KR102100460B1 - Syringic acid derivatives, and uses of the same - Google Patents

Abstract

여기서, X는 S 또는 NH; n은 0 내지 4의 정수; R₁은 수소, C₁-C₈의 알킬기, C₁-C₈의 히드록시알킬기, C₁-C₈의 싸이오알킬기, C₁-C₈의 알콕시기, C₆-₁₀의 아릴기, C₂-C₈의 헤테로아릴기, 아미노기, 아마이드기, 카르복실기, 카바메이트기 또는 카보네이트기로 이루어진 군에서 선택된 어느 하나이고, 아미노기, 아마이드기, 우레아기, 카바메이트기 등은 각각 C₁-C₄의 알킬기, C₁-C₄의 싸이오알킬기, C₆-C₁₀의 아릴, C₂-C₈의 헤테로아릴기로 더 치환될 수 있으며;R₂ 는 수소 또는 C₁-C₃ 알킬기이다.The present invention relates to the use of a useful compound having skin moisturizing and anti-inflammatory effects, ultimately promoting skin barrier function, improving skin homeostasis, and improving health and appearance. More specifically, the compound represented by Formula 1 below, It relates to a skin moisturizing factor active composition or an anti-inflammatory active composition comprising an isomer thereof, or a salt thereof.
[Formula 1]

Where X is S or NH; n is an integer from 0 to 4; R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, C ₂ -C ₈ Heteroaryl group, amino group, amide group, carboxyl group, carbamate group or any one selected from the group consisting of carbonate groups, amino group, amide group, urea group, carbamate group, etc. C ₁ -C ₄ the alkyl group, an arylthio group of C ₁ -C _4, C ₆ may be further substituted with a heteroaryl -C ₁₀ aryl, C ₂ -C ₈ in which; R ₂ Is hydrogen or a C ₁ -C ₃ alkyl group.

Description

SYRINGIC ACID DERIVATIVES, AND USES OF THE SAME}

The present invention relates to the use of useful compounds having skin moisturizing and anti-inflammatory effects, ultimately promoting skin barrier function, improving skin homeostasis, and improving health and appearance.

Disorders or damages to the skin barrier that occur naturally with aging of the skin can result in a small irritation due to a decrease in the ability to recover the skin, skin dryness and loss of transparency, reduced sensitivity to irritating contact dermatitis, or increased sensitivity to external irritations. It appears as a phenomenon that easily causes dermatitis. In addition, it is introduced that materials that are generally used and required to improve skin barriers and maintain homeostasis should satisfy moisturizing and anti-inflammatory properties.

First, the moisturizing material is a compound that helps to maintain the moisture of the stratum corneum by reducing or supplementing the amount of evaporation of moisture in the skin to prevent the skin from drying out. However, it is not enough to improve the weakened skin barrier by simply adjusting the amount of moisture. Therefore, unlike the previous one, the development of a basic and effective moisturizing material should be made, and for this, the development of the moisturizing material described below can be considered. The material compound 1) the effect of promoting the synthesis of hyaluronic acid 2) the effect of helping the maintenance of the natural moisturizing factor created by the skin 3) the effect of ceramide and filagrin related to the moisturizing action of the skin If it shows an effect to help biosynthesis, it can be said that it has sufficient potential as a material to help moisturize to improve the skin barrier.

In addition, the factors described above are explained as follows.

[Hyaluronic Acid]

Hyaluronic acid is a natural component that is present in animal and human joint fluids, cartilage, and skin. It is combined with a large amount of water to become a gel and is involved in skin flexibility. It is known for its excellent ability to contain water, which can hold about 6000 times more than its own weight, and is widely used as a material to help maintain moisture balance. If the content in the stratum corneum increases, it can be expected to improve the skin barrier restoration ability. In detail, as our age increases, the amount of hyaluronic acid decreases and the production capacity decreases. In particular, when the age is 40 years or less, it is only about half of its peak, so the skin decreases moisturizing ability and elasticity decreases. It is of interest as an anti-aging material because wrinkles can be accelerated. However, since the molecular weight is so large that it cannot penetrate into the skin, skin cells directly increase the production of hyaluronic acid or use a procedure that dermatology supplies directly to the skin as an injection.

[Filaggrin]

Filagreen is known as a substance that helps fibrous tissue called keratin of skin keratinocytes to clump together. Recently, natural moisturizing factors (NMFs) in keratinocytes are made from decomposition products of filaggrin.As a result of research findings of lack of filaggrin and mutation of chromosomes in atopic dermatitis and dry skin, filagrin Interest is increasing. Due to the lack of filaggrin, the bond between keratinocytes in the stratum corneum is weakened, and external stimuli can be easily transferred to the inside of the skin to exacerbate various inflammatory diseases. Since it does not contain, it may cause dryness.

Therefore, a material that aids in the production of pilar green may be a fundamental solution.

[Aquaporin-3]

Aquaporin-3, which is present in the keratinocyte cell membrane of the skin, is a membrane protein that plays an important role in transporting moisture and glycerol to the skin epidermal layer. When they are activated, moisture and active ingredients can be effectively absorbed by our skin. It is said to help. In particular, skin without aquaporin-3 is known to have poor skin elasticity and reduced recovery of skin barrier function, and is known to act on moisturizing, wound healing, and maintaining skin epidermal homeostasis (Reference: Boury -Jamot M, Handb Exp Pharmacol., 2009 (190), 205-217). Therefore, the material compound that helps the biosynthesis of aquaporin-3 is expected to have a very high utilization value as a basic moisturizing material for various skin diseases that cause skin dryness such as atopy.

Second, atopic dermatitis is a chronic inflammatory skin disease with severe itching and overall dry skin, and eczema lesions recur repeatedly on a specific site, accompanied by an inflammatory response and inflammatory disease mediated by immune cells, T-cells, deteriorating skin barrier function. It is recurred repeatedly and inflammation is easily caused by various stimuli. In order to treat such inflammatory diseases, steroidal anti-inflammatory agents are used, but steroidal agents have various side effects and limit their use.

Inflammatory responses are induced by infections of bacteria and viruses, local ischemia, immune responses by antigen-antibodies, and toxins or chemicals, resulting in symptoms such as erythema, edema, fever, and pain. Various immune cells such as polymorphonuclear leucocyte, marcrophage, lymphocyte, and mast cell are involved in this inflammatory reaction, and they act in slightly different forms depending on the stage of inflammation. In the early stage of inflammation, pro-inflammatory cytokine, mainly IL-1βIL-6, and TNF-α, which they secrete, act as important mediators, and in the next stage, PAF produced through the phospholipase A2 pathway and the cyclooxygenase pathway, It is known that many mediators such as lipid mediators such as prostaglandins and leucotriene, free radicals and nitric oxide interact with pro-inflammatory cytokines to form a complex network.

Therefore, we are trying to develop an anti-inflammatory agent as a method of inhibiting pro-inlammatory cytokine from being secreted by external stimulation and a method of controlling cytokine stimulating target cells, but are insufficient to expect the effect and cause inflammatory diseases. Research to understand the mechanism of action of these cytokines and to explore their modulators is urgently needed.

Since it is synthesized through the reaction of phenolic acids and amino acids existing in nature, such as syringic acid, the pharmacological and toxic mechanisms for the decomposing substances produced are well known even if the bonds are separated by metabolism or decomposition. Compounds having the advantage of being easy to secure safety in using synthetic derivative compounds are known (the following Patent Document 1; all of the contents of Patent Document 1 are within the scope of the contents of the present invention and are not in disposition). All of them are cited and incorporated as part of the specification of the present invention). For this substance, only a higher melanin-inhibiting effect than arbutin, that is, a whitening effect is known, and no moisturizing or anti-inflammatory effect is known at all.

WO 2017/099531 A1 (2017.06.15.)

In the present invention, it helps to produce hyaluronic acid and aquaporin-3 related to the maintenance of the epidermis and the improvement of the skin barrier, and decreases the secretion of interleukin-6 (IL-6) and prostaglandin E2, PGE2 ) It is intended to be proposed as a compound having anti-inflammatory efficacy through the inhibition of expression and the inhibition of production of NO, one of free radicals. In other words, it is intended to provide a composition for external application for skin or an anti-aging material excellent in moisturizing and atopy improvement. In addition, it is intended to provide a novel method for synthesizing these compounds.

The present invention to solve the problems of the prior art described above,

It provides a skin moisturizing factor active composition comprising a compound represented by the formula (1), isomers thereof, or salts thereof.

[Formula 1]

Where X is S or NH;

n is an integer from 0 to 4;

R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, C ₂ -C ₈ Heteroaryl group, amino group, amide group, carboxyl group, carbamate group or any one selected from the group consisting of carbonate groups, amino group, amide group, urea group, carbamate group, etc. C ₁ -C ₄ May be further substituted with an alkyl group, C ₁ -C ₄ thioalkyl group, C ₆ -C ₁₀ aryl, or C ₂ -C ₈ heteroaryl group;

R ₂ is hydrogen or a C ₁ -C ₃ alkyl group.

In addition, in the present invention, the formula (1) provides a skin moisturizing factor active composition comprising a compound represented by the following formula 2, or formula 3, an isomer thereof, or a salt thereof.

[Formula 2]

[Formula 3]

Here, R ₂ is hydrogen or a C ₁ -C ₃ alkyl group;

R ₃ to R ₅ are each independently hydrogen, a C ₁ -C ₈ alkyl group, a C ₁ -C ₈ hydroxyalkyl group, a C ₁ -C ₈ thioalkyl group, or a C ₁ -C ₈ alkoxy group, C ₆ - ₁₀ aryl group, a heteroaryl group of C ₂ -C _8, an amino group, an amide group, a carboxyl group, a carbamate group or any one selected from the group consisting of a carbonate group, amino group, amide group, urea group, a carbamate group etc. each of the alkyl thio group of _{C 1 -C 4, C 1 -C} 4, C 6 may be further substituted with a heteroaryl -C ₁₀ aryl, C ₂ -C ₈ in which;

The R ₃ and R ₄ may be the same or different from each other.

In addition, it provides a cosmetic composition for moisturizing skin comprising the active ingredient of the skin moisturizing factor of the present invention as an active ingredient.

In addition, it provides a pharmaceutical composition for improving or treating symptoms of skin dryness or atopic dermatitis comprising the active ingredient of the skin moisturizing factor of the present invention as an active ingredient.

In addition, it provides an anti-inflammatory active composition characterized in that it comprises a compound represented by the following formula (1), isomers thereof, or salts thereof.

[Formula 1]

Where X is S or NH;

n is an integer from 0 to 4;

R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, C ₂ -C ₈ Heteroaryl group, amino group, amide group, carboxyl group, carbamate group or any one selected from the group consisting of carbonate groups, amino group, amide group, urea group, carbamate group, etc. C ₁ -C ₄ May be further substituted with an alkyl group, C ₁ -C ₄ thioalkyl group, C ₆ -C ₁₀ aryl, or C ₂ -C ₈ heteroaryl group;

R ₂ is hydrogen or a C ₁ -C ₃ alkyl group.

In addition, in the present invention, the formula (1) provides an anti-inflammatory active composition characterized in that it comprises a compound represented by the following formula 2, or formula 3, isomers thereof, or salts thereof.

[Formula 2]

[Formula 3]

Here, R ₂ is hydrogen or a C ₁ -C ₃ alkyl group;

R ₃ to R ₅ are each independently hydrogen, a C ₁ -C ₈ alkyl group, a C ₁ -C ₈ hydroxyalkyl group, a C ₁ -C ₈ thioalkyl group, or a C ₁ -C ₈ alkoxy group, C ₆ - ₁₀ aryl group, a heteroaryl group of C ₂ -C _8, an amino group, an amide group, a carboxyl group, a carbamate group or any one selected from the group consisting of a carbonate group, amino group, amide group, urea group, a carbamate group etc. each of the alkyl thio group of _{C 1 -C 4, C 1 -C} 4, C 6 may be further substituted with a heteroaryl -C ₁₀ aryl, C ₂ -C ₈ in which;

The R ₃ and R ₄ may be the same or different from each other.

In addition, it provides a skin trouble-improving cosmetic composition comprising the anti-inflammatory active composition of the present invention as an active ingredient.

In addition, it provides a pharmaceutical composition for improving or treating symptoms of an inflammatory disease comprising the anti-inflammatory active composition of the present invention as an active ingredient.

In addition, in the present invention, the inflammatory disease provides psoriasis, or a pharmaceutical composition characterized by atopic dermatitis.

Further, 4-acetoxy-3,5-dimethoxybenzoic acid and alkyl haloformate are reacted to prepare (4-acetoxy-3,5-dimethoxybenzoic) (alkylcarbonic) unhydride intermediate. ; And

It provides a method for producing a compound of Formula 1, characterized in that it comprises the step of reacting the anhydride intermediate with a standard amino acid or an ester compound thereof.

[Formula 1]

Where X is S or NH;

n is an integer from 0 to 4;

R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, C ₂ -C ₈ Heteroaryl group, amino group, amide group, carboxyl group, carbamate group or any one selected from the group consisting of carbonate groups, amino group, amide group, urea group, carbamate group, etc. C ₁ -C ₄ May be further substituted with an alkyl group, C ₁ -C ₄ thioalkyl group, C ₆ -C ₁₀ aryl, or C ₂ -C ₈ heteroaryl group;

R ₂ is hydrogen or a C ₁ -C ₃ alkyl group.

In addition, in the present invention, the standard amino acid provides a method for preparing a compound of Formula 1, characterized in that it is phenylalanine, cysteine, or serine.

In addition, an intermediate compound represented by the following Chemical Formula 4 is provided.

[Formula 4]

Here, R is hydrogen, a C ₁ -C ₃ alkyl group, or a phenyl group.

The present invention, while being able to be safely used on the skin, compounds can help restore or improve the skin barrier by using the activity of increasing the production of hyaluronic acid (HA) and aquaporin 3 (AQP3), which are skin moisturizing factors in keratinocytes. It can be used as a fundamental improvement agent. Furthermore, the compounds show anti-inflammatory activity through the activity of inhibiting the production of NO, which plays an important role in inducing inflammation, the inhibition of PGE2, an inflammatory factor, and the inhibition of IL-6, an inflammatory cytokine. It can be used for improvement purposes, and can be usefully used for improving and treating inflammatory diseases such as atopic dermatitis.

1 and 2 are graphs showing the results of measuring NO (Nitric Oxide) production inhibitory activity of the compounds of the present invention.
Figure 3 is a graph of the results of measuring the inhibitory effect of inflammatory cytokine production of the compounds of the present invention.
4 is a graph showing the results of measuring the inhibitory effect of inflammatory factors of the compounds of the present invention.
5 is a graph showing the results of measuring the hyaluronic acid synthesis promoting effect of the compounds of the present invention.
6 is a graph showing the results of measuring the effect of promoting the synthesis of aquaporin-3 of the compounds of the present invention.

Hereinafter, the present invention will be described in detail.

The present invention,

It characterized in that it comprises a compound represented by the formula (1), isomers thereof, or salts thereof.

[Formula 1]

Where X is S or NH;

n is an integer from 0 to 4;

R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, C ₂ -C ₈ Heteroaryl group, amino group, amide group, carboxyl group, carbamate group or any one selected from the group consisting of carbonate groups, amino group, amide group, urea group, carbamate group, etc. C ₁ -C ₄ May be further substituted with an alkyl group, C ₁ -C ₄ thioalkyl group, C ₆ -C ₁₀ aryl, or C ₂ -C ₈ heteroaryl group;

R ₂ is hydrogen or a C ₁ -C ₃ alkyl group.

Specifically, Chemical Formula 1 may include a compound represented by Chemical Formula 2 or Chemical Formula 3, an isomer thereof, or a salt thereof.

[Formula 2]

[Formula 3]

Here, R ₂ to R ₅ are the same as defined in Formula 1, R _{3 and} R ₄ They may be the same or different from each other.

As a more specific example, the formula 2 or 3 may be any one of the following compounds 1 to 6.

The compound of Formula 1 may be prepared through the steps summarized in Scheme 1 below.

[Scheme 1]

That is, 4-acetoxy-3,5-dimethoxybenzoic acid and alkyl haloformate are reacted to prepare (4-acetoxy-3,5-dimethoxybenzoic) (alkyl carbonic) unhydride intermediate. step; And

It may be prepared through a step of reacting the intermediate with a standard amino acid or an ester compound thereof.

Here, the standard amino acid is 22 kinds of amino acids, glycine, glutamine, glutamic acid, lysine, leucine, methionine, valine, serine, selenocysteine, cysteine, arginine, asparagine, aspartic acid, isoleucine, alanine, tyrosine, threonine, Tryptophan, phenylalanine, proline, pyrrolysine, and histidine.

As can be seen through Reaction Scheme 1, the compounds represented by Chemical Formula 1 are phenolic acid derivatives obtained through the reaction of syringic acid with amino acids, which are produced when bonds are separated by metabolism or decomposition. The pharmacological and toxic mechanisms for decomposition substances are already well-known, which has the advantage of ensuring safety.

 In WO 2017/099531 (Patent Document 1), the method of formula (1) is disclosed, but the method disclosed herein has a high reactivity of the intermediate compound and is unstable, making it very difficult to react under normal conditions of normal temperature and atmospheric pressure in the atmosphere. There was, and accordingly, it was not possible to mass-produce, and it was difficult to confirm the degree of reaction or to measure the purity through an intermediate compound. In addition, the reaction time for preparing the intermediate compound is also very long, thereby increasing the overall process time, and there is also a problem in that production efficiency is deteriorated.

Thus, the present invention can solve this problem by preparing the (4-acetoxy-3,5-dimethoxybenzoic) (alkyl carbonic) unhydride intermediate compound represented by the following formula (4).

[Formula 4]

(Here, R is hydrogen, a C ₁ -C ₃ alkyl group, or a phenyl group.)

That is, in obtaining the syringic acid derivatives represented by Formula 1 obtained in Scheme 1, an intermediate hybrid anhydride represented by Formula 4 was prepared so that the shirring acid had reactivity with the amino acid ester, and the intermediate compound was contained. In the reaction between the intermediate product reaction solution and the amino acid ester, a separate separation and purification of the intermediate compound is not required in the intermediate product reaction solution, and reaction is possible within one container to minimize process steps. Through the reaction of time, it was confirmed a method capable of producing a syringic acid derivative with high yield and high purity.

Since the compound of Formula 1 has a skin moisturizing factor activity, it may be used as a cosmetic composition for skin moisturizing, or for improving or treating symptoms of diseases such as skin dryness, psoriasis, or atopic dermatitis. That is, it was confirmed that the compounds of the present invention have an effect of promoting the skin moisturizing factors hyaluronic acid, aquaporin-3 synthesis, and the like. A more detailed understanding of this may be made through examples and test examples to be described later.

In addition, since the compound of Formula 1 also has anti-inflammatory activity, skin problems (e.g., rash, pruritus, erythema, acne, occurrence of swelling, etc.) are improved cosmetic compositions or inflammatory diseases (e.g., psoriasis, atopy) Dermatitis, etc.) may be used as a pharmaceutical composition for improving or treating symptoms. That is, the compounds of the present invention, as well as the activity of inhibiting the production of NO (nitrogen monoxide), one of the free radicals that cause skin inflammation, secretion of the inflammatory mediator, Interleukin-6 (IL-6) It was confirmed as a material exhibiting anti-inflammatory activity through reduction and inhibition of prostaglandin E2 (PGE2) expression. A more detailed understanding of this may be made through examples and test examples to be described later.

In the present invention, the siring acid derivative compound, an isomer thereof, or a salt thereof may be 0.0001 to 10% by weight, preferably 0.001 to 5% by weight of the total weight of the cosmetic composition. If it is less than 0.0001%, efficacy and effect may be insignificant, and if it exceeds 10% by weight, there may be a stability problem of the formulation.

In addition, the syringic acid derivative compound represented by Chemical Formula 1, an isomer thereof, or a salt thereof may be used for anti-inflammatory and prevention, improvement or treatment of atopic disease.

The pharmaceutical composition may further contain a pharmaceutical adjuvant and other therapeutically useful substances, such as a preservative, stabilizer, hydration or emulsifying accelerator, salt and / or buffer for osmotic pressure control. The pharmaceutical composition may be formulated as a lotion, cream, ointment or gel, but is not limited thereto.

In the present invention, the syringic acid derivative compound, isomers thereof, or salts thereof may be 0.0001 to 10% by weight, preferably 0.001 to 5% by weight of the total weight of the pharmaceutical composition. If it is less than 0.0001%, efficacy and effectiveness may be negligible, and if it exceeds 10% by weight, there may be stability problems of the formulation.

Hereinafter, the present invention will be described in more detail through Examples and Test Examples. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.

Example

Example  1-Compound 1, (R)- methyl -2- Acetamido -3- (4-hydroxy-3,5- Dimethoxy Preparation of benzoylthio) propanoate

[Example 1-1] Preparation of 4-acetoxy-3,5-dimethoxybenzoyl anhydride

In a 1.0L flask, 4-acetoxy-3,5-dimethoxybenzoic acid (47.0 g, 196 mmol) was added, dichloromethane (376 mL) and triethylamine (29 mL) were added, and the mixture was stirred at low temperature (5 degrees). To this, ethyl chloroformate (20 mL) was added dropwise. Stir at 5 ° C for 30 minutes and filter the solution to obtain a 4-acetoxy-3,5-dimethoxybenzoyl anhydride solution.

[Example 1-2] Preparation of (R) -methyl-2-acetamido-3- (4-hydroxy-3,5-dimethoxybenzoylthio) propanoate

In a 1.0L flask, L-cysteine methyl ester (34.7g, 196mmol) and anhydrous potassium carbonate (27g, 196mmol) were added, dichloromethane (139mL) was added, and the mixture was stirred at low temperature (5 ° C). To this, a solution of 4-acetoxy-3,5-dimethoxybenzoyl anhydride obtained in Example 1-1 was added dropwise. The reaction solution was stirred at room temperature for 2 hours, and water (139 mL) was added to terminate the reaction. The organic layer obtained by separation is wiped with an aqueous 1N-hydrochloric acid solution, a saturated aqueous sodium hydrogen carbonate solution, and a saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and then concentrated under reduced pressure. The obtained solid was purified with methyl T-butyl ether (tert-Butyl methyl ether) to obtain a solid 64g (82%). The obtained solid (64 g, 160 mmol) was placed in a 1 L flask, dichloromethane (320 mL) was added, and the mixture was stirred at low temperature (5 degrees). To this, a solution of hydrazine hydrate (80%, 14 mL, 320 mmol) diluted with methanol (64 mL) is added dropwise.

The reaction solution was stirred at room temperature for 2 hours, and dichloromethane (320 mL) was added. The solution is wiped with water and a 1N-hydrochloric acid aqueous solution, dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and the solution is concentrated under reduced pressure. The obtained solid was purified with ethyl acetate to obtain a white solid (45 g, 79%).

Example  2-Compound 2, (R) -ethyl-2- Acetamido -3- (4-hydroxy-3,5- Dimethoxy Preparation of benzoylthio) propanoate

[Example 2-1] Preparation of 4-acetoxy-3,5-dimethoxybenzoyl anhydride

In a 1.0L flask, 4-acetoxy-3,5-dimethoxybenzoic acid (40.0g, 167mmol) was added, dichloromethane (320mL) and triethylamine (26mL) were added, and the mixture was stirred at low temperature (5 degrees). Ethyl chloroformate (17 mL) was added dropwise. Stir at 5 ° C for 30 minutes and filter the solution to obtain a 4-acetoxy-3,5-dimethoxybenzoyl anhydride solution.

[Example 2-1] Preparation of (R) -ethyl-2-acetamido-3- (4-hydroxy-3,5-dimethoxybenzoylthio) propanoate

In a 1.0L flask, L-cysteine ethyl ester (341.8g, 167mmol) and anhydrous potassium carbonate (23g, 167mmol) were added, dichloromethane (127mL) was added, and the mixture was stirred at low temperature (5 degrees). To this, a solution of 4-acetoxy-3,5-dimethoxybenzoyl anhydride obtained in Example 2-1 was added dropwise. The reaction solution was stirred at room temperature for 2 hours, and water (127 mL) was added to terminate the reaction. The organic layer obtained by separation is wiped with an aqueous 1N-hydrochloric acid solution, a saturated aqueous sodium hydrogen carbonate solution, and a saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and then concentrated under reduced pressure. The obtained solid was purified with methyl T-butyl ether (tert-Butyl methyl ether) to obtain a solid 56g (81%). The obtained solid (56 g, 135 mmol) was placed in a 1 L flask, dichloromethane (392 mL) was added, and the mixture was stirred at low temperature (5 degrees). A solution of hydrazine hydrate (80%, 12 mL, 271 mmol) diluted with ethanol (22 mL) is added dropwise.

The reaction solution was stirred at room temperature for 2 hours, and dichloromethane (330 mL) was added. The solution is wiped with water and a 1N-hydrochloric acid aqueous solution, dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and the solution is concentrated under reduced pressure. The obtained solid was purified with isopropyl ether to obtain a white solid (38g, 75%).

Example  3-compound 4, ( S )- methyl -3-hydroxy-2- (4-hydroxy-3,5- Dimethoxybenzo Ilthio) Preparation of propanoate

[Example 3-1] Preparation of 4-acetoxy-3,5-dimethoxybenzoyl anhydride

Anhydrous solution was prepared using 4-acetoxy-3,5-dimethoxybenzoic acid (4.8g, 20mmol) by the method described in Example 1-1 and used in the next reaction.

[Example 3-2] Preparation of ( S ) -methyl-3-hydroxy-2- (4-hydroxy-3,5-dimethoxybenzoylthio) propanoate

 Compound (S) -serineol methyl ester (2.4g, 20mmol) was used to carry out the binding reaction and deprotection reaction in the manner described in Example 1-2, followed by separation-purification to obtain Compound 4, white solid (3.0g, 50%). ).

Example  4-compound 6, ( S ) -Ethyl-3-phenyl-2- (4-hydroxy-3,5- Dimethoxybenzami Figure) Preparation of propanoate

[Example 4-1] Preparation of 4-acetoxy-3,5-dimethoxybenzoyl anhydride

Anhydrous solution was prepared using 4-acetoxy-3,5-dimethoxybenzoic acid (2.4 g, 10 mmol) by the method described in Example 1-1 and used in the next reaction.

[Example 4-2] Preparation of ( S ) -ethyl-3-phenyl-2- (4-acetoxy-3,5-dimethoxybenzamido) propanoate

(S) -Phenyl alanine ethyl ester (1.9g, 9.8mmol), anhydrous potassium carbonate (1.4g, 10mmol) was added to a 100mL flask, dichloromethane (15mL) was added, and the mixture was stirred at low temperature (5 ° C). To this, a solution of 4-acetoxy-3,5-dimethoxybenzoyl anhydride obtained in Example 4-1 was added dropwise. The reaction solution was stirred at room temperature for 2 hours, and water (20 mL) was added to terminate the reaction. The organic layer obtained by separation is wiped with an aqueous 1N-hydrochloric acid solution, a saturated aqueous sodium hydrogen carbonate solution, and a saturated aqueous sodium chloride solution. The organic layer was dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and then concentrated under reduced pressure. The obtained solid was purified by column chromatography to obtain the target product (3.6 g, 93%).

¹ H NMR (400MHz, CDCl ₃ ) δ 1.29 (t, J = 6.8 Hz, 3H), 2.34 (s, 3H), 3.21-3.32 (m, 2H), 3.83 (s, 6H), 4.20-4.26 (m , 2H), 5.01-5.06 (m, 1H), 6.48 (d, J = 6.8 Hz, 1H), 6.93 (s, 2H), 7.14-7.16 (m, 2H), 7.23-7.31 (m, 3H).

[Example 4-3] Preparation of ( S ) -ethyl-3-phenyl-2- (4-hydroxy-3,5-dimethoxybenzamido) propanoate

In a 100 mL flask, add the compound obtained in Example 4-2 (1.6 g, 3.85 mmol) and dichloromethane (20 mL) and stir at low temperature (5 degrees). To this, a solution of hydrazine hydrate (80%, 0.34 mL, 7.77 mmol) diluted with ethanol (5 mL) is added dropwise. The reaction solution was stirred at room temperature for 2 hours, and dichloromethane (20 mL) was added. The solution is wiped with water and a 1N-hydrochloric acid aqueous solution, dried over anhydrous magnesium sulfate (MgSO ₄ ), filtered, and the solution is concentrated under reduced pressure. The obtained solid was purified by column chromatography to obtain the target product (1.1 g, 78%).

¹ H NMR (400MHz, CDCl ₃ ) δ 1.23-1.30 (m, 3H), 3.24-3.27 (m, 2H), 3.90 (s, 6H), 4.20-4.25 (m, 2H), 5.02-5.07 (m, 1H), 6.51 (d, J = 7.6 Hz, 1H), 6.96 (s, 2H), 7.15-7.17 (m, 2H), 7.25-7.31 (m, 3H).

Test example

Test example  1-of compounds NO (Nitric Oxide) generation  Inhibition activity measurement

It was measured whether the compounds have the activity of inhibiting the production of NO (nitric oxide) induced by lipopolysaccharide (LPS) in macrophage cells, and 2-Amino-4-methylpyridine (10 μM) was used as a positive control. Did. In RAW 264.7 macrophage cells, LPS was treated to express NO synthase enzyme, and the amount of NO produced was measured by the Griess method. Griess reagent (1% sulfanylamine, 0.1% N- (1-naphthyl) -ethylene diamine dihydrochloride, 2.5% H ₃ PO ₄ ) oxidizes NO to change to NO ₂ and the generated NO ₂ is 540 nm The absorbance was measured at and the concentration was determined using a calibration curve of NaNO ₂ . That is, RAW 264.7 macrophages were cultured to be 1 X 10 ⁵ cells per 1 ml of DMEM medium, placed in 48 wells, cultured for 18 hours, and then adhered to the cells, followed by 20 μl of LPS (1 μg / ml), After 100 μl of each sample containing 2-Amino-4-methylpyridine as a positive control was incubated for 20 hours, the amount of NO generated in the culture medium was quantified using a Griess reagent. And FIG. 2.

As can be seen from the schematic results, in the anti-inflammatory experiment conducted at a concentration of 1-50 μg / mL, which showed a survival rate of 95% or higher in the cellular MTT reaction, the NO production inhibitory activity of the compounds was compared to the positive control 2-Amino-4-methylpyridine. It showed little activity, but showed concentration-dependent NO production inhibitory activity during sample treatment.

Test example  2-inflammatory properties of compounds cytokine  Production inhibition evaluation

It was measured whether the compounds have the activity of inhibiting the production of IL-6 and PGE ₂ induced by LPS (lipopolysaccharide) in macrophage cells, and 2-Amino-4-methylpyridine (10 μM) was used as a positive control. Used. IL-6 and PGE2 are produced by macrophages that are activated by stimulators, such as lipopolysaccharide (LPS), in various cellular signaling systems. Anti-inflammatory activity can be confirmed by suppressing IL-6 and PGE2, which are excessively produced by LPS, when they are produced in excess, causing an inflammatory reaction. Raw264.7 macrophage was dispensed in a 24-well plate to be 3 × 10 ⁵ cells / well and cultured for 24 hours. Subsequently, the samples were treated by concentration, and after incubation for 1 hour, LPS was treated at 1 μg / Ml concentration in each well, and supernatant cultured for 24 hours was taken to measure the amount of IL-6 and PGE2 produced (FIGS. 3 and 4). ).

3 and 4, the compounds showed less activity than the positive control 2-Amino-4-methylpyridine, but showed concentration-dependent IL-6 and PGE2 production inhibitory activity in concentration-dependent sample processing.

Test example  3-Experimental effect of promoting hyaluronic acid synthesis of compounds

The effect of promoting the biosynthesis of hyaluronic acid in human keratinocytes HaCaT culture was confirmed. HaCaT cells were adjusted to 1.0 × 10 ⁵ cells / ml using DMEM medium, inoculated into 24 well plates, and cultured for 18 hours. A new medium containing the test substance is treated and cultured for 24 hours. Then, take the cell culture supernatant and measure the HA content in the cell culture solution using Hyaluronic Acid (HA) ELISA kit. At this time, retinoic acid (10 μM), which is known as a substance that promotes hyaluronic acid biosynthesis, was tested together, and is illustrated in FIG. 5.

As can be seen from the results, the compound represented by Chemical Formula 1 showed less activity than the positive control retinoic acid, but increased concentration-dependently in the amount of hyaluronic acid, a moisturizing-related factor, in keratinocytes (human kerationcyte) induced by dryness. It was confirmed that it is a material that can be fundamentally improved in relation to moisturizing of the skin.

Test example  4-of compounds Aquaporin -3 Synthesis promotion effect experiment

HaCaT cells were adjusted to 1.0 × 10 ⁵ cells / ml using DMEM medium, inoculated into 24 well plates, and cultured for 18 hours. Then, a new medium containing the test substance is treated and cultured for 24 hours. After obtaining the lysate of the cultured cells, the Aquaporin 3 (AQP3) ELISA kit is used to measure the AQP3 content in the cell and test together Retinoic acid (10 μM), which is known as a substance that promotes biosynthesis of Aquaporin-3 , In Figure 6, it is schematic.

As can be seen from the results, as a result of treating the compounds represented by Formula 1 with concentrations in the keratinocytes (human kerationcyte) in which dryness was induced, it was confirmed that the amount of aquaporin-3, a moisturizing-related factor, was increased in a concentration-dependent manner. Did.

Claims (12)

A cosmetic composition for moisturizing skin comprising a compound represented by the following Chemical Formula 1, an isomer thereof, or a salt thereof.
[Formula 1]

Where X is S or NH;
n is an integer from 0 to 4;
R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, Any one selected from the group consisting of C ₂ -C ₈ heteroaryl group, amino group, amide group, carboxyl group, carbamate group or carbonate group;
R ₂ is hydrogen or a C ₁ -C ₃ alkyl group. The method according to claim 1, wherein the formula 1 is a cosmetic composition for moisturizing skin, characterized in that it comprises a compound represented by the following formula 2, or formula 3, isomers thereof, or salts thereof.
[Formula 2]

[Formula 3]

Here, R ₂ is hydrogen or a C ₁ -C ₃ alkyl group;
R ₃ to R ₅ are each independently hydrogen, a C ₁ -C ₈ alkyl group, a C ₁ -C ₈ hydroxyalkyl group, a C ₁ -C ₈ thioalkyl group, or a C ₁ -C ₈ alkoxy group, C ₆ - ₁₀ aryl group, a heteroaryl group of C ₂ -C _8, an amino group, an amide group, a carboxyl group, a carbamate group, or is any one selected from the group consisting of carbonate groups;
The R ₃ and R ₄ may be the same or different from each other. delete A pharmaceutical composition for improving or treating symptoms of skin dryness or atopic dermatitis comprising the compound represented by Formula 1 of claim 1 as an active ingredient. An anti-inflammatory active composition comprising a compound represented by the following Chemical Formula 1, isomers thereof, or salts thereof.
[Formula 1]

Where X is S or NH;
n is an integer from 0 to 4;
R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, Any one selected from the group consisting of C ₂ -C ₈ heteroaryl group, amino group, amide group, carboxyl group, carbamate group or carbonate group;
R ₂ is hydrogen or a C ₁ -C ₃ alkyl group. The method according to claim 5, wherein the formula 1 is an anti-inflammatory active composition characterized in that it comprises a compound represented by the following formula 2, or formula 3, isomers thereof, or salts thereof.
[Formula 2]

[Formula 3]

Here, R ₂ is hydrogen or a C ₁ -C ₃ alkyl group;
R ₃ to R ₅ are each independently hydrogen, a C ₁ -C ₈ alkyl group, a C ₁ -C ₈ hydroxyalkyl group, a C ₁ -C ₈ thioalkyl group, or a C ₁ -C ₈ alkoxy group, C ₆ - ₁₀ aryl group, a heteroaryl group of C ₂ -C _8, an amino group, an amide group, a carboxyl group, a carbamate group, or is any one selected from the group consisting of carbonate groups;
The R ₃ and R ₄ may be the same or different from each other. A skin trouble-improving cosmetic composition comprising the compound represented by Formula 1 of claim 5 as an active ingredient. A pharmaceutical composition for improving or treating symptoms of an inflammatory disease comprising the compound represented by Formula 1 of claim 5 as an active ingredient. The pharmaceutical composition according to claim 8, wherein the inflammatory disease is psoriasis or atopic dermatitis. Reacting 4-acetoxy-3,5-dimethoxybenzoic acid with an alkyl haloformate to prepare (4-acetoxy-3,5-dimethoxybenzoic) (alkylcarbonic) unhydride intermediate; And
A method of preparing a compound of Formula 1, comprising reacting the anhydride intermediate with a standard amino acid or an ester compound thereof.
[Formula 1]

Where X is S or NH;
n is an integer from 0 to 4;
R ₁ is hydrogen, C ₁ -C ₈ alkyl, C ₁ -C ₈ hydroxy alkyl, C ₁ -C arylthio group, an alkoxy group of C ₁ -C ₈ of _8, a C ₆ - ₁₀ aryl group, Any one selected from the group consisting of C ₂ -C ₈ heteroaryl group, amino group, amide group, carboxyl group, carbamate group or carbonate group;
R ₂ is hydrogen or a C ₁ -C ₃ alkyl group. The method of claim 10, wherein the standard amino acid is phenylalanine, cysteine, or serine. An intermediate compound represented by the following formula (4).
[Formula 4]

Here, R is hydrogen, a C ₁ -C ₃ alkyl group, or a phenyl group.

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