KR20100059050A - Composition for preventing or treating ceramide metabolism-related enzyme mediated disease comprising lignan derivatives as an active ingredient - Google Patents

Abstract

PURPOSE: A composition containing lignin derivative is provided to treat atopic dermatitis or xerosis to also treat diseases mediated by ceramide metabolic enzyme. CONSTITUTION: A composition for preventing or treating diseases mediated by ceramide metabolic enzyme contains rosmarinic acid of chemical formula 1, lithospermic acid of chemical formula 2, 9"-methyl lithospermate of chemical formula 2, or lignin derivative of 9'-methyl lithospermate of chemical formula 2 as an active ingredient. The composition enhances ceramide content on the epidermis.

Description

Composition for preventing or treating ceramide metabolism-related enzyme mediated disease comprising lignan derivatives as an active ingredient

The present invention relates to a composition for use in the prophylaxis or treatment of diseases mediated by ceramide metabolism-related enzymes. More specifically, the present invention relates to a composition for the prevention or treatment of diseases mediated by enzymes related to ceramide metabolism by containing a soy lignan derivative as an active ingredient.

The epidermis, the outermost layer of skin, is an organ that is always in contact with the external environment and serves as a protective barrier. The skin barrier in the stratum corneum of the epidermis suppresses the loss of water and electrolytes, protects the body from external physical damage and chemicals, and prevents bacteria and viruses from penetrating the skin. (Feingold KR. Cosmet Toilet. 112: 49-59, 1997). Keratinocyte intercellular lipids are intercellular lamellae structures made of lipid mixtures such as ceramides, cholesterol, and free fatty acids. In particular, ceramide, which accounts for 40-50% of the epidermal sebaceous membrane, is composed of spingosine, amide-linked nonhydroxy acid, alpha-hydroxy acid or omega hydroxy acid. ω-hydroxy acid), ester-linked fatty acid, etc., and has a complex structure, the polarity formed by these structural factors is important for maintaining the sebaceous membrane layer structure ( Choi MJ et al. Am. J. Clin. Dermatol. 6: 215-223, 2005). There are two known enzymes responsible for ceramide synthesis and ceramide degradation associated with ceramide metabolism, serine-palmitoyl trasnferase and ceramidase, respectively. Therefore, in order to increase the total amount of ceramide, it is necessary to increase the synthesis of ceramide through serine-palmitoyl trasnferase (SPT), or to reduce ceramide degradation by ceramidase.

Root of the domestic plant (Radix lithospermi) is the root of the weary, and contains components such as lithosperman, a polysaccharide together with shikonin and acetylshikonin, and promotes blood circulation. Detoxification, antibacterial and anti-inflammatory action is known as a herbal medicine that is commonly used to treat a variety of wounds, boils, burns, eczema. Korean Patent Laid-Open Publication No. 2000-0038904 discloses that the herbaceous plant promotes angiogenesis of wound and burned cells, and has anti-inflammatory and wound healing promoting effects, and in 2004-0011984, the fat layer of the skin epidermal barrier. Licorice extract has been reported for use in treating atopic dermatitis or dryness in which the structure is impaired. However, there have been no reports of the structure of the active substance exhibiting physiological activity in the herbaceous plant, and there is no known detailed mechanism for treating atopic dermatitis or dryness caused by the compound.

In general, among the various methods for the development of drugs with new ingredients, the use of natural medicines used in traditional medicine, rather than the effort by experimental modification of existing drugs, is very likely to find new active ingredients and has been used for a long time. Therefore, there is less advantage of toxicity caused by the developed drugs. Therefore, the present inventors investigated various native plants and herbal medicines. Among them, the present study was conducted by confirming that the soybean lignan derivative obtained by purely separating and purifying the herb extract from chromatography increased the expression level of serine-palmitoyl transferase. The invention was completed.

Accordingly, an object of the present invention is to provide a compound that increases the expression level of serine-palmitoyl transferase obtained by purely separating and purifying from porcelain.

In addition, another object of the present invention is to provide a composition for preventing or treating a mediated disease by a ceramide metabolism-related enzyme containing the compound as an active ingredient.

In order to achieve the above object, the present invention is a rosemary acid, Lithospermic acid, 9 ”-Methyl lithospermate and 9'-methyl resource isolated from Provided is a composition for preventing or treating a disease mediated by a ceramide metabolism related enzyme containing at least one lignan derivative selected from the group consisting of permate (9'-Methyl lithospermate) as an active ingredient.

The present invention can improve the epidermal condition of the skin and treat atopic dermatitis or dryness in which the fat layer structure of the epidermal barrier is impaired, thereby ultimately reducing the damage of diseases mediated by enzymes related to ceramide metabolism.

The composition of the present invention is a rosemary acid (Rosmarinic acid, Compound 1), Lithospermic acid (Lithospermic acid, Compound 2) represented by the following formula 1 to 2 isolated from the licorice, 9 "-methyl resource permate (9" -Methyl At least one lignan derivative selected from the group consisting of lithospermate, compound 3) and 9'-methyl lysospermate (9'-Methyl lithospermate, compound 4) is contained as an active ingredient.

Lignan is a phenol compound composed of phenylpropanoid units, which are distributed in small amounts and in all parts of the roots, leaves, stems, seeds, and fruits of the plant. Lignan, like estrogen, is a substance that enters the gut cycle and is released in the urine as a glucuronide conjugate or a sulfate conjugate.

The licorice extract of the present invention can be obtained by a conventional extraction and separation method, first, the dried licorice is pulverized and placed in an extractor with 4 to 5 times the extraction solvent and left for 2 days to extract and concentrated by a concentrator. And dry to obtain an extract. In the present invention, the extraction solvent for separation may be water or an organic solvent of C ₁ ~ C ₆ , preferably purified water, methanol (methanol), ethanol (ethanol), propanol, isopropanol (isopropanol) , Butanol, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride, hexane, cyclohexane ) And various solvents such as petroleum ether may be used alone or in combination, and more preferably 70% ethanol may be used.

The composition of the present invention contains such an active ingredient in an amount of 0.01 to 50% by weight based on the total weight of the composition. If it is less than 0.01% by weight, effective efficacy is difficult to realize, and if it is more than 50% by weight, it is difficult to formulate.

The lignan derivatives according to the present invention can be used as pharmaceuticals and additives because of their high stability as well as separation from various plants. When applied to pharmaceutical products, the lignan derivative according to the present invention for the purpose of improving the epidermal condition of the skin and treating atopic dermatitis, dry skin, psoriasis, etc., in which the fatty layer structure of the skin epidermal barrier is damaged due to the reduction of the skin epidermal ceramide. Compounds can be formulated as oral or parenteral dosage forms in solid, semisolid or liquid form with the addition of a commercially available inorganic or organic carrier as the active ingredient.

Formulations for oral administration include tablets, pills, granules, soft / hard capsules, powders, fine granules, powders, emulsions, syrups, pellets, and the like. have. Formulations for parenteral administration include injections, drops, ointments, lotions, sprays, suspensions, emulsions, suppositories, and the like.

In order to formulate the active ingredient of the present invention, it can be easily formulated according to the conventional method, and surfactants, excipients, coloring agents, spices, preservatives, stabilizers, buffers, suspensions, and other commonly used auxiliaries can be suitably used.

When ingesting the composition of the present invention can be ingested 1 ~ 12000mg per day as an active ingredient the lignan derivative represented by the formula (1) -2.

In addition, the present invention provides a food containing the lignan derivative represented by the formula (1) to (2) as an active ingredient.

When the ingredient is contained in a food, the soybean extract or the lignan derivative represented by Chemical Formulas 1 to 2 may be added as it is, or used together with other food or food ingredients, and may be appropriately used according to a conventional method.

Hereinafter, the specific method of the present invention will be described step by step with reference to Examples, but the scope of the present invention is not limited only to these test examples.

Test Example 1 Isolation and Structure Verification of Lignan Derivatives

In order to confirm the structure of the active compound having a prophylactic or therapeutic effect of the enzyme-mediated diseases related to ceramide metabolism and the preparation of the soybean lignan fractions, the experiment was carried out as follows.

Step 1: Isolation and Purification of Lignan Derivatives from Licorice

5 kg of dried roots of the vinegar were finely crushed and then immersed in 70% ethanol for 2 days and extracted at room temperature. The ethanol extract was concentrated under reduced pressure, extracted using chloroform, and then diion HP-20 column chromatography was performed using the remaining aqueous layer, and the mixture was divided into five fractions. The fraction (17.3 g) eluting in the 60% methanol solvent showing the strongest activity was collected and the active fraction (LE-A, 3.2 g) was obtained by RP-18 column chromatography using 30% methanol. Got. Finally, some LE-A fractions were used to perform high pressure liquid chromatography (HPLC; Gilson system, YMC column (20 × 150 mm, particle size 4 μm), UV detection at 280 and 320 nm) to increase SPT expression. Compound 1 (16.0 mg), Compound 2 (10.2 mg), Compound 3 (15.0 mg), and Compound 4 (21.0 mg) were isolated.

Step 2: Physical and chemical characterization and chemical structure analysis of 4 compounds

First, the chemical structure of Compound 2 was estimated for structural analysis of the compounds isolated from the porcelain. Compound 2 was obtained as a dark brown powder and showed a maximum absorption peak at 251, 289 and 306 nm as measured by UV spectra with methanol solvent. In addition, the [M + Na] + peak was observed at 561 by FAB mass spectrometry. ¹ H NMR analysis showed a 6.72 ppm double proton, 6.61 ppm, 6.68 ppm and 6.80 ppm trisubstituted benzene ring and a pair of other signals at 6.82 ppm and 7.20 ppm The peaks of ppm, 6.77 ppm and 6.80 ppm were confirmed. In addition, proton peaks ( J = 16.2 Hz) which were trans at 6.32 ppm and 7.84 ppm were confirmed. ¹³ C NMR showed three carboxyl carbons near 170 and six hydroxyl peaks near 145 ppm. In particular, we observed oxidized methine peaks appearing near 4.36 ppm (δ _C 89.3 ppm) in the ¹ H and ¹³ C-NMR spectral data and based on the partial structural information of these molecules, The compound was identified as a lithospermic acid, and each compound was found to have a structural analysis similar to the above. Compound 1 was rosmarinic acid, and Compound 3 was 9 ”-methyl resource permate (9” -methyl. lithospermate), Compound 4 was confirmed to be 9'-methyl lithospermate (9'-methyl lithospermate) and the results of the nuclear magnetic resonance spectra of the compounds ¹ H, ¹³ C are shown in Table 1 below.

¹ H- and ¹³ C-NMR data of compounds 1-4 in CD ₃ OD location One 2 3 4 δ _H (mult., J in Hz) δ _c δ _H (mult., J in Hz) δ _c δ _H (mult., J in Hz) δ _c δ _H (mult., J in Hz) δ _c One 127.7 124.8 124.7 124.7 2 7.17 (s) 115.2 128.4 127.1 127.9 3 146.8 149.0 149.0 149.0 4 149.7 145.2 145.4 145.4 5 6.78 (d, 8.4) 116.3 6.82 (d, 8.4) 118.2 6.81 (d, 8.4) 118.5 6.82 (d, 8.7) 118.4 6 6.92 (d, 8.4) 123.2 7.20 (d, 8.4) 121.7 7.19 (d, 8.4) 122.1 7.22 (d, 8.4) 122.1 7 7.85 (d, 16.2) 147.7 7.84 (d, 16.2) 144.0 7.72 (d, 15.9) 144.0 7.83 (d, 15.6) 144.4 8 6.32 (d, 16.2) 114.5 6.32 (d, 16.2) 116.4 6.30 (, 15.9) 116.8 6.34 (d, 15.9) 116.2 9 168.5 168.4 168.3 168.2 One' 129.4 129.8 129.4 128.9 2' 6.78 (br s) 117.6 6.8 (br s) 113.6 6.74 (br s) 113.6 6.74 113.6 3 ' 146.2 145.3 145.4 145.4 4' 145.3 146.2 146.3 146.3 5 ' 6.72 (d, 8.4) 116.5 6.68 (d, 8.1) 116.5 6.68 (d, 8.1) 116.5 6.68 (d, 8.1) 116.5 6 ' 6.65 (d, 8.4) 121.9 6.61 (dd, 8.1, 1.8) 118.5 6.68 (dd, 8.1, 2.1) 118.5 118.4 7 ' 3.06 (dd, 14.1, 8.4)
3.01 (dd, 14.1, 3.6) 38.0 3.05 (dd, 14.1, 8.4)
2.99 (dd, 14.1, 3.6) 38.3 3.09 dd (14.4, 3.9)
2.99 dd (14.4, 8.7) 38.0 3.03 (br s)
3.07 (br s) 38.0 8' 5.13 (dd, 8.4, 3.6) 74.8 5.12 (dd, 8.0, 3.6) 75.1 5.19 dd (8.7, 3.9) 75.0 5.12 (dd, 7.2, 6.0) 74.9 9 ' 173.6 174.2 173.8 172.3 One" 134.2 133.5 133.9 2" 6.80 (s) 118.0 6.78 (d, 1.8) 117.6 117.7 3 ” 146.7 146.8 146.8 4" 146.8 147.0 146.9 5 ” 6.77 (d, 8.1) 116.4 6.77 (d, 8.1) 116.5 6.77 (d, 8.1) 116.5 6 ” 6.72 (d, 8.1) 121.9 6.61 (dd, 8.1, 1.8) 121.9 6.58 (dd, 8.1, 1.8) 121.8 7 ” 5.90 (d, 4.8) 58.5 5.89 (d, 4.8) 57.4 5.91 (d, 4.9) 57.8 8" 4.36 (d, 4.5) 89.3 4.43 (d, 4.8) 88.6 4.36 (d, 4.5) 89.0 9 ” 176.2 173.6 175.5 OCH ₃ 3.70 s 53.4 3.68 52.8

(NMR spectra are recorded on the Varian Inova 300 spectrometer)

In order to measure the prophylactic or therapeutic effect of the ceramide metabolism-related enzyme-mediated disease of the porcine lignan fraction of Test Example 1, the experiments of Test Examples 2 to 5 were performed.

[Test Example 2] Effect on the expression of enzyme related to ceramide metabolism of lignan derivatives (observation of SPT mRNA expression)

Serine-palmitoyl transferase (SPT) mRNA expression levels were observed in the following manner to determine the effect of lignan derivatives on ceramide metabolism-related enzyme expression. Example 1 was treated with the amount of 100% by weight of 70% ethanol extract of Licorice, Example 2 was treated with 100 ppm by weight of the total fraction of licorice lignan, which is the LE-A fraction obtained in Test Example 1, Example 3 in Test Example 1 Treatment of the extracted soybean lignan derivative 1 (compound 1) in an amount of 100 ppm, Example 4 treated with the extraction of the soybean lignan derivative 2 (compound 2) in an amount of 100 ppm, Example 5 in Example 1 The extracted soybean lignan derivative 3 (compound 3) was treated with an amount of 100 ppm, and Example 6 was treated with the extracted soybean lignan derivative 4 (compound 4) in an amount of 100 ppm. In addition, Comparative Example 1 (negative control group), in which nothing was treated as a control, and Comparative Example 2 (positive control group) treated with 30 μM of nicotinamide were prepared.

All of the samples of licorice lignan fractions of Examples 1 to 6 were dissolved in DMSO at a concentration of 1000 ×, diluted with medium, and added to the test plates for each experimental group. To observe Serine-palmitoyl transferase (SPT) mRNA expression, 12 hours after treatment with the test substance, washed twice with cold PBS and trizol (Gibco Laboratories, USA) After extracting the total RNA using the Fermentas First Strand cDNA Synthesis Kit (# K1612) was synthesized cDNA, and then PCR was performed using Qiagen's Quantitect SYBR green PCR kit (cat # 204343). Primers for SPT and primers for GAPDH for mRNA expression comparison were purchased from Applied Biosystems' TaqMan gene expression assay, and real-time PCR was performed using Corbett (Applied Biosystems).

The experiment for observing the serine-palmitoyl transferase (SPT) mRNA expression was performed twice independently, the average value is shown in Table 2 below.

Serine-palmitoyl transferase (SPT LC2) mRNA expression level division Average (%) Standard Deviation (%) Comparative Example 1 Negative Control (No Processing) 100 9.1 Comparative Example 2 Positive control group (nicotinamide, 30 μM) 128 17.0 Example 1 Licorice 70% EtOH Extract (100ppm) 145.7 21.1 Example 2 Total fraction of licorice lignans (100 ppm) 165 4.2 Example 3 Zacholignan derivative 1 (100ppm) 146.3 5.5 Example 4 Zacholignan derivatives 2 (100 ppm) 150.5 10.8 Example 5 Zacholignan derivatives 3 (100 ppm) 151.0 7.1 Example 6 Zacholignan derivatives 4 (100 ppm) 159.0 2.8

As can be seen in Table 2, it can be seen that the expression of SPT mRNA is increased by about 145 ~ 165% by treatment of the extract or lignan derivatives (Examples 1 to 6), in particular, compound 4 among (Example 6) 9'-methyl lithospermate (9'-Methyl lithospermate) was confirmed that the most effective increase in the expression of SPT mRNA.

[Test Example 3] Effect on the expression of enzymes related to ceramide metabolism of lignan derivatives (observation of SPT protein expression)

Serine-palmitoyl transferase (SPT) protein expression levels were observed to determine the effect of lignan derivatives on ceramide metabolism-related enzyme expression. Example 7 was treated with 100 ppm of 70% ethanol extract of Licorice, Example 8 was treated with 100 ppm of the total fraction of licorice lignan, which is the LE-A fraction obtained in Test Example 1, and Example 9 was tested in Example 1 Treating the extracted soybean lignan derivative 1 in an amount of 100ppm, Example 10 was treated with the amount of the soybean lignan derivative 2 extracted in Test Example 1 in an amount of 100ppm, Example 11 was 100ppm of the soybean lignan derivative 3 extracted in Test Example 1 In the amount, Example 12 was treated with 100 ppm of the soybean lignan derivative 4 extracted in Test Example 1. In addition, Comparative Example 3 (negative control group), in which nothing was treated as a control, and Comparative Example 4 (positive control group) treated with 30 μM of nicotinamide were prepared.

After 24 hours of treatment with the test material to observe Serine-palmitoyl transferase (SPT) protein expression, the test plates are washed with cold PBS and then rapidly frozen until used for analysis. Stored in a deep freezer. After crushing at 4 ℃ with a cell lysate solution, the protein amount was quantified by BCA solution. Samples were prepared by mixing the same amount of protein and LDS sample buffer, and then separated by sodium dodesyl sulfate (SDS) polyacrylamide gel electrophoresis. Acrylamide gel containing proteins isolated for Western blot analysis was transferred to nitrocellulose membrane by electroblotting, and then stained with Ponceus S solution. Confirmed metastasis. Blocking of nonspecific proteins by incubating the protein-transformed membrane with TBS-T (0.1% Tween 20 in TBS) containing 5% non-fat milk at room temperature for 1 hour. ) And washed twice with TBS-T. After washing, abcam's SPT primary antibody (Cat. # Ab23696) was treated at a concentration of 1: 500, overnight at 4 ° C, and then at room temperature using a secondary antibody (1: 2000) suitable for the primary antibody. Reaction was carried out for 1 hour. After washing with TBS-T, ECL (enhanced chemiluminoesence) solution was applied to the X-ray film in the dark and analyzed for the expression of SREBP-1 protein.

The experiment for observing the serine-palmitoyl transferase (SPT) protein expression was carried out independently twice, the analysis of the results was used ImageJ 1.34s provided by NIH. The average value is shown in Table 3 below, and the results of Western blot analysis are shown in FIG. 1.

Serine-palmitoyl transferase (SPT) protein expression level division Average (%) Standard Deviation (%) Comparative Example 3 Negative Control (No Processing) 100.0 19.0 Comparative Example 4 Positive control group (nicotinamide, 30 μM) 149.3 17.2 Example 7 Licorice 70% EtOH Extract (100ppm) 120.1 18.5 Example 8 Total fraction of licorice lignans (100 ppm) 127.3 14.7 Example 9 Zacholignan derivative 1 (100ppm) 142.3 10.1 Example 10 Zacholignan derivatives 2 (100 ppm) 151.1 26.6 Example 11 Zacholignan derivatives 3 (100 ppm) 132.4 23.0 Example 12 Zacholignan derivatives 4 (100 ppm) 167.0 10.9

As can be seen in Table 3, it can be seen that the expression of the SPT protein is increased by 125 to 170% by treatment of the extract or lignan derivatives (Examples 7 to 12), especially derivative 4 of the licorice lignan derivatives (Example 12) The 9'-methyl lithospermate (9'-Methyl lithospermate) was the most effective in increasing the SPT protein expression, it was confirmed that this is similar to the mRNA expression pattern.

It can be seen that the composition according to the present invention can increase the expression of SPT to increase the production of ceramide.

[Test Example 4] Effect of dietary supply of functional dietary materials on the change of total ceramide content of epidermis

Laboratory Animal Preparation

Four-week-old NC / Nga mice and BALB / c mouse males, the normal control mice, were purchased (central laboratory animals) and maintained in a normal breeding room without air filtration for natural induction of inflammation and drying. After dividing the control group and each experimental group by the ingot method, each experimental group was reared in two cages with 10 birds per cage, so that the feed and water were ingested freely, and maintained at a temperature of 22 ± 1 ℃ and a humidity of 60 ± 5% at intervals of 12 hours. Changed the contrast.

Dietary composition

The mice were adapted to solid blended feed for 1 week before breeding into the experimental diet. The experimental group was divided into five groups (n = 20) by the ingot method according to the weight, and then the experimental diet was supplied for 5 or 10 weeks. The dietary composition of each experimental group is shown in Table 4 below, and dietary fat was fixed at 10% of the dietary weight and was supplied with corn oil. The dietary feeding group fed 1% of the total dietary weight to the diet with the powder provided as an upper limit. Differences in dry meal weight resulting from the use of soybean dietary materials were adjusted by subtracting corn starch. The BALB / c mouse group (control group A), which received a diet containing no self-contained dietary material for 10 weeks, was placed as a positive control group and the NC / Nga Tnd mouse group (control group B) as a negative control group. Experimental groups 1 to 5 were provided by mixing a mixture of the extract of Japonica and Japonignan fraction.

Determination of total content of epidermal ceramide

The epidermis was pulverized with polytron, extracted with lipid using Chloroform / Methanol (2: 1, v / v), dried over N ₂ gas and dissolved in 100 μl of chloroform. Lipids extracted from the epidermis were separated by ceramide using high performance liquid chromatography (Waters 1525 Binary HPLC pump, Waters 2487 Dual λ Absorbance Detector, Waters Co., USA). In the HPLC analysis, the column used UG 120 (250 × 4.60 mm; 5 μm) from Shiseido Co. The mobile phase was a mixture of hexane and 3% isopropanol (IPA), and the flow rate was 1 ml / min. The ceramide fraction isolated at absorbance 254 nm was quantified using an external ceramides standard and expressed as ceramides μg / μg protein along with the measurement of the protein content of the epidermis used, and the results are shown in FIG. 2.

As can be seen from Figure 2, the total ceramide content measured in the epidermis of each group after 5 weeks of supply of the soybean-derived dietary material was 3.14 ± 0.4 (control A), 0.84 ± 0.2 (control B), 1.73 ± 0.2 (Experimental group 1 ), 1.45 ± 0.3 (Experiment 2), 0.90 ± 0.2 (Experiment 3), 1.22 ± 0.3 (Experiment 4), 1.20 ± 0.3 (Experiment 5) (μg / μg protein). According to the measured ceramide content, the skin barrier injury group (Experimental Group 1 ~ 5 and Control Group B) has lower epidermal ceramide content than the normal control group (Control Group A). It can be seen that the content is high. In particular, the ceramide content was significantly increased compared to the control group B of atopic soldiers when dietary supplementation of 70% ethanol extract of various functional materials, and the ceramide content was slightly increased even when diet was supplied to each fraction of lignan.

After 10 weeks of dietary feeding of each functional dietary material, the total ceramide content measured in the epidermis of each group was 0.07 ± 0.01 (control A), 0.02 ± 0.003 (control B), 0.06 ± 0.01 (experiment 1), 0.04 ± 0.015 ( Experimental group 2), 0.05 ± 0.02 (experimental group 3), 0.05 ± 0.02 (experimental group 4), 0.05 ± 0.02 (experimental group 5) (μg / μg protein) was absolutely lower than the ceramide content measured at week 5. This is believed to be due to the absolute decrease in the ceramide content of the skin as each animal ages. However, changes in dietary supply of each functional dietary material for 10 weeks were more pronounced. The increase in ceramide content of Experimental Group 1 containing 70% ethanol extract of Licorice was more pronounced, showing a level of ceramide content similar to that of Control A, which was the normal control group. In addition, significant increase in ceramide was also seen in groups 3 to 5, which means that the increase in ceramide increase was also achieved by the addition of the respective lignan derivative compounds of the licorice lignan fraction.

Test Example 5 Effect of Dietary Supply of Functional Dietary Materials on Epidermal Growth

Since the increase in DNA is an indicator of cell proliferation, epidermal proliferation was measured using the incorporation of the DNA construct [ ³ H] thymidine (Thymidine Incorporation Assay). Epidermal tissue of 4 mm ² was collected from the control groups A, B and Experimental groups 1 to 5 in Test Example 4, and then incubated at 37 ° C. with [ ³ H] thymidine, and then heated at 95 ° C. with 0.5 M NaOH to release DNA. It was. Fluorescence emitted by reacting DNA with EtBr was measured by using β-counter (Beckmann Instruments). The measurement results are shown in FIG. 3.

It has been reported that epidermal cell proliferation is secondaryly increased when skin is dried by decreasing ceramide content and water retention in the epidermis. 3, the epidermal hyperplasia of the skin barrier injury group (control group B) measured by thymidine incorporation after 5 weeks of feeding was significantly higher than that of the normal control group (experiment group A). It can be seen that there is an inverse correlation.

Epidermal hyperproliferation was significantly suppressed to the normal control level in all experimental groups 1-5 after five weeks of dietary supplementation of several functional dietary materials, suggesting that all functional materials had an inhibitory effect on epidermal hyperproliferation regardless of changes in ceramide content of epidermis. do.

1 is a result of Western blot analysis comparing the increase in the expression level of SPT protein.

Figure 2 shows the experimental results comparing the total amount of epidermal ceramide.

Figure 3 shows the experimental results comparing the degree of epidermal hyperproliferation.

Claims (4)

Rosemary acid represented by Chemical Formula 1, Rosmarinic acid represented by Chemical Formula 2, Lithospermic acid represented by Chemical Formula 2, 9 ”-Methyl lithospermate represented by Chemical Formula 2, and represented by Chemical Formula 2. 9'-Methyl lithospermate (9'-Methyl lithospermate) is a composition for the prevention or treatment of diseases mediated by a ceramide metabolism-related enzyme containing at least one lignan derivative selected from the group consisting of. [Formula 1]

[Formula 2]

(However, R ₁ and R ₂ is a resource permic acid, R ₁ is H, R ₂ is H, 9 ”-methyl resource permate is R ₁ is H, R ₂ is CH ₃ , and 9′-methyl resource is Mate is R ₂ is CH ₃ and R ₁ is H) According to claim 1, wherein the active ingredient is a composition for the prevention or treatment of diseases mediated by the ceramide metabolism-related enzyme, characterized in that it is contained in an amount of 0.01 to 50% by weight relative to the total weight of the composition. According to claim 1, wherein the composition is a composition for the prevention or treatment of diseases mediated by the ceramide metabolism-related enzyme, characterized in that to increase the expression level of serine-palmitoyl transferase. Rosemary acid represented by Chemical Formula 1, Rosmarinic acid represented by Chemical Formula 2, Lithospermic acid represented by Chemical Formula 2, 9 ”-Methyl lithospermate represented by Chemical Formula 2, and represented by Chemical Formula 2. A composition for enhancing the ceramide content of the epidermis containing at least one lignan derivative selected from the group consisting of 9'-methyl lithospermate as an active ingredient. [Formula 1]

[Formula 2]

(However, R ₁ and R ₂ is a resource permic acid, R ₁ is H, R ₂ is H, 9 ”-methyl resource permate is R ₁ is H, R ₂ is CH ₃ , and 9′-methyl resource is Mate is R ₂ is CH ₃ and R ₁ is H)

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