Phytosphingosine-analogous ceramide

Abstract

The present invention relates to a novel pseudo-ceramide compound and a method for preparing same, the compound being represented by chemical formula 1. The pseudo-ceramide of the present invention not only has a moisturizing effect, increased filaggrin production, and an anti-inflammatory effect, but also has a simple and economical synthesis process, and is suitable for use in a topical dermatologic composition for skin moisturization and cosmetic applications. [Chemical Formula 1] wherein A* is hydrogen or an acyl group of a C7-C16 saturated or unsaturated aliphatic chain, which is unsubstituted or substituted with a hydroxyl group, R1 is a C7-C15 saturated or unsaturated aliphatic chain which is unsubstituted or substituted with a hydroxyl group, and the difference in the number of carbon atoms between A* and R1 is at least 7.

Description

Phytosphingosine analogue ceramide

[0001] The present invention relates to a novel ceramide-like compound and a method for producing the same.

[0002] The skin is largely divided into three layers: the epidermis, the dermis, and the subcutaneous layer. Among these, the epidermis layer is responsible for the skin's barrier function with keratinocytes and skin lipids, and controls the cell division and differentiation of keratinocytes to protect the skin from external harmful substances, prevent the leakage of internal substances, and prevent the evaporation of skin moisture.

[0003] Ceramide is the main lipid component of the stratum corneum, the upper layer of the skin, and acts as an important barrier by preventing external compounds from entering the epidermis and preventing moisture loss. When the concentration of ceramide in the skin decreases, the protective barrier function of the stratum corneum decreases, resulting in various dermatological symptoms, such as atopic dermatitis and psoriasis.

[0004] The addition of ceramides to cosmetics has been found to improve the skin's barrier function, restore lost moisture and have an "anti-wrinkle" effect, and ceramides have also been used in medications for the treatment of, for example, hypersensitivity eczema (Kerscher et al, (1991) Eur J Dermatol, 1, 39-43).

[0005] Furthermore, recent studies have revealed that the diverse structures of ceramides are also involved in unique signaling roles in cells. That is, ceramides and their metabolites, including sphingosine-1-phosphate, serve as signaling lipids that regulate multiple cellular functions, such as proliferation, differentiation, and apoptosis, as well as the epidermal antimicrobial barrier. Along with fatty acids, cholesterol, and phospholipids, ceramides are structural components of cell membranes and are considered lipid signaling molecules involved in various cellular processes.

[0006] However, natural ceramides have great difficulty in producing high-content products due to problems such as low solubility, difficulty in mass production through extraction methods, and high price.

[0007] Accordingly, most cosmetics manufacturers are working on research and development to synthesize economical and easy-to-use pseudo-ceramides, and pseudo-ceramides currently developed domestically include Amorepacific Corporation's Republic of Korea Publication No. 2017-0076561 (Patent Document 1) and Koreana Cosmetics Corporation's Republic of Korea Registration Patent No. 10-0539965 (Patent Document 2), but their general use is still limited due to the unit price issue due to the difficult manufacturing method (therefore, the entire contents of Patent Documents 1 and 2 are cited and incorporated as background technology of this specification).

[0008] In addition, it is known that not only commercially available natural ceramides but also synthetic ceramide compounds still have limitations in producing high-content products due to low hydrophilicity and solubility.

[0009] Therefore, there is a continuous need for research and development of pseudo-ceramides that improve the properties of existing pseudo-ceramides, while also having a simple production process and being economically advantageous.

[0010] [Prior Art Literature]

[0011] (Patent Document 1) KR10-2017-0076561A (July 4, 2017)

[0012] (Patent Document 2) KR10-0539965B1 (December 23, 2005)

[0013] To address the economical issues of the natural ceramide compounds described above and the formulation difficulties due to their low hydrophilicity, the present invention seeks to provide a novel pseudo-ceramide. Furthermore, the synthesized pseudo-ceramide not only promotes bioactivity to enhance skin elasticity and moisturizing effects, but also increases the production of filaggrin, a natural moisturizing factor known to play a key role in skin moisturizing, and exhibits the effect of suppressing the inflammatory substance PGE2.

[0014] The present invention has been devised to solve the problems of the above-described prior art.

[0015] A compound represented by the following chemical formula 1 is provided.

[0016] [Chemical Formula 1]

[0017]

[0018] Here, A * is hydrogen, or an acyl group of a saturated or unsaturated aliphatic chain having 7 to 16 carbon atoms, which is unsubstituted or substituted with a hydroxy group, and R1 is a saturated or unsaturated aliphatic chain having 7 to 15 carbon atoms, which is unsubstituted or substituted with a hydroxy group, and A * The difference in the number of carbon atoms between R1 and R2 is at least 7.

[0019] In addition, a composition for external application to the skin containing the compound of the present invention as an active ingredient is provided.

[0020] The pseudoceramide of the present invention not only has a moisturizing effect, increases filaggrin production, and has an anti-inflammatory effect, but is also suitable for use in external skin moisturizing compositions and cosmetics while having a simple and economical synthesis process.

[0021] Figure 1 shows the results of cytotoxicity evaluation in skin keratinocytes (HaCaT cells).

[0022] Figure 2 shows the results of evaluating the amount of hyaluronic acid produced in skin keratinocytes (HaCaT cells).

[0023] Figure 3 shows the results of evaluating the amount of filaggrin produced in skin keratinocytes (HaCaT cells).

[0024] Figure 4 shows the results of measuring the amount of PGE2 produced in macrophages to confirm the anti-inflammatory effect of pseudo-ceramide.

[0025] Hereinafter, the present invention will be described in detail.

[0026]

[0027] One aspect of the present invention is a compound represented by the following chemical formula 1.

[0028] [Chemical Formula 1]

[0029]

[0030] Here, A * is an acyl group of a saturated or unsaturated aliphatic chain having 7 to 16 carbon atoms, which is hydrogen, or unsubstituted or substituted with a hydroxy group,

[0031] R1 may be a saturated or unsaturated aliphatic chain of 7 to 15 carbon atoms, unsubstituted or substituted with a hydroxy group, provided that A * The difference in the number of carbon atoms between R1 and R2 is preferably at least 7, and more preferably at least 8. The range of carbon atoms or A * Depending on the difference in the number of carbon atoms between R1 and R2, there may be differences in physical or chemical properties such as solubility and skin migration, or in pharmacological effects such as moisturizing, skin barrier improvement, production of natural moisturizing factors such as hyaluronic acid and filaggrin, and anti-inflammation.

[0032] More specifically, the compound of formula 1 may be a compound represented by formula 2 or formula 3.

[0033] [Chemical Formula 2]

[0034]

[0035] [Chemical Formula 3]

[0036]

[0037] Here, R1 and R2 are each independently a saturated or unsaturated aliphatic chain having 3 to 30 carbon atoms, which is unsubstituted or substituted with a hydroxy group.

[0038]

[0039] Another aspect of the present invention is a composition for external application to the skin containing the compounds of the above-described chemical formulas 1 to 3 as an active ingredient. Here, the composition includes not only the compound represented by the chemical formula 1 itself, but also all pharmaceutically acceptable salts, hydrates, or solvates thereof. That is, unless otherwise specified, the term "compound according to the present invention" or "compound of the chemical formula 1" is used as a concept encompassing the compound itself, its pharmaceutically acceptable salts, hydrates, solvates, isomers, and prodrugs.

[0040] Non-limiting examples of the pharmaceutically acceptable salts include those formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, Acid addition salts formed with organic acids such as tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid can be mentioned.

[0041] Furthermore, as will be described later, the compound according to the present invention is non-toxic, promotes bioactivity, and enhances skin elasticity and moisturizing effects. In addition, it exhibits the effect of increasing the production of hyaluronic acid and filaggrin, which are known to play an important role in skin moisturizing by forming natural moisturizing factors. Based on these effects, it is suitable for use in skin moisturizing external skin compositions and skin moisturizing cosmetics.

[0042] In the composition of the present invention, the effective ingredient may be contained in an amount of 0.01 wt% to 20 wt% based on the total weight of the composition. Specifically, it may be 0.1 wt% to 10 wt%, and more specifically, 0.5 wt% to 5 wt%. When the effective ingredient is contained in the above range, it is not only appropriate for exhibiting the intended effects of the present invention, but also satisfies both the stability and solubility of the composition, and it may also be appropriate to include it in the above range from the perspective of cost effectiveness.

[0043] The topical skin preparation or cosmetic composition according to the present invention may be provided in any formulation suitable for topical application. For example, the composition may be provided in the form of a solution, an emulsion obtained by dispersing an oil phase in an aqueous phase, an emulsion obtained by dispersing an aqueous phase in an oil phase, a suspension, a solid, a gel, a powder, a paste, a foam, or an aerosol composition. Such compositions may be prepared according to conventional methods in the art.

[0044] The skin external preparation or cosmetic composition according to the present invention may, in addition to the above-mentioned substances, contain other ingredients that preferably have a synergistic effect on the main effect, within a range that does not impair the main effect. Specifically, the cosmetic composition according to the present invention may further contain arbutin or an ascorbic acid derivative, which can provide a skin whitening effect. In addition, the cosmetic composition according to the present invention may further contain a moisturizer, an emollient, a surfactant, an ultraviolet absorber, a preservative, a bactericide, an antioxidant, a pH adjuster, an organic and inorganic pigment, a fragrance, a cooling agent, or an antiperspirant.

[0045]

[0046]

[0047] The present invention will be described in more detail below using examples. However, it should be made clear that the following examples are intended only to provide a more detailed explanation of the invention and are not intended to limit the scope of the rights herein.

[0048]

[0049] Example

[0050] Example 1: Synthesis of N-(1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)octanamide [Pseudo Sphingosine-1]

[0051]

[0052] Dissolve trishydroxymethylaminomethane hydrochloride (5 g) in a mixture of tetrahydrofuran and distilled water (THF 40 ml + H_2 O 15 ml), add magnesium oxide (6.55 g), and stir at a temperature below 20°C. Dissolve octanoyl chloride (99%) (5.675 g) in 20 ml of tetrahydrofuran, add slowly dropwise over 8 minutes, and react at 20°C for 2 hours. After confirming that all reactants have reacted by TLC, add an additional 75 ml of a mixture of tetrahydrofuran and distilled water (4:1) and stir to sufficiently dissolve the product. After removing magnesium oxide through a filter, work up twice with distilled water (100 ml). The aqueous layer was separated, the organic layer was dried with anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and then recrystallized with EA and hexane to obtain 8.66 g (81.12% yield) of a white solid.

[0053]

[0054] Example 2: Synthesis of N-(1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)palmitamide [Pseudo Sphingosine-2]

[0055]

[0056] Dissolve trishydroxymethylaminomethane hydrochloride (5 g) in a mixture of tetrahydrofuran and distilled water (THF 40 ml + H_2 O 15 ml), add magnesium oxide (6.55 g), and stir at a temperature below 20°C. Dissolve palmitoyl chloride (98%) (9.827 g) in 20 ml of tetrahydrofuran, add slowly dropwise over 8 minutes, and react at 20°C for 2 hours. After confirming that all reactants have reacted by TLC, add an additional 75 ml of a mixture of tetrahydrofuran and distilled water (4:1) and stir to sufficiently dissolve the product. After removing magnesium oxide through a filter, work-up twice with distilled water (100 ml). The aqueous layer was separated, the organic layer was dried with anhydrous magnesium sulfate, filtered, concentrated under reduced pressure, and recrystallized with chloroform to obtain 10.10 g (80.42% yield) of a white solid.

[0057]

[0058] Example 3: Synthesis of 3-hydroxy-2-(hydroxymethyl)-2-octanamidopropyl palmitate [Sphingosine Ceramide-1]

[0059]

[0060] Take N-(1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)octanamide (1 g) and dissolve it cleanly in tetrahydrofuran (20 ml) and stir at below 20℃. Then, add triethylamine (98%) (0.549 ml) dropwise and stir for 15 minutes. Dissolve palmitoyl chloride (98%) (1.057 g) in 5 ml of tetrahydrofuran and slowly add dropwise over 2 minutes and react at 20℃ for 3 hours. After confirming that all reactants have reacted by TLC, filter and remove the produced TEA·HCl salt. After evaporating the tetrahydrofuran organic layer completely using a rotary evaporator, dissolve the product in 30 ml of ethyl acetate. After working up twice with distilled water (30 ml), dry the organic solution layer over anhydrous magnesium sulfate and filter. 1.61 g (78.2% yield) of product was obtained through recrystallization from EA:hexane at low temperature.

[0061]

[0062] Example 4: Synthesis of 3-hydroxy-2-(hydroxymethyl)-2-palmitamidopropyl octanoate [Sphingosine Ceramide-2]

[0063]

[0064] Take N-(1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)palmitamide (1 g) and dissolve it cleanly in tetrahydrofuran (28 ml) and stir at below 20°C. Then, add triethylamine (98%) (0.549 ml) dropwise and stir for 15 minutes. Dissolve octanoyl chloride (99%) (0.537 g) in 5 ml of tetrahydrofuran and slowly add dropwise over 2 minutes and react at 20°C for 3 hours. After confirming that all reactants have reacted by TLC, filter and remove the produced TEA·HCl salt. After evaporating the tetrahydrofuran organic layer completely using a rotary evaporator, dissolve the product in 30 ml of chloroform. After working up twice with distilled water (30 ml), dry the organic solution layer over anhydrous magnesium sulfate and filter. 2.267 g (81.31% yield) of the product was obtained through recrystallization from chloroform at low temperature.

[0065]

[0066] Comparative example (control group)

[0067] - Ceramide NP: ceramide 3

[0068] - D-Sphingosine

[0069] - Probarrier ceramide: Hydroxypropyl Bispalmitamide MEA (PC104)

[0070] - CBO skin barrier enhancer: Oleamide serino

[0071] - NAG: N-acetylglucosamine

[0072] - Dexamethasone

[0073]

[0074] Experimental example

[0075] In order to determine the benefits and effects of the novel ceramide analogue compared to Ceramide NP, a natural ceramide, cytotoxicity, moisturizing factor (Hyaluronic acid) production effect, and Filaggrin production amount were evaluated.

[0076]

[0077] Experimental Example 1: Cytotoxicity Evaluation of Novel Ceramide-like Agents

[0078] We conducted a cytotoxicity evaluation of the ceramides described in the previous examples. Ceramide NP and D-Sphingosine were used as comparison groups. EZ-Cytox measures the extent to which WST, a highly sensitive water-soluble tetrazolium salt, reacts with dehydrogenase in living cells to produce orange, water-soluble formazan, thereby determining the number of living cells. Dehydrogenase, which reacts with WST, is an enzyme present in the mitochondrial electron transport chain of metabolically active cells and is only effective in living cells. Therefore, formazan production is linearly correlated with the number of living cells, which can be determined by measuring absorbance (450 nm). The measurement method is as follows: cells are seeded on plates at a certain concentration using DMEM medium, cultured for 24 hours, and then treated with test substances, followed by 24 hours of culture. After adding EZ-Cytox (10% of the medium volume), incubate at 37°C for 1 hour, transfer 100 μl of the reacted medium to a 96-well plate, and measure the absorbance at 450 nm.

[0079] Cell viability (%) Control 100.00 Ceramide NP 103.31 D-Sphingosine 10.82 Pseudo Sphingosine-1 (Example 1) 106.10 Pseudo Sphingosine-2 (Example 2) 104.41 Sphingosine Ceramide-1 (Example 3) 105.04 Sphingosine Ceramide-2 (Example 4) 104.55

[0080] The results were as shown in Table 1 and Figure 1.

[0081] Based on the above results, additional experiments were conducted at a concentration of 20 μg / mL for the remaining substances, excluding D-sphingosine 20 μg / mL, which exhibited cytotoxicity.

[0082]

[0083] Experimental Example 2: Evaluation of the moisturizing factor (hyaluronic acid (HA) production effect) of a novel ceramide-like substance.

[0084] The hyaluronic acid production effect of the similar ceramide described in the previous examples was evaluated. HA was measured in a keratinocyte culture using DMEM medium. The measurement method was HaCaT 1.0 × 10 5 After seeding cells / ml in a 24-well plate using DMEM medium, culture for 24 hours and then treating with new medium containing the test substance and culture for 24 hours. Obtain the supernatant of the cultured cells and measure the amount of HA (using the Hyaluronan Quantikine ELISA kit (R&D System, USA) according to the protocol).

[0085] * Positive control: N-Acetylglucosamine (10 mM) (polymerizes with glucuronic acid on the cell surface to form HA)

[0086] The results were as shown in Fig. 2.

[0087] As can be seen from the results shown in Fig. 2, the example compounds showed a significantly higher hyaluronic acid production effect than the comparative group Ceramide NP, and Sphingosine Ceramide-1 and Sphingosine Ceramide-2 showed an even higher production effect.

[0088]

[0089] Experimental Example 3: Evaluation of Filaggrin Production in Human Keratinocytes (HaCaT) by Novel Ceramide-like Agents

[0090] The Filaggrin production amount of the pseudo-ceramide described in the previous examples was evaluated.

[0091] FLG exists in the stratum corneum of the epidermis and accounts for most of the intercellular lipids. It is known to play an important role in skin hydration by forming natural moisturizing factors through the decomposition process. The amount of FLG produced, which is involved in skin barrier formation, is measured within keratinocytes. The measurement method is HaCaT 1.0 × 10 5 After seeding cells / ml in a 24-well plate using DMEM medium, culture for 24 hours and then treating with new medium containing the test substance and culture for 24 hours. Lysate of cultured cells is obtained and the FLG content in the cells is measured (using Human Filaggrin (FLG) ELISA kit (CUSABIO, USA) according to the protocol).

[0092] The results were as shown in Fig. 3.

[0093] As can be seen from the results shown in Fig. 3, the example compounds showed an effect of increasing the amount of Filaggrin produced, except for Pseudo Sphingosine-1, compared to the comparative group Ceramide NP, and Pseudo Sphingosine-2 showed an effect of increasing the amount of Filaggrin produced by approximately 31%.

[0094]

[0095] Experimental Example 4: Evaluation of the inhibition of PGE2 production in macrophages (RAW264.7 cells) by a novel ceramide-like substance.

[0096] The inhibition of PGE2 production by similar ceramides described in the previous examples was evaluated.

[0097] The anti-inflammatory effect is confirmed by measuring the excessive PGE2 produced by LPS. The confirmation method is as follows: RAW264.7 cells are seeded at 1.5 × 105 cells / well in a 24-well plate, cultured for 24 hours, and then treated simultaneously with a medium containing the test substance and LPS (1 μg / ml) and cultured for 24 hours. The PGE2 content in the cell culture is measured (Positive control: Dexamethasone 5 μM) using a PGE2 Parameter assay kit (R&D System, USA).

[0098] The results were as shown in Fig. 4.

[0099] As can be seen from the results shown in Figure 4, the example compounds showed a PGE2 production reduction effect, except for Pseudo Sphingosine-1, compared to the comparative group Ceramide NP.

Claims

1. A compound represented by the following chemical formula 1. [Chemical Formula 1] Here, A * is an acyl group of a saturated or unsaturated aliphatic chain having 7 to 16 carbon atoms, which is hydrogen, or unsubstituted or substituted with a hydroxy group, R1 is a saturated or unsaturated aliphatic chain of 7 to 15 carbon atoms, which is unsubstituted or substituted with a hydroxy group, A * The difference in the number of carbon atoms between R1 and R2 is at least 7.

2. In claim 1, a compound represented by the following chemical formula 2. [Chemical Formula 2] Here, R1 is a saturated or unsaturated aliphatic chain having 7 to 15 carbon atoms, which is unsubstituted or substituted with a hydroxy group.

3. In claim 1, a compound represented by the following chemical formula 3. [Chemical Formula 3] Here, R1 and R2 are each independently a saturated or unsaturated aliphatic chain having 7 to 15 carbon atoms, which is unsubstituted or substituted with a hydroxy group.

4. In claim 2, a compound represented by the following chemical formula 4 or chemical formula 5. [Chemical Formula 4] [Chemical Formula 5] 5. In claim 3, a compound represented by the following chemical formula 6 or chemical formula 7. [Chemical Formula 6] [Chemical Formula 7] 6. A composition for external application to the skin containing the compound of claims 1 to 5 as an active ingredient.

7. A composition for external application to the skin, wherein the effective ingredient is contained in an amount of 0.01 to 20 wt% based on the total weight of the composition, in claim 6.

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