A healing soothing modified protein composition and method of making same

Abstract

The present application relates to the technical field of skin care, and particularly discloses a modified protein composition for repairing and soothing and a preparation method thereof. The composition is a multi-phase emulsified compound. The silk fibroin is modified by amphoteric modification. The modified silk fibroin is combined with a plant oil component to have hydrophobic and charge properties, can remove skin sensitizers and resist bacterial adhesion, and can form a protective lipoprotein layer to provide a protective barrier for damaged skin. In addition, the composition contains rich penetration enhancers and potential substances, which can make the modified protein form a positive charge in the plant extraction complex liquid, penetrate and absorb. The small-molecule polypeptides and active ingredients in the composition can deeply penetrate into the skin and enter the dermis layer to repair damaged cells, so that the soothing and anti-inflammatory effects are achieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of skin care, in particular to a modified protein composition for repairing and soothing and a preparation method thereof. BACKGROUND

[0002] Most people have skin problems. With the change of seasons, stress, irregular life and external damage, the skin is prone to sensitive damage. Data shows that one out of every three women has sensitive skin. The skin barrier is composed of sebum film and stratum corneum. After damage, the physiological activities of the skin are hindered, which can cause dryness, redness and itching, and trigger various skin problems. The current repair products may not cause further sensitive damage, but the repair effect is weak, only with general moisturizing effect, and the effect is not obvious. They cannot repair and soothe damaged skin immediately. Some products have certain repair effect, but the effective components are difficult to reach the dermis for deep repair. The inflammation at the bottom of the skin still exists, and the damaged skin is still in a state of being easily infected. Some irritating components stay on the face for a long time, which can cause further damage to the skin.

[0003] With the prevalence of green and natural concept, plant oil and plant extract are increasingly used in skin care products because they have various active ingredients and are less irritating than chemical reagents. However, some plant oils and plant extracts are not pure or are easily oxidized, which can cause irritation. Protein components such as collagen are effective ingredients in skin care. However, they are difficult to absorb because they are macromolecular substances, and only stay on the surface and surface layer of the skin, which cannot exert their efficacy. The current common solution is to add a penetration enhancer, but the effect of the penetration enhancer is not obvious. Excessive penetration can cause skin irritation. Small molecule proteins are also a common method. Although they can improve absorption, the absorption capacity is limited, and the utilization rate of the skin is low. Therefore, it is necessary to solve the above problems in the current effective skin care products and develop a stable and non-irritating composition product with comprehensive functions such as antibacterial repair and soothing. SUMMARY

[0004] In view of the above problems, the present application provides a preparation method of a modified protein composition for repairing and soothing.

[0005] The technical content of the present application is as follows:

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

[0007] Step 1: preparing a modified protein;

[0008] The preparation method of the modified protein is to add the protein to PBS buffer at a solid-liquid ratio of 1:10 g / mL, then mix 3-5 wt% of sodium dextran sulfate salt, freeze-thaw once at -10--20℃, then add 6-9 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and thiol compound with a mass ratio of 11-15:7-9:3-6, adjust the pH to 7-9, then add cross-linking enzyme, heat at a temperature of 45-55℃ for 3-5h under an inert atmosphere, dialyze with a protein dialysis bag, then freeze-dry to obtain; the cross-linking enzyme is transglutaminase with an addition amount of 35-40 U / g of protein;

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

[0010] By adding protein to sodium salt freeze-thaw to increase protein solubility and reduce molecular weight, open the inside, increase the surface group, thiol compound can reduce and adjust the molecular weight of the protein, introduce the amphoteric substance taurine for surface modification, introduce primary amino and hydrophobic group to improve the repair function of the protein while endowing it with amphoteric properties, have self-emulsifying function in aqueous solution, can be promoted to be absorbed by the skin, and its hydrophobic and charge properties enable it to be complexed with plant oil to stabilize the active ingredients therein, while having antibacterial and anti-protein properties on the skin surface; sodium salt improves the anti-inflammatory soothing ability of the protein and improves the modification efficiency of the protein by increasing enzyme activity;

[0011] Step 2: After treating the plant oil, mix it with the modified protein and plant extract complex liquid at a temperature of 50-60℃ for 30-50min to prepare a modified protein composition;

[0012] The plant oil is a complex of olea europaea fruit oil and sapium sebiferum seed oil with a mass ratio of 10-18:11-15; olea europaea fruit oil and sapium sebiferum seed oil contain fat-soluble substances such as fatty acids and other moisturizing ingredients and polyphenols, vitamins and other antioxidant repair ingredients that are easily absorbed by the skin;

[0013] The treatment is to treat the plant oil with plasma for 3-6min; by increasing the hydrogen ion acceptor to enhance its antioxidant and antibacterial properties, the contained hydrogen ions can combine with the modified protein to enhance the activity and antibacterial properties of the modified protein, while promoting the mutual solubility of the two;

[0014] The preparation of the plant extract complex liquid is to mix 6-8 wt% of lactic acid with pepper and eucalyptus leaves at a mass ratio of 13-15:3-5, grind for 60-80min, then add an aqueous solvent at a solid-liquid ratio of 1:6-10 g / mL to heat and reflux for 2-4h, then filter with a macromolecular resin to obtain; the aqueous solvent is a urea solution with a mass fraction of 14-19%;

[0015] The prepared plant extraction composite liquid is rich in tetrahydro piperine, eucalyptus oil and esterate and the like which have penetration promoting effect, can replace the use of chemical solvents, improve the activity of carbon and hydrogen parts of cell membranes, promote the absorption of nutrients, and the lactic acid added for grinding promotes the analysis of small molecules, urea can improve the dissolution of esterate and purify by reducing macromolecular impurities; the contained lactic acid can disperse modified proteins, the modified proteins change the zeta potential to positive charge through modification of the protein surface, and then combine with the cell membrane with negative charge, and play the role of silk fibroin and active ingredients.

[0016] The application further provides a modified protein composition for repairing and soothing, comprising the following components in parts by weight: modified protein 8-12 parts, plant composite extraction liquid 19-26 parts, and plant oil 6-10 parts.

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

[0018] The application has the following beneficial effects: the preparation method of the modified protein composition for repairing and soothing provided by the application can modify proteins in amphoteric mode, and the modified proteins have hydrophobic and charge properties after being combined with the plant oil component, can remove skin sensitizers and resist bacterial adhesion, can form a lipoprotein layer with protective effect, can enhance the protective function of the sebum membrane and thicken the epidermis and the stratum corneum, can provide a protective barrier for damaged skin and repair the skin surface barrier, the prepared plant extraction composite liquid contains rich penetration promoting substances, and the potential substances can promote the modified proteins to form positive charges in the plant extraction composite liquid, can be penetrated and absorbed, and the active ingredients in the plant oil and the modified proteins can penetrate into the dermis along with the plant extraction composite liquid with high penetration performance to repair damaged cells;

[0019] The modified protein composition for repairing and soothing provided by the application contains modified proteins with antibacterial protection and penetration repair, plant oil with protection and repair, and plant active ingredients with anti-inflammatory, antioxidant, anti-allergic and antihistamine effects, and various components have synergistic effects to achieve the effect of soothing and anti-inflammatory. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The modified proteins have penetration and absorption performance for the skin;

[0021] Figure 2 The fluorescence detection diagram of LOR proteins and FLG proteins;

[0022] Figure 3 The relative OD average value of FLG proteins;

[0023] Figure 4 The relative OD average value of LOR proteins;

[0024] Figure 5 Epidermal viable cell layer thickness;

[0025] Figure 6 Claudin-1 expression level;

[0026] Figure 7 Claudin-5 expression level

[0027] Figure 8 Skin keratin layer moisture content after 0 days, 7 days and 14 days of using the modified protein composition for repair and relief;

[0028] Figure 9 Transdermal water loss rate of the skin after 0 days, 7 days and 14 days of using the modified protein composition for repair and relief;

[0029] Figure 10 Facial redness of the volunteers;

[0030] Figure 11 Skin erythema amount. DETAILED DESCRIPTION

[0031] The present application will be further described in the following specific examples, which should not be construed as limiting the scope of the present application. After reading the present application, those skilled in the art will be able to modify various equivalent forms of the present application, which are within the scope of the appended claims.

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

[0033] Example 1

[0034] A method for preparing a modified protein composition for repair and relief, comprising the following steps:

[0035] Step 1: Preparation of modified protein: silk fibroin is added to PBS buffer at a solid-liquid ratio of 1:10 g / mL, mixed with 4 wt% dextran sulfate sodium salt, frozen and thawed once at -15°C, then 8 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine and glutathione with a mass ratio of 13:8:5 is added, the pH is adjusted to 7, then 38 U / g of transglutaminase is added, heated at 50°C for 4 h under an inert atmosphere, dialyzed with a protein dialysis bag, and then freeze-dried to obtain;

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

[0037] The preparation of the plant extraction composite liquid is as follows: pepper and eucalyptus leaves are mixed and ground for 70 min at a mass ratio of 14:4 with 7 wt% lactic acid, then heated and refluxed for 3 h at a solid-liquid ratio of 1:8 g / mL with a 17 wt% urea solution, and then filtered with a macromolecular resin to obtain the plant extraction composite liquid.

[0038] Example 2

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

[0040] Step 1: Preparation of modified protein: keratin is mixed with 3 wt% dextran sulfate sodium salt in PBS buffer at a solid-liquid ratio of 1:10 g / mL, then frozen and thawed once at -10°C, then a composition of 6 wt% pentaerythritol tetra(ethylhexanoate), taurine and glutathione at a mass ratio of 11:7:3 is added, the pH is adjusted to 7, then 35 U / g protein of transglutaminase is added, heated at 45°C for 3 h in an inert atmosphere, dialyzed with a protein dialysis bag, and then freeze-dried to obtain;

[0041] Step 2: 6 parts of a composite of olive fruit oil and Chinese tallow seed oil at a mass ratio of 10:11 are treated with plasma for 3 min, then mixed with 8 parts of the modified protein and 19 parts of the plant extraction composite liquid at 50°C for 30 min to obtain the modified protein composition;

[0042] The preparation of the plant extraction composite liquid is as follows: pepper and eucalyptus leaves are mixed and ground for 60 min at a mass ratio of 13:3 with 6 wt% lactic acid, then heated and refluxed for 2 h at a solid-liquid ratio of 1:6 g / mL with a 14 wt% urea solution, and then filtered with a macromolecular resin to obtain the plant extraction composite liquid.

[0043] Example 3

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

[0045] Step 1: Preparation of modified protein: silk fibroin is mixed with 5 wt% dextran sulfate sodium salt in PBS buffer at a solid-liquid ratio of 1:10 g / mL, then frozen and thawed once at -20°C, then a composition of 9 wt% pentaerythritol tetra(ethylhexanoate), taurine and glutathione at a mass ratio of 15:9:6 is added, the pH is adjusted to 9, then 40 U / g protein of transglutaminase is added, heated at 55°C for 5 h in an inert atmosphere, dialyzed with a protein dialysis bag, and then freeze-dried to obtain;

[0046] Step 2: 10 parts of a complex of olive fruit oil and Chinese tallow oil in a mass ratio of 18:15 is treated by plasma for 6 min, and then mixed with 12 parts of modified protein and 26 parts of plant extraction complex liquid at 60°C for 50 min to prepare a modified protein composition;

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

[0048] Example 4

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

[0050] Step 1: preparing modified protein: silk fibroin is added to PBS buffer in a solid-liquid ratio of 1:10 g / mL, mixed with 3 wt% dextran sulfate sodium salt, frozen and thawed once at -20°C, and then added with a combination of pentaerythritol tetra(ethylhexanoate), taurine and glutathione in a mass ratio of 12:8:4 at 8 wt%, adjusted to pH 7, added with transglutaminase in an amount of 35 U / g protein, heated at 50°C for 5 h in an inert atmosphere, dialyzed with a protein dialysis bag, and then freeze-dried to obtain;

[0051] Step 2: 7 parts of a complex of olive fruit oil and Chinese tallow oil in a mass ratio of 11:12 is treated by plasma for 3 min, and then mixed with 9 parts of modified protein and 20 parts of plant extraction complex liquid at 50°C for 35 min to prepare a modified protein composition;

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

[0053] Example 5

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

[0055] Step 1: Preparation of modified protein: keratin was mixed with 5wt% sodium sulfate dextran in PBS buffer at a solid-liquid ratio of 1:10 g / mL, and then subjected to freeze-thawing once at -15°C. Then, a composition of 6wt% pentaerythritol tetra(ethylhexanoate), taurine and glutathione with a mass ratio of 14:8:5 was added. After adjusting the pH to 7, 40U / g protein of transglutaminase was added. The mixture was heated at 55°C for 5h under an inert atmosphere. After dialysis with a protein dialysis bag and freeze-drying, a modified protein was obtained;

[0056] Step 2: A compound composed of 9 parts of oil olive fruit oil and Chinese tallow oil with a mass ratio of 17:14 was treated with plasma for 5min, and then mixed with 11 parts of modified protein and 25 parts of plant extraction complex at 55°C for 45min to obtain a modified protein composition.

[0057] The plant extraction complex was prepared by mixing pepper and eucalyptus leaves with a mass ratio of 14:5 with 7wt% lactic acid and grinding for 75min. Then, 18% urea solution was added at a solid-liquid ratio of 1:9 g / mL and heated to reflux for 4h. After filtration with a macromolecular resin, the plant extraction complex was obtained.

[0058] Example 6

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

[0060] Step 1: Preparation of modified protein: silk protein was mixed with 3wt% sodium sulfate dextran in PBS buffer at a solid-liquid ratio of 1:10 g / mL, and then subjected to freeze-thawing once at -10°C. Then, a composition of 7wt% pentaerythritol tetra(ethylhexanoate), taurine and glutathione with a mass ratio of 15:9:6 was added. After adjusting the pH to 7, 38U / g protein of transglutaminase was added. The mixture was heated at 45°C for 5h under an inert atmosphere. After dialysis with a protein dialysis bag and freeze-drying, a modified protein was obtained;

[0061] Step 2: A compound composed of 6 parts of oil olive fruit oil and Chinese tallow oil with a mass ratio of 18:11 was treated with plasma for 6min, and then mixed with 8 parts of modified protein and 26 parts of plant extraction complex at 50°C for 50min to obtain a modified protein composition.

[0062] The plant extraction complex was prepared by mixing pepper and eucalyptus leaves with a mass ratio of 15:3 with 8wt% lactic acid and grinding for 60min. Then, 14% urea solution was added at a solid-liquid ratio of 1:10 g / mL and heated to reflux for 4h. After filtration with a macromolecular resin, the plant extraction complex was obtained.

[0063] Comparative Example 1

[0064] Comparative Example 1 differs from Example 1 in that the protein of Comparative Example 1 is not modified.

[0065] Comparative Example 2

[0066] Comparative Example 2 differs from Example 1 in that the plant oil of Comparative Example 2 is not subjected to plasma treatment.

[0067] Comparative Example 3

[0068] Comparative Example 3 differs from Example 1 in that the plant composite extract solution of Comparative Example 3 is prepared 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 h, and filtering with a macromolecular resin; other conditions are the same.

[0069] Comparative Example 4

[0070] Comparative Example 4 differs from Example 1 in that the protein of Comparative Example 4 is not subjected to freezing and thawing in the presence of sodium salt during modification; other conditions are the same.

[0071] I. The zeta potentials of the modified protein prepared in the examples and the unmodified protein of Comparative Example 1 in plant oil (ξ1) and in plant extract solution (ξ2) are detected respectively, and the results are shown in Table 1. It can be seen that the modified protein prepared in the present application has amphoteric properties, is negatively charged in plant oil, and is positively charged in plant extract solution, while the unmodified protein does not have amphoteric properties, and the number of negatively charged amino acids on the surface is greater than that of positively charged amino acids.

[0072] Table 1 Zeta potential of modified protein surface

[0073]

[0074] II. Neutrophils in the surface layer of the skin are the first line of defense for cell defense, and fight against pathogens and participate in the development of inflammation in wound infection; mechanical damage causes structural damage to skin tissue, and zebrafish tail fins have self-renewing function, and the observation of the regeneration rate of the tail fins can evaluate the efficacy of the test substance in promoting tissue regeneration.

[0075] The zebrafish to be tested is divided into 11 groups, 5 zebrafish in each group, and placed in a six-hole plate, 0.2 mg of sodium dodecyl sulfate is added to establish a zebrafish skin soothing model, the composition prepared by the examples and the comparative examples is given by water-soluble administration, the dosage is 10 mg, and an equal amount of normal saline is used as a blank control, the number of neutrophils of the zebrafish is detected after 24 h, and the detection results are recorded; after a period of feeding, the tail fins of the zebrafish in each group are cut off to establish a zebrafish repair model, and an equal amount of the composition prepared by the examples and the comparative examples is added to the hole plate containing the zebrafish, the tail fin area of the zebrafish is detected after 24 h to judge the repair effect of the zebrafish, and the detection results are shown in Table 2.

[0076] Table 2 Data of zebrafish test

[0077]

[0078]

[0079] As shown in Table 2, the composition prepared by the examples has better anti-inflammatory and wound healing effects than the comparative examples 1-4, by adding the modified protein capable of penetrating repair and skin anti-inflammatory protection, the antioxidant plant oil containing hydrogen ion acceptor, and the plant composite extract liquid with strong penetration soothing effect, wherein the modified protein is modified by adding antibacterial and repair components such as sodium salt and the like to improve the repair performance of the protein, and the prepared composition has strong repair and soothing performance, and can obviously repair the wound and soothe the inflammatory site in a short time.

[0080] III. Repair and soothing modified protein composition 3D skin model repair detection

[0081] (1) Penetration performance of modified protein

[0082] According to the 3D skin model, the penetration and absorption of the repair and soothing modified protein composition on the skin are detected, the plant composite extract liquid and the modified protein prepared in Example 1 are mixed at a solid-liquid ratio of 1:10 g / mL and added to PBS buffer, then applied to the 3D epidermis model, the modified protein is fluorescently labeled, and HPLC method is used for detection, and the detection results are shown in Table 3. Figure 1 As shown in Table 3, the penetration and absorption of the modified protein mixed with the plant composite extract liquid is better than that of the unmodified protein.

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

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

[0085] Loricrin-LOR is a key component in the CE assembly process of the protein membrane sheath, accounting for about 80% of the CE content, and is located in the upper part of the stratum spinosum, and is cross-linked with other barrier-related proteins to form an abnormally insoluble keratin sheath film, which wraps the keratin fiber bundles to form a unique stratum corneum barrier structure of the epidermis, and plays a reinforcing role on the skin barrier;

[0086] The repair performance of the repair and soothing modified protein composition was detected by 3D skin barrier-related protein content determination, and the results are shown in Figures 2-4 BC, NC, PC and EG are normal skin, damaged skin, positive control and damaged skin after 6h of EG1 prepared according to the repair and soothing modified protein composition of the embodiment 1 respectively, it can be seen that the repair and soothing modified protein composition prepared according to the application can repair damaged skin, and has a fast effect, compared with the NC group, it can improve the content of LOR protein and FLG protein in the damaged skin.

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

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

[0089] The detection model used is 3D epidermis damage model, the detection method is H&E staining, the stimulation condition is 0.1% SLS, EG1 and EG2 are respectively the damaged model treated once and treated 5 times with the repair and soothing modified protein composition prepared according to the embodiment 1, and the results are shown in Figure 5 It can be seen that compared with the NC group, the stratum corneum of the damaged model treated once with the repair and soothing modified protein composition becomes a little tighter, the thickness of the living cell layer increases, the uniformity of the cell nucleus staining (deep purple) improves, and the damage state of the cells is recovered; and the thickness of the living cell layer of the damaged model treated 5 times with the repair and soothing modified protein composition prepared according to the embodiment 1 increases significantly, the damage is obviously repaired, and the cell vitality is full.

[0090] (4) Expression of Claudins of the skin barrier

[0091] Claudins are present in all epithelial and endothelial cells. Claudins are four-transmembrane proteins. The PDZ site at the carboxyl terminus in 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 methods: After irradiating HaCaT with UVB, the modified protein composition for repair and soothing prepared in Example 1 was cultured for 24 h. Claudin-1 and Claudin-5 were detected by qRT-PCR, and the results are as follows. Figure 6 and Figure 7 As shown, the repair 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 in the face of UV damage.

[0093] IV. Detection of skin moisture content, transepidermal water loss, and facial redness and erythema.

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

[0095] 32 volunteers (Chinese with sensitive skin - screened via questionnaire) used the repairing and soothing modified protein composition prepared in Example 1 twice daily for 28 days. Facial skin moisture content was measured before use and at 7 and 14 days after use. CM825) and transdermal water loss test ( TM Hex), the results are as follows Figure 8 and Figure 9 As shown, facial redness of subjects was detected before use and at 14 and 28 days later. The results of facial redness in subjects 7 and 13 are as follows. Figure 10 As shown, the amount of skin erythema in volunteers with UV erythema before and after using the composition was assessed using a colorimetric method. The amount of skin erythema before and 2 hours after using the composition is as follows: Figure 11 As shown, the skin's moisture content increases, transepidermal water loss decreases, facial redness is significantly improved, and the amount of erythema decreases after using the repairing and soothing modified protein composition of the present invention.

[0096] 5. The repair and soothing modified protein compositions prepared in Example 1 and Comparative Example 1 were applied to the skin of the test subjects. Bacterial tests were performed on the applied areas of the skin after 6 hours and 12 hours, respectively, to obtain the percentage of bacteria on the skin before application and to test the adhesion strength of the skin. The test results are shown in Table 3.

[0097] Table 3 Germicidal and adhesion strength detection of the composition

[0098]

[0099] As shown in Table 3, the modified protein with amphoteric properties is added in the repair and soothing modified protein composition of the present application, 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-inflammation, the surface amphoteric substance taurine improves the antibacterial property, compared with the unmodified protein, which can form a protective effect on the skin, and can inhibit bacteria and prevent further inflammation of the skin.

[0100] In summary, the repair and soothing modified protein composition prepared by the present application has the effects of penetrating the dermis layer, repairing and protecting the skin surface, and anti-inflammation, can perform multiple repair and soothing on the damaged skin, and has obvious effect, and can be added in skin care products as an efficacy ingredient.

Claims

1. A method for preparing a repairing and soothing modified protein composition, characterized in that, Includes the following steps: Step 1: Prepare the modified protein; The modified protein is prepared by adding the protein to PBS buffer at a solid-liquid ratio of 1:10 g / mL, mixing it with 3-5 wt% sodium dextran sulfate, performing a freeze-thaw cycle at -10 to -20°C, adding 6-9 wt% of a composition of pentaerythritol tetra(ethylhexanoate), taurine, and thiol compounds in a mass ratio of 11-15:7-9:3-6, adjusting the pH to 7-9, adding cross-linking enzyme, heating at 45-55°C for 3-5 hours in an inert atmosphere, dialyzing with a protein dialysis bag, and then freeze-drying to obtain the protein. The protein is one or more of fibroin, keratin and silk protein; The cross-linking enzyme is a transglutaminase added at an amount of 35-40 U / g protein; Step 2: Process the vegetable oil, then mix it with the modified protein and plant extract compound at a temperature of 50-60℃ for 30-50 minutes to obtain the modified protein composition; The vegetable oil is a complex composed of olive fruit oil and tallow tree seed oil in a mass ratio of 10-18:11-15. The process involves treating the vegetable oil with plasma for 3-6 minutes. The preparation of the plant extract compound solution involves mixing and grinding pepper and eucalyptus leaves with 6-8 wt% lactic acid at a mass ratio of 13-15:3-5 for 60-80 minutes, then adding them to an aqueous solvent at a solid-liquid ratio of 1:6-10 g / mL and heating under reflux for 2-4 hours. The solution is then filtered through a macromolecular resin.

2. The method for preparing a repairing and soothing modified protein composition according to claim 1, characterized in that, The aqueous solvent is a urea solution with a mass fraction of 14-19%.

3. A repairing and soothing modified protein composition prepared by the method according to claim 1 or 2, characterized in that, It contains the following components by weight: 8-12 parts modified protein, 19-26 parts plant compound extract, and 6-10 parts plant oil.

4. The repairing and soothing modified protein composition according to claim 3, characterized in that, It contains the following components by weight: 10 parts modified protein, 23 parts plant compound extract, and 8 parts plant oil.

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