wherein L is a substituted or unsubstituted C _1-2 alkylene, a compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof.
According to claim 1,
The compound represented by Formula 1 is a compound represented by Formula 1-1 below, a compound, a stereoisomer, a hydrate thereof, or a pharmaceutically acceptable salt thereof:
[Formula 1-1]

.
A composition for strengthening the skin barrier comprising a compound represented by the following formula (1), a stereoisomer thereof, a hydrate thereof, or a pharmaceutically acceptable salt thereof as an active ingredient:
[Formula 1]

In Formula 1,
R ¹ , R ² , and R ⁴ are each independently substituted or unsubstituted C _1-10 alkyl;
R ³ is substituted or unsubstituted C _1-10 alkoxy;
L is substituted or unsubstituted C _1-5 alkylene; and
The substituted C _1-10 alkyl, C _1-10 alkoxy, and C _1-5 alkylene are each independently substituted with one or more substituents selected from the group consisting of C _1-5 alkyl and halogen.
6. The method of claim 5,
wherein R ¹ and R ² are each independently substituted or unsubstituted C _1-8 alkyl;
wherein R ³ is substituted or unsubstituted C _1-8 alkoxy;
wherein R ⁴ is substituted or unsubstituted C _1-8 alkyl;
L is substituted or unsubstituted C _1-3 alkylene; and
The substituted C _1-8 alkyl, C _1-8 alkoxy, and C _1-3 alkylene are each independently substituted with one or more substituents selected from the group consisting of C _1-3 alkyl and halogen. A composition for strengthening the barrier.
6. The method of claim 5,
wherein R ¹ and R ² are each independently substituted or unsubstituted C _1-6 alkyl;
wherein R ³ is substituted or unsubstituted C _1-3 alkoxy;
wherein R ⁴ is substituted or unsubstituted C _1-5 alkyl; and
Wherein L is a substituted or unsubstituted C _1-2 alkylene, a composition for strengthening the skin barrier.
6. The method of claim 5,
The compound represented by the formula (1) is a compound represented by the following formula (1-1), a composition for strengthening the skin barrier:
[Formula 1-1]

.
6. The method of claim 5,
The compound is to strengthen the cell-to-cell tight junction (tight junction), skin barrier strengthening composition.
6. The method of claim 5,
The compound increases the expression of one or more proteins selected from the group consisting of claudin-1, claudin-4, occludin, and filaggrin. , a composition for strengthening the skin barrier.
An external preparation for skin barrier strengthening comprising the composition for skin barrier strengthening according to claim 5.

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