Ceramide derived from linolenic acid and its preparation method and application

Abstract

The present invention belongs to the field of biomedicine and discloses a ceramide derived from linolenic acid, which has a structure of general formula I or an isomer of general formula I: #imgabs0# wherein R 1 Selected from #imgabs1#R 2 Selected from one of the following structures: #imgabs2# The present invention also discloses its preparation method and use. The present invention reacts long-chain linolenic acid with sphingosine and phytosphingosine to obtain a class of novel ceramide compounds, which have a significant inhibitory effect on tyrosinase and can effectively prevent the formation of melanin, thereby having a good skin whitening effect and can be used in the fields of cosmetics.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to ceramide derived from linolenic acid and a preparation method and application thereof. Background Art

[0002] Ceramide is a compound composed of a sphingosine molecule and a fatty acid molecule, in which the carbon chain length, unsaturation and number of hydroxyl groups of the sphingosine and fatty acid parts can vary. Ceramides are involved in a variety of physiological functions, including apoptosis, cell growth arrest, differentiation, cell senescence, cell migration and adhesion. The role of ceramide and its downstream metabolites have also been mentioned in several pathological conditions, including cancer, neurodegenerative diseases, diabetes, microbial pathology, obesity and inflammation.

[0003] Ceramide is the main component of lipids in the extracellular matrix of the human skin epidermis. Together with cholesterol and saturated fatty acids, ceramide creates an impermeable protective structure to prevent excessive water evaporation and also prevent the entry of microorganisms. Currently, 15 types of ceramides have been detected in the skin, which can be roughly divided into three categories: short-chain ceramides, long-chain ceramides, and ceramides that bind to keratinocytes.

[0004] Due to the importance of ceramides, there is a widespread demand for functional ceramides in the market. However, the extraction and purification of natural ceramides are economically problematic. Therefore, it is very necessary to quickly construct new ceramides through chemical synthesis and explore the functional uses and efficacy differences of the ceramides. Summary of the invention

[0005] The object of the present invention is to provide a class of ceramides with novel structures, namely ceramides derived from linolenic acid.

[0006] Another object of the present invention is to provide a method for synthesizing ceramide derived from linolenic acid.

[0007] Another object of the present invention is to provide the use of ceramide derived from linolenic acid.

[0008] In order to achieve one of the above purposes, the present invention adopts the following technical solution:

[0009] In the first aspect of the present invention, the ceramide derived from linolenic acid has a structure of the general formula I or an isomer of the general formula I:

[0010]

[0011] Among them, R 1 Selected from R 2 Select one of the following structures:

[0012]

[0013] Furthermore, R 2 Select from one of the following structures: Isomers include enantiomers, diastereomers, and cis-trans isomers.

[0014] Furthermore, the linolenic acid-derived ceramide is selected from

[0015]

[0016]

[0017] Further, the linolenic acid-derived ceramide is gamma-linolenic acid sphingosine.

[0018] The second aspect of the present invention is a method for preparing ceramide derived from linolenic acid, comprising the following steps:

[0019]

[0020] Compound S1 reacts with p-nitrobenzenesulfonyl chloride and an organic base to obtain compound S2;

[0021] Compound S2 reacts with a sphingoid base to give compound I.

[0022] Furthermore, the molar ratio of the compound S1, p-nitrobenzenesulfonyl chloride, organic base and sphingosine base is (1-2):(1-2):(2-6):1.

[0023] Furthermore, the organic base is triethylamine.

[0024] Furthermore, the solvent of the reaction is ethyl acetate.

[0025] The third aspect of the present invention is the use of α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine in cosmetics and medicines.

[0026] Specifically, α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine, or γ-linolenic acid phytosphingosine has a whitening effect.

[0027] Specifically, α-linolenic acid sphingosine inhibits tyrosinase, and the inhibition rate of tyrosinase is more than 5% at a concentration of 1mM, and / or the inhibition rate of tyrosinase is more than 8% at a concentration of 2mM, and / or the inhibition rate of tyrosinase is more than 10% at a concentration of 3mM, and / or the inhibition rate of tyrosinase is more than 13% at a concentration of 4mM, and / or the inhibition rate of tyrosinase is more than 16% at a concentration of 5mM, and / or the inhibition rate of tyrosinase is more than 18% at a concentration of 6mM.

[0028] Specifically, α-linolenic acid phytosphingosine inhibits tyrosinase, and the inhibition rate of tyrosinase is more than 7% at a concentration of 1mM, and / or the inhibition rate of tyrosinase is more than 9% at a concentration of 2mM, and / or the inhibition rate of tyrosinase is more than 9% at a concentration of 3mM, and / or the inhibition rate of tyrosinase is more than 12% at a concentration of 4mM, and / or the inhibition rate of tyrosinase is more than 11% at a concentration of 5mM, and / or the inhibition rate of tyrosinase is more than 14% at a concentration of 6mM.

[0029] Specifically, γ-linolenic acid sphingosine inhibits tyrosinase, and the inhibition rate of tyrosinase is more than 27% at a concentration of 1mM, and / or the inhibition rate of tyrosinase is more than 33% at a concentration of 2mM, and / or the inhibition rate of tyrosinase is more than 40% at a concentration of 3mM, and / or the inhibition rate of tyrosinase is more than 35% at a concentration of 4mM, and / or the inhibition rate of tyrosinase is more than 33% at a concentration of 5mM, and / or the inhibition rate of tyrosinase is more than 33% at a concentration of 6mM.

[0030] Specifically, γ-linolenic acid phytosphingosine inhibits tyrosinase, with an inhibition rate of more than 6% at a concentration of 1mM, and / or an inhibition rate of more than 12% at a concentration of 2mM, and / or an inhibition rate of more than 18% at a concentration of 3mM, and / or an inhibition rate of more than 24% at a concentration of 4mM, and / or an inhibition rate of more than 31% at a concentration of 5mM, and / or an inhibition rate of more than 35% at a concentration of 6mM.

[0031] Specifically, α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine have anti-aging effects.

[0032] Specifically, α-linolenic acid sphingosine inhibits elastase, and the inhibition rate of tyrosinase is more than 10% at a concentration of 3 mg / L, and / or the inhibition rate of tyrosinase is more than 15% at a concentration of 6 mg / L, and / or the inhibition rate of tyrosinase is more than 16% at a concentration of 9 mg / L, and / or the inhibition rate of tyrosinase is more than 23% at a concentration of 15 mg / L, and / or the inhibition rate of tyrosinase is more than 24% at a concentration of 18 mg / L.

[0033] Specifically, α-linolenic acid phytosphingosine inhibits elastase, and the inhibition rate of tyrosinase is more than 7% at a concentration of 3 mg / L, and / or the inhibition rate of tyrosinase is more than 10% at a concentration of 9 mg / L, and / or the inhibition rate of tyrosinase is more than 11% at a concentration of 12 mg / L, and / or the inhibition rate of tyrosinase is more than 13% at a concentration of 18 mg / L.

[0034] Specifically, γ-linolenic acid sphingosine inhibits elastase, and the inhibition rate of tyrosinase is more than 24% at a concentration of 3 mg / L, and / or the inhibition rate of tyrosinase is more than 16% at a concentration of 6 mg / L, and / or the inhibition rate of tyrosinase is more than 16% at a concentration of 9 mg / L, and / or the inhibition rate of tyrosinase is more than 12% at a concentration of 15 mg / L, and / or the inhibition rate of tyrosinase is more than 23% at a concentration of 18 mg / L.

[0035] Specifically, γ-linolenic acid phytosphingosine inhibits elastase, and the inhibition rate of tyrosinase is more than 8% at a concentration of 3 mg / L, and / or the inhibition rate of tyrosinase is more than 9% at a concentration of 6 mg / L, and / or the inhibition rate of tyrosinase is more than 13% at a concentration of 9 mg / L, and / or the inhibition rate of tyrosinase is more than 14% at a concentration of 12 mg / L, and / or the inhibition rate of tyrosinase is more than 17% at a concentration of 18 mg / L.

[0036] α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine has at least one of the following effects: tissue repair, anti-inflammation, and tissue healing.

[0037] Specifically, α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine inhibit the production of interstitial collagenase MMP1 and have anti-photoaging effects.

[0038] A composition comprises at least one of α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine, wherein the composition has at least one of whitening, anti-aging, anti-photoaging, barrier, repair and anti-inflammatory effects.

[0039] The composition contains acceptable excipients, including one or more of solubilizers, preservatives, antioxidants, pH regulators, penetration enhancers, liposomes, moisturizers, thickeners, chelating agents, skin feel regulators, surfactants, emulsifiers, flavors and pigments; the composition is in the form of cream, emulsion, solution, film, aerosol or spray.

[0040] α-linolenic acid is one of the essential nutrients for people. It has the functions of enhancing intelligence, improving memory, protecting eyesight and improving sleep. α-linolenic acid is stronger and safer than DHA and other nutrients. α-linolenic acid can be converted into DHA, DPA, EPA, etc. in the body, while supplementing DHA and other nutrients can only play a partial role. α-linolenic acid is only a part of its function as a source of energy for growth, cell metabolism and muscle movement. It is more of a structural substance and metabolic regulatory substance, playing a structural and regulatory function. γ-linolenic acid is one of the essential polyunsaturated fatty acids for the human body. It is a precursor substance for the synthesis of human prostaglandins and can be used in the fields of medicine and food.

[0041] The present invention has the following beneficial effects:

[0042] The present invention reacts long-chain linolenic acid with sphingosine and phytosphingosine to obtain a class of ceramide compounds with novel structures. The compounds have a significant inhibitory effect on tyrosinase and can effectively prevent the production of melanin, thereby having a good skin whitening effect. The prior art has not systematically studied the whitening effect of ceramide derived from linolenic acid. The present invention has experimentally discovered and verified that ceramide derived from linolenic acid has a high inhibition rate on tyrosinase, can reduce melanin production, prevent melanin deposition, and has a whitening effect; the present invention also finds that ceramide derived from linolenic acid has the effect of inhibiting elastase, can reduce elastin degradation, and promotes skin to be firmer and elastic, has a more excellent anti-aging effect, and can be used in the fields of cosmetics, medicines, etc. In addition, ceramide derived from linolenic acid also has tissue repair, anti-inflammatory, and tissue healing effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] Figures 1 to 4 is a bar graph of the tyrosinase inhibition rate of ceramide in Example 2;

[0044] Figures 5-6 This is a bar graph of the cell proliferation activity detection results in Example 3;

[0045] Figures 7-8This is a bar graph of the anti-inflammatory and repair efficacy test results of Example 4;

[0046] Fig. 9 The healing ability test results of Example 5 are shown in FIG.

[0047] Figures 10 to 13 This is a bar graph of the elastase inhibition rate of Example 6;

[0048] Fig.14 This is a bar graph of MMP1 expression levels in Example 7. DETAILED DESCRIPTION

[0049] The present invention will be further described below in conjunction with specific embodiments.

[0050] Unless otherwise stated, chemicals were purchased from commercial products and were not further purified. Ethyl acetate was used as an anhydrous solvent in the experiments. Thin layer chromatography (TLC) used 60F254 silica gel plates. Silica gel column chromatography used Qingdao Ocean Silica Gel (particle size 0.040-0.063 mm). TLC color development used UV light (254 nm) or iodine. NMR spectra were characterized using a Bruker DPX 400 nuclear magnetic resonance instrument. 1 H NMR is at 400 MHz, the solvent is deuterated methanol, deuterated DMSO or deuterated tetrahydrofuran, and tetramethylsilane (TMS) is used as the internal standard. The unit of chemical shift is ppm, and the unit of coupling constant is Hz. 1 In H NMR, δ represents chemical shift, s represents singlet, d represents doublet, t represents triplet, q represents quartet, and m represents multiplet.

[0051] Example 1

[0052] Synthesis of ceramides derived from linolenic acid

[0053]

[0054] Dissolve 1.2eq (60mmol) of p-nitrobenzenesulfonyl chloride in 70ml of ethyl acetate, 1.32eq (66mmol) of compound S1 (α-linolenic acid or γ-linolenic acid) and 3eq (150mmol) of triethylamine in 30ml of ethyl acetate, add dropwise to the ethyl acetate solution of p-nitrobenzenesulfonyl chloride, react at 40°C for 12h, and TLC shows that the reaction of p-nitrobenzenesulfonyl chloride is complete. Add 2eq (100mmol) of triethylamine and 1eq (50mmol) of sphingosine or phytosphingosine, react at 40°C overnight, and TLC shows that the reaction of sphingosine base is complete.

[0055] Post-treatment: add 100 mL of water, adjust pH to 5-6 with 2N dilute hydrochloric acid, extract, extract twice with 100 mL of saturated brine, dry over anhydrous sodium sulfate, filter and concentrate in vacuo. The residue thus obtained is purified by silica gel column to obtain the product (50-70% yield).

[0056]

[0057] 1 H NMR (400 MHz, Methanol-d4) δ5.42–5.22(m,6H),4.07(td,J=5.8,4.3 Hz,1H),3.79–3.65(m,3H),3.58(t,J=5.8 Hz,1H),3.52(ddd,J=9.6,5.6,2.4 Hz,1H),2.80(t,J=6.0 Hz,4H),2.22(t,J=7.5 Hz,2H),2.12–2.04(m,4H),1.91–1.80(m,1H),1.67–1.52(m,4H),1.35–1.28(d,J=22.7 Hz,31H),0.97(t,J=7.6 Hz, 3H), 0.89 (t, J = 7.6 Hz, 3H).

[0058]

[0059] 1 H NMR (400 MHz, Methanol-d4) δ5.74–5.63(m,1H),5.49–5.42(m,1H),5.41–5.24(m,6H),4.04(t,J=7.4 Hz,1H),3.85(dt,J=7.5,5.0 Hz,1H),3.68(d,J=5.0 Hz,2H),2.80(t,J=6.0Hz,4H),2.24–2.14(m,2H),2.14–1.97(m,6H),1.59(dt,J=8.0,3.9 Hz, 2H), 1.41–1.22 (m, 30H), 0.97 (t, J = 7.6 Hz, 3H), 0.89 (t, J = 7.6 Hz, 3H).

[0060]

[0061] 1H NMR (400 MHz, Methanol-d4) δ5.52–5.33(m,6H),4.09–4.02(m,1H),3.72–3.68(m,3H),3.53(t,J=5.8 Hz,1H),3.50–3.44(m,1H),2.76(t,J=6.2 Hz,4H),2.18(t,J=7.6 Hz,2H),2.10–2.02(m,4H),1.94–1.82(m,1H),1.69–1.54(m,4H),1.32–1.25(d,J=22.4 Hz,30H),0.92(t,J=7.6 Hz, 3H), 0.83 (t, J = 7.6 Hz, 3H).

[0062]

[0063] 1 H NMR (400MHz, Methanol-d4) δ5.76–5.66 (m, 1H), 5.49 (ddt, J = 15.3, 7.4, 1.5Hz, 1H), 5. 45–5.30(m,6H),4.07(t,J=7.4Hz,1H),3.88(dt,J=7.5,5.0Hz,1H),3.71(dd,J=5.0,1. 2Hz,2H),2.84(td,J=5.7,3.3Hz,4H),2.23(t,J=7.5Hz,2H),2.18–1.99(m,6H),1.64(t t,J=9.2,6.9Hz,2H),1.46–1.24(m,30H),0.93(t,J=7.6Hz,3H),0.90(t,J=7.6Hz,3H).

[0064] Example 2

[0065] Comparison of the whitening efficacy of the compound of Example 1 by intracellular tyrosinase inhibition method

[0066] Intracellular tyrosinase method to detect whitening effect: B16 cells were seeded in 96-well plates (density 1×10 4 / well), incubate in the incubator for 24 hours, discard the supernatant, add 100 μL of compounds of different concentrations diluted with DMEM medium, and the DMEM medium without drugs is the negative control group. After incubation for 24 hours, discard the drug-containing culture medium, wash twice with PBS, add 100 μL, 1wt% Triton X-100, place at -80℃, freeze for 1 hour, and then thaw at room temperature. After thawing, add 20 μL, 0.5wt% L-dopa to each well and shake horizontally at room temperature for 4 hours. After the reaction, measure the absorbance at a wavelength of 490nm, and calculate the cell tyrosinase inhibition rate = A给药孔 / A 空白孔 ×100%.

[0067] The results are as follows Figures 1 to 4 As shown in Table 1, Figure 1 As a result of alpha-linolenic acid phytosphingosine, Figure 2 As a result of γ-linolenic acid phytosphingosine, Figure 3 As a result of α-linolenic acid sphingosine, Figure 4 The results are for γ-linolenic acid sphingosine. α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine have good inhibitory effects on tyrosinase at different concentrations, especially γ-linolenic acid sphingosine still has a high inhibition rate at a very low concentration (1mM), while γ-linolenic acid phytosphingosine requires a higher concentration (4mM) to have a high inhibition rate.

[0068] Table 1

[0069] Concentration (mM) 1 2 3 4 5 6 α-linolenic acid sphingosine (%) 5.1 8.2 10.4 13.5 16.1 18.6 α-linolenic acid phytosphingosine (%) 7.1 9.3 9.1 12.6 11.6 14.3 γ-linolenic acid sphingosine (%) 27.7 33.6 40.4 35.2 33.8 33.4 γ-linolenic acid phytosphingosine (%) 6.8 12.6 18.2 24.0 31.6 35.6

[0070] Example 3

[0071] MTT assay to detect the effects of compounds on cell proliferation

[0072] HaCaT cells were cultured at 1×10 4 The cells were seeded at a density of 100 μL / well in a 96-well plate and incubated overnight in an incubator. After 24 hours, the supernatant was discarded, and 100 μL of culture medium containing different concentrations of compounds (α-linolenic acid phytosphingosine, γ-linolenic acid phytosphingosine) was added. After incubation for another 24 hours, the culture medium was removed, and 100 μL of thiazolyl blue (MTT) was added to each well. The absorbance at 450 nm was measured, and the cell survival rate was calculated as A 给药孔 / A 空白孔 ×100%.

[0073] The results are as follows Figure 5 , 6 As shown, Figure 5 As a result of alpha-linolenic acid phytosphingosine, Figure 6 The results for γ-linolenic acid phytosphingosine show that α-linolenic acid phytosphingosine and γ-linolenic acid phytosphingosine have promoting effects on cell viability at lower concentrations (0.0625mmol / L, 0.125mmol / L), showing obvious effects in promoting cell proliferation and having good tissue repair capabilities. Even at very high concentrations (2mmol / L), the cell viability is still above 50%, indicating that the cytotoxicity of the compound is controllable and exhibits good biosafety.

[0074] Example 4

[0075] LPS-induced cell assay to detect anti-inflammatory and repair effects

[0076] B16 mouse melanoma cells were cultured at a density of 1×10 4 100 μL of compounds (α-linolenic acid phytosphingosine, γ-linolenic acid phytosphingosine) diluted in DMEM medium were added. The model group was a cell without drug addition, and the negative control group was a DMEM medium without drug. Each group had 3 replicates and was incubated in an environment of 5% CO2 and 37°C. 2 hours after administration, 10 μg / mL LPS was added to the lipopolysaccharide model group and the experimental group and incubated together for 24 hours. After the reaction, 50 μL of the cell supernatant was taken and the intracellular IL-6 gene expression was detected using an IL-6 ELISA kit.

[0077] The results are as follows Figure 7 , 8 As shown, Figure 7 As a result of alpha-linolenic acid phytosphingosine, Figure 8 The results of γ-linolenic acid phytosphingosine showed that both α-linolenic acid phytosphingosine and γ-linolenic acid phytosphingosine had excellent inhibitory effects on IL-6 cytokine expression. Under the stimulation of LPS at a working concentration of 10 μg / mL, the IL-6 level was 40.7 times the basal level. Under the action of α-linolenic acid phytosphingosine at concentrations of 0.0625, 0.125, 0.25, and 0.5 mM, the IL-6 factor level was significantly reduced, which was 29.6, 24.8, 19.5, and 14.7 times the basal level, respectively, in a dose-dependent manner; under the action of γ-linolenic acid phytosphingosine at concentrations of 0.0625, 0.125, 0.25, and 0.5 mM, the IL-6 factor level was 30.2, 26.1, 18.2, 17.2, and 18.8 times the basal level, respectively. Therefore, these two compounds have good anti-inflammatory effects and can promote the repair of skin damaged by inflammation.

[0078] Example 5

[0079] Scratch assay to detect tissue healing ability

[0080] Principle: When cells grow to a fused monolayer state, a scratch tool is used to create a blank area on the fused monolayer of cells. The cells in the blank area are removed by mechanical force. After a period of culture, the migration of cells to the cell-free area is observed, and the migration distance of cells is measured to reflect the migration ability of cells.

[0081] Steps:

[0082] 1. Marking the culture plate. First, use a marker pen to evenly mark horizontal lines on the back of the 6-well plate with a ruler, approximately every 0.5 to 1 cm, across the holes, with at least 5 lines in each hole. Be careful not to make the lines too thick.

[0083] 2. Add about 5×10 5 cells (different cell numbers vary and are adjusted according to the growth rate of the cells). The inoculation principle is that the fusion rate reaches 100% after overnight.

[0084] 3. Cell streaking. On the second day, use the tip of the pipette to scratch the cell layer perpendicular to the cell plane along the line drawn on the back of the plate the day before (it is best to use the same pipette tip between different wells).

[0085] 4. Wash the cells. After the scratch is completed, wash the cells three times with sterile PBS to wash away the cells that are not attached to the wall, that is, the cells that are scratched during the scratching process. The gaps left after the scratching are clearly visible, and then replace with fresh serum-free culture medium.

[0086] 5. Cell culture and observation. The compound (α-linolenic acid phytosphingosine, γ-linolenic acid phytosphingosine) was diluted with culture medium (concentration of 0.08 mM) and added to the cell culture dish. The cells were cultured in a 37°C, 5wt% CO2 incubator. The cells were taken out after 24h and 48h, and the width of the scratch was measured under a microscope and photographed.

[0087] The results are as follows Fig. 9 As shown, compared with the solvent control group, the scratch width of the experimental group was narrower, and α-linolenic acid phytosphingosine and γ-linolenic acid phytosphingosine had better tissue healing ability.

[0088] Example 6

[0089] Elastase inhibition assay to test anti-aging effects

[0090] Elastase inhibition method: Take 2 mL of 2 mg / mL elastase solution, add different concentrations of compounds, vortex mix thoroughly, shake at 37°C, 400 r / min for 20 min, immediately add 5 mL of 0.5 mol / L phosphate buffer at pH 6.0, vortex mix, take an appropriate amount of the mixture into a 2 mL centrifuge tube, centrifuge at 9 391×g for 10 min, accurately pipette 200 μL of the supernatant into a 96-well plate, measure the absorbance at a wavelength of 495 nm with an enzyme reader, and simultaneously perform a spectral scan from 400 to 800 nm.

[0091] The substrate plus enzyme solution was used as the blank control group, the substrate plus enzyme and sample solution was used as the enzyme inhibition group, and the substrate plus sample without enzyme solution was used as the background subtraction group. Each group was set up with 3 replicates. Inhibition rate (%) = [1–(An–An′) / (A0–A0′)]×100%, where A0 is the absorbance of adding enzyme without sample, A0′ is the absorbance of adding substrate without sample and enzyme, An is the absorbance of adding sample solution only, and An′ is the absorbance of adding sample without enzyme. If An′>An, it shows a promoting effect, and the promotion rate (%) = [1–(An′–An) / (A0–A0′)]×100%.

[0092] The results are as follows Figures 10 to 13 As shown in Table 2, Fig.10 As a result of alpha-linolenic acid phytosphingosine, Fig.11 As a result of γ-linolenic acid phytosphingosine, Fig.12 As a result of α-linolenic acid sphingosine, Fig.13 The results are for γ-linolenic acid sphingosine. α-linolenic acid sphingosine, α-linolenic acid phytosphingosine, γ-linolenic acid sphingosine or γ-linolenic acid phytosphingosine all have good inhibitory effects on elastase at different concentrations, especially γ-linolenic acid sphingosine has a high inhibition rate at a very low concentration (3 mg / L), and α-linolenic acid sphingosine requires a higher concentration (15 mg / L) to have a high inhibition rate.

[0093] Table 2

[0094] Concentration (mg / L) 3 6 9 12 15 18 α-linolenic acid sphingosine (%) 10.6 15.8 16.7 — 23.7 24.5 α-linolenic acid phytosphingosine (%) 7.1 — 10.3 11.9 — 13.2 γ-linolenic acid sphingosine (%) 24.6 16.1 16.7 — 12.3 23.7 γ-linolenic acid phytosphingosine (%) 8.2 9.3 13.7 14.4 — 17

[0095] Elastin is a component of elastic fibers in connective tissue, which can make tissues elastic and maintain tissue structural stability together with other extracellular matrices such as collagen. Elastase can degrade elastin under conditions such as inflammation or long-term ultraviolet radiation, thereby causing aging of skin tissue. The compounds of the present invention have good anti-aging ability by inhibiting the activity of elastase.

[0096] Example 7

[0097] MMP1, also known as interstitial collagenase and matrix metalloproteinase, belongs to the matrix metalloproteinase family. Its main substrate is fibrous collagen. It can degrade collagen fibers and gelatin in the extracellular matrix and change the cell microenvironment. MMP1 plays an important role in elastin. Inhibition of MMP1 can increase the synthesis of collagen and elastin in fibroblasts, and reduced MMP activity can increase the rate of collagen synthesis.

[0098] HaCaT cells were cultured at 1×10 5The cells were seeded at a density of 100 μL / well in a 96-well plate and incubated overnight in an incubator. After 24 hours, the supernatant was discarded and 100 μL of culture medium containing different concentrations of α-linolenic acid phytosphingosine (no drug was added to the model group) was added. The negative control group was a DMEM culture medium without drugs. Each group had 3 replicates. After incubation for 2 hours in an environment of 5% CO2 and 37°C, UVA ultraviolet rays were irradiated. The distance between the ultraviolet radiation source and the cells was 15 cm, and the UVA intensity was 200 mJ / cm 2 The irradiation time was 1.5 h. After the irradiation, the cells were incubated in the incubator for 12 h. The MMP-1 ELISA kit was used to detect the expression of MMP-1 gene in the cells.

[0099] The results are as follows Fig.14 As shown, the MMP1 expression level of the negative control group was set to 1, the expression level of the model group was 1.64, when the concentration of α-linolenic acid phytosphingosine was 50 mg / L, the expression level of MMP1 was 1.41, when the concentration was 100 mg / L, the expression level of MMP1 was 1.23, when the concentration was 150 mg / L, the expression level of MMP1 was 1.26, and when the concentration was 200 mg / L, the expression level of MMP1 was 1.13, showing concentration dependence. The higher the concentration, the more significant the inhibitory effect on MMP1.

[0100] After UVA radiation, keratinocytes promote the increase of MMP1 expression in fibroblasts, which causes the degradation of skin extracellular matrix and skin collagen, leading to skin photoaging. The above results show that α-linolenic acid phytosphingosine can inhibit the production of MMP1 in fibroblasts caused by UV radiation, and has a certain effect on preventing skin photoaging.

[0101] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. Ceramide derived from linolenic acid, having the structure of general formula I:

2. A method for preparing the linolenic acid-derived ceramide according to claim 1, comprising the following steps: Compound S1 reacts with p-nitrobenzenesulfonyl chloride and an organic base to obtain compound S2; Compound S2 reacts with sphingoid base to obtain compound I; The molar ratio of the compound S1, p-nitrobenzenesulfonyl chloride, organic base and sphingosine base is (1-2): (1-2): (2-6): 1; The organic base is triethylamine; The solvent for the reaction is ethyl acetate.

3. Use of the linolenic acid-derived ceramide according to claim 1 in the preparation of cosmetics or medicines, wherein the use is whitening or anti-aging.

4. Use of the linolenic acid-derived ceramide according to claim 1 in the preparation of cosmetics or medicines, wherein the linolenic acid-derived ceramide inhibits tyrosinase, and the inhibition rate of tyrosinase at a concentration of 1 mM is more than 27%, and / or the inhibition rate of tyrosinase at a concentration of 2 mM is more than 33%, and / or the inhibition rate of tyrosinase at a concentration of 3 mM is more than 40%, and / or the inhibition rate of tyrosinase at a concentration of 4 mM is more than 35%, and / or the inhibition rate of tyrosinase at a concentration of 5 mM is more than 33%, and / or the inhibition rate of tyrosinase at a concentration of 6 mM is more than 33%.

5. A composition comprising the ceramide derived from linolenic acid according to claim 1, wherein the composition has whitening or anti-aging effects.

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