Modified protein composition for repairing and relieving and preparation method thereof

Abstract

The invention relates to the technical field of skin care products, and particularly discloses a repairing and soothing modified protein composition and a preparation method thereof.The composition is a multi-phase emulsified compound, silk fibroin is subjected to amphoteric modification, the composition has hydrophobic and charge performance when being compounded with vegetable oil components, skin allergens can be removed, bacterial attachment can be resisted, and the repairing and soothing effects are achieved. A lipoprotein layer with a protective effect can be formed to provide a protective barrier for damaged skin, and rich permeation enhancing substances and contained potential substances can promote the modified protein to form positive charges in the plant extraction compound liquid, so that the plant extraction compound liquid can permeate and absorb; the micromolecule polypeptide and the active ingredients in the composition can deeply permeate into the skin and enter the corium layer along with the plant extract composite liquid with high permeability to repair damaged cells, so that the soothing and anti-inflammatory effects are achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and in particular to a repairing and soothing modified protein composition and a preparation method thereof. Background Art

[0002] Most people have skin problems. With the change of seasons, stress, irregular lifestyle and external damage, the skin is prone to sensitivity and damage. Data shows that one in every three women has sensitive skin problems. The skin barrier is composed of the sebum membrane and the stratum corneum. Once damaged, the skin's various physiological activities are hindered, and it is prone to dryness, redness and itching, causing various skin problems. Although some of the current repair products will not cause further sensitivity and damage to the skin, their repair effect is weak and only has a general moisturizing effect. The effect is not obvious and cannot immediately repair and soothe damaged skin. Although some have a certain repair effect, the effective ingredients are difficult to reach the dermis for deep repair. The inflammation in the bottom layer of the skin is still there and it cannot provide protection and repair. The damaged skin is still in a state of being susceptible to infection and is prone to damage and deterioration. In addition, some irritating ingredients stay on the face for too long, causing further damage to the skin.

[0003] With the prevalence of green and natural concepts, plant oils and plant extracts are increasingly used in skin care products because they have a variety of active ingredients and the raw materials are natural and less irritating than chemical reagents. However, some popular plant oils and plant extracts are not extracted purely or are easily oxidized and thus contain irritating ingredients, or are not breathable when applied to the face and can easily cause further irritation to the skin. Protein ingredients such as collagen are a major functional ingredient in skin care. As a large molecule, it is difficult for the skin to absorb. It only stays on the surface and top layer of the skin and cannot exert its efficacy. It can only protect the skin on the surface. The most commonly used solution is to add an absorption promoter, but the absorption effect of the preparation is not obvious. Excessive penetration can easily cause skin irritation. Small molecule protein is also a common method. Although it can improve absorption, its absorption capacity is limited and the utilization rate of the skin is low. Therefore, it is necessary to solve the above problems existing in current functional skin care products, and to develop a stable and non-irritating composite product with comprehensive functions of antibacterial, repair and soothing. It has certain research and development prospects. Summary of the invention

[0004] In view of the above problems, the object of the present invention is to provide a method for preparing a repairing and soothing modified protein composition.

[0005] The technical contents of the present invention are as follows:

[0006] The present invention provides a method for preparing a repairing and soothing modified protein composition, comprising the following steps:

[0007] Step 1: preparing modified protein;

[0008] The preparation method of the modified protein comprises the following steps: adding the protein to a PBS buffer at a solid-liquid ratio of 1:10 g / mL, mixing with 3-5 wt% of sodium dextran sulfate, freezing and thawing once at -10--20°C, adding 6-9 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and a thiol compound at a mass ratio of 11-15:7-9:3-6, adjusting the pH to 7-9, adding a cross-linking enzyme, heating at a temperature of 45-55°C for 3-5h in an inert atmosphere, dialyzing with a protein dialysis bag, and then freeze-drying to obtain the modified protein; the cross-linking enzyme is a transglutaminase added in an amount of 35-40 U / g protein;

[0009] The protein is one or more of fibroin, keratin and silk protein;

[0010] By adding sodium salt to the protein and freezing and thawing it, the solubility of the protein is increased and the molecular weight is reduced, the interior is opened, the surface groups are increased, the thiol compound can reduce the protein and adjust the molecular weight, the amphoteric substance taurine is introduced for surface modification, the primary amino group and the hydrophobic group are introduced to improve the repair function of the protein while giving it amphoteric properties, it has a self-emulsifying function in aqueous solution and can be absorbed by the skin, and its hydrophobic and charge properties make it able to stabilize the active ingredients in the compound with vegetable oil, and it has antibacterial and anti-protein properties on the skin surface; sodium salt improves the anti-inflammatory and soothing ability of the protein and improves the modification efficiency of the protein by increasing the enzyme activity;

[0011] Step 2: The vegetable oil is treated, and then stirred and mixed with the modified protein and the plant extract composite liquid at a temperature of 50-60° C. for 30-50 minutes to obtain a modified protein composition;

[0012] The vegetable oil is a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 10-18:11-15; olive oil and Chinese tallow tree oil contain fat-soluble substances that are easily absorbed by the skin, such as moisturizing components such as fatty acids, and antioxidant repair components such as polyphenols and vitamins;

[0013] The treatment is to treat the vegetable oil with plasma for 3-6 minutes; by increasing hydrogen ion receptors, the antioxidant and antibacterial properties of the vegetable oil are enhanced; the hydrogen ions contained in the vegetable oil can combine with the modified protein to enhance the activity and antibacterial properties of the modified protein, and at the same time promote the mutual solubility of the two.

[0014] The plant extract composite liquid is prepared by adding 6-8wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 13-15:3-5, mixing and grinding for 60-80 minutes, then adding the mixture to an aqueous solvent at a solid-liquid ratio of 1:6-10g / mL, heating and refluxing for 2-4 hours, and filtering with a macromolecular resin to obtain the obtained solution; the aqueous solvent is a urea solution with a mass fraction of 14-19%;

[0015] The prepared plant extract composite liquid is rich in tetrahydropiperine, eucalyptus oil and esters that promote penetration, can replace the use of chemical solvents, improve the activity of hydrocarbon sites of cell membranes, and promote the absorption of nutrients. The addition of lactic acid for grinding can promote the separation of small molecular components, and urea can improve the dissolution of esters and purify by reducing large molecular impurities. The lactic acid contained in it can disperse the modified protein, which can induce the zeta potential of the modified protein to change to a positive charge by modifying the protein surface, and then combine with the negatively charged cell membrane to exert the effects of silk core protein and active ingredients.

[0016] The present invention also provides a repairing and soothing modified protein composition, comprising the following components in parts by weight: 8-12 parts of modified protein, 19-26 parts of plant composite extract, and 6-10 parts of vegetable oil;

[0017] Preferably, the modified protein composition comprises the following components in parts by weight: 10 parts of modified protein, 23 parts of plant composite extract, and 8 parts of vegetable oil.

[0018] Beneficial effects of the present invention: The preparation method of the repairing and soothing modified protein composition provided by the present invention, wherein the protein is modified to be amphiphilic, and the protein is compounded with the treated plant oil component to have hydrophobic and charge properties, can remove skin allergens and resist bacterial attachment, and can form a lipoprotein layer with a protective effect, enhance the protective function of the sebum film and thicken the epidermis and stratum corneum, provide a protective barrier for damaged skin and repair the skin surface barrier, the prepared plant extract composite liquid contains rich penetration-promoting substances, and the potential substances contained can promote the modified protein to form a positive charge in the plant extract composite liquid, which can penetrate and absorb, and the active ingredients in the plant oil and modified protein deeply penetrate into the skin and enter the dermis along with the plant extract composite liquid with high penetration performance to repair damaged cells;

[0019] The repairing and soothing modified protein composition provided by the present invention contains a modified protein with both antibacterial protection and penetration repair, a protective and repairing plant oil, and anti-inflammatory, antioxidant, anti-allergic and anti-histamine plant active ingredients. Various components work synergistically to achieve the soothing and anti-inflammatory effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 To improve the permeability and absorption of proteins into the skin;

[0021] Figure 2 It is the fluorescence detection diagram of LOR protein and FLG protein;

[0022] Figure 3 is the average relative OD of FLG protein;

[0023] Figure 4 is the average relative OD of LOR protein;

[0024] Figure 5 is the thickness of the epidermal living cell layer;

[0025] Figure 6 is the expression level of Claudin-1;

[0026] Figure 7 is the expression level of Claudin-5

[0027] Figure 8 The moisture content of the stratum corneum of the skin after using the repairing and soothing modified protein composition at 0 days, 7 days and 14 days;

[0028] Fig. 9 The transepidermal water loss rate of the skin after using the repairing and soothing modified protein composition for 0 days, 7 days and 14 days;

[0029] Fig.10 The facial redness of the volunteers;

[0030] Fig.11 The amount of skin erythema. DETAILED DESCRIPTION

[0031] The present invention is further described in detail below through specific implementation cases. It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of protection of the present invention. After reading the present invention, various equivalent forms of modifications to the present invention by those skilled in the art are all within the scope of the claims attached to this application.

[0032] Unless otherwise specified, all raw materials and reagents of the present invention are raw materials and reagents from the conventional market.

[0033] Example 1

[0034] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0035] Step 1: preparing modified protein: adding fibroin to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 4 wt% of sodium dextran sulfate, freeze-thaw once at -15°C, adding 8 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 13:8:5, adjusting the pH to 7, adding 38 U / g of transglutaminase, heating at 50°C for 4 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0036] Step 2: 8 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 14:13 were treated with plasma for 4 minutes, and then stirred and mixed with 10 parts of modified protein and 22 parts of plant extract composite liquid at a temperature of 55° C. for 40 minutes to obtain a modified protein composition;

[0037] The plant extract composite liquid is prepared by adding 7wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 14:4, mixing and grinding for 70 minutes, then adding a 17% mass fraction urea solution at a solid-liquid ratio of 1:8g / mL, heating and refluxing for 3 hours, and filtering with a macromolecular resin to obtain the composite liquid.

[0038] Example 2

[0039] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0040] Step 1: preparing modified protein: adding keratin to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 3 wt% of sodium dextran sulfate, freeze-thaw once at -10°C, adding 6 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 11:7:3, adjusting the pH to 7, adding 35 U / g of transglutaminase, heating at 45°C for 3 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0041] Step 2: treating 6 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 10:11 with plasma for 3 minutes, and then stirring and mixing with 8 parts of modified protein and 19 parts of plant extract composite liquid at a temperature of 50° C. for 30 minutes to obtain a modified protein composition;

[0042] The plant extract composite liquid is prepared by adding 6wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 13:3, mixing and grinding for 60 minutes, then adding a 14% mass fraction urea solution at a solid-liquid ratio of 1:6g / mL, heating and refluxing for 2 hours, and filtering with a macromolecular resin to obtain the obtained solution.

[0043] Example 3

[0044] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0045] Step 1: preparing modified protein: adding silk protein to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 5 wt% of sodium dextran sulfate, freeze-thaw once at -20°C, adding 9 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 15:9:6, adjusting the pH to 9, adding 40 U / g of transglutaminase, heating at 55°C for 5 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0046] Step 2: 10 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 18:15 were treated with plasma for 6 minutes, and then stirred and mixed with 12 parts of modified protein and 26 parts of plant extract composite liquid at a temperature of 60° C. for 50 minutes to obtain a modified protein composition;

[0047] The plant extract composite liquid is prepared by adding 8wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 15:5, mixing and grinding for 80 minutes, then adding a 19% urea solution at a solid-liquid ratio of 1:10g / mL, heating and refluxing for 4 hours, and filtering with a macromolecular resin to obtain the composite liquid.

[0048] Example 4

[0049] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0050] Step 1: preparing modified protein: adding fibroin to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 3 wt% sodium dextran sulfate, freeze-thaw once at -20°C, adding 8 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 12:8:4, adjusting the pH to 7, adding 35 U / g protein of transglutaminase, heating at 50°C for 5 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0051] Step 2: treating 7 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 11:12 with plasma for 3 minutes, and then stirring and mixing with 9 parts of modified protein and 20 parts of plant extract composite liquid at a temperature of 50° C. for 35 minutes to obtain a modified protein composition;

[0052] The plant extract composite liquid is prepared by adding 7wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 14:3, mixing and grinding for 65 minutes, then adding a 15% mass fraction urea solution at a solid-liquid ratio of 1:7g / mL, heating and refluxing for 3 hours, and filtering with a macromolecular resin to obtain the obtained solution.

[0053] Example 5

[0054] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0055] Step 1: preparing modified protein: adding keratin to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 5 wt% of sodium dextran sulfate, freeze-thaw once at -15°C, adding 6 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 14:8:5, adjusting the pH to 7, adding 40 U / g of transglutaminase, heating at 55°C for 5 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0056] Step 2: 9 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 17:14 were treated with plasma for 5 minutes, and then stirred and mixed with 11 parts of modified protein and 25 parts of plant extract composite liquid at a temperature of 55° C. for 45 minutes to obtain a modified protein composition;

[0057] The plant extract composite liquid is prepared by adding 7wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 14:5, mixing and grinding for 75 minutes, then adding a urea solution with a mass fraction of 18% at a solid-liquid ratio of 1:9g / mL, heating and refluxing for 4 hours, and filtering with a macromolecular resin to obtain the obtained solution.

[0058] Example 6

[0059] A method for preparing a repairing and soothing modified protein composition comprises the following steps:

[0060] Step 1: preparing modified protein: adding silk protein to PBS buffer at a solid-liquid ratio of 1:10 g / mL and mixing with 3 wt% of sodium dextran sulfate, freeze-thaw once at -10°C, adding 7 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 15:9:6, adjusting the pH to 7, adding 38 U / g of transglutaminase, heating at 45°C for 5 h in an inert atmosphere, dialyzing with a protein dialysis bag and freeze-drying to obtain;

[0061] Step 2: treating 6 parts of a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 18:11 with plasma for 6 minutes, and then stirring and mixing with 8 parts of modified protein and 26 parts of plant extract composite liquid at a temperature of 50° C. for 50 minutes to obtain a modified protein composition;

[0062] The plant extract composite liquid is prepared by adding 8wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 15:3, mixing and grinding for 60 minutes, then adding a 14% mass fraction urea solution at a solid-liquid ratio of 1:10g / mL, heating and refluxing for 4 hours, and filtering with a macromolecular resin to obtain the obtained solution.

[0063] Comparative Example 1

[0064] The difference between Comparative Example 1 and Example 1 is that the protein in Comparative Example 1 is not modified, and the other properties remain unchanged.

[0065] Comparative Example 2

[0066] The difference between Comparative Example 2 and Example 1 is that the vegetable oil in Comparative Example 2 is not subjected to plasma treatment.

[0067] Comparative Example 3

[0068] The difference between Comparative Example 3 and Example 1 is that the plant composite extract of Comparative Example 3 is obtained by adding equal amounts of pepper and eucalyptus leaves to water at a solid-liquid ratio of 1:6-10 g / mL, heating and refluxing for 2-4 hours, and filtering with a macromolecular resin; the rest remains unchanged.

[0069] Comparative Example 4

[0070] The difference between Comparative Example 4 and Example 1 is that in the protein modification of Comparative Example 4, no sodium salt is added for freeze-thawing, and the other conditions remain unchanged.

[0071] 1. The potential ξ1 of the modified protein prepared in Example 1 and the potential ξ2 of the unmodified protein in Comparative Example 1 in vegetable oil and plant extract were respectively detected. The results are shown in Table 1. It can be seen that the modified protein prepared in the present invention has amphoteric properties, carrying a negative charge in vegetable oil and a positive charge in plant extract, while the unmodified protein does not have amphoteric properties, and the negatively charged amino acids on the surface are more than the positively charged amino acids.

[0072] Table 1 Zeta potential of modified protein surface

[0073]

[0074] Second, the surface neutrophils of the skin are the first line of defense for cellular defense. They fight pathogens in wound infections and participate in the development of inflammation. Mechanical damage will cause structural damage to skin tissue. The zebrafish tail fin has a self-regeneration function. By observing the regeneration speed of the tail fin, the efficacy of the test substance in promoting tissue regeneration can be evaluated.

[0075] The zebrafish to be tested were divided into 11 groups, 5 in each group, and placed in a six-well plate. 0.2 mg of sodium dodecyl sulfate was added to establish a zebrafish skin soothing model. The composition prepared in the embodiment and the comparative example was administered by water dissolution at a dosage of 10 mg, and an equal amount of physiological saline was used as a blank control. After 24 hours, the number of neutrophils in the zebrafish was detected and the test results were recorded. After feeding for a period of time, the tail fins of the zebrafish in each group were cut off to establish a zebrafish repair model. An equal amount of the composition prepared in the embodiment and the comparative example was added to the well plate containing the zebrafish. After 24 hours, the tail fin area of ​​the zebrafish was detected to determine the repair effect of the zebrafish. The test results are shown in Table 2.

[0076] Table 2 Data of zebrafish test

[0077]

[0078]

[0079] As can be seen from Table 2, the compositions prepared in the embodiments of the present invention have better anti-inflammatory and wound healing effects than those in Comparative Examples 1-4, by adding modified proteins capable of penetrating repair and skin anti-inflammatory protection, antioxidant plant oils containing hydrogen ion receptors, and plant composite extracts with strong penetrating soothing effects, wherein the modified proteins are amphiphilically modified by adding antibacterial and repairing ingredients such as sodium salts and other substances to improve the repair properties of the proteins, and the obtained compositions have stronger repairing and soothing properties, which can significantly repair wounds and soothe inflammatory sites in a short time.

[0080] 3. 3D skin model repair test of the repair and soothing modified protein composition

[0081] (1) Permeability of modified protein

[0082] The penetration and absorption of the repairing and soothing modified protein composition on the skin was tested according to the 3D skin model. The plant composite extract prepared in Example 1 and the modified protein were added to PBS buffer at a solid-liquid ratio of 1:10 g / mL, mixed and applied to the 3D epidermal model, and the modified protein was fluorescently labeled and tested by HPLC. The test results are as follows: Figure 1 As shown, it can be seen that the osmotic absorption of the modified protein mixed with the plant composite extract is better than that of the unmodified protein.

[0083] (2) Barrier protein content in damaged skin

[0084] Differentiation proteins in the skin include filaggrin (FLG) and loricrin (LOR); FLG is a key component in the CE assembly process, interacting with keratin intermediate filaments to form dense keratin fiber bundles, thereby forming a flat and tough scaffold structure for keratinocytes. In addition to being a structural component of the skin barrier, FLG can also be hydrolyzed by Caspase-14 to form a natural moisturizing factor, thus also playing a moisturizing role;

[0085] Loricrin-LOR is a key component in the assembly process of the protein membrane CE, accounting for about 80% of the CE content. It is located in the middle and upper part of the spinous cell layer and cross-links with other barrier-related proteins to form an abnormally insoluble keratin membrane. This keratin membrane wraps the keratin fiber bundles to form a unique stratum corneum barrier structure of the epidermis, which strengthens the skin barrier.

[0086] The repair performance of the repair and soothing modified protein composition was tested by measuring the content of 3D skin barrier related proteins. The results are as follows Figure 2-4 As shown, BC, NC, PC and EG are normal skin, damaged skin, positive control and damaged skin after applying the repairing and soothing modified protein composition prepared in Example 1 for 6 hours, respectively. It can be seen that the repairing and soothing modified protein composition prepared by the present invention can repair damaged skin with rapid effect. Compared with the NC group, it can increase the content of LOR protein and FLG protein in damaged skin.

[0087] (3) Improve the health and vitality of the epidermis

[0088] In HE staining, cells and nuclei of the epidermis are stained purple, and other tissues and structures show different colors and morphologies;

[0089] The detection model used is 3D epidermal injury model, the detection method is H&E staining, the stimulation condition is 0.1% SLS, EG1 and EG2 are respectively treated with the repairing and soothing modified protein composition prepared in Example 1 after the damaged model is treated once and after treatment 5 times, the results are as follows Figure 5 As shown, compared with the NC group, the stratum corneum of the damaged model treated once with the repairing and soothing modified protein composition of EG1 became tighter, the thickness of the living cell layer increased, the uniformity of the cell nucleus staining (dark purple) improved, and the damaged state of the cells was restored; while the thickness of the living cell layer of the damaged model treated 5 times with the repairing and soothing modified protein composition prepared in Example 1 increased significantly, the damage was significantly repaired, and the cell vitality was full.

[0090] (4) Expression of claudins in the skin barrier

[0091] Claudins exist in all epithelial and endothelial cells. Claudins are four transmembrane proteins. The PDZ site at the carboxyl end of their structure can form a tight junction structure with proteins containing PDZ domains in the cytoplasm, such as ZO-1, and play the role of "fence" and "barrier".

[0092] Experimental method: After irradiating HaCaT with UVB, the repairing and soothing modified protein composition prepared in Example 1 was used for 24 hours, and Claudin-1 and Claudin-5 were detected by qRT-PCR. The results are as follows: Figure 6 and Figure 7 As shown, it can be seen that for ultraviolet damage, the repairing and soothing modified protein composition of the present invention can increase the expression levels of Claudin-1 and Claudin-5 and repair the skin barrier.

[0093] 4. Skin moisture content, transepidermal water loss, facial redness and erythema detection

[0094] Maintaining a certain ratio between transepidermal water loss and skin moisture content is an important criterion for evaluating healthy skin. The moisture of healthy skin can maintain the operation of various physiological activities of the skin. The transepidermal water loss rate is the amount of water that passes through the skin to the outside of the human body. The barrier function of damaged skin is low and the transepidermal water loss is high.

[0095] 32 volunteers (Chinese sensitive skin - screened by questionnaire) used the repairing and soothing modified protein composition prepared in Example 1 twice a day for 28 days, and measured the facial skin moisture content ( CM825) and transepidermal water loss test ( TM Hex), the result is as follows Figure 8 and Fig. 9 As shown, the facial redness of the subjects was tested before use and 14 days and 28 days later. The facial redness of subjects 7 and 13 was as shown in Fig.10 As shown, the amount of skin erythema of volunteers with ultraviolet erythema before and after using the composition was evaluated by colorimetry. The amount of skin erythema before and 2 hours after using the composition is as shown in Fig.11 As shown; it can be seen that after using the repairing and soothing modified protein composition of the present invention, the moisture content of the skin is increased, the transepidermal water loss rate of the skin is reduced, the facial redness is significantly improved, and the amount of skin erythema is reduced.

[0096] 5. The repairing and soothing modified protein composition prepared in Example 1 and Comparative Example 1 was applied to the skin of the subjects, and bacteria were detected on the applied area of ​​the skin surface after 6 hours and 12 hours, respectively, to obtain the percentage of bacteria on the skin compared with that before application, and the adhesion strength of the skin was tested. The test results are shown in Table 3.

[0097] Table 3 Bactericidal and adhesion strength test of the composition

[0098]

[0099] As can be seen from Table 3, the repairing and soothing modified protein composition of the present invention contains modified proteins with amphoteric properties, which can provide hydrophobic protection on the skin surface. The added sodium salt can promote the modification and preparation of the protein and soothe and anti-inflammatory effects. The surface amphoteric substance taurine improves the antibacterial properties. Compared with the unmodified protein, it can form a protective effect on the skin, and can have antibacterial and anti-adhesion effects on the skin, which can prevent further inflammation of the skin.

[0100] In summary, the repairing and soothing modified protein composition prepared by the present invention has the effects of penetrating the dermis to repair and protecting the skin surface and anti-inflammatory effects, can perform multiple repairs and soothing on damaged skin, has obvious effects, and can be added to skin care products as an effective ingredient.

Claims

1. A method for preparing a repairing and soothing modified protein composition, characterized in that: The following steps are involved: Step 1: preparing modified protein; Step 2: The vegetable oil is treated, and then stirred and mixed with the modified protein and plant extract composite liquid at a temperature of 50-60° C. for 30-50 minutes to obtain a modified protein composition.

2. The method for preparing a repairing and soothing modified protein composition according to claim 1, characterized in that: The preparation method of the modified protein comprises the following steps: adding the protein to a PBS buffer at a solid-liquid ratio of 1:10 g / mL, mixing with 3-5 wt% of sodium dextran sulfate, freezing and thawing once at -10--20°C, adding 6-9 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and a thiol compound at a mass ratio of 11-15:7-9:3-6, adjusting the pH to 7-9, adding a cross-linking enzyme, heating at a temperature of 45-55°C for 3-5 hours in an inert atmosphere, dialyzing with a protein dialysis bag and then freeze-drying the mixture.

3. The method for preparing a repairing and soothing modified protein composition according to claim 2, characterized in that: The protein is one or more of fibroin, keratin and silk protein.

4. The method for preparing a repairing and soothing modified protein composition according to claim 2, characterized in that: The cross-linking enzyme is transglutaminase with an addition amount of 35-40 U / g protein.

5. The method for preparing a repairing and soothing modified protein composition according to claim 1, characterized in that: The vegetable oil is a complex composed of olive oil and Chinese tallow tree oil in a mass ratio of 10-18:11-15.

6. The method for preparing a repairing and soothing modified protein composition according to claim 1, characterized in that: The plant extract composite liquid is prepared by adding 6-8wt% lactic acid to pepper and eucalyptus leaves at a mass ratio of 13-15:3-5, mixing and grinding for 60-80 minutes, then adding it to an aqueous solvent at a solid-liquid ratio of 1:6-10g / mL, heating and refluxing for 2-4 hours, and filtering with a macromolecular resin to obtain the obtained solution.

7. The method for preparing a repairing and soothing modified protein composition according to claim 6, characterized in that: The aqueous solvent is a urea solution with a mass fraction of 14-19%.

8. The method for preparing a repairing and soothing modified protein composition according to claim 1, characterized in that: The treatment in step 2 is to treat the vegetable oil with plasma for 3-6 minutes.

9. A repairing and soothing modified protein composition prepared by the preparation method according to any one of claims 1 to 8, characterized in that: The invention comprises the following components in parts by weight: 8-12 parts of modified protein, 19-26 parts of plant compound extract and 6-10 parts of vegetable oil.

10. A repairing and soothing modified protein composition according to claim 9, characterized in that: The invention comprises the following components in parts by weight: 10 parts of modified protein, 23 parts of plant compound extract and 8 parts of vegetable oil.

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