Composition containing photolyase and application of composition in preparation of product with light repairing and / or anti-aging effects

Abstract

The invention relates to the technical field of biological medicine, and particularly discloses a composition containing photolyase and application of the composition in preparation of a product with light repairing and / or anti-aging effects. The composition containing the photolysis synthase is prepared from the photolysis synthase and an evening primrose root extract. Researches show that the light repairing effect of the composition containing the photolyase disclosed by the invention is equivalent to that of the photolyase; therefore, when the composition containing the photolysis synthase is used as an effective component for developing a product with a light repairing effect, compared with the composition adopting the photolysis synthase, the composition containing the photolysis synthase is beneficial to reducing the development cost of the product. In addition, the composition containing the photolyase, disclosed by the invention, has relatively good light repairing effect; therefore, the anti-aging effect can be further realized through the photoremediation effect; a product taking the compound as an active component also has an anti-aging effect.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a composition containing photolyase and application thereof in preparing products with photorepair and / or anti-aging effects. Background Art

[0002] Skin photoaging is the damage caused by long-term exposure to light, which leads to aging phenomena such as roughness, thickening, dryness, sagging, and deepening and thickening of wrinkles. Among them, sunlight and blue light emitted by electronic product screens are the main factors that cause skin cell damage and aging due to their short wavelength, high energy and strong penetration. Therefore, it is of great application value to develop a product that can repair skin cell damage caused by blue light.

[0003] Photolyase is an enzyme that can absorb blue light energy and use this energy to repair DNA; it has a good repair effect; and it is currently widely used in the field of skin care products. For example, Chinese invention patent 202410591317.0 discloses a sunscreen composition containing photolyase and collagen, and its preparation method and application; the sunscreen composition includes the following components and their mass percentages: photolyase 0.01~5%, collagen 0.01~5%, sunscreen 20~50%, auxiliary materials 20.91~53.6%, and the balance deionized water, etc.; wherein the collagen is composed of type I collagen, type III collagen, and other collagens in a mass ratio of 12~15:1~5:87-80. The present invention combines the function of photolyase to repair DNA and the film-forming performance of collagen on the skin surface to provide a composition containing photolyase and collagen to synergistically enhance sunscreen performance.

[0004] However, the raw materials of photolyase are currently expensive; therefore, developing a composition containing photolyase that has other components and has the same repairing effect as photolyase is of great application value in providing more raw material choices for the development of skin care products and reducing the production cost of corresponding skin care products. Summary of the invention

[0005] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a composition containing photolyase and its use in preparing products with photorepair and / or anti-aging effects.

[0006] The present invention firstly provides a composition containing photolyase, which comprises photolyase and evening primrose root extract.

[0007] The inventors found in their research that a composition consisting of photolyase and evening primrose root extract has a photorepair effect.

[0008] Preferably, the weight ratio of photolyase to evening primrose root extract is 1:1-10.

[0009] More preferably, the weight ratio of photolyase to evening primrose root extract is 1:3-6.

[0010] Most preferably, the weight ratio of photolyase to evening primrose root extract is 1:4.

[0011] Preferably, the evening primrose root extract is prepared by the following method: Take evening primrose root, add solvent and perform extraction, after the extraction is completed, concentrate and dry the extract to obtain evening primrose root solvent extract; take the evening primrose root solvent extract to obtain the evening primrose root extract.

[0012] Preferably, the solvent consists of n-propanol, tetrahydrofuran and water.

[0013] In further research, the inventors found that in the preparation process of evening primrose root extract, the selection of extraction solvent is very critical. Only the composition obtained by combining the evening primrose root extract obtained by extraction with a solvent composed of n-propanol, tetrahydrofuran and water with photolyase can have a photorepair effect equivalent to that of photolyase; however, the photorepair effect of the composition obtained by combining the evening primrose root extract obtained by extraction with other solvents or a combination of solvents with photolyase cannot achieve a photorepair effect equivalent to that of photolyase.

[0014] Preferably, the usage ratio of evening primrose root to solvent is 1kg:5~15L.

[0015] Most preferably, the ratio of evening primrose root to solvent is 1 kg:8 L.

[0016] More preferably, the solvent is n-propanol, tetrahydrofuran and water in a volume ratio of 5-10:2-4:3-5.

[0017] Most preferably, the solvent is n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:4.

[0018] Preferably, the preparation method of the evening primrose root extract further comprises the following steps: applying the evening primrose root solvent extract to a macroporous resin column, first eluting with 5 to 10 times the column volume of an ethanol aqueous solution with a volume fraction of 12 to 15% to remove impurities; then eluting with 5 to 10 times the column volume of an ethanol aqueous solution with a volume fraction of 27 to 30%, collecting the eluate eluted with the ethanol aqueous solution with a volume fraction of 27 to 30%, and concentrating and drying to obtain the evening primrose root extract.

[0019] Most preferably, the preparation method of the evening primrose root extract also comprises the following steps: subjecting the evening primrose root solvent extract to a macroporous resin column, first eluting with 8 times the column volume of an ethanol aqueous solution with a volume fraction of 13% to remove impurities; then eluting with 8 times the column volume of an ethanol aqueous solution with a volume fraction of 29%, collecting the eluate eluted with the 29% ethanol aqueous solution, and concentrating and drying to obtain the evening primrose root extract.

[0020] Preferably, the macroporous resin column is a D101 macroporous resin column.

[0021] The inventors surprisingly found in further research that the photorepair effect of the composition obtained by combining the evening primrose root extract prepared by further eluting the evening primrose root solvent extract through the above-mentioned macroporous resin column of the present invention with photolyase is significantly higher than that of photolyase.

[0022] The inventors also found in the research that the elution conditions of the macroporous resin column of the present invention are very critical; only when the evening primrose root solvent extract prepared under the above-mentioned macroporous resin column elution conditions of the present invention is combined with photolyase to obtain a composition, its photorepair effect is significantly higher than that of photolyase; however, the photorepair effect of the composition obtained by combining the evening primrose root extract prepared under other macroporous resin column elution conditions with photolyase is not significantly higher than that of photolyase.

[0023] Preferably, the photolyase-containing composition refers to a photolyase-containing composition having the effect of increasing the activity of skin fibroblasts.

[0024] The skin fibroblast activity refers to the skin fibroblast activity after being irradiated with blue light.

[0025] The present invention also provides an application of the above-mentioned composition containing photolyase in preparing products with photorepair effects.

[0026] Preferably, the light repair specifically refers to the repair of skin cell damage caused by blue light.

[0027] Preferably, the skin cells specifically refer to skin fibroblasts.

[0028] The present invention also provides an application of the composition containing photolyase in preparing a product having the function of improving the activity of skin fibroblasts.

[0029] The skin fibroblast activity refers to the skin fibroblast activity after being irradiated with blue light.

[0030] The present invention also provides an application of the above-mentioned composition containing photolyase in preparing a product with anti-aging effect.

[0031] Preferably, the product is a skin care product or a medicine.

[0032] Beneficial effects: The present invention provides a new composition containing photolyase; studies have shown that the photolyase-containing composition of the present invention has a photorepair effect equivalent to that of photolyase; therefore, using the photolyase-containing composition of the present invention as an effective ingredient for developing products with photorepair effects is beneficial to reducing product development costs compared to using photolyase; and also provides more raw material options for the development of products with photorepair effects. In addition, since the photolyase-containing composition of the present invention has good photorepair effects; therefore, it can also further achieve anti-aging effects through photorepair effects; products using it as an active ingredient also have anti-aging effects. DETAILED DESCRIPTION

[0033] The present invention is further explained below in conjunction with specific examples, but the examples do not limit the present invention in any form.

[0034] Example 1 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 2 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:4.

[0035] Example 2 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; Wherein, the evening primrose root extract is prepared by the following method: (1) taking evening primrose root, adding a solvent, heating to boiling, and then reflux extraction for 2 hours. After the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; wherein the ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:4; (2) The evening primrose root solvent extract is loaded onto a D101 macroporous resin column, and impurities are first eluted with 8 times the column volume of a 13% ethanol aqueous solution; then, the impurities are eluted with 8 times the column volume of a 29% ethanol aqueous solution, and the eluate eluted with the 29% ethanol aqueous solution is collected, concentrated, and dried to obtain the evening primrose root extract.

[0036] Example 3 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly at a weight ratio of 1:10 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 1 hour, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:15L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 5:4:5.

[0037] Example 4 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed uniformly in a weight ratio of 1:1 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 1.5 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:5L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 10:2:3.

[0038] Comparative Example 1 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 2 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of n-propanol and water in a volume ratio of 8:4.

[0039] Comparative Example 2 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 2 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of tetrahydrofuran and water in a volume ratio of 3:4.

[0040] Comparative Example 3 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 2 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:8L; and the solvent is methanol.

[0041] Comparative Example 4 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; The evening primrose root extract is prepared by the following method: taking evening primrose roots, adding a solvent, heating to boiling, and then reflux extraction for 2 hours, and after the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; taking the evening primrose root solvent extract to obtain the evening primrose root extract; The dosage ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of methanol, tetrahydrofuran and water in a volume ratio of 8:3:4.

[0042] Comparative Example 5 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; Wherein, the evening primrose root extract is prepared by the following method: (1) taking evening primrose root, adding a solvent, heating to boiling, and then reflux extraction for 2 hours. After the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; wherein the ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:4; (2) The evening primrose root solvent extract is loaded onto a D101 macroporous resin column, and impurities are first eluted with 8 times the column volume of a 5% ethanol aqueous solution; then, the impurities are eluted with 8 times the column volume of a 15% ethanol aqueous solution, and the eluate eluted with the 15% ethanol aqueous solution is collected, concentrated, and dried to obtain the evening primrose root extract.

[0043] Comparative Example 6 Preparation of a composition containing photolyase The photolyase and evening primrose root extract are mixed evenly in a weight ratio of 1:4 to obtain the photolyase-containing composition; Wherein, the evening primrose root extract is prepared by the following method: (1) taking evening primrose root, adding a solvent, heating to boiling, and then reflux extraction for 2 hours. After the extraction, concentrating and drying the extract to obtain an evening primrose root solvent extract; wherein the ratio of evening primrose root to solvent is 1kg:8L; the solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:4; (2) The evening primrose root solvent extract is loaded onto a D101 macroporous resin column, and impurities are first eluted with 8 times the column volume of a 45% ethanol aqueous solution; then, the impurities are eluted with 8 times the column volume of a 60% ethanol aqueous solution, and the eluate eluted with the 60% ethanol aqueous solution is collected, concentrated, and dried to obtain the evening primrose root extract.

[0044] Experimental Example 1 HFF-1 human skin fibroblasts were collected and cultured at 1×10 4 The cells were seeded into 96-well plates at a density of 100 cells / well. The cells were divided into an experimental group, a blank control group, and a blue light control group; then RPMI-1640 medium was added for culture, and after culturing in a cell culture incubator at 37°C and 5% CO2 for 12 hours, the experimental group and the blue light control group were placed under a blue light lamp for 12 hours, and the blank control group was not irradiated with a blue light lamp; then the experimental group was replaced with a fresh RPMI-1640 medium containing 100 μg / mL of the sample to be tested, and the blank control group and the blue light control group were replaced with a fresh RPMI-1640 medium without the sample to be tested; the culture was continued for 24 hours; after the culture was completed, 200 μL of 1 mg / mL MTT solution was added to each well, and the incubation was continued for 3 hours; finally, 200 μL of DMSO was added to each well, and the OD value was measured at 490 nm using an enzyme marker after oscillation and dissolution; 5 parallel wells were set up for each group; the cell activity was calculated according to the OD value; the cell activity of the experimental group = (OD 实验组 / OD 空白对照组 )*100%; Cell activity in blue light control group = (OD 蓝光对照组 / OD 空白对照组)*100%; the test results are shown in Table 1.

[0045]

[0046] From the experimental results in Table 1, it can be seen that the cell activity of human skin fibroblasts in the blue light control group was only 48.6% after being irradiated with blue light, which shows that blue light irradiation caused damage to human skin fibroblasts. However, the cell activity of human skin fibroblasts in experimental group 9 after adding photolyase to culture reached 82.2%, which is much higher than that of the blue light control group without photolyase, which shows that photolyase can significantly repair the damage of skin fibroblasts caused by blue light and has excellent light repair effect.

[0047] It can be seen from the experimental results in Table 1 that the cell survival rate of experimental group 1 is equivalent to that of experimental group 9; this shows that the composition composed of photolyase and evening primrose root extract prepared according to the method of the present invention has a photorepair effect; and its photorepair effect is equivalent to that of photolyase.

[0048] From the experimental results in Table 1, it can be seen that the cell survival rates of experimental groups 3 to 6 are lower than or even significantly lower than those of experimental groups 1 and 9; this indicates that in the preparation process of evening primrose root extract, the selection of extraction solvent is very critical. Only the evening primrose root extract obtained by extraction with a solvent consisting of n-propanol, tetrahydrofuran and water and the photolyase-derived composition can have a photorepair effect equivalent to that of photolyase; however, the evening primrose root extract obtained by extraction with other solvents or a combination of solvents and the photolyase-derived composition cannot achieve a photorepair effect equivalent to that of photolyase.

[0049] It can be seen from the experimental results in Table 1 that the cell survival rate of experimental group 2 is further significantly higher than that of experimental group 1 and experimental group 9; this indicates that the photorepair effect of the composition obtained by combining the evening primrose root extract prepared by further eluting the evening primrose root solvent extract through the macroporous resin column described in the present invention with photolyase is significantly higher than that of photolyase.

[0050] It can be seen from the experimental results in Table 1 that the cell survival rate of experimental group 7 was not further significantly higher than that of experimental group 1 and experimental group 9; the cell survival rate of experimental group 8 was even lower than that of experimental group 1 and experimental group 9; this shows that: the elution conditions of the macroporous resin column of the present invention are very critical; only when the composition obtained by combining the evening primrose root extract prepared by the evening primrose root solvent extract under the above-mentioned macroporous resin column elution conditions of the present invention with photolyase can its photorepair effect be significantly higher than that of photolyase; however, the photorepair effect of the composition obtained by combining the evening primrose root extract prepared by the evening primrose root solvent extract under other macroporous resin column elution conditions with photolyase is not significantly higher than that of photolyase; it has a very excellent photorepair effect.

Claims

1. A composition containing photolyase, characterized in that Contains photolyase and evening primrose root extract.

2. The composition containing photolyase according to claim 1, characterized in that The weight ratio of photolyase to evening primrose root extract is 1:1-10.

3. The composition containing photolyase according to claim 1, characterized in that The weight ratio of photolyase to evening primrose root extract is 1:3-6.

4. The composition containing photolyase according to claim 3, characterized in that The weight ratio of photolyase to evening primrose root extract was 1:

4.

5. The composition containing photolyase according to claim 1, characterized in that The evening primrose root extract is prepared by the following method: Take evening primrose root, add solvent and perform extraction, after the extraction is completed, concentrate and dry the extract to obtain evening primrose root solvent extract; take the evening primrose root solvent extract to obtain the evening primrose root extract.

6. The composition containing photolyase according to claim 1, characterized in that The solvent consists of n-propanol, tetrahydrofuran and water.

7. The composition containing photolyase according to claim 1, characterized in that The solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 5-10:2-4:3-5.

8. The composition containing photolyase according to claim 1, characterized in that The solvent is composed of n-propanol, tetrahydrofuran and water in a volume ratio of 8:3:

4.

9. Use of the composition containing photolyase according to any one of claims 1 to 8 in the preparation of a product with photorepair effect.

10. Use of the composition containing photolyase according to any one of claims 1 to 8 in the preparation of a product with anti-aging effect.

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