Composition and application thereof in anti-aging

Abstract

The invention provides a composition and application thereof in anti-aging, and relates to the technical field of cosmetics. It is proved for the first time that the 5-aminolevulinic acid or the salt thereof and the vitriol can achieve a synergistic effect, so that the composition has a more excellent effect of promoting the expression of the type I collagen. According to the invention, 5-aminolevulinic acid or salt thereof is compounded with glassine, and the synergistic effect of 5-aminolevulinic acid or salt thereof and glassine is utilized, so that the I-type collagen expression promoting effect of the product is improved, and the content of 5-aminolevulinic acid or salt thereof in the product is reduced at the same time; the problem that the product is toxic or poor in safety due to too high content of 5-aminolevulinic acid or salt thereof is solved, and the problem that the I-type collagen expression promoting effect is poor due to low content of 5-aminolevulinic acid or salt thereof is also solved; the composition which is better in effect, higher in safety and lower in cost and has excellent anti-wrinkle and firming effects is provided for the field of cosmetics.

Description

Technical Field

[0001] This application relates to the field of cosmetic technology, and more particularly to a composition and its application in anti-aging. Background Technology

[0002] 5-Aminolevulinic acid (5-ALA) or its salts (5-ALA or ALA) are widely used in agriculture as plant growth regulators. 5-ALA is a precursor to chloroplast proteins, promoting chlorophyll synthesis and accumulation, thereby improving crop photosynthetic efficiency. Simultaneously, when applied to the skin, ALA effectively promotes collagen production and cell metabolism, thus improving skin elasticity, reducing wrinkles, and providing anti-aging effects. However, due to its high toxicity, ALA is difficult to use in skincare product formulations.

[0003] Pro-Xylane is a naturally occurring polypeptide in the human body that regulates cell proliferation, migration, and differentiation. Studies have shown that Pro-Xylane can promote collagen synthesis by influencing different signaling pathways. For example, it can activate the PI3K / Akt and JNK signaling pathways, thereby promoting fibroblast proliferation and collagen synthesis. Furthermore, Pro-Xylane can inhibit the TGF-β1 signaling pathway, thus reducing collagen degradation, and shows great promise in the field of skin anti-aging. Summary of the Invention

[0004] To address the issue that ALA is difficult to use in skincare formulations due to its high toxicity, this application demonstrates for the first time that when 5-aminolevulinic acid or its salt is combined with BOXER in a mass ratio of 1:1000-1:300000, the composition exhibits excellent effects in promoting type I collagen expression, ensuring both safety and anti-wrinkle and firming effects.

[0005] On one hand, this application provides a composition comprising 5-aminolevulinic acid or a salt thereof and bosine.

[0006] In the composition, 5-aminolevulinic acid or its salt and bosine synergistically promote the expression of type I collagen.

[0007] Further, the 5-aminolevulinic acid or its salts include 5-aminolevulinic acid, 5-aminolevulinic acid salt, 5-aminolevulinic acid sulfate, 5-aminolevulinic acid carbonate, 5-aminolevulinic acid nitrate, 5-aminolevulinic acid acetate, or 5-aminolevulinic acid phosphate.

[0008] Further, the mass ratio of the 5-aminolevulinic acid or its salt to Bosein is 1:(1000-300000).

[0009] The mass ratio of 5-aminolevulinic acid or its salt to Bosein can be 1:1000, 1:1100, 1:1200, 1:1300, 1:1400, 1:1499, 1:1500, 1:1600, 1:1700, 1:1800, 1:1900, 1:2000, 1:2100, 1:2200, 1:2300, 1:2400, 1:2500, 1:2600, 1:2700, 1:2800, 1:2900, 1:2999, 1:3000, 1:4000, 1:5000, 1:6000, or 1:7. Any value from 000, 1:8000, 1:9000, 1:10000, 1:11000, 1:12000, 1:13000, 1:14000, 1:15000, 1:16000, 1:17000, 1:18000, 1:19000, 1:20000, 1:29999, 1:30000, 1:40000, 1:50000, 1:60000, 1:70000, 1:80000, 1:90000, 1:100000, 1:200000, 1:299999, 1:300000.

[0010] Preferably, the mass ratio of 5-aminolevulinic acid or its salt to Bosein is 1:(1499-30000).

[0011] Preferably, the composition is an anti-aging composition; more preferably, the composition is a highly safe anti-aging composition.

[0012] Further, the mass content of the 5-aminolevulinic acid or its salt, calculated as a percentage by mass of the composition, is 0.00000001%-0.002%.

[0013] Wherein, calculated by mass percentage of the composition, the mass content of 5-aminolevulinic acid or its salt may be 0.00000001%, 0.00000002%, 0.00000003%, 0.00000004%, 0.00000005%, 0.00000006%, 0.00000007%, 0.00000008%, 0.00000009%, 0.0000001%, 0.0000002%, 0.0000003%, 0.0000004%, 0.0000005%, 0.0000006%, 0.0000007%, 0.0000008%, 0.0000009%, 0.000001%, 0.0 0.00002%, 0.000003%, 0.000004%, 0.000005%, 0.000006%, 0.000007%, 0.000008%, 0.000009%, 0.00001%, 0.00002%, 0.00003%, 0.00004%, 0.00005%, 0.0000 Any value from 6%, 0.00007%, 0.00008%, 0.00009%, 0.0001%, 0.0002%, 0.0003%, 0.0004%, 0.0005%, 0.0006%, 0.0007%, 0.0008%, 0.0009%, 0.001%, 0.0015%, and 0.002%.

[0014] Furthermore, the Bosein content is 0.01%-3% by mass percentage of the composition.

[0015] The Bosein content, calculated as a percentage by mass of the composition, can be 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09%, 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%, 1.7%, 1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, or 3.0%.

[0016] This application may also add excipients, which may be appropriate solvents, propellants, solubilizers, co-solvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, flow aids, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesion agents, binding agents, penetration promoters, pH adjusters, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, encapsulating agents, humectants, absorbents, diluents, flocculants and anti-flocculation agents, filter aids, release inhibitors, etc.

[0017] In a preferred embodiment, the solvent may be water or a culture medium. Those skilled in the art can select appropriate solvents to formulate the above composition according to actual needs, and no specific type of solvent is limited herein.

[0018] In addition, one or more diluents or carriers may be added to the composition of this application to prepare commonly used cosmetic dosage forms such as ointments, creams, emulsions, patches, masks, liquids, and tinctures.

[0019] The compositions of this application can be prepared by common methods. In a preferred embodiment, the preparation method of the composition includes: dissolving 5-aminolevulinic acid or its salt (ALA) and Bosein in a solvent and mixing them.

[0020] On the other hand, this application also provides the use of the composition in anti-aging.

[0021] The anti-aging effect is achieved by synergistically increasing the collagen content in cells.

[0022] On the other hand, this application also provides the use of the composition in increasing collagen content.

[0023] Furthermore, the method of increasing collagen content is to synergistically promote collagen synthesis.

[0024] Furthermore, the collagen is type I collagen.

[0025] Preferably, the method of increasing collagen content is to increase the expression level of type I collagen gene.

[0026] In a preferred embodiment, human fibroblasts were used as a model in this application to conduct experiments, demonstrating the synergistic promoting effect of the composition on the expression level of type I collagen gene in cells, thereby demonstrating the anti-aging efficacy of the composition.

[0027] The present invention has the following beneficial effects:

[0028] This application demonstrates for the first time that 5-aminolevulinic acid or its salt and Bosein can synergistically enhance each other when combined in a mass ratio of 1:1000 to 1:300000, resulting in a more superior effect on promoting type I collagen expression.

[0029] This application combines 5-aminolevulinic acid or its salt with BOXER to achieve a synergistic effect. This enhances the product's ability to promote type I collagen expression while reducing the content of 5-aminolevulinic acid or its salt in the product. This avoids the problems of toxicity or poor safety caused by excessive 5-aminolevulinic acid or its salt content, and also solves the problem of poor type I collagen expression promotion caused by low 5-aminolevulinic acid or its salt content. This provides the cosmetics industry with a composition that is more effective, safer, and less costly, and has excellent anti-wrinkle and firming effects. Detailed Implementation

[0030] To more clearly illustrate the overall concept of this application, a detailed description is provided below by way of embodiments. Numerous specific details are set forth in the following description to provide a more thorough understanding of the invention. However, it will be apparent to those skilled in the art that the invention can be practiced without one or more of these details. In other instances, certain technical features well-known in the art have not been described to avoid confusion with the invention.

[0031] Unless otherwise specified in the examples, the conditions shall be performed according to the standard conditions or the conditions recommended by the manufacturer.

[0032] Unless otherwise specified, all reagents or instruments used in the following embodiments, unless otherwise indicated by the manufacturer, are commercially available products. Specifically, the RNA reverse transcription kit and RNA extraction kit were purchased from Vazyme, and the cell culture medium was 90% low-glucose DMEM medium + 10% fetal bovine serum. The low-glucose DMEM medium was purchased from VivaCell, and the fetal bovine serum was purchased from Gibco.

[0033] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in the fields of microbiology, biochemistry, analytical chemistry, cell culture, and related areas.

[0034] In addition, the "water" mentioned in this invention includes any feasible water that can be used in the art, such as deionized water, distilled water, ion-exchanged water, double-distilled water, high-purity water, and purified water.

[0035] In the following examples, unless otherwise specified, % means wt%, i.e., weight percentage.

[0036] Example 1

[0037] 5-Aminolevulinate and Bosein were dissolved in water, wherein the mass content of 5-aminolevulinate was 0.0002% and the mass content of Bosein was 0.2998%.

[0038] Example 2

[0039] 5-Aminolevulinate and Bosein were dissolved in water, wherein the mass content of 5-aminolevulinate was 0.0001% and the mass content of Bosein was 0.2999%.

[0040] Example 3

[0041] 5-Aminolevulinate and Bosein were dissolved in water, wherein the mass content of 5-aminolevulinate was 0.00001% and the mass content of Bosein was 0.29999%.

[0042] Example 4

[0043] 5-Aminolevulinate and Bosein were dissolved in water, wherein the mass content of 5-aminolevulinate was 0.000001% and the mass content of Bosein was 0.299999%.

[0044] Example 5

[0045] 5-Aminolevulinate and Bosein were dissolved in water, wherein the mass content of 5-aminolevulinate was 0.001% and the mass content of Bosein was 0.299%.

[0046] Comparative Example 1

[0047] 5-Aminolevulinate salt was dissolved in water, and the mass content of 5-aminolevulinate salt was 0.3%.

[0048] Comparative Example 2

[0049] Bosein was dissolved in water, and the mass content of Bosein was 0.3%.

[0050] Test case

[0051] The above embodiments and comparative examples were subjected to cell anti-aging tests. The test method included the following steps:

[0052] (1) Cell seeding: at 8×10 4 Human fibroblasts were seeded into 24-well plates at a density of / wells and incubated overnight in an incubator (37°C, 5% CO2).

[0053] (2) Experimental grouping: The experiment included a blank control group, a negative control group, a positive control group, and a sample group. The sample group had 5 concentration gradients (i.e., Examples 1-5).

[0054] (3) Solution preparation: Prepare working solutions of different concentrations of test substances according to the test concentration setting table (Table 1).

[0055] (4) Irradiation: Irradiation was carried out when the cell deposition rate in the 24-well plates reached 40%-60%. The total dose received by the negative control group, positive control group, and sample group was 9 J / cm². 2 The control group was subjected to UVA irradiation, while the blank control group was placed in the same environment but did not receive UVA irradiation.

[0056] (5) Drug administration: After irradiation, 1 mL of cell culture medium was added to each well of the blank control group and the negative control group; 1 mL of culture medium containing 100 μg / mL VC and 7 μg / mL VE was added to each well of the positive control group; and 1 mL of the example and comparative sample containing the corresponding concentration of the test substance was added to each well of the sample group.

[0057] (6) Cell collection: After incubation for 24 hours, add 0.5 mL of lysis buffer to each well, incubate at room temperature for 5 minutes to allow for complete lysis, transfer to 1.5 mL RNase-free Eppendorf tubes, and extract RNA according to the operating procedure.

[0058] (7) Reverse transcription: Synthesize cDNA according to the instructions of the RNA reverse transcription kit.

[0059] (8) The CollagenⅠ (type I collagen) gene was detected by qRT-PCR. The results are shown in Tables 2 and 3. The data were analyzed for significant differences.

[0060] Table 1 Test Groups

[0061]

[0062]

[0063] Table 2 Results of Collagen I gene expression level assay

[0064]

[0065] Note: Compared with group BC, p < 0.001 is indicated as ###; compared with group NC, p < 0.001 is indicated as ***. Compared with group ALA (Comparative Example 1), p < 0.01 is indicated as ▲▲; compared with group Bosein (Comparative Example 2), p < 0.05 is indicated as ▼.

[0066] Table 3. Results of Collagen I gene expression level assay

[0067]

[0068] Note: Compared with group BC, p < 0.001 is indicated as ###; compared with group NC, p < 0.001 is indicated as ***. Compared with group ALA (Comparative Example 1), p < 0.001 is indicated as ▲▲▲; compared with group Bosein (Comparative Example 2), p < 0.01 is indicated as ▼▼.

[0069] As shown in Table 2, compared with the NC group, ALA and Bosein monomers at a concentration of 0.3% significantly promoted the expression level of Collagen I gene in human fibroblasts (p<0.001). The ALA and Bosein composition prepared using the method of Example 2 also significantly promoted the expression level of Collagen I gene in human fibroblasts (p<0.001). Furthermore, compared with Comparative Examples 1 and 2, the ALA and Bosein composition prepared using the method of Example 2 at a mass ratio of 1:2999 significantly enhanced the expression level of Collagen I gene (p<0.05).

[0070] As shown in Table 3, compared with the NC group, ALA and Bosein monomers at a concentration of 0.3% significantly promoted the production of Collagen I in human fibroblasts (p<0.001). The ALA and Bosein compositions prepared using the methods of Examples 1 and 3 also significantly promoted Collagen I gene expression in human fibroblasts (p<0.001). Furthermore, compared with Comparative Examples 1 and 2, the ALA and Bosein compositions prepared using the methods of Examples 1 and 3, when mixed at mass ratios of 1:1499 and 1:29999, significantly increased Collagen I gene expression (p<0.01).

[0071] in conclusion:

[0072] Based on a UVA-stimulated human fibroblast model, this application verified that monomers of 0.3% ALA, 0.3% BOXER, and combinations of ALA and BOXER all significantly enhanced the gene expression of type I collagen.

[0073] Furthermore, compared to the monomers, the composition, when compounded at a mass ratio of 1:1499-1:29999, significantly promotes the production of type I collagen. This indicates that the combination of ALA and BOXER has anti-wrinkle and firming effects, and that the two have a synergistic effect.

[0074] The above description is merely an embodiment of this application and is not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. A composition, characterized in that, The composition comprises 5-aminolevulinic acid or a salt thereof and bosine.

2. The composition according to claim 1, characterized in that, The mass ratio of 5-aminolevulinic acid or its salt to Bosein is 1:(1000-300000).

3. The composition according to claim 2, characterized in that, The mass ratio of 5-aminolevulinic acid or its salt to Bosein is 1:(1499-30000).

4. The composition according to claim 1, characterized in that, The 5-aminolevulinic acid or its salt, calculated as a percentage by mass of the composition, has a content of 0.00000001%-0.002%.

5. The composition according to claim 1, characterized in that, The Bosein content is 0.01%-3% by mass percentage of the composition.

6. The use of the composition according to any one of claims 1-5 in anti-aging.

7. The use of the composition according to any one of claims 1-5 in increasing collagen content.

8. The application according to claim 7, characterized in that, The method to increase collagen content is to promote collagen synthesis.

9. The application according to claim 7, characterized in that, The collagen is type I collagen.

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