Novel ceramides, their preparation and use

Abstract

The present invention belongs to the field of biomedical technology and discloses a novel ceramide having the structure of Formula I: The present invention further discloses a method for preparing and using ceramides. The present invention uses naturally occurring, readily available 10-hydroxycarboxylic acids as starting materials to construct ceramides with novel structures. The synthetic route is simple and convenient, and they are suitable for large-scale production. The resulting ceramides exhibit excellent effects in tissue repair, anti-inflammatory repair, tissue healing, moisturizing, and anti-aging, and can be used in the fields of supplements, cosmetics, and pharmaceuticals.

Description

[Technical Field]

[0001] The present invention belongs to the field of biomedical technology, and specifically relates to a novel ceramide, its preparation method and use. [Background technology]

[0002] Ceramides (also known as molecular nails) are naturally present in the skin and are a very important component of the skin barrier (stratum corneum), accounting for 40-50% by weight. From a chemical structural perspective, ceramides are sphingosine lipids composed of a long-chain sphingosine base and a fatty acid. Because the carbon chain length, degree of unsaturation, and number of hydroxyl groups in the sphingosine and fatty acid moieties are all variable, a ceramide molecule refers to a group of compounds rather than a single compound.

[0003] Ceramides are widely distributed, especially in the skin. Currently, nine types of natural ceramides are known, and various types of ceramides are used in skin care products. Ceramides have excellent properties in regulating the skin barrier function, restoring skin moisture, and strengthening adhesion between skin keratinocytes.

[0004] A decrease in ceramide levels leads to dry skin, a loss of skin surface defenses, and increased susceptibility to foreign substances, leading to secondary skin infections and skin rejection. Specifically, invaders induce the release of cytokines from surface cells such as keratinocytes, lanocytes, and melanocytes, causing inflammation. Therefore, moisturizing the skin is important for maintaining and improving the skin barrier. Compared to conventional moisturizers, physiological lipid mixtures containing ceramide compounds can promote the recovery of damaged skin barrier function. Clinical trials have shown that they are similar in efficacy to moderate or severe topical steroid preparations in improving symptoms in patients with atopic dermatitis.

[0005] Due to the importance of ceramides, many cosmetic and pharmaceutical companies are researching and developing corresponding products. However, natural ceramides are difficult to extract, and are expensive and unsuitable for commercialization, making it difficult to produce natural ceramides on a large scale. Therefore, some companies are working on developing ceramides that are structurally similar to ceramides present in the skin and can provide similar functional effects.

[0006] Considering the wide demand for functional ceramides in the market, it is necessary to solve the problem of their shortage and enhance their efficacy by rapidly synthesizing ceramides using short-chain fragments that are naturally derived and easily available. Summary of the Invention

[0007] An object of the present invention is to provide a ceramide having a novel structure.

[0008] Another object of the present invention is to provide a method for synthesizing ceramide using easily available 10-hydroxydecanoic acid as a starting material.

[0009] Another object of the present invention is to provide uses of ceramides.

[0010] In order to achieve one of the above objectives, the present invention adopts the following technical solutions. A ceramide having the structure of Formula I, or an enantiomer or diastereomer thereof. JPEG0007818877000001.jpg32170

[0011] The method for synthesizing ceramide includes the following steps: JPEG0007818877000002.jpg80170Compound S1 is reacted with TBDPSCl to obtain compound S2. Compound S2 is reacted with phenyl p-nitrotrifluoroacetate to obtain compound S3. Compound S3 is deprotected to give compound S4. Compound S4 is reacted with linoleoyl chloride to obtain compound S5. Compound S5 is reacted with phytosphingosine to give ceramide.

[0012] Furthermore, the molar ratio of the compound S1 to TBDPSCl is 1:(1 to 3).

[0013] Furthermore, the molar ratio of the compound S2 to phenyl p-nitrotrifluoroacetate is 1:(1 to 3).

[0014] Furthermore, the molar ratio of the compound S4 to linoleoyl chloride is 1:(1 to 3).

[0015] Furthermore, the molar ratio of the compound S5 to phytosphingosine is 1:(1 to 2).

[0016] Ceramides can be used in the fields of cosmetics, supplements or medicines.

[0017] In a first aspect, ceramide has a tissue repair effect, and when the ceramide concentration is 0.0625 mM, the cell activity is 114% or more, and when the ceramide concentration is 0.5 mM, the cell activity is 120% or more.

[0018] In a second embodiment, ceramide has an anti-inflammatory repair effect. The principle is to inhibit the expression of IL-6 cytokine. When the ceramide concentration is 0.1 mM, the inhibition rate of the cytokine IL-6 is 46% or more; when the ceramide concentration is 0.2 mM, the inhibition rate of the cytokine IL-6 is 73% or more; when the ceramide concentration is 0.4 mM, the inhibition rate of the cytokine IL-6 is 86% or more; when the ceramide concentration is 0.8 mM, the inhibition rate of the cytokine IL-6 is 94% or more; and when the ceramide concentration is 1.0 mM, the inhibition rate of the cytokine IL-6 is 97% or more. The anti-inflammatory repair effect is a unique effect discovered by the present inventors as a result of studying the biological activity of ceramide; there have been no reports to date of ceramide having an anti-inflammatory repair effect.

[0019] In a third aspect, ceramide has a tissue healing effect: when the concentration of ceramide is 0.08 mM, the cell healing rate is 74% or more.

[0020] In a fourth aspect, ceramide has a moisturizing effect. The principle behind this is to increase the expression level of AQP3 aquaporin. When the ceramide concentration is 0.4 mM, the expression level of AQP3 aquaporin is 112% or more compared to the solvent control group. When the ceramide concentration is 0.8 mM, the expression level of AQP3 aquaporin is 133% or more compared to the solvent control group. When the ceramide concentration is 1.6 mM, the expression level of AQP3 aquaporin is 147% or more compared to the solvent control group.

[0021] In a fifth aspect, ceramide has an anti-aging effect. The principle is to improve the inhibition rate against elastase. When the ceramide concentration is 0.4 mM, the inhibition rate against elastase is 18% or more, when the ceramide concentration is 0.8 mM, the inhibition rate against elastase is 22% or more, and when the ceramide concentration is 1.0 mM, the inhibition rate against elastase is 25% or more. The anti-aging effect is a unique effect discovered by the present inventors as a result of studying the biological activity of ceramide, and there have been no reports to date that ceramide has an anti-aging effect.

[0022] The present invention provides a composition comprising a ceramide, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient, which has tissue repair, anti-inflammatory repair, tissue healing, moisturizing, or anti-aging effects.

[0023] As used herein, the term "pharmaceutically acceptable salt" refers to a salt of one embodiment of the present invention that is pharmaceutically acceptable and possesses the desired pharmacological activity of the parent compound. Such salts include (1) acid addition salts formed with inorganic or organic acids or (2) salts formed by substitution of an acidic proton present in the parent compound. Examples of inorganic acids include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid. Examples of organic acids include acetic acid, propionic acid, caproic acid, cyclopentylpropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, hydroxysuccinic 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-ethanedisulfonic acid, 2-hydroxyethylsulfonic 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, tert-butylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid.

[0024] As used herein, "hydrate" refers to a compound bound to water, wherein the bond between the compound and the water comprises a non-covalent bond.

[0025] As used herein, the term "solvate" refers to a combination formed by solute molecules or ions with solvent molecules or ions.

[0026] As used herein, the term "isomer" refers to a compound of the present invention or a salt thereof that has the same chemical or molecular formula but different optical or steric properties.

[0027] Unless otherwise specified, the term "compounds of the invention" or "ceramide" includes the compound itself, its pharmaceutically acceptable salts, its hydrates, its solvates, and its isomers.

[0028] The present invention has the following beneficial effects: The present invention constructs a structurally novel ceramide using naturally occurring, commercially available, and inexpensive 10-hydroxycarboxylic acid as a starting material, thereby avoiding the use of commercially difficult raw materials, and the synthetic route is simple and convenient, making it suitable for large-scale production. The resulting ceramide exhibits excellent effects in tissue repair, anti-inflammatory repair, tissue healing, moisturizing, and anti-aging, and is more effective than the known ceramides 3 and 3B. In particular, the unique anti-inflammatory and anti-aging effects have rarely been reported for this type of compound, and it can be used in the fields of supplements, cosmetics, and pharmaceuticals. [Brief explanation of the drawings]

[0029] [Figure 1] 1 is a vertical bar graph showing the test results of the cell proliferation activity of ceramide according to the present invention. [Figure 2] 1 is a vertical bar graph showing the test results of the cell proliferation activity of ceramide 3. [Figure 3] 1 is a vertical bar graph showing the test results of the cell proliferation activity of ceramide 3B. [Figure 4] 1 is a vertical bar graph showing test results of the anti-inflammatory and repair effect of ceramide according to the present invention. [Figure 5] 1 is a vertical bar graph showing the test results of the anti-inflammatory and repair effects of different ceramides in Example 3. [Figure 6] 10 is a vertical bar graph showing the test results of tissue healing ability in Example 4. [Figure 7] 1 is a vertical bar graph showing test results of the moisturizing effect of ceramide according to the present invention. [Figure 8] 1 is a vertical bar graph showing the test results of the moisturizing effect of different ceramides in Example 5. [Figure 9] 1 is a vertical bar graph showing test results of the anti-aging effect of the ceramide according to the present invention. [Figure 10] 1 is a vertical bar graph showing the test results of the anti-aging effects of different ceramides in Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0030] The present invention will be further described below with reference to specific examples.

[0031] All reactions were carried out under a nitrogen atmosphere. Unless otherwise specified, reagents were purchased commercially and used without further purification. Dichloromethane, tetrahydrofuran, pyridine, and N,N-dimethylformamide were all anhydrous solvents used in the experiments. Thin-layer chromatography (TLC) was performed using 60F254 silica gel plates. Qingdao Haiyang silica gel (particle size 0.040–0.063 mm) was used for silica gel column chromatography. UV light (254 nm) or iodine was used for TLC color development. NMR spectra were characterized using a Bruker DPX 400 nuclear magnetic resonance spectrometer. 1 H NMR was performed at 400 MHz using deuterated methanol, deuterated DMSO, or deuterated tetrahydrofuran as the solvent, with tetramethylsilane (TMS) as the internal standard. Chemical shifts are in ppm, and coupling constants are in Hz. 1 In H NMR, δ indicates chemical shift, s indicates single peak, d indicates double peak, t indicates triple peak, q indicates quadruple peak, and m indicates multiple peak.

[0032] EA is ethyl acetate, DCM is dichloromethane, DMF is N,N-dimethylformamide, Py. is pyridine, Im. is imidazole, TBDPSCl is tert-butyldiphenylchlorosilane, and TBAF is 4-n-butylammonium fluoride.

[0033] Example 1 Ceramide synthesis JPEG0007818877000003.jpg161701.0 eq (160 mmol) of S1 and 2.0 eq (320 mmol) of imidazole were dissolved in 100 mL of DMF, cooled in an ice bath, and a mixture of 2.0 eq (320 mmol) of TBDPSCl and 50 mL of DMF was added dropwise. After the addition was complete, the temperature was raised to 40°C and the reaction was continued until TLC detection confirmed that the raw material S1 had completely reacted.

[0034] Workup: Diluted with 400 mL of EA, washed with 250 mL of saturated NH4Cl twice, washed with 300 mL of saturated NaCl twice, dried over anhydrous Na2SO4, filtered and concentrated in vacuo, and the resulting residue was purified by silica gel column to give 79.4 g of product S2. JPEG0007818877000004.jpg231701.0 eq (160 mmol) of S2 and 1.0 eq (160 mmol) of phenyl p-nitrotrifluoroacetate were dissolved in 150 mL of pyridine, and the reaction was continued at room temperature with stirring until TLC detection confirmed that the raw material S2 had completely reacted.

[0035] Work-up: Dilute with 300 mL of EA, wash with 150 mL of diluted hydrochloric acid twice, wash with 300 mL of water twice, and wash with 200 mL of saturated NaCl once. The organic phase was dried with anhydrous NaSO, filtered, and concentrated in vacuo to obtain an oily product, which was purified by silica gel column to obtain 67.4 g of product S3. JPEG0007818877000005.jpg171701.0 eq (124 mmol) of S3 was dissolved in 100 mL of THF and cooled in an ice bath. 1.2 eq (149 mmol) of TBAF was dissolved in 40 mL of THF and added dropwise to the above reaction solution. After the addition was completed, the reaction was continued at room temperature until TLC detection confirmed that the raw material S3 had completely reacted.

[0036] Work-up: Dilute with 200 mL of EA, wash with 100 mL of diluted hydrochloric acid once, wash with 100 mL of saturated NaCl once, add anhydrous NaSO to the organic phase for drying, filter and concentrate under vacuum to obtain an oily product, which was purified by silica gel column to obtain 21.6 g of product S4.

[0037] 1 H NMR(400MHz,Methanol-d4)δ8.34-8.23(m,2H),7.39-7.30(m,2H),3.54(t,J=6.6Hz,2H) ,2.63(t,J=7.4Hz,2H),1.73(t,J=6.8Hz,2H),1.53(t,J=6.8Hz,2H),1.46-1.26(m,10H). JPEG0007818877000006.jpg361701.0 eq (71 mmol) of S4 was dissolved in 200 mL of pyridine, cooled in an ice bath, and 4.0 eq (284 mmol) of linoleoyl chloride was added dropwise. After the addition was completed, the reaction was continued at room temperature until TLC detection confirmed that the raw material S4 had completely reacted.

[0038] Workup: Diluted with 400 mL of DCM, washed with 600 mL of dilute HCl twice, washed with 500 mL of saturated NaCl twice, and dried over anhydrous NaSO. Rotary evaporation gave a black oil, which was purified on a silica gel column to give 30 g of product S5.

[0039] 1 H NMR(400MHz,Methanol-d4)δ8.32-8.25(m,2H),7.37-7.29(m,2H),5.41-5.31(m,4H),3.64(t,J=6.6Hz,2H),2.53(t,J=7.4Hz,2H),2.29(t,J= 7.6Hz,2H),2.26-2.19(m,4H),2.08(q,J=6.9Hz,4H),1.63(t,J=6.8Hz,4H),1.43(t,J=6.8Hz,4H),1.46-1.26(m,20H),0.86(t,J=6.9Hz,3H). JPEG0007818877000007.jpg321701.0 eq (53 mmol) of S5 and 1.0 eq (53 mmol) of phytosphingosine were dissolved in 100 mL of pyridine, heated to 45°C, and the reaction was continued until TLC detection confirmed that the raw material phytosphingosine had completely reacted.

[0040] Workup: Dilute with 200 mL of THF, wash twice with 200 mL of dilute hydrochloric acid, wash twice with 200 mL of saturated NaCl, and dry the organic phase with anhydrous Na2SO4. Rotary evaporation gave the crude product, which was then purified on a silica gel column to give 15 g of product.

[0041] 1 H NMR(400MHz,Chloroform-d)δ6.37(d,J=7.5Hz,1H),5.36(tddt,J=10.7,5.4,3.7,1.6Hz,4H),4.1 5(dt,J=7.2,2.6Hz,1H),4.05(t,J=6.8Hz,2H),3.91(t,J=6.8Hz,2H),3.73(s,2H),3.67-3.51(m,2 H),2.88-2.70(m,3H),2.29(t,J=7.6Hz,2H),2.26-2.19(m,2H),2.05(q,J=6.9Hz,4H),1.80-1.70 (m,1H),1.61(p,J=6.9Hz,6H),1.55-1.44(m,2H),1.39-1.21(m,47H),0.88(td,J=6.9,4.0Hz,6H).

[0042] Example 2 Testing cell proliferation activity by MTT assay: HaCaT cells 1 × 10 4 The cells were seeded into a 96-well plate at a density of 100 cells / well and cultured overnight in an incubator. After 24 hours, the supernatant was removed, and 100 μL of medium containing different concentrations of the sample (product obtained in Example 1) or blank medium was added. After further incubation for 24 hours, the medium was removed, and 100 μL of thiazole blue (MTT) was added to each well. The absorbance at 450 nm was measured, and the cell viability = A 投与ウェル / A ブランクウェル ×100% was calculated.

[0043] The results are shown in Figure 1. At a compound concentration of 0.0625 mM, the cell activity reached 114.7%, and at a concentration of 0.5 mM, the cell activity reached 122.3%, demonstrating significant cell proliferation-promoting effects and tissue repair potential. Furthermore, within the high concentration range of 1 mM, the compound exhibited no cytotoxicity and demonstrated good biological safety.

[0044] The test results of the cell proliferation activity of ceramide 3 and ceramide 3B are shown in Figures 2 and 3. The ceramide and ceramide 3B of the present invention have excellent cell healing and tissue repair capabilities.

[0045] Example 3 Evaluation of anti-inflammatory and repair effects by LPS cell induction method B16 mouse melanoma cells were grown at a density of 1 × 10 4 Cells were seeded at 100 μg / well into a 96-well plate, placed in an incubator, and cultured overnight as adherent cells. After 24 h, the supernatant was discarded, and 100 μL of different concentrations of sample (product obtained in Example 1) diluted with DMEM medium was added. A negative control group contained DMEM medium without sample. Three duplicate wells were prepared for each group, and the wells were incubated in an environment of 5% CO2 and 37°C. Two hours after administration, 10 μg / mL LPS was added to the lipopolysaccharide model group and experimental group, and both were incubated for 24 h. After the reaction was completed, 50 μL of the cell supernatant was sampled, and intracellular IL-6 gene expression was detected using an IL-6 ELISA kit.

[0046] The results are shown in Figure 4. A cellular inflammation model was successfully established using LPS. Expression of IL-6 inflammatory factors in the model cells was 23.14-fold higher than in the control group. The compounds of the present invention have excellent inhibitory effects on IL-6 cytokine expression. At concentrations of 0.1, 0.2, 0.4, 0.8, and 1.00 mm, IL-6 factor expression was 12.37, 6.15, 3.04, 1.31, and 0.65 times higher than in the control group, respectively. Compared to the LPS model, these compounds were inhibited by 46.54%, 73.42%, 86.86%, 94.30%, and 97.19%, respectively. Therefore, the compounds have excellent anti-inflammatory effects and are expected to be effective in repairing inflammatory skin damage.

[0047] Ceramide 3, ceramide 3B, and the ceramide of the present invention were compared at 1 mmol / L. The results are shown in Figure 5. All three ceramides effectively inhibited the expression of the IL-6 inflammatory factor, with the ceramide of the present invention showing the most significant inhibitory effect on the IL-6 inflammatory factor.

[0048] Example 4 Evaluation of tissue healing ability by scratch assay Principle: When cells proliferated and fused into a monolayer, a blank area was created on the fused monolayer using a scratch tool. The cells in the blank area were removed by mechanical force. After culturing for a period of time, the migration status of the cells into the cell-free area was observed, and the migration distance of the cells was measured to evaluate the cell migration ability.

[0049] operation 1. Lines were drawn on the culture plate. First, horizontal lines were drawn on the back of a 6-well plate using a marker pen and a ruler, passing through the wells at equal intervals of approximately 0.5 to 1 cm. At least five lines passed through each well. When drawing the lines, care was taken not to make them too thick.

[0050] 2. Cell plating. Approximately 5 x 10 cells per well. 5 (Different cell numbers were added depending on the cell growth rate.) The seeding principle is that the confluence rate will reach 100% overnight.

[0051] 3. Cell scratching: The next day, a pipette tip was used to scratch the cell layer perpendicular to the cell plane along the line drawn on the back of the plate the previous day (it is advisable to use the same pipette tip for different wells).

[0052] 4. Cell washing. After scratching, cells that were not attached to the wall were washed three times with sterile PBS to remove cells that had been scratched during scratching. After the gap created after scratching became clearly visible, the medium was replaced with fresh serum-free medium.

[0053] 5. Cell culture and observation. Samples (the compound of Example 1, ceramide 3, and ceramide 3B) were diluted with culture medium (concentration: 0.08 mM) and added to cell culture dishes. The cells were cultured in a 37°C, 5 wt% CO2 incubator. After 24 hours, the cells were removed and the width of the scratch was observed and measured under a microscope, photographed, and the healing rate was calculated using ImageJ software.

[0054] The results are shown in Figure 6. The self-healing rate after 24 hours in the solvent control group was 35.74%, while the self-healing rate after 24 hours in the ceramide group was 74.48%, demonstrating a higher tissue healing ability than ceramide 3 and ceramide 3B. The compounds of the present invention significantly improved the cell healing rate and had good skin tissue repair activity.

[0055] Example 5 Evaluation of moisturizing effect using AQP3 cell method Aquaporin 3 (AQP3) is a protein transporter responsible for the transport of water, glycerol, urea, and other substances across the cell membrane and is primarily expressed in keratinocytes and dermal fibroblasts. AQP3 is not only involved in skin hydration and barrier function, but also plays an important role in skin injury, repair, and healing, making it an important factor for maintaining normal skin morphology and function.

[0056] HaCat cells were cultured at a density of 1 x 10 4Cells were seeded at 100 μg / well into a 96-well plate, placed in an incubator, and cultured overnight as adherent cells. After 24 h, the supernatant was discarded, and 100 μL of different concentrations of the sample (product obtained in Example 1) diluted with DMEM medium was added. A negative control group contained DMEM medium without sample. Three duplicate wells were prepared for each group, and the wells were incubated in an environment of 5% CO2 and 37°C. Two hours after administration, 10 μg / mL of LPS was added to the lipopolysaccharide model group and experimental group, and both were incubated for 24 h. After the reaction was completed, 50 μL of the cell supernatant was sampled, and intracellular AQP3 gene expression was detected using an AQP3 kit.

[0057] The results are shown in Figure 7. The compounds of the present invention effectively increased the expression level of AQP3 aquaporin in a concentration-dependent manner. At concentrations of 0.4, 0.8, and 1.6 mM, the expression levels were 1.12, 1.33, and 1.47 times higher than those of the solvent control group, respectively.

[0058] The results of comparing 2.5 mmol / L ceramide 3, ceramide 3B, and the ceramide of the present invention are shown in Figure 8. These three ceramides all increased the expression level of AQP3 aquaporin to different levels, with the ceramide of the present invention showing the most pronounced effect and the highest moisturizing performance.

[0059] Example 6 Evaluation of anti-aging effects using elastase inhibition method 2 mL of 2 mg / mL elastase solution was taken, and samples of different concentrations were added, vortexed thoroughly, and shaken at 37°C and 400 r / min on a shaker for 20 minutes. Immediately, 5 mL of 0.5 mol / L phosphate buffer solution at pH 6.0 was added, and vortexed uniformly. An appropriate amount of the mixture was placed in a 2 mL centrifuge tube and centrifuged at 9391 × g for 10 minutes. 200 μL of the supernatant was precisely aspirated and transferred to a 96-well plate. The absorbance at a wavelength of 495 nm was measured using a microplate reader, and a spectral scan was performed from 400 to 800 nm.

[0060] The substrate + enzyme solution served as the blank control group, the substrate + enzyme + sample (product obtained in Example 1) solution served as the enzyme inhibition group, and the substrate + sample (without enzyme) solution served as the background. Three replicate wells were prepared for each group. Inhibition rate (%) = [1-(An-An') / (A0-A0')] × 100%. In the formula, A0 is the absorbance when the enzyme was added but not the sample, A0' is the absorbance when only the substrate was added without the sample or enzyme, An is the absorbance when only the sample solution was added, and An' is the absorbance when the sample was added but not the enzyme. When An' > An, it indicated a promotion effect, and the promotion rate (%) = [1-(An'-An) / (A0-A0')] × 100%.

[0061] The results are shown in Figure 9. The compounds of the present invention effectively improve the elastase inhibition rate, which shows concentration dependency. At concentrations of 0.4, 0.8, and 1.0 mM, the inhibition rates are 18.21%, 22.07%, and 25.37%, respectively, indicating that they can inhibit the expression of elastase, protect elastin, and maintain or improve skin elasticity.

[0062] The results of comparing 1 mmol / L ceramide 3, ceramide 3B, and the ceramide of the present invention are shown in Figure 10. The effect of the ceramide of the present invention is far greater than that of ceramide 3 and ceramide 3B, and it exhibits the highest elastase inhibitory effect.

[0063] In the present invention, the effects of ceramides on cell repair, proliferation, anti-inflammation, moisturizing, and elastase inhibition were comprehensively tested. As a result, compared to commercially available ceramide 3 and ceramide 3B, the ceramide of the present invention showed the best performance in all indicators, especially in anti-inflammation, tissue healing, and elastase inhibition.

[0064] The above are only specific embodiments of the present invention, and the scope of protection of the present invention is not limited thereto. Any modifications or replacements that can be easily thought of by those skilled in the art within the technical scope described in the present invention should be included in the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be based on the scope of protection of the claims.

Claims

1. A ceramide having the structure of Formula I.

2. 10. A method for synthesizing ceramide according to claim 1, comprising the steps of: Compound S1 is reacted with TBDPSCl to obtain compound S2; Compound S2 is reacted with phenyl p-nitrotrifluoroacetate to obtain compound S3; Deprotection of compound S3 to obtain compound S4; Compound S4 is reacted with linoleoyl chloride to obtain compound S5, A synthesis method comprising reacting compound S5 with phytosphingosine to obtain ceramide.

3. The method of claim 2, wherein the molar ratio of the compound S1 to TBDPSCl is 1:(1-3), the molar ratio of the compound S2 to phenyl p-nitrotrifluoroacetate is 1:(1-3), the molar ratio of the compound S4 to linoleoyl chloride is 1:(1-3), and the molar ratio of the compound S5 to phytosphingosine is 1:(1-2).

4. A supplement composition or pharmaceutical composition containing the ceramide described in claim 1 as an active ingredient.

5. The ceramide has a tissue repair effect, and when the concentration of the ceramide is 0.0625 mM, the cell activity is 114% or more, and / or when the concentration of the ceramide is 0.5 mM, the cell activity is 120% or more, Here, the cell activity refers to cell viability, i.e., the ratio of the absorbance of the cell culture medium to which the sample has been added to the absorbance of the cell culture medium to which the sample has not been added, as described in claim 4.

6. the ceramide has an anti-inflammatory repair effect, and when the concentration of the ceramide is 0.1 mM, the inhibition rate against cytokine IL-6 is 46% or more, and / or when the concentration of the ceramide is 0.2 mM, the inhibition rate against cytokine IL-6 is 73% or more, and / or when the concentration of the ceramide is 0.4 mM, the inhibition rate against cytokine IL-6 is 86% or more, and / or when the concentration of the ceramide is 0.8 mM, the inhibition rate against cytokine IL-6 is 94% or more, and / or when the concentration of the ceramide is 1.0 mM, the inhibition rate against cytokine IL-6 is 97% or more, The supplement composition or pharmaceutical composition according to claim 4, wherein the inhibition rate against the cytokine IL-6 = 1 - (expression level of IL-6 gene in experimental group - expression level of IL-6 gene in blank group) / (expression level of IL-6 gene in model group - expression level of IL-6 gene in blank group) x 100%.

7. The ceramide has a tissue healing effect, and when the concentration of the ceramide is 0.08 mM, the cell healing rate is 74% or more; The supplement composition or pharmaceutical composition of claim 4, wherein the cell healing rate is calculated as follows in a scratch injury experiment: (initial scratch injury area of ​​cells - scratch injury area after culturing cells and sample for 24 hours) / initial scratch injury area of ​​cells x 100%.

8. The ceramide has a moisturizing effect, and when the concentration of the ceramide is 0.4 mM, the expression level of AQP3 aquaporin is 112% or more compared to the solvent control group, when the concentration of the ceramide is 0.8 mM, the expression level of AQP3 aquaporin is 133% or more compared to the solvent control group, and when the concentration of the ceramide is 1.6 mM, the expression level of AQP3 aquaporin is 147% or more compared to the solvent control group, Here, the expression level of the AQP3 aquaporin refers to the actual abundance or relative content of aquaporin 3 protein or its encoding gene AQP3 mRNA in cells or tissues, as described in claim 4.

9. The ceramide has an anti-aging effect, and when the concentration of the ceramide is 0.4 mM, the inhibition rate against elastase is 18% or more, when the concentration of the ceramide is 0.8 mM, the inhibition rate against elastase is 22% or more, and when the concentration of the ceramide is 1.0 mM, the inhibition rate against elastase is 25% or more, The elastase inhibition rate refers to the degree to which a sample inhibits elastase activity, and is equal to [1-(An-An') / (A0-A0')] x 100%, where A0 is the absorbance when elastase is added but no sample is added, A0' is the absorbance when only substrate is added but no sample or elastase is added, An is the absorbance when only sample solution is added, and An' is the absorbance when sample is added but no elastase is added. The supplement composition or pharmaceutical composition of claim 4.

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