Composition for relieving and repairing skin and application thereof

Abstract

The invention relates to the technical field of skin care, in particular to a composition for relieving and repairing skin and application thereof. The invention provides the composition containing the animal placenta protein, the fibronectin and the recombinant collagen, the composition contains few components which are allowed to be added in the current cosmetic raw material catalog in China, the safety is guaranteed, the effects of all the components are clear, and the composition is verified by systematic cell experiments and human body experiments and has a good application prospect. The composition can be used as a cosmetic raw material in cosmetics for relieving and repairing skin.

Description

Technical Field

[0001] The invention relates to the technical field of skin care, in particular to a composition for soothing and repairing skin and application thereof. Background Art

[0002] The entire surface of the human body is covered with skin. The skin is a soft tissue that is flexible and elastic and can be pushed and stretched within a certain range. The skin is the largest and most important organ of the human body, accounting for about 8% of the total weight of the human body. The skin barrier constitutes the largest protective layer between the human body and the outside world, which can not only reduce the loss of subcutaneous moisture and active substances, but also effectively alleviate the stimulation caused by external ultraviolet rays, dust particles and other factors. In recent years, with the growth of the economy, people have higher and higher requirements for living standards, and they have paid more and more attention to their facial skin problems. However, the air pollution and excessive ultraviolet radiation that can be seen everywhere will cause damage to our skin, and skin problems that can be seen everywhere will also appear, such as spots, sensitive skin, acne, etc., which seriously affect people's pursuit of beauty.

[0003] In recent years, the research on animal placenta has been increasing, and the physiological activities of many animal placentas such as cattle, sheep, deer, donkeys, etc. have been continuously discovered. Animal placenta extract is a kind of compound active ingredients derived from animal placenta, rich in amino acids, proteins, lecithin, polysaccharides, vitamins, mineral elements, etc., and has the effects of enhancing memory, resisting stress, regulating endocrine, enhancing immunity, and delaying aging.

[0004] Fibronectin (FN) is a high molecular weight non-collagenous glycoprotein, mainly produced by liver and vascular endothelial cells, widely present in plasma, various cell surfaces and cell matrix, and is one of the main components of the extracellular matrix. FN plays a very critical role in regulating cell adhesion, migration, growth, proliferation and differentiation, and participates in wound healing, embryonic development, immune regulation and other processes, and has multiple biological activities and biological functions of cell regeneration and repair.

[0005] Collagen is a very important connective tissue protein in human skin. Collagen fibers formed by collagen combine with elastic fibers in the dermis to form a mesh structure, which can support the skin structure, maintain skin elasticity and toughness, retain skin moisture, make the skin smoother, tender and elastic, and is one of the most important proteins for skin beauty. Recombinant collagen is a high-molecular biological protein produced by reverse transcription of human collagen mRNA into cDNA through bioengineering technology, and then a gene segment after enzyme cutting is recombined in a vector, and then produced through high-density fermentation, separation, renaturation and purification processes. Yeast recombinant collagen, which uses yeast as a genetic engineering vector, has a protein modification ability similar to that of the human body. After collagen synthesis, it is modified to give a folded conformation and a specific amino acid structure, becoming a physiologically active collagen with a similarity of more than 98% to the human collagen sequence. Recombinant collagen has the advantages of no virus hidden dangers, low rejection reaction, good water solubility, neutral pH, high denaturation temperature, and strong processability, making it a cosmetic raw material with good application prospects.

[0006] The ingredients of the compositions currently disclosed in the market for soothing and repairing effects are too complex. For example, a composition with soothing and repairing effects developed by a certain company includes squalane, jojoba seed oil, shea butter, soothing complex (shea butter, argan oil, waste malt meal, wax and vitamin E), and ceramide. The evaluation method for its soothing and repairing effect is too simple, and there is a lack of systematic efficacy evaluation and efficacy verification research from cell experiments to human trials. Therefore, it is of great significance to develop a composition for skin care with simple ingredients and comprehensive and systematic efficacy verification results in the field of cosmetics. Summary of the invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a composition for soothing and repairing skin and its application. The composition provided by the present invention includes animal placental protein, fibronectin and recombinant collagen, and has good skin soothing and repairing effects after optimized formula.

[0008] The present invention provides a composition for soothing and repairing skin, which comprises animal placental protein, fibronectin and recombinant collagen;

[0009] The mass ratio of the animal placental protein, fibronectin and recombinant collagen is (5-500): (0.001-0.1): (2.5-250).

[0010] Furthermore, the mass ratio of the animal placental protein, fibronectin and recombinant collagen is (5-50): (0.001-0.01): (2.5-25).

[0011] Specifically, in some embodiments, the mass ratio of the animal placental protein, fibronectin and recombinant collagen is 50:0.01:25.

[0012] The present invention designs 7 groups of compositions through experiments, each group having different components. Through the cell efficacy evaluation test, it is found that when only one or two of animal placental protein, fibronectin and recombinant collagen are in the composition, the cell proliferation, repair and anti-inflammatory effects are not optimally achieved. Only when the composition contains animal placental protein, fibronectin and recombinant collagen at the same time and the mass ratio is 50:0.01:25, can the combination with other ratios play a more effective synergistic effect.

[0013] The invention provides a method for preparing the composition, which comprises weighing animal placental protein, fibronectin and recombinant collagen according to concentrations, mixing them evenly, and sealing them for standby use.

[0014] The present invention provides use of the composition in preparing a product for soothing and / or repairing skin.

[0015] The present invention also provides a product for soothing and / or repairing skin, which comprises the composition of the present invention and an acceptable auxiliary material or adjuvant.

[0016] Furthermore, the mass fraction of the composition is 3% to 30%.

[0017] Specifically, in an embodiment of the present invention, the mass fraction of the composition is 3%.

[0018] The present invention sets up a test group and a control group, and uses products containing the composition of the present invention and products not containing the composition of the present invention on human bodies for efficacy evaluation. The results show that after subjects of different age ranges use the product containing the composition of the present invention, the erythema area is reduced and the epidermal water loss is reduced, indicating that the product containing the composition of the present invention has soothing and repairing effects.

[0019] Furthermore, in the product, the auxiliary material or adjuvant is selected from one or more of butylene glycol, 1,3-propylene glycol, C20-22 alcohol, caprylyl glycol, phenoxyethanol, tert-butyl alcohol, tromethamine, polydimethylsiloxane, acrylic acid (ester) copolymer, polyacrylamide, C13-14 isoparaffin, laureth-7, caprylhydroxamic acid, C20-22 alcohol phosphate, p-hydroxyacetophenone, xanthan gum, ethylhexylglycerin, glucose and water.

[0020] In the product, the mass fraction of butanediol is 8%, the mass fraction of 1,3-propylene glycol is 1%, the mass fraction of C20-22 alcohol is 0.8%, the mass fraction of caprylyl glycol is 0.5%, the mass fraction of phenoxyethanol is 0.5%, the mass fraction of tert-butyl alcohol is 0.2%, the mass fraction of tromethamine is 0.2%, the mass fraction of polydimethylsiloxane is 1%, the mass fraction of acrylic acid (ester) copolymer is 1.5%, the mass fraction of polyacrylamide is 1.4%, the mass fraction of C13-14 isoparaffin is 1.4%, the mass fraction of laureth-7 is 1.4%, the mass fraction of caprylhydroxamic acid is 1%, the mass fraction of C20-22 alcohol phosphate is 0.8%, the mass fraction of p-hydroxyacetophenone is 0.3%, the mass fraction of xanthan gum is 0.5%, the mass fraction of ethylhexylglycerol is 0.5%, the mass fraction of glucose is 0.5%, the mass fraction of the composition is 3%, and the balance is water.

[0021] In an embodiment of the present invention, the auxiliary materials or adjuvants include butylene glycol, 1,3-propylene glycol, C20-22 alcohol, caprylyl glycol, phenoxyethanol, tert-butyl alcohol, tromethamine, polydimethylsiloxane, acrylic acid (ester) copolymers, polyacrylamide, C13-14 isoparaffin, laureth-7, caprylhydroxamic acid, C20-22 alcohol phosphate, p-hydroxyacetophenone, xanthan gum, ethylhexylglycerin, macadamia ternifolia seed oil, glucose and water. The auxiliary materials can be better matched with the composition. Compared with other auxiliary materials, these auxiliary materials are more conducive to the soothing and repairing effects, and are conducive to the stable storage of the product.

[0022] The product described in the present invention has a dosage form including one or more of creams, emulsions, aqueous solutions, gels, oils, powders, loose powders, aerosols, films, and freeze-dried products, but the present invention does not limit this.

[0023] The present invention also provides a method for soothing and / or repairing skin, comprising administering the product of the present invention. The administration method includes smearing, applying, spraying, and introduction, which are not limited by the present invention.

[0024] The present invention systematically optimizes the combination of animal placental protein, fibronectin and recombinant collagen, and screens out a composition with good soothing and repairing effects through systematic cell experimental verification. After verification through human trials, the composition has a significant soothing and repairing effect on the skin of the subjects, and can be used as a cosmetic raw material in cosmetics with soothing and repairing effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 The figure shows the results of the cell proliferation promotion experiment;

[0026] Figure 2 The scratch test results are shown in FIG.

[0027] Figure 3 Shows the scratch repair rate statistics chart;

[0028] Figure 4 The statistical diagram of relative expression of inflammatory factor IL-6 is shown;

[0029] Figure 5 The statistical diagram of relative expression of inflammatory factor TNF-α is shown;

[0030] Figure 6 The figure shows the results of the skin soothing effect test;

[0031] Figure 7 Graph showing skin repair efficacy test results. DETAILED DESCRIPTION

[0032] The present invention provides a composition for soothing and repairing skin and its application. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application of this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0033] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0034] The present invention is further described below in conjunction with the embodiments: Preparation Example 1 Composition formula and product composition

[0035] The concentrations of the various components in the examples and comparative examples are shown in Table 1.

[0036] Table 1

[0037]

[0038] The ingredients of the test group and the control group of the combined product are shown in Table 2.

[0039] Table 2

[0040]

[0041]

[0042] The preparation method of the composite product is as follows:

[0043] (1) Put part of phase A (raw materials 1, 2, 12, 13, 14) into a water pot, heat to 80-85°C, stir until completely dissolved, and stir at constant temperature for 20 minutes before adding the rest of phase A and stirring until completely dissolved;

[0044] (2) Mix and stir phase E (raw materials 1 and 5) and phase G (raw materials 2 and 8) until uniformly mixed;

[0045] (3) Add phase B (raw materials 9 and 10) into an emulsifying pot, heat to 80-85°C, and stir to dissolve completely;

[0046] (4) Add the pre-treated phase A and phase C (raw materials 3 and 6) into an emulsifying pot, stir homogenously and cool down;

[0047] (5) When the temperature reaches 60°C, add phase D (raw material 7) and stir homogenously for 2 to 4 minutes until the mixture is uniformly mixed;

[0048] (6) Add pre-treatment phase E, stir and mix evenly, and continue stirring to cool down;

[0049] (7) Cool down to 45°C, add all ingredients in phase F (the rest of the raw materials) and the pre-treated phase G one by one, and stir until completely uniform;

[0050] (8) When the temperature drops below 38°C, filter with filter cloth and discharge the material after inspection.

[0051] Experimental Example 1 Evaluation of Cell Efficacy of Composition

[0052] 1. Evaluation of cell proliferation promoting efficacy

[0053] The CCK8 method was used to detect the proliferation-promoting effect of each composition formula of the embodiment / comparative example in Table 1 on human skin fibroblasts (HSF). The experimental steps are as follows: after culturing human skin fibroblasts for 24 hours, the composition of the embodiment / comparative example was added to the skin fibroblast culture medium at a specified concentration, NC was a negative control without adding the composition, and the culture medium was replaced and cultured for 48 hours, and then CCK8 reagent was added and incubated for 2 hours, and then the OD was obtained by using an enzyme marker. 450 , calculate cell viability.

[0054] 2. Evaluation of damage repair efficacy

[0055] The specific experimental steps are as follows:

[0056] 1) After HSF is revived, subculture 1-2 times, every 3-4 days, to ensure that the cells are in good condition;

[0057] 2) Select cells in good condition and inoculate 96-well plates: discard the old culture medium, wash twice with PBS, digest with trypsin substitute for 2-3 minutes, add 5 mL of complete culture medium to terminate, centrifuge and count, and adjust the cell density to 1×10 5 / mL, and 100 μL of cell suspension was added to each well after mixing. The final inoculation density was 1×10 4 cells / well;

[0058] 3) After culturing for about 48 hours, when the cell confluence is about 95%, a 96-well wound scratching tool Wound Maker is used to make a scratch model, and then the cells are washed once with PBS to remove floating cell clusters;

[0059] 4) Dilute the composition to a specified concentration with DMEM high-glucose basal medium, add 100 μL / well of the sample dilution solution for culture, and incubate at 37° C. and 5% carbon dioxide for 48 to 72 hours, and observe the cell proliferation and migration every day.

[0060] 3. Evaluation of anti-inflammatory efficacy

[0061] Select RAW264.7 cells in good condition and inoculate them in 6-well plates. Discard the old culture medium, wash twice with PBS, digest with trypsin, resuspend and count. The inoculation density is 8×10 5 / well, add 2 mL of complete medium (high-glucose DMEM + 10% FBS + 1% double antibody) to each well, put it in an incubator and incubate for 24 hours; then, discard the old medium and wash the cells twice with PBS. Add high-glucose DMEM basal medium containing 100 ng / mL lipopolysaccharide (LPS) and the composition of the embodiment / comparative example, the negative control group only uses high-glucose DMEM basal medium, and the positive control group only uses high-glucose basal DMEM medium containing 100 ng / mL LPS, and continue to incubate for 24 hours; discard the old medium, wash twice with PBS, directly add 1 mL / well TransZolUp to lyse the cells, collect the cells, extract RNA, reverse transcribe and qPCR to detect the expression levels of IL-6 and TNF-α.

[0062] Experimental Example 2 Evaluation of the human efficacy of the composition used in the product

[0063] 1. Human soothing effect test

[0064] Ten subjects aged between 18 and 55 were randomly selected and divided into an experimental group and a control group. There was no statistically significant difference in gender and age between the subjects in each group (P>0.05), indicating that the subjects were comparable.

[0065] Each group of subjects applied the same amount of the composition prepared by the test group and the control group on the same part of the face after cleansing in the morning and evening every day for 4 weeks. The erythema area was measured at the 0th, 2nd and 4th week, and the experimental data were statistically analyzed.

[0066] 2. Human body repair efficacy test

[0067] Ten subjects aged between 18 and 55 were randomly selected. The test areas met the skin repair efficacy test conditions and were randomly divided into an experimental group and a control group. There was no statistically significant difference in age between each group of subjects (P>0.05), so the groups were comparable.

[0068] Each group of subjects used it twice a day, once in the morning and once in the evening, for a total of 1 week. The dosage was based on daily usage habits while maintaining the original skin care habits, and the same dosage products in the original skin care steps were replaced with the test products.

[0069] MPA580 was used for skin testing on days 0, 1, 3, and 7, and the experimental data were statistically analyzed.

[0070] Effect Example 1

[0071] 1. Cell efficacy evaluation

[0072] The results of the cell proliferation promotion experiment showed that the effect of promoting the proliferation of human fibroblasts in Example 1 was the most ideal (see Figure 1 ); The scratch test results show that the damage repair effect of Example 1 is the best (see Figure 2 , Figure 3 ); The results of the inflammatory factor inhibition experiment showed that Example 1 had the most obvious effect in inhibiting the expression levels of inflammatory factors IL-6 and TNF-α (see Figure 4 , Figure 5 ).

[0073] 2. Human efficacy evaluation

[0074] The soothing efficacy test showed that, compared with the control group, the erythema area of ​​the test group supplemented with the composition of Example 1 was significantly reduced at the 2nd and 4th weeks (see Figure 6 and Table 3); the repair efficacy test showed that compared with the control group, the test group supplemented with the composition of Example 1 had significantly reduced epidermal water loss on days 1, 3, and 7 (see Figure 7 and Table 4).

[0075] Table 3 Analysis of erythema area

[0076]

[0077] Table 4 Analysis of water loss

[0078]

[0079] The above are only preferred embodiments of the present invention. It should be pointed out that, for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A composition for soothing and repairing skin, characterized in that: It includes animal placental protein, fibronectin and recombinant collagen; The mass ratio of the animal placental protein, fibronectin and recombinant collagen is (5-500): (0.001-0.1): (2.5-250).

2. The composition according to claim 1, characterized in that The mass ratio of the animal placenta protein, fibronectin and recombinant collagen is (5-50): (0.001-0.01): (2.5-25).

3. The composition according to claim 1, characterized in that The mass ratio of the animal placenta protein, fibronectin and recombinant collagen is 50:0.01:

25.

4. Use of the composition according to any one of claims 1 to 3 in the preparation of a product for soothing and / or repairing the skin.

5. A product for soothing and / or repairing the skin, characterized in that: The invention comprises the composition according to any one of claims 1 to 3 and an acceptable auxiliary material or adjuvant.

6. The product according to claim 5, characterized in that in, The mass fraction of the composition according to any one of claims 1 to 3 is 3%.

7. The product according to claim 5, characterized in that The auxiliary material or adjuvant is selected from one or more of butylene glycol, 1,3-propylene glycol, C20-22 alcohol, caprylyl glycol, phenoxyethanol, tert-butyl alcohol, tromethamine, polydimethylsiloxane, acrylic acid (ester) copolymer, polyacrylamide, C13-14 isoparaffin, laureth-7, caprylhydroxamic acid, C20-22 alcohol phosphate, p-hydroxyacetophenone, xanthan gum, ethylhexylglycerin, glucose and water.

8. The product according to any one of claims 5 to 7, characterized in that: The mass fraction of butanediol is 8%, the mass fraction of 1,3-propylene glycol is 1%, the mass fraction of C20-22 alcohol is 0.8%, the mass fraction of caprylyl glycol is 0.5%, the mass fraction of phenoxyethanol is 0.5%, the mass fraction of tert-butyl alcohol is 0.2%, the mass fraction of tromethamine is 0.2%, the mass fraction of polydimethylsiloxane is 1%, the mass fraction of acrylic acid (ester) copolymer is 1.5%, the mass fraction of polyacrylamide is 1.4%, the mass fraction of C13-14 isoparaffin is 1.4%, the mass fraction of laureth-7 is 1.4%, the mass fraction of caprylhydroxamic acid is 1%, the mass fraction of C20-22 alcohol phosphate is 0.8%, the mass fraction of p-hydroxyacetophenone is 0.3%, the mass fraction of xanthan gum is 0.5%, the mass fraction of ethylhexylglycerol is 0.5%, the mass fraction of glucose is 0.5%, the mass fraction of the composition according to any one of claims 1 to 3 is 3%, and the balance is water.

9. The product according to any one of claims 5 to 8, characterized in that: The dosage forms include one or more of creams, emulsions, aqueous solutions, gels, oils, powders, loose powders, aerosols, films, and freeze-dried products.

10. A method for soothing and / or repairing skin, characterized in that The method comprises administering the product according to any one of claims 5 to 9.

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