Moisturizing anti-wrinkle polypeptide composition as well as preparation method and application thereof

Abstract

The invention discloses a moisturizing anti-wrinkle polypeptide composition as well as a preparation method and application thereof, and relates to the technical field of daily chemical materials. The invention discloses a preparation method of a moisturizing and anti-wrinkle polypeptide composition, which comprises the following steps: uniformly stirring meadowfoam seed oil, caprylic acid / capric acid / succinic triglyceride and polypeptide liposome to obtain the moisturizing and anti-wrinkle polypeptide composition, wherein the polypeptide liposome is prepared by taking a combination of soybean lecithin, modified cholesterol and cholesterol as a capsule material and taking acetyl hexapeptide-8, snake venom peptide and a blue copper peptide compound as a core material through an ethanol injection method. The modified cholesterol is obtained by modifying cholesterol, the modified cholesterol is added into the capsule material, the defects that an existing liposome material is prone to aggregation and leakage, poor in stability and the like are effectively overcome, and the prepared material is good in stability, capable of being slowly released, high in skin retention amount and capable of being more easily absorbed by skin cells; the function of obviously delaying skin aging is realized.

Description

Technical Field

[0001] The invention relates to the technical field of daily chemical materials, and in particular to a moisturizing and anti-wrinkle polypeptide composition and a preparation method and application thereof. Background Art

[0002] Skin is the first barrier between the human body and the outside world. With the continuous stimulation of the environment and the passage of time, the effectiveness of some defense mechanisms tends to gradually weaken, causing a variety of skin injuries, which are manifested as skin aging. Skin aging is a complex process with multiple factors and mechanisms, characterized by skin injuries such as dryness, sagging, roughness and wrinkles. Wrinkles are the result of skin aging, which not only directly affects the normal structure and physiological function of skin tissue, but also affects the stability of the internal and external environment of the function, leading to different degrees of imbalance in metabolism, which may promote and accelerate the aging of the matrix. Under normal circumstances, the muscle contraction process is: nerve impulse-acetylcholine (Ach)-end plate potential-muscle action potential-muscle contraction. Wrinkles are formed by repeated contraction of facial muscles over the years. The formation of skin aging is divided into endogenous aging and exogenous aging. Among them, endogenous aging is mainly controlled by genetic factors and individual genes. With the increase of age, epidermal cells are multi-differentiated, collagen synthesis capacity is reduced, and cytoplasmic fluidity is reduced. This is an irreversible process that cannot be avoided and can only be delayed. It is physiological aging. Exogenous aging is the result of the skin being affected by a variety of external factors, including ultraviolet radiation, extreme temperatures, air pollution, dry environment and other factors. It can be prevented and treated through physical intervention and local application of active ingredients.

[0003] The barrier effect of the stratum corneum, the outermost layer of the skin, makes it difficult for the ingredients in ordinary cosmetics, especially water-soluble skin care factors, to reach the dermis to exert their skin care effects. Liposomes are a double-layer closed vesicle structure formed by amphiphilic molecules with polar heads and non-polar tails dispersed in the aqueous phase. The formation mechanism is the hydrophilic-hydrophobic interaction between external water molecules and amphiphilic molecules. The hydrophilic head is exposed to the aqueous phase to form a hydrophilic core, and the hydrophobic tail gathers to avoid the aqueous phase to form a hydrophobic interlayer, which is used to encapsulate fat-soluble, water-soluble or amphiphilic substances. The structure of liposomes is similar to that of cell membranes, with strong affinity and easier fusion with cell membranes, which increases the ability of drugs to penetrate cell membranes and enter cells, thereby achieving the delivery of active ingredients. However, liposomes have defects such as easy aggregation, easy leakage and poor stability, which seriously affect their application in the field of skin care products. In the prior art, liposomes are coated with polymer materials such as chitosan to improve the poor stability of liposomes. However, the coating of polymer materials significantly reduces the interaction between liposomes and cell membranes, which reduces the efficiency of liposome endocytosis and makes it difficult for active ingredients to efficiently enter cells to exert their effects. Summary of the invention

[0004] The purpose of the present invention is to provide a moisturizing and anti-wrinkle polypeptide composition and a preparation method and application thereof, to solve the following technical problems: Existing liposomes have defects such as easy aggregation, easy leakage and poor stability, which seriously affect their application in the field of skin care products.

[0005] The purpose of the present invention can be achieved through the following technical solutions: A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: Evenly stirring the meadowfoam seed oil, caprylic / capric / succinic triglyceride, and polypeptide liposomes to obtain a moisturizing and anti-wrinkle polypeptide composition; The preparation method of the polypeptide liposome comprises the following steps: S1: mixing the polypeptide complex and PBS buffer to obtain a core material solution; S2: blending soybean lecithin, modified cholesterol, cholesterol and anhydrous ethanol to obtain a lipid ethanol solution; S3: The core material solution and the lipid ethanol solution are mixed, subjected to ultrasonic treatment at 30-50°C for 0.5-1h, and then subjected to vacuum rotary evaporation to obtain polypeptide liposomes.

[0006] As a further embodiment of the present invention: the moisturizing and anti-wrinkle polypeptide composition comprises the following raw materials in parts by weight: 1-10 parts of meadowfoam seed oil, 84-96 parts of caprylic / capric / succinic triglycerides, and 3-6 parts of polypeptide liposomes.

[0007] As a further embodiment of the present invention: the polypeptide complex is obtained by mixing acetyl hexapeptide-8, snake venom peptide, and blue copper peptide in a mass ratio of 0.05-1:0.05-1:0.05-1; 0.3-0.5 g of the polypeptide complex is added to every 10 mL of PBS buffer.

[0008] As a further embodiment of the present invention: the mass ratio of soybean lecithin, modified cholesterol and cholesterol in the lipid ethanol solution is 6-10:1:1; the total mass of soybean lecithin, modified cholesterol and cholesterol accounts for 10-15% of the lipid ethanol solution; the volume ratio of the core material solution to the lipid ethanol solution is 1:1-1.2.

[0009] As a further embodiment of the present invention: the preparation method of modified cholesterol comprises the following steps: A1: Cholesterol, succinic anhydride, n-heptane and butanone were added to a reaction kettle in a nitrogen atmosphere, and the temperature was raised to 40-60°C. The reaction was kept warm for 2-4 hours, and n-heptane and butanone were removed. The mixture was treated with acetone and filtered under reduced pressure to obtain component 1. A2: Add component 1, N,N-dimethylformamide, N,N'-dimethyl 1,3-propylenediamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate into a reaction kettle and disperse evenly. Control the temperature to 50-60°C, keep the temperature for reaction for 6-9h, add ethanol to terminate the reaction, filter, and obtain component 2. A3: Reflux component 2, distilled water, aqueous hydrochloric acid solution, and 3-chloro-1,2-propanediol at elevated temperature for 6-8 hours, wash with water and ethanol, centrifuge, and dry to obtain modified cholesterol.

[0010] As a further embodiment of the present invention: the addition ratio of cholesterol, succinic anhydride, n-heptane and butanone in A1 is 10 g: 0.1-0.4 g: 10-20 mL: 10-20 mL.

[0011] As a further embodiment of the present invention, the addition ratio of component 1, N,N-dimethylformamide, N,N'-dimethyl 1,3-propylenediamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate in A2 is 1:100-200mL:10-20mL:0.1-0.5g.

[0012] As a further embodiment of the present invention: the hydrochloric acid aqueous solution in A3 is a 30-36.5wt% hydrochloric acid aqueous solution; the addition ratio of component 2, distilled water, hydrochloric acid aqueous solution, and 3-chloro-1,2-propanediol is 1g: 100-500mL: 10-12mL: 10-20g.

[0013] A moisturizing and anti-wrinkle polypeptide composition is prepared by any one of the above preparation methods.

[0014] The moisturizing and anti-wrinkle polypeptide composition is used in the field of daily chemical skin care.

[0015] As a further solution of the present invention: the skin care products in the field of daily skin care include creams, lotions, essences, and essence water.

[0016] Beneficial effects of the present invention: (1) The present application firstly uses succinic anhydride to react with cholesterol to obtain component one; and uses the carboxyl group on component one to react with the amino group of N,N'-dimethyl 1,3-propylenediamine to graft a tertiary amine group on cholesterol to obtain component two; finally, 3-chloro-1,2-propanediol is grafted with the tertiary amine group on component two to obtain modified cholesterol. The present application lipidates the polypeptide complex, and uses the modified cholesterol prepared in the present application to synergistically act with soybean lecithin and cholesterol as liposome capsule materials to encapsulate the polypeptide complex to obtain polypeptide liposomes.

[0017] The polypeptide complex of the present application is added to skin care products through liposome preparations, which can effectively improve the existence state of the polypeptide components, improve the stability and cellular absorption of the polypeptide active ingredients, and the effective ingredients in the skin care products will not have quality problems due to excessive temperature during transportation. Liposomes have the characteristics of slow release, and the in vitro release is significantly lower than that of free polypeptide compositions. The liposome preparation of the present application has the effect of improving the water absorption and moisturizing ability of the material, giving the effective ingredients good stability, slow release, and easier absorption by skin cells. The moisturizing and anti-wrinkle polypeptide composition prepared in the present application has the effect of significantly delaying skin aging, easier penetration into the internal cells of the skin, enhancing the skin's immune repair activity, and improving the skin's texture and anti-wrinkle effect.

[0018] (2) The present application adds modified cholesterol to liposomes to modify the structure of liposomes, increase the skin penetration depth and the degree of adhesion to the skin, increase the water solubility and adhesion to the skin, and thus improve the cell absorption rate; it can provide sufficient viscosity for the topically applied liposomes to prevent the liposomes from being quickly cleared from the skin cells, with a high skin retention amount, and prevent the rapid reduction of effective ingredients on the skin surface by maintaining membrane integrity; improve the stability of the polypeptide complex and the concentration of the polypeptide complex at the site of action, and effectively improve the bioavailability of the ingredients. In addition, the unique antibacterial group of modified cholesterol gives the liposomes excellent antibacterial and bactericidal effects.

[0019] (3) This application adds acetyl hexapeptide-8, snake venom peptide, and blue copper peptide as active peptide ingredients. After the combination of several peptides, a synergistic effect is produced, which has the effect of smoothing dynamic lines, static lines and fine lines; its unique liposome structure can promote the penetration of active substances in the skin, so that it can reach the target point and be slowly released under safe conditions to obtain a more lasting effect. The peptide complex works synergistically with white pondweed seed oil and caprylic / capric / succinic acid triglyceride, among which white pondweed seed oil and caprylic / capric / succinic acid triglyceride are used as moisturizers to provide moisturizing effect, improve skin texture, enhance product stability, and make the skin softer and smoother. DETAILED DESCRIPTION

[0020] The following will be described clearly and completely in conjunction with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0021] Example 1 The preparation method of modified cholesterol comprises the following steps: A1: In a nitrogen atmosphere, 10 g cholesterol (produced by J&K Company), 0.1 g succinic anhydride, 10 mL n-heptane, and 20 mL butanone were added to a reaction kettle, the temperature was raised to 40°C, and the reaction was kept warm for 2 h. After removing n-heptane and butanone, the mixture was treated with acetone and filtered under reduced pressure to obtain component 1; A2: Add 10g of component 1, 1000mL of N,N-dimethylformamide, 100mL of N,N'-dimethyl 1,3-propylenediamine, and 1g of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate into a reaction kettle and disperse them evenly. Control the temperature at 50°C and keep the reaction for 6h. Add ethanol to stop the reaction and filter to obtain component 2. A3: 10 g of component 2, 1000 mL of distilled water, 120 mL of 30 wt% hydrochloric acid aqueous solution, and 100 g of 3-chloro-1,2-propanediol were heated to reflux for 6 h, washed with water and ethanol, centrifuged, and dried to obtain modified cholesterol.

[0022] Example 2 The preparation method of modified cholesterol comprises the following steps: A1: In a nitrogen atmosphere, 10 g cholesterol (produced by J&K Company), 0.3 g succinic anhydride, 20 mL n-heptane, and 10 mL butanone were added to a reaction kettle, the temperature was raised to 50°C, and the reaction was kept warm for 3 h. After removing n-heptane and butanone, the mixture was treated with acetone and filtered under reduced pressure to obtain component 1; A2: Add 10g of component 1, 1500mL of N,N-dimethylformamide, 150mL of N,N'-dimethyl 1,3-propylenediamine, and 3g of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate into a reaction kettle and disperse them evenly. Control the temperature at 55°C and keep the reaction for 7h. Add ethanol to stop the reaction and filter to obtain component 2. A3: 10 g of component 2, 3000 mL of distilled water, 120 mL of 30 wt% hydrochloric acid aqueous solution, and 150 g of 3-chloro-1,2-propanediol were heated to reflux for 7 h, washed with water and ethanol, centrifuged, and dried to obtain modified cholesterol.

[0023] Example 3 The preparation method of modified cholesterol comprises the following steps: A1: In a nitrogen atmosphere, 10 g cholesterol (produced by J&K Company), 0.4 g succinic anhydride, 20 mL n-heptane, and 20 mL butanone were added to a reaction kettle, the temperature was raised to 60°C, and the reaction was kept warm for 4 h. After removing n-heptane and butanone, the mixture was treated with acetone and filtered under reduced pressure to obtain component 1; A2: 10 g of component 1, 2000 mL of N,N-dimethylformamide, 200 mL of N,N'-dimethyl 1,3-propylenediamine, and 5 g of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate were added to a reaction kettle and dispersed evenly. The temperature was controlled at 60°C and the reaction was kept warm for 9 hours. Ethanol was added to terminate the reaction and the mixture was filtered to obtain component 2. A3: 10 g of component 2, 5000 mL of distilled water, 120 mL of 30 wt% hydrochloric acid aqueous solution, and 200 g of 3-chloro-1,2-propanediol were heated to reflux for 8 h, washed with water and ethanol, centrifuged, and dried to obtain modified cholesterol.

[0024] Example 4 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 10 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g modified cholesterol prepared in Example 1, 1 g cholesterol (produced by J&K Company), and anhydrous ethanol were mixed to obtain a 12 wt % lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain polypeptide liposomes; S4: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0025] Example 5 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g modified cholesterol prepared in Example 2, 1 g cholesterol (produced by J&K Company), and anhydrous ethanol were mixed to obtain a 12 wt % lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain polypeptide liposomes; S4: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0026] Example 6 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g modified cholesterol prepared in Example 3, 1 g cholesterol (produced by J&K Company), and anhydrous ethanol were mixed to obtain a 12 wt % lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain polypeptide liposomes; S4: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0027] Comparative Example 1 The preparation method of modified cholesterol comprises the following steps: In a nitrogen atmosphere, 10 g of cholesterol (produced by J&K Company), 0.3 g of succinic anhydride, 20 mL of n-heptane, and 10 mL of butanone were added to a reaction kettle, the temperature was raised to 50°C, and the reaction was kept warm for 3 hours. After removing n-heptane and butanone, the product was treated with acetone and filtered under reduced pressure to obtain modified cholesterol.

[0028] Comparative Example 2 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g modified cholesterol prepared in Comparative Example 1, 1 g cholesterol (produced by J&K Company), and anhydrous ethanol were blended to obtain a 12 wt % lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain polypeptide liposomes; S4: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0029] Comparative Example 3 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 2 g cholesterol (produced by J&K Pharmaceutical Co., Ltd.), and 100 mL anhydrous ethanol were mixed to obtain a 12 wt% lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain polypeptide liposomes; S4: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0030] Comparative Example 4 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g cholesterol (produced by J&K Pharmaceutical Co., Ltd.), and 100 mL anhydrous ethanol were mixed to obtain a 12 wt% lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain a polypeptide liposome pre-product; S4: After PEG-2000 and anhydrous ethanol are mixed, the polypeptide liposome pre-product is added, ultrasonically combined for 20 minutes, and filtered with a 0.45um microporous membrane to obtain polypeptide liposomes; the mass ratio of PEG-2000 and cholesterol added during the preparation process is 1:1; S5: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0031] Comparative Example 5 A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: A method for preparing a moisturizing and anti-wrinkle polypeptide composition comprises the following steps: S1: 0.1 g of acetyl hexapeptide-8, 0.1 g of snake venom peptide, and 0.1 g of blue copper peptide were mixed and added into 100 mL of PBS buffer (pH 6.5 Na2HPO4-KH2PO4 buffer) to obtain a core material solution; S2: Add 1 g chitosan, 3 g 2,3-epoxypropyltrimethylammonium chloride and 100 mL isopropanol into a reaction kettle, control the temperature to 80°C, keep the reaction for 9 h, wash with ethanol, wash with acetone, filter and dry in sequence to obtain quaternary ammonium salt chitosan; S3: 10 g soybean lecithin (produced by Shanghai Taiwei Pharmaceutical Co., Ltd.), 1 g cholesterol (produced by J&K Pharmaceutical Co., Ltd.), and 100 mL anhydrous ethanol were mixed to obtain an 11 wt% lipid ethanol solution; S3: 30 mL of the core material solution was mixed with 100 mL of the lipid ethanol solution, and ultrasonic treatment was performed at 50°C for 1 h, and ethanol was removed by vacuum rotary evaporation at 50°C to obtain a polypeptide liposome pre-product; S4: After blending quaternary ammonium salt chitosan and anhydrous ethanol, the polypeptide liposome pre-product is added, ultrasonically combined for 20 minutes, and filtered through a 0.45 um microporous membrane to obtain polypeptide liposomes; during the preparation process, the mass ratio of quaternary ammonium salt chitosan to cholesterol is 1:1; S5: 6 g of meadowfoam seed oil, 90.1 g of caprylic / capric / succinic triglyceride, and 3.9 g of polypeptide liposomes were mixed evenly to obtain a moisturizing and anti-wrinkle polypeptide composition.

[0032] Performance Testing 1. Drug loading and encapsulation efficiency 1. Materials 1. Sample selection The polypeptide liposomes prepared in the process of preparing the moisturizing and anti-wrinkle polypeptide composition in Examples 4-6 and Comparative Examples 2-5; 2. Test methods Transfer 3 mL of polypeptide liposomes to a dialysis bag (Mw=4.5 kD), place in 500 mL of distilled water for 6 h, take 1 mL of the liquid in the dialysis bag, add 3 mL of 3% Triton X-100 to break the emulsion and absorb moisture, and use HPLC to determine the amount of liposome-encapsulated polypeptide complexes. Calculate the encapsulation efficiency and drug loading based on the mass of the detected polypeptide complexes. The calculation method is shown in the following formula: Encapsulation efficiency = (liposome encapsulated polypeptide complex amount / peptide complex dosage) × 100% Drug loading = (liposome-encapsulated peptide complex amount / liposome mass) × 100% (II) Experimental results Table 1: Statistical table of polypeptide liposome test results in Examples 4-6 and Comparative Examples 2-5

[0033] (III) Experimental conclusion Adding modified cholesterol to polypeptide liposomes can effectively increase the amount of polypeptide complex encapsulated by liposomes.

[0034] 2. Stable performance 1. Materials 1. Sample selection The polypeptide liposomes prepared in the process of preparing the moisturizing and anti-wrinkle polypeptide composition in Examples 4-6 and Comparative Examples 2-5; 2. Test methods ① Blend the polypeptide liposomes with deionized water to form an emulsion, place it at -10°C for 24 hours, centrifuge it at 2500rpm for 15 minutes, calculate the mass of the precipitated solid at the bottom as m1, and calculate the preservation rate according to the following formula; ② Place the polypeptide liposomes in a water bath at 30℃, 50℃, and 70℃ for 5h, take out the liposomes and shake them fully with petroleum ether, centrifuge at 2500rpm for 15min, calculate the mass of the precipitated solid at the bottom and record it as m1, and calculate the preservation rate according to the following formula; the test results are shown in Table 2; Preservation rate / %=[(m0-m1) / m0]×100% (II) Experimental results Table 2: Statistical table of the stability test results of polypeptide liposomes in Examples 4-6 and Comparative Examples 2-5

[0035] (III) Experimental conclusion Adding modified cholesterol to polypeptide liposomes can effectively improve the structural stability and temperature resistance of liposomes.

[0036] 3. In vitro drug release experiment 1. Materials 1. Ex vivo skin The rats were killed by cervical dislocation, and the abdominal skin was removed to remove subcutaneous fat, rinsed with saline, and stored in 4°C saline for later use. Before use, they were soaked in pH 7.4 PBS solution for 30 min, rinsed, and the surface moisture was dried with filter paper for later use. 2. Sample Preparation The cream was used as a matrix, and the moisturizing and anti-wrinkle polypeptide compositions prepared in Examples 4-6 and Comparative Examples 2-5 were added thereto, 18 g of white mineral oil, 3 g of emulsifier S-305, 0.3 g of xanthan gum, 5 g of glycerol, 0.2 g of preservative caprylhydroxamic acid, and 0.05 g of the moisturizing and anti-wrinkle polypeptide composition were added thereto, and water was added to 100 g to prepare a moisturizing and anti-wrinkle facial cream.

[0037] 3. Test methods The test was performed using a Franz drug transdermal diffusion tester with a receiving cell volume of 6.5 mL and a transdermal diffusion area of ​​3 cm 2 PBS (pH 7.4) was used as a diffusion graft, and the moisturizing anti-wrinkle cream was evenly applied on the ex vivo skin surface. The test was carried out at 200 rpm and 37 ± 0.5 °C. 0.5 mL of samples were taken at 4 h, 8 h, 12 h, and 24 h (while adding an equal amount of isothermal PBS). The absorbance per unit area (cm 2 ) Transmission Qm, the test results are shown in Table 3: (II) Experimental results Table 3: Statistical table of the results of the polypeptide liposome transdermal test in Examples 4-6 and Comparative Examples 2-5

[0038] (III) Experimental conclusion The polypeptide liposomes prepared in the present application effectively increase the amount of polypeptide components absorbed by skin cells.

[0039] 4. Human skin patch test 1. Materials and methods 1. Test material: Using cream as a base, add the moisturizing and anti-wrinkle polypeptide composition prepared in Example 5, wherein 18g of white mineral oil, 3g of emulsifier S-305, 0.3g of xanthan gum, 5g of glycerol, 0.2g of preservative caprylhydroxamic acid, 0.05g of the moisturizing and anti-wrinkle polypeptide composition are added, and water is added to 100g to prepare a moisturizing and anti-wrinkle facial cream; 2. Negative control: blank control.

[0040] 3. Subjects: A total of 60 subjects, including 21 males and 39 females, aged 24 to 59 years old, with an average age of 48.6±13.5 years old. All subjects met the voluntary inclusion criteria.

[0041] 4. Test method: Select qualified patch test equipment and use the closed patch test method. Place about 0.020g-0.025g of the test substance (white cream) in the patch tester and apply it to the back of the subject with low-allergenic tape. Remove the test substance after 24 hours and observe the skin reaction at 0.5, 24, and 48 hours after removal. Record the results according to the skin reaction grading standards in the "Technical Specification for Safety of Cosmetics" (2015 edition).

[0042] (II) Experimental results Table 4: Statistics of cosmetics human skin patch test results

[0043] Note: The skin reactions of 60 subjects at different observation times are shown in Table 4.

[0044] (III) Experimental conclusion The results of human skin patch tests showed that no adverse skin reactions occurred among 60 people.

[0045] The above is a detailed description of an embodiment of the present invention, but the content is only a preferred embodiment of the present invention and cannot be considered to limit the scope of implementation of the present invention. All equivalent changes and improvements made within the scope of the present invention should still fall within the scope of the patent coverage of the present invention.

Claims

1. A method for preparing a moisturizing and anti-wrinkle polypeptide composition, characterized in that: The steps include: Evenly stirring the meadowfoam seed oil, caprylic / capric / succinic triglyceride and polypeptide liposomes to obtain a moisturizing and anti-wrinkle polypeptide composition; The preparation method of the polypeptide liposome comprises the following steps: S1: mixing the polypeptide complex and PBS buffer to obtain a core material solution; S2: blending soybean lecithin, cholesterol, modified cholesterol and anhydrous ethanol to obtain a lipid ethanol solution; S3: The core material solution and the lipid ethanol solution are mixed, subjected to ultrasonic treatment at 30-50°C for 0.5-1h, and then subjected to vacuum rotary evaporation to obtain polypeptide liposomes.

2. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 1, characterized in that: The moisturizing and anti-wrinkle polypeptide composition comprises the following raw materials in parts by weight: 1-10 parts of meadowfoam seed oil, 84-96 parts of caprylic / capric / succinic triglycerides, and 3-6 parts of polypeptide liposomes.

3. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 1, characterized in that: The polypeptide complex is obtained by mixing acetyl hexapeptide-8, snake venom peptide and blue copper peptide in a mass ratio of 0.05-1:0.05-1:0.05-1; 0.3-0.5 g of the polypeptide complex is added to every 10 mL of PBS buffer.

4. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 1, characterized in that: The mass ratio of soybean lecithin, modified cholesterol and cholesterol in the lipid ethanol solution is 6-10:1:1; the total mass of soybean lecithin, modified cholesterol and cholesterol accounts for 10-15% of the lipid ethanol solution; and the volume ratio of the core material solution to the lipid ethanol solution is 1:1-1.

2.

5. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 1, characterized in that: The preparation method of the modified cholesterol comprises the following steps: A1: Cholesterol, succinic anhydride, n-heptane and butanone were added to a reaction kettle in a nitrogen atmosphere, and the temperature was raised to 40-60°C. The reaction was kept warm for 2-4 hours, and n-heptane and butanone were removed. The mixture was treated with acetone and filtered under reduced pressure to obtain component 1. A2: Add component 1, N,N-dimethylformamide, N,N'-dimethyl 1,3-propylenediamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate into a reaction kettle and disperse evenly. Control the temperature to 50-60°C, keep the temperature for reaction for 6-9h, add ethanol to terminate the reaction, filter, and obtain component 2. A3: Reflux component 2, distilled water, aqueous hydrochloric acid solution, and 3-chloro-1,2-propanediol at elevated temperature for 6-8 hours, wash with water and ethanol, centrifuge, and dry to obtain modified cholesterol.

6. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 5, characterized in that: The addition ratio of cholesterol, succinic anhydride, n-heptane and butanone in A1 is 10 g: 0.1-0.4 g: 10-20 mL: 10-20 mL.

7. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 5, characterized in that: The addition ratio of component 1, N,N-dimethylformamide, N,N'-dimethyl 1,3-propylenediamine, and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate in A2 is 1g:100-200mL:10-20mL:0.1-0.5g.

8. The method for preparing a moisturizing and anti-wrinkle polypeptide composition according to claim 5, characterized in that: The hydrochloric acid aqueous solution in A3 is a 30-36.5wt% hydrochloric acid aqueous solution; the addition ratio of component 2, distilled water, hydrochloric acid aqueous solution, and 3-chloro-1,2-propanediol is 1g: 100-500mL: 10-12mL: 10-20g.

9. A moisturizing and anti-wrinkle polypeptide composition, characterized in that: Prepared by the preparation method described in any one of claims 1 to 8.

10. A moisturizing and anti-wrinkle polypeptide composition as claimed in claim 9 for use in the field of daily chemical skin care.

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