Polypeptide composition containing ceramide E and application thereof

Abstract

The invention belongs to the field of cosmetics, and discloses a ceramide E-containing polypeptide composition and application thereof. The polypeptide composition containing ceramide E is prepared from the following components in parts by mass: 5 to 10 parts of ceramide E, 3 to 6 parts of fibronectin, 3 to 6 parts of palmitoyl pentapeptide-4, 1.5 to 3.5 parts of ergothioneine, 20 to 40 parts of oil-phase solvent, 1.5 to 4.5 parts of caprylic / capric triglyceride, 0.5 to 2.5 parts of soybean lecithin, 0.5 to 1.5 parts of polysorbate-80, 0.1 to 0.5 part of cholesterol and the balance of water, wherein the total mass part of the water is 100. In the polypeptide composition disclosed by the invention, ceramide E, fibronectin, ergothioneine and palmitoyl pentapeptide-4 are mutually promoted and have synergistic interaction, and a final product shows very good repairing and antioxidant effects.

Description

Technical Field

[0001] The present invention belongs to the field of cosmetics, and in particular relates to a polypeptide composition containing ceramide E and application thereof. Background Art

[0002] Skin is the largest organ in the human body, covering the surface of the human body. It has the functions of protection, sensation, and temperature regulation. Its structure from top to bottom is epidermis, dermis, and hypodermis. As humans grow, their skin ages in different stages. At the same time, there are various problems with the skin, such as the loss of moisture in the stratum corneum and the destruction of the skin barrier.

[0003] Ceramide, one of the sphingolipids, exists in the stratum corneum of the skin and accounts for about 50% of the intercellular lipids in the stratum corneum. It has long been known that sphingolipids such as ceramide, glucosylceramide, and galactosylceramide have the effects of promoting the moisture retention of the stratum corneum and improving skin roughness.

[0004] In practical applications, ceramide is a highly crystalline substance. When used in cosmetics, it has problems such as low solubility, poor water dispersibility, and easy crystallization. It is not easy to add to aqueous and water-in-oil cosmetic systems, which affects its skin care effect and limits its application in cosmetics. On the other hand, the applicant found that in various types of ceramide-containing cosmetics sold on the market, the stability and transdermal performance of ceramide are poor, and it cannot be well absorbed and utilized on the skin. Summary of the invention

[0005] In order to overcome the above-mentioned shortcomings and deficiencies of the prior art, the primary purpose of the present invention is to provide a polypeptide composition containing ceramide E. The polypeptide composition containing ceramide E has good repairing and anti-oxidation effects.

[0006] Another object of the present invention is to provide a method for preparing the above-mentioned polypeptide composition containing ceramide E.

[0007] Another object of the present invention is to provide an application of the above-mentioned polypeptide composition containing ceramide E.

[0008] The purpose of the present invention is achieved through the following solutions: In a first aspect, the present invention provides a polypeptide composition containing ceramide E, which comprises, by weight: 5-10 parts of ceramide E, 3-6 parts of fibronectin, 3-6 parts of palmitoyl pentapeptide-4, 1.5-3.5 parts of ergothioneine, 20-40 parts of oil phase solvent, 1.5-4.5 parts of caprylic / capric triglyceride, 0.5-2.5 parts of soy lecithin, 0.5-1.5 parts of polysorbate-80, 0.1-0.5 parts of cholesterol and water to make up the total weight of 100 parts.

[0009] In some embodiments of the present invention, the ceramide E-containing polypeptide composition includes, by mass, 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soy lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0010] In some embodiments of the present invention, the ceramide E-containing polypeptide composition is a liposome.

[0011] In some embodiments of the present invention, the oil phase solvent includes at least one of glycerol, propylene glycol, and butylene glycol.

[0012] In some embodiments of the present invention, the oil phase solvent includes glycerol, propylene glycol and butylene glycol, and the mass ratio of glycerol, propylene glycol and butylene glycol is (10-40): (2-15): (2-15); preferably 20:5:5.

[0013] Ceramide E, also known as cetyl-pg hydroxyethyl palmitamide, is a modified and optimized molecular structure of natural ceramide 2. It can improve the skin barrier, enhance the cohesion of epidermal cells, promote epidermal hydration, and improve the skin's water retention capacity.

[0014] Fibronectin (Fn) is a high molecular weight glycoprotein in the extracellular matrix and a protein with a wide range of biological activities. Fibronectin is a high molecular weight glycoprotein composed of two similar subunits, each of which has six functional regions and an RGD sequence. It can bind to heparin, collagen, fiber and other substances, and has the function of promoting cell adhesion, migration, proliferation and differentiation. It plays a very important role in all stages of tissue repair.

[0015] Palmitoyl Pentapeptide-4 is a pentapeptide synthetic chain protein produced by connecting five amino acid peptides. It is a signal peptide that can maintain the stability of skin structure, increase skin thickness, prevent the formation of wrinkles, and delay cell aging by stimulating the synthesis of substances such as collagen and sucrose amine.

[0016] Ergothioneine (EGT) is a natural amino acid derivative derived from histidine. As a rare natural chiral amino acid antioxidant, ergothioneine has multiple functions such as anti-aging, cancer prevention, maintaining DNA biosynthesis, maintaining normal cell growth, and maintaining cell immune function. It is widely used in food, medicine, cosmetics and other fields.

[0017] The present invention integrates four active substances, namely, ceramide E, which is insoluble in water, fibronectin, which is difficult to be absorbed through the skin, and palmitoyl pentapeptide-4, which is soluble in water, and ergothioneine, with the help of wall materials, emulsifiers, solvents and auxiliary agents, to form a liposome type polypeptide composition. Liposomes are cell membrane-like structures, closed microvesicles with a bilayer structure formed by amphiphilic molecules. Among them, fibronectin with a larger molecular weight and ceramide E, which are insoluble in water, are wrapped in the innermost chamber of the liposome, and palmitoyl pentapeptide-4 and ergothioneine are wrapped between the phospholipid bilayers. The liposomes containing four active substances formed by the present invention have good repair, anti-oxidation and light damage repair effects.

[0018] The invention selects glycerol, propylene glycol and butylene glycol as oil phase solvents, which can well dissolve caprylic / capric triglyceride, soybean lecithin and polysorbate-80. At the same time, glycerol, propylene glycol and butylene glycol have a certain solubility in water, which can promote the formation of liposomes and are conducive to the formation of liposomes with small particle size and high encapsulation rate.

[0019] In the present invention, soybean lecithin is used as the wall material of the polypeptide composition, which can encapsulate water-insoluble ceramide E and fibronectin in the inner cavity of the liposome, thereby improving the transdermal absorption performance and stability of ceramide E and fibronectin.

[0020] In the present invention, cholesterol, as a ligand of soybean lecithin, can adjust the fluidity of the membrane, facilitate the orderly arrangement of lecithin in the membrane, and improve the encapsulation rate and stability of the polypeptide composition.

[0021] In the present invention, caprylic acid / capric acid triglyceride is used as a stabilizer to improve the stability of the polypeptide liposome and prevent the polypeptide composition from deteriorating and stratifying.

[0022] In the present invention, polysorbate-80 is used as an emulsifier for the polypeptide composition, and plays an emulsifying role in the process of mixing the aqueous phase and the oil phase, which is conducive to forming a polypeptide composition with a small particle size, avoiding the rupture of the polypeptide composition, and preventing the outflow of active substances such as ceramide E, fibronectin, palmitoyl pentapeptide-4, and ergothioneine.

[0023] In a second aspect, the present invention provides a method for preparing the above-mentioned polypeptide composition containing ceramide E, comprising: (1) Evenly mixing ceramide E, fibronectin, caprylic / capric triglyceride, soybean lecithin, polysorbate 80, cholesterol and an oil phase solvent to obtain an oil phase; (2) mixing water, palmitoyl pentapeptide-4 and ergothioneine to obtain an aqueous phase; (3) adding the water phase to the oil phase, stirring and reacting to obtain a water-oil phase mixture; (4) subjecting the water-oil phase mixture to a high-speed shear dispersion treatment to obtain crude liposomes; (5) The crude liposomes are subjected to high-pressure homogenization to obtain the ceramide E-containing polypeptide composition.

[0024] In some embodiments of the present invention, the adding method in step (3) is preferably dropwise addition, and the dropping speed is 1-4 mL / min; preferably, the dropping speed is 2 mL / min.

[0025] In some embodiments of the present invention, the stirring reaction time in step (3) is 20-50 min, the stirring reaction speed is 150-400 rpm, and the stirring reaction temperature is 20-40°C.

[0026] Preferably, the stirring reaction time in step (3) is 30 min, the stirring reaction speed is 250 rpm, and the stirring reaction temperature is 25°C.

[0027] In some embodiments of the present invention, the rotation speed of the shear dispersion treatment in step (4) is 6000-12000 rpm, and the time of the shear dispersion treatment is 10-20 min.

[0028] Preferably, the rotation speed of the shear dispersion treatment is 9000 rpm, and the time of the shear dispersion treatment is 15 min.

[0029] In some embodiments of the present invention, the homogenization pressure in step (5) is 800-1200 bar, and the number of homogenization cycles is 1-5 times.

[0030] Preferably, the homogenization pressure is 1000 bar, and the number of homogenization cycles is 2 times.

[0031] In a third aspect, the present invention provides use of the above-mentioned ceramide E-containing polypeptide composition in the preparation of cosmetics.

[0032] Preferably, the ceramide E-containing polypeptide composition can be prepared into any preparation in the field of cosmetics, such as solution, suspension, emulsion, paste, gel, cream, lotion, spray, oil, etc., but the present invention is not limited thereto.

[0033] Preferably, the amount of the ceramide E-containing polypeptide composition of the present invention added to the cosmetic is 1-30% by mass.

[0034] In a fourth aspect, the present invention provides a repair lotion, comprising, by mass percentage: 4-10% of the above-mentioned polypeptide composition containing ceramide E, 2.1-10% of a moisturizer, 0.1-0.5% of an antioxidant, 2-7% of an emulsifier, 2.52-13.1% of a skin conditioner, 0.33-1.4% of a thickener, 0.02-0.08% of a chelating agent, 0.8-5% of a preservative and the remainder of water.

[0035] In some embodiments of the present invention, the repair lotion includes, by mass percentage: 6% of a polypeptide composition containing ceramide E, 7% of a moisturizer, 0.3% of an antioxidant, 4.5% of an emulsifier, 8.05% of a skin conditioner, 0.85% of a thickener, 0.05% of a chelating agent, 1% of a preservative and 69.25% of water.

[0036] In some embodiments of the present invention, the moisturizing agent includes at least one of glycerin and panthenol.

[0037] In some embodiments of the present invention, the antioxidant comprises tocopherol acetate.

[0038] In some embodiments of the present invention, the emulsifier includes at least one of coconut glucoside, cetyl alcohol, and cetearyl alcohol.

[0039] In some embodiments of the present invention, the skin conditioning agent includes at least one of squalane, polydimethylsiloxane, shea butter, and tert-butyl alcohol.

[0040] In some embodiments of the present invention, the thickener includes at least one of polyacrylate crosspolymer-6, xanthan gum, and ammonium acryloyldimethyltaurate / VP copolymer.

[0041] In some embodiments of the invention, the chelating agent comprises disodium EDTA.

[0042] In some embodiments of the present invention, the preservative includes at least one of p-hydroxyacetophenone, butylene glycol, pentylene glycol, hexylene glycol, phenoxyethanol, benzyl alcohol, imidazolidinyl urea, sodium benzoate, methylisothiazolinone, caprylyl glycol, paeonol, caprylylhydroxamic acid, caprylic glyceride, hydantoin, methylparaben, caprylyl glycol, hydroxyacetone, and methylparaben.

[0043] In a fifth aspect, the present invention provides a method for preparing the above-mentioned repairing emulsion, comprising: S1. Mix water, humectant, thickener and chelating agent, heat and stir to obtain phase A; S2, mixing an antioxidant, an emulsifier, and a skin conditioner, heating and stirring to obtain phase B; S3, under heating conditions, adding the phase B to the phase A, mixing and homogenizing to obtain a mixture; S4. After cooling, add a preservative and a polypeptide composition containing ceramide E to the mixture, and stir evenly to obtain a repairing lotion.

[0044] In some embodiments of the present invention, the heating temperatures in steps S1-S3 are all relatively independently 75-85°C.

[0045] In some embodiments of the present invention, the speed of the homogenization treatment in step S3 is 12000-18000 r / min, and the time of the homogenization treatment is 20-40 min.

[0046] In some embodiments of the present invention, the cooling temperature in step S4 is 35-45°C.

[0047] Compared with the prior art, the present invention has the following beneficial effects: (1) Ceramide E has low solubility in water and poor compatibility in the formula. The molecular weight of fibronectin is 440-450KDa, which makes it difficult to directly penetrate into the stratum corneum to play a role in repairing the skin barrier, affecting its addition to skin care products and the manifestation of its efficacy. The present invention encapsulates the water-insoluble ceramide E and the difficultly transdermally absorbed fibronectin in the form of liposomes, thereby enhancing the transdermal absorption effect of ceramide E and fibronectin and enhancing the compatibility of the two in the system. Ceramide E and fibronectin can better play the role of moisturizing, repairing the barrier and anti-wrinkle.

[0048] (2) The polypeptide composition of the present invention adds water-soluble active substances of ergothioneine and palmitoyl pentapeptide-4. The water-soluble active substances are wrapped in the hydrophilic layer of the phospholipid bilayer and play a skin care effect together with ceramide E and fibronectin. The final product has good repair, antioxidant and efficacy. When the polypeptide composition containing ceramide E is used as an active substance to prepare a repair lotion, the obtained repair lotion has a good photodamage repair effect.

[0049] (3) Ergothioneine has strong antioxidant properties. Adding ergothioneine to the system of the present invention can not only enhance the skin care effect of the liposome as a whole, but also enhance the overall antioxidant performance of the system, improve the stability of the system, and extend the shelf life of the product.

[0050] (4) In the liposome system of the present invention, ceramide E, fibronectin, ergothioneine and palmitoyl pentapeptide-4 promote each other and synergistically enhance each other, and the final product exhibits excellent repair and antioxidant effects. DETAILED DESCRIPTION

[0051] The present invention is further described in detail below in conjunction with the examples, but the embodiments of the present invention are not limited thereto. If the specific conditions are not specified in the examples, they are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0052] Unless otherwise specified, all percentages mentioned in the present invention are by mass. Unless otherwise specified, all unspecified temperatures in the present invention refer to room temperature (15-40°C).

[0053] The fibronectin and mussel protein used in the present invention are from Dongguan Juwei New Material Technology Co., Ltd.

[0054] Example 1 A polypeptide composition containing ceramide E.

[0055] A polypeptide composition containing ceramide E, comprising, by weight: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0056] Among them, the oil phase solvent is glycerol, propylene glycol, and butylene glycol, and the mass ratio is 20:5:5.

[0057] The preparation method of the polypeptide composition containing ceramide E is as follows: (1) Evenly mixing ceramide E, fibronectin, caprylic / capric triglyceride, soybean lecithin, polysorbate 80, cholesterol and an oil phase solvent to obtain an oil phase; (2) mixing water, palmitoyl pentapeptide-4 and ergothioneine to obtain an aqueous phase; (3) adding the aqueous phase to the oil phase at a rate of 2 mL / min, stirring the mixture for 30 min at 25°C and 250 rpm to obtain a water-oil phase mixture; (4) subjecting the water-oil phase mixture to a high-speed shear dispersion treatment at a speed of 9000 rpm for 15 min to obtain crude liposomes; (5) The crude liposomes were subjected to high-pressure homogenization treatment at a homogenization pressure of 1000 bar for 2 times to obtain a polypeptide composition containing ceramide E.

[0058] Example 2-6 A polypeptide composition containing ceramide E.

[0059] Examples 2-6 all provide a polypeptide composition containing ceramide E, the preparation method of which is the same as that of Example 1, and the components and their mass fractions are shown in Table 1.

[0060] Table 1 Among them, the oil phase solvents of Examples 1-4 and 6 are glycerol, propylene glycol, and butylene glycol in a mass ratio of 20:5:5; the oil phase solvent of Example 5 is glycerol and propylene glycol in a mass ratio of 15:15.

[0061] Example 7 A polypeptide composition containing ceramide E.

[0062] A polypeptide composition containing ceramide E comprises, by weight: 10 parts of ceramide E, 3 parts of fibronectin, 3 parts of palmitoyl pentapeptide-4, 2 parts of ergothioneine, 40 parts of oil phase solvent, 4.5 parts of caprylic / capric triglyceride, 2.5 parts of soy lecithin, 0.5 parts of polysorbate-80, 0.1 parts of cholesterol and 34.4 parts of water.

[0063] Among them, the oil phase solvent is glycerol and butanediol, and the mass ratio is 30:10.

[0064] A method for preparing a polypeptide composition containing ceramide E is as follows: (1) Evenly mixing ceramide E, fibronectin, caprylic / capric triglyceride, soybean lecithin, polysorbate 80, cholesterol and an oil phase solvent to obtain an oil phase; (2) mixing water, palmitoyl pentapeptide-4 and ergothioneine to obtain an aqueous phase; (3) adding the aqueous phase to the oil phase at a rate of 4 mL / min, stirring the mixture for 50 min at a temperature of 20°C and a stirring speed of 400 rpm to obtain a water-oil phase mixture; (4) subjecting the water-oil phase mixture to a high-speed shear dispersion treatment at a speed of 6000 rpm for 20 min to obtain crude liposomes; (5) The crude liposomes were subjected to high-pressure homogenization treatment at a homogenization pressure of 800 bar for 5 times to obtain a polypeptide composition containing ceramide E.

[0065] Example 8 A polypeptide composition containing ceramide E.

[0066] A polypeptide composition containing ceramide E comprises, by weight: 7 parts of ceramide E, 5 parts of fibronectin, 6 parts of palmitoyl pentapeptide-4, 3 parts of ergothioneine, 20 parts of oil phase solvent, 1.5 parts of caprylic / capric triglyceride, 0.5 parts of soy lecithin, 1.5 parts of polysorbate-80, 0.5 parts of cholesterol and 55 parts of water.

[0067] Among them, the oil phase solvent is propylene glycol and butylene glycol, and the mass ratio is 10:10.

[0068] A method for preparing a polypeptide composition containing ceramide E is as follows: (1) Evenly mixing ceramide E, fibronectin, caprylic / capric triglyceride, soybean lecithin, polysorbate 80, cholesterol and an oil phase solvent to obtain an oil phase; (2) mixing water, palmitoyl pentapeptide-4 and ergothioneine to obtain an aqueous phase; (3) adding the aqueous phase to the oil phase at a rate of 1 mL / min, stirring the mixture for 50 min at 40°C and 150 rpm to obtain a water-oil phase mixture; (4) subjecting the water-oil phase mixture to a high-speed shear dispersion treatment at a speed of 12,000 rpm for 10 min to obtain crude liposomes; (5) The crude liposomes are subjected to high-pressure homogenization treatment at a homogenization pressure of 1200 bar and once to obtain a polypeptide composition containing ceramide E.

[0069] Comparative Examples 1-6 A composition.

[0070] Comparative Examples 1-6 all provide a composition, the components contained in the composition and the mass fraction of each component are specifically shown in Table 2, wherein the type and ratio of the oil phase solvent and the preparation method of the composition are the same as those in Example 1.

[0071] Table 2 Comparative Example 7 A polypeptide composition.

[0072] A polypeptide composition comprises, by weight: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0073] Wherein, the oil phase solvent is hydrogenated castor oil.

[0074] The preparation method of the polypeptide composition of Comparative Example 7 is the same as that of Example 1.

[0075] Comparative Example 8: a polypeptide composition.

[0076] A polypeptide composition comprises, by weight: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-60, 0.3 parts of cholesterol and 45.7 parts of water.

[0077] The type and ratio of the oil phase solvent and the preparation method of the polypeptide composition are the same as those in Example 1.

[0078] The difference between Comparative Example 8 and Example 1 is that in Comparative Example 8, polysorbate 60 is used instead of polysorbate 80.

[0079] Comparative Example 9 A polypeptide composition.

[0080] A polypeptide composition comprises, by weight: 8 parts of ceramide 1 (ceramide EOS), 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soy lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0081] The type and ratio of the oil phase solvent and the preparation method of the polypeptide composition are the same as those in Example 1.

[0082] The difference between Comparative Example 9 and Example 1 is that, in Comparative Example 9, ceramide 1 is used instead of ceramide E of the present invention.

[0083] Comparative Example 10 A polypeptide composition.

[0084] A polypeptide composition comprises, by weight, 8 parts of ceramide E, 4 parts of mussel protein, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0085] The type and ratio of the oil phase solvent and the preparation method of the polypeptide composition are the same as those in Example 1.

[0086] The difference between Comparative Example 10 and Example 1 is that, in Comparative Example 10, mussel protein is used instead of the fibronectin of the present invention.

[0087] Comparative Example 11 A polypeptide composition.

[0088] A polypeptide composition comprises, by weight: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of oligopeptide-1, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0089] The type and ratio of the oil phase solvent and the preparation method of the polypeptide composition are the same as those in Example 1.

[0090] The difference between Comparative Example 11 and Example 1 is that Comparative Example 11 uses oligopeptide-1 instead of palmitoyl pentapeptide-4 of the present invention.

[0091] Comparative Example 12: a polypeptide composition.

[0092] A polypeptide composition comprises, by weight: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of vitamin C, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soybean lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

[0093] The type and ratio of the oil phase solvent and the preparation method of the polypeptide composition are the same as those in Example 1.

[0094] The difference between Comparative Example 12 and Example 1 is that Comparative Example 12 replaces ergothioneine of the present invention with vitamin C.

[0095] Application Example 1 A repairing lotion.

[0096] Application Example 1 is a repairing lotion, which includes, by mass percentage, 6% of the ceramide E-containing polypeptide composition of Example 1, 7% of a moisturizer, 0.3% of an antioxidant, 4.5% of an emulsifier, 8.05% of a skin conditioner, 0.85% of a thickener, 0.05% of a chelating agent, 1% of a preservative and 72.25% of water.

[0097] Among them, the 7% moisturizer includes: 6% glycerin and 1% panthenol; 0.3% Antioxidants include: 0.3% Tocopheryl Acetate; 4.5% emulsifiers include: 1.4% coco-glucoside, 0.6% cetyl alcohol, and 2.5% cetearyl alcohol; 8.05% skin conditioning agents including: 2.5% squalane, 2.5% dimethicone, 3% shea butter, and 0.05% t-butyl alcohol; 0.85% Thickener including: 0.15% Polyacrylate Crosspolymer-6, 0.2% Xanthan Gum, and 0.5% Ammonium Acryloyldimethyltaurate / VP Copolymer; 0.05% chelating agent includes: 0.05% disodium EDTA; 1% preservatives include: 0.5% p-hydroxyacetophenone and 0.5% 1,2-hexanediol.

[0098] The preparation method of the repairing emulsion of Application Example 1 comprises: S1. Mix water, humectant, thickener and chelating agent, heat to 80°C and stir evenly to obtain phase A; S2, mixing the antioxidant, emulsifier and skin conditioner, heating to 80° C. and stirring evenly to obtain phase B; S3. Add phase B to phase A at 80°C, mix and homogenize at a speed of 15000 r / min for 30 min to obtain a mixture; S4. After cooling to 40° C., add a preservative and the polypeptide composition containing ceramide E of Example 1 to the mixture, and stir evenly to obtain a repairing lotion containing ceramide E.

[0099] Application Example 2-4 A repairing lotion.

[0100] The proportions of the raw materials and the preparation methods of the repairing lotions of Application Examples 2-4 are the same as those of Application Example 1, except that the polypeptide composition containing ceramide E is different. The polypeptide composition containing ceramide E used in the repairing lotions of Application Examples 1-4 is shown in Table 3.

[0101] Table 3 Test Example 1: Transdermal absorption effect test.

[0102] Test samples: polypeptide compositions of Examples 1-8 and Comparative Examples 1-12; unencapsulated control group: 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, and 81.5 parts of 50% (v / v) DMSO aqueous solution, mixed evenly; the above test samples were all prepared into a solution with a mass concentration of 10% using physiological saline.

[0103] Test steps: RJY-6B drug transdermal diffusion tester was used for the test. Take the skin from the back of the pig ear, remove the hair and fat, cut a 3*3cm skin disc, soak it in saline for 0.5h, wipe it dry with absorbent cotton, and fix it in a vertical permeation test diffusion cell. Add saline as the receiving solution to the receiving cell, add the test sample to the supply cell, seal the supply cell with a sealing film after adding, turn on the magnetic stirring at a speed of 300r / min, and slowly extract all the receiving solution with a syringe after 8h. After ultrasonic demulsification, use HPLC to determine the content of ceramide E and fibronectin in the receiving solution (standard curve method), and calculate the permeability of ceramide E and fibronectin (or mussel protein) based on the original content of ceramide E and fibronectin (or mussel protein). The test results are shown in Table 4 below.

[0104] Table 4 The test results show that, compared with the unwrapped active substance, the polypeptide composition of the embodiment of the present invention is more easily transdermal absorption after ceramide E, fibronectin, palmitoyl pentapeptide-4 and ergothioneine are wrapped, and the ceramide E of the embodiment has a permeability of more than 85%, and the permeability of fibronectin is more than 80%; the permeability of unwrapped ceramide E and fibronectin is only 5.8% and 3.2%. The encapsulation technology of the present invention can greatly improve the transdermal absorption effect of ceramide E and fibronectin. From the comparison of comparative examples 1-4, comparative examples 9-12 and embodiments 1-4, it can be seen that the active components ceramide E, fibronectin, palmitoyl pentapeptide-4 and ergothioneine affect each other, which will affect the transdermal absorption effect of the final product. When the ratio of the four changes, the permeability of ceramide E and fibronectin will also change. Lack of one of the components, or replacing the components with components with similar functions, the transdermal absorption effect of the final product becomes worse. From the results of Example 1 and Comparative Examples 5-8, it can be seen that the addition of the ligand of the polypeptide composition, the addition of the emulsifier, the type of the emulsifier, and the change of the oil phase solvent will affect the transdermal absorption effect of the polypeptide composition. In the present invention, the emulsifier is preferably polysorbate-80, and the oil phase solvent is preferably at least one of glycerol, propylene glycol, and butylene glycol.

[0105] Test Example 2: Repair performance test.

[0106] Test samples: the polypeptide compositions of Examples 1-8, Comparative Examples 1-12, and the unencapsulated control group (see Test Example 1), were prepared into a 120 ppm solution using H-DMEM culture medium containing 10% FBS.

[0107] Test steps: The culture insert was attached to a 24-well plate, and HaCaT cells in the logarithmic growth phase (purchased from the Cell Resource Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences) were cultured at 4.5×10 5 The cells were inoculated into the plug at a density of 10 cells / mL. A negative control group and a test sample group were set up, with 3 plugs in each group. One plug had 2 holes, and 70 μL of cell suspension was inoculated into each hole. The cells were cultured at 5% CO2 and 37°C for 24 h.

[0108] When the cell coverage rate is 95%-100%, use sterile tweezers to vertically remove the scratch plug in the 24-well plate, take 1mL PBS solution to each well and wash once, add 1mL H-DMEM medium containing 10% FBS to the blank control group, and add 1mL of the test sample to the test sample group. Observe under a microscope at 0h and 24h time points. Use ImageJ software to measure the scratch area of ​​each group and calculate the cell migration rate (%). Each group of samples was tested in parallel 3 times, and the test results were averaged.

[0109] The test results are shown in Table 5 below.

[0110] Table 5 The test results show that, compared with the negative control group to which the test sample was not added, after adding the polypeptide composition of the embodiment of the present invention and the comparative example, the cell migration rate is improved to a certain extent, and is better than the unwrapped composition, indicating that the polypeptide composition of the embodiment of the present invention and the comparative example can promote the migration of cells and show a repair effect. The cell migration rate of Example 1 is higher than that of Comparative Examples 1-4, Comparative Examples 9-12 and Examples 2-4, indicating that the active components ceramide E, fibronectin, palmitoyl pentapeptide-4 and ergothioneine have a synergistic effect, and the four affect each other, which will affect the cell migration effect of the final product. When the ratio of the four changes, the cell migration rate of ceramide E and fibronectin will also change, lacking one of the components, or replacing the components therein, and the repair of the final product is deteriorated. From the results of Example 1 and Comparative Examples 5-8, it can be seen that the addition of the ligand of the polypeptide composition, the addition of the emulsifier, the type of the emulsifier, and the change of the oil phase solvent will affect the repair effect of the polypeptide composition. In the present invention, the emulsifier is preferably polysorbate-80, and the oil phase solvent is preferably at least one of glycerol, propylene glycol, and butylene glycol.

[0111] Test Example 3: Antioxidant performance test.

[0112] Superoxide dismutases (SOD) are widely present and are the first line of defense against oxidative stress. SOD has important physiological functions in scavenging free radicals and protecting cells from oxidative stress damage. This test uses keratinocytes HaCaT (purchased from the Cell Resource Center of the Institute of Basic Medicine, Chinese Academy of Medical Sciences) induced with 0.1μg / mL H2O2 for 1 hour as an in vitro antioxidant model, and measures the relative expression of SOD1 (Cu / Zn-SOD) mRNA to evaluate the antioxidant effect of the sample to be tested.

[0113] Test samples: the polypeptide compositions of Examples 1-8, Comparative Examples 1-12, and the unencapsulated control group (see Test Example 1), were prepared into a 60 ppm solution using H-DMEM culture medium containing 10% FBS.

[0114] Test steps: HaCaT cells in the logarithmic phase were added to each well of a 6-well culture plate. 5cells, added H-DMEM medium containing 10% FBS, and cultured for 24 hours at 37°C, 5% CO2, and saturated humidity; the medium was discarded, 1 mL of H-DMEM medium containing 10% FBS was added to the negative control group, and 1 mL of 0.1 μg / mL H2O2 was added to the model control group and the sample group to be tested for induction for 1 hour, the medium was discarded, 1 mL of H-DMEM medium containing 10% FBS was added to the negative control group and the model control group, and 1 mL of sample solution was added to each sample group, and culture was continued for 24 hours. After the culture was completed, the total RNA of HaCaT cells in each well was extracted and reverse transcribed for fluorescence quantitative PCR. Real-time fluorescence quantitative results were obtained using 2 －△△CT Each group of samples was tested 3 times in parallel and the test results were averaged.

[0115] The test results are shown in Table 6 below.

[0116] Table 6 Compared with the negative control group, the model control group did not add any active substance, and after H2O2 modeling, the SOD1 gene expression of the cell decreased, and after adding active substances, the relative expression of the SOD1 gene was increased, showing an antioxidant effect. The peptide compositions of the embodiments of the present invention and the comparative examples can promote the expression of the SOD1 gene, showing an antioxidant effect. The expression of the SOD1 gene of Example 1 is higher than that of Comparative Examples 1-4, Comparative Examples 9-12 and Example 2-4, indicating that the active components ceramide E, fibronectin, palmitoyl pentapeptide-4 and ergothioneine have synergistic effects, and the four influence each other, which will affect the cell antioxidant effect of the final product, and when the ratio of the four changes, the cell mobility of ceramide E and fibronectin will also change, lacking one of the components, or replacing the components therein, and the antioxidant variation of the final product. From the results of Example 1 and Comparative Examples 5-8, it can be seen that the addition of the ligand of the polypeptide composition, the addition of the emulsifier, the type of the emulsifier, and the change of the oil phase solvent will affect the antioxidant effect of the polypeptide composition. In the present invention, the emulsifier is preferably polysorbate-80, and the oil phase solvent is preferably at least one of glycerol, propylene glycol, and butylene glycol.

[0117] Test Example 4: Photodamage repair efficacy test.

[0118] Test samples: the repair lotion and matrix lotion of Application Examples 1-4 (the formula and preparation method are the same as those of Application Example 1, except that the polypeptide composition containing ceramide E is replaced with deionized water of the same mass ratio).

[0119] Test steps: Use a solar simulator to irradiate the test area once at a dose of 1.5 times the MED (minimal erythemadose, MED; the minimum ultraviolet dose that makes the skin red is called 1 MED).

[0120] Twenty volunteers with intact skin on the inner forearms were selected, and two test areas were divided on the left and right forearms, for a total of 80 test areas, with 16 test areas for each test sample. A daylight simulator was used to irradiate the test area once at a dose of 1.5 times the MED to cause UV damage. 24 hours after UV damage, the skin hemoglobin value was measured; then the volunteers applied 0.2±0.03g of the sample on the test area, and after applying it evenly, the skin hemoglobin value of the test area was tested again 12 hours after the sample was applied, and the rate of change of the skin hemoglobin value before and after the sample was applied was calculated to evaluate the photodamage repair effect of the sample.

[0121] The test results are shown in Table 7 below.

[0122] Table 7 The test results show that compared with the matrix essence, the repair lotion containing the polypeptide composition of the embodiment of the present invention has a good light damage repair effect. When light is irradiated on the surface of the skin, a variety of active oxygen species are easily produced in the cells, causing damage to the skin cells, causing skin redness, swelling, and sunburn. The polypeptide composition containing ceramide E of the present invention can promote the migration of cells in the stratum corneum and can also remove ROS (peroxides) generated by light. Therefore, after adding it, it can reduce the hemoglobin value of damaged skin and play a role in repairing light-damaged skin.

[0123] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. A polypeptide composition containing ceramide E, characterized in that The invention comprises, by weight: 5-10 parts of ceramide E, 3-6 parts of fibronectin, 3-6 parts of palmitoyl pentapeptide-4, 1.5-3.5 parts of ergothioneine, 20-40 parts of oil phase solvent, 1.5-4.5 parts of caprylic / capric triglyceride, 0.5-2.5 parts of soy lecithin, 0.5-1.5 parts of polysorbate-80, 0.1-0.5 parts of cholesterol and water to make up the total weight of 100 parts.

2. The ceramide E-containing polypeptide composition according to claim 1, characterized in that : In parts by mass, it includes 8 parts of ceramide E, 4 parts of fibronectin, 4 parts of palmitoyl pentapeptide-4, 2.5 parts of ergothioneine, 30 parts of oil phase solvent, 3 parts of caprylic / capric triglyceride, 1.5 parts of soy lecithin, 1 part of polysorbate-80, 0.3 parts of cholesterol and 45.7 parts of water.

3. The ceramide E-containing polypeptide composition according to claim 1 or 2, characterized in that: The oil phase solvent includes at least one of glycerol, propylene glycol and butylene glycol; preferably, it is a mixture of glycerol, propylene glycol and butylene glycol, and the mass ratio of glycerol, propylene glycol and butylene glycol is (10-40): (2-15): (2-15).

4. A method for preparing a polypeptide composition containing ceramide E according to any one of claims 1 to 3, characterized in that The following steps are involved: (1) Evenly mixing ceramide E, fibronectin, caprylic / capric triglyceride, soybean lecithin, polysorbate 80, cholesterol and an oil phase solvent to obtain an oil phase; (2) mixing water, palmitoyl pentapeptide-4 and ergothioneine to obtain an aqueous phase; (3) adding the water phase to the oil phase, stirring and reacting to obtain a water-oil phase mixture; (4) subjecting the water-oil phase mixture to a high-speed shear dispersion treatment to obtain crude liposomes; (5) The crude liposomes are subjected to high-pressure homogenization to obtain the ceramide E-containing polypeptide composition.

5. The method for preparing a polypeptide composition containing ceramide E according to claim 4, characterized in that: The method of adding in step (3) is dropwise addition, and the speed of the dropwise addition is 1-4 mL / min; The stirring reaction time in step (3) is 20-50 min, the stirring reaction speed is 150-400 rpm, and the stirring reaction temperature is 20-40° C.; The rotation speed of the shearing and dispersing treatment in step (4) is 6000-12000 rpm, and the time of the shearing and dispersing treatment is 10-20 min; The homogenization pressure in step (5) is 800-1200 bar, and the number of homogenization cycles is 1-5 times.

6. Use of the ceramide E-containing polypeptide composition according to any one of claims 1 to 3 in the preparation of cosmetics.

7. A repairing lotion, characterized in that Calculated by mass percentage, it comprises: 4-10% of the ceramide E-containing polypeptide composition according to any one of claims 1 to 3, 2.1-10% of a moisturizer, 0.1-0.5% of an antioxidant, 2-7% of an emulsifier, 2.52-13.1% of a skin conditioner, 0.33-1.4% of a thickener, 0.02-0.08% of a chelating agent, 0.8-5% of a preservative and the remainder of water.

8. The repairing emulsion according to claim 7, characterized in that: The moisturizing agent includes at least one of glycerin and panthenol; The antioxidant includes tocopherol acetate; The emulsifier includes at least one of coconut glucoside, cetyl alcohol, and cetearyl alcohol; The skin conditioning agent includes at least one of squalane, polydimethylsiloxane, shea butter, and tert-butyl alcohol; The thickener includes at least one of polyacrylate cross-linked polymer-6, xanthan gum, and ammonium acryloyldimethyltaurate / VP copolymer; The chelating agent includes disodium EDTA; The preservative includes at least one of p-hydroxyacetophenone, butylene glycol, pentylene glycol, hexylene glycol, phenoxyethanol, benzyl alcohol, imidazolidinyl urea, sodium benzoate, methylisothiazolinone, caprylyl glycol, paeonol, caprylylhydroxamic acid, caprylyl glyceride, hydantoin, methyl parahydroxybenzoate, caprylyl glycol, hydroxyacetone, and methylparaben.

9. A method for preparing the repairing emulsion according to claim 7 or 8, characterized in that The following steps are involved: S1. Mix water, humectant, thickener and chelating agent, heat and stir to obtain phase A; S2, mixing an antioxidant, an emulsifier, and a skin conditioner, heating and stirring to obtain phase B; S3, under heating conditions, adding the phase B to the phase A, mixing and homogenizing to obtain a mixture; S4. After cooling, add a preservative and a polypeptide composition containing ceramide E to the mixture, and stir evenly to obtain a repairing lotion.

10. The method for preparing the repairing emulsion according to claim 9, characterized in that: The heating temperatures in steps S1-S3 are all relatively independent of each other and are 75-85°C; The speed of the homogenization treatment in step S3 is 12000-18000 r / min, and the time of the homogenization treatment is 20-40 min; The temperature of the cooling step S4 is 35-45°C.

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