Nanoliposome for repairing skin's natural barrier and its preparation method and application

Abstract

The present application discloses a nanoliposome for repairing the natural barrier of the skin and its preparation method and application, which belongs to the field of liposome preparation technology. The nanoliposome for repairing the natural barrier of the skin provided by the present application comprises, according to the total mass percentage of the raw materials, ceramide 6-15%, madecassoside 3-5%, solvent 5-15%, co-emulsifier 40-60%, emulsifier 3-10%, chitosan 0.1-0.3% and the balance is water to 100%; the average particle size distribution after being placed at a temperature of 18°C to 45°C for 6 months is 200-300nm. Phase A obtained by dissolving ceramide and madecassoside is mixed with phase B obtained by mixing the emulsifier and co-emulsifier, and then subjected to high-pressure homogenization to obtain a liposome emulsion, and chitosan is added to the reaction to obtain the product, thereby giving the nanoliposome excellent product stability, strengthening the skin barrier, enhancing the self-repair ability of the skin, regulating the pathological microenvironment of the skin, and promoting wound healing.

Description

Technical Field

[0001] The present application belongs to the technical field of liposome preparation, and in particular relates to a nanoliposome for repairing the natural barrier of the skin, and a preparation method and application thereof. Background Art

[0002] Liposome technology is a method for encapsulating, preserving, and controlling the release of bioactive ingredients. Liposomes are spherical vesicles composed of a phospholipid bilayer and an aqueous core, ranging in size from nanometers to micrometers. However, because ceramides are oil-soluble, they are difficult to incorporate into solubilization systems. Adding ceramide NPs to traditional liposomes can easily cause crystallization, limiting their application and development in the food, pharmaceutical, and cosmetic sectors.

[0003] The prior art with announcement number CA 117398293 B discloses a method for preparing liposomes encapsulating soothing and repairing ingredients using a microjet homogenizer, wherein ceramide NP, Centella asiatica extract complex, stearyl glycyrrhetinate, phosphatidylcholine, cholesterol, phytosphingosine, and tocopherol are dissolved in an organic solvent and mixed to obtain an oil phase; glycerin, Dendrobium nobile extract, and water are mixed to obtain an aqueous phase; the aqueous phase is poured into the oil phase and mixed to obtain a pre-emulsion, which is then passed through a microjet homogenizer to obtain liposomes encapsulating soothing and repairing ingredients.

[0004] However, the liposomes prepared as described above have the following problems: first, the binding between the components is weak, resulting in poor stability of the liposomes; second, there are many types of components, the effective ingredients are unclear, and there are also problems of strong irritation and poor skin absorption. Summary of the Invention

[0005] The present application discloses a nanoliposome for repairing the natural barrier of the skin, and its preparation method and application, aiming to solve the technical problems of existing liposomes such as poor stability, unclear effective ingredients, strong irritation, and poor skin absorption.

[0006] In order to achieve the above objectives, the technical solution of this application is:

[0007] The first aspect of the present application provides a nanoliposome for repairing the natural barrier of the skin, wherein the components thereof include, by weight percentage of the total raw materials: 6-15% ceramide, 3-5% madecassoside, 5-15% solvent, 40-60% co-emulsifier, 3-10% emulsifier, 0.1-0.3% chitosan, and the balance being water to 100%;

[0008] The average particle size distribution of the nanoliposomes for repairing the natural barrier of the skin is 200-300 nm after being placed at a temperature of -18°C to 45°C for 6 months.

[0009] In combination with the first aspect, preferably, the solvent is one or more of caprylyl glyceryl ether, butylene glycol laureth-23, PEG / PPG-dimethyl ether, trideceth-12, and n-octyl ether.

[0010] In combination with the first aspect, preferably, the co-emulsifier is one or more of glycerol, cholesterol, hexyldecanol, polyethylene glycol-400, 1,3-propylene glycol, polypropylene glycol, 1,2-pentanediol, phytosphingosine, and octyldodecanol.

[0011] In combination with the first aspect, preferably, the emulsifier is one or more of hydrogenated lecithin, soy lecithin, triglyceride, polysorbate, polyglyceryl-10 laurate, PEG-60 hydrogenated castor oil, and caprylic / capric triglyceride.

[0012] The second aspect of the present application provides a method for preparing the nanoliposome for repairing the natural barrier of the skin according to the first aspect, the method comprising:

[0013] Dissolving ceramide and madecassoside in a solvent to obtain phase A;

[0014] Mix the emulsifier and the co-emulsifier to obtain phase B;

[0015] Add phase A to phase B, mix well, and homogenize under high pressure to obtain liposome emulsion;

[0016] Chitosan is added into the liposome emulsion for compounding to obtain the nano liposome for repairing the natural barrier of the skin.

[0017] In combination with the second aspect, preferably, when dissolving ceramide and madecassoside in the solvent, the temperature is 20-30°C.

[0018] In combination with the second aspect, preferably, the conditions of the high-pressure homogenization treatment are: pressure of 20-100 MPa, temperature of 40-80° C., and number of cycles of 1-5 times.

[0019] In combination with the second aspect, preferably, when chitosan is added to the liposome emulsion for reaction, the reaction temperature is 20-50° C. and the reaction time is 10-20 min.

[0020] The third aspect of the present application provides the use of nanoliposomes for repairing the natural barrier of the skin prepared by the preparation method of the second aspect in the preparation of medicines and cosmetics.

[0021] Compared with the prior art, the advantages or beneficial effects of the embodiments of the present application include at least:

[0022] The nanoliposomes for repairing the natural barrier of the skin provided in the present application are nanoliposomes for repairing the natural barrier of the skin with a particle size of 200-300 nm made from raw materials including: ceramide, hydroxy-Centella glycoside, solvent, co-emulsifier, emulsifier, chitosan and water. On the one hand, chitosan is compounded on the surface of the nanoliposomes of hydroxy-Centella glycoside and ceramide by electrostatic attraction and hydrogen bonding, which can give the nanoliposomes excellent product stability, so that the nanoliposomes for repairing the natural barrier of the skin maintain an average particle size of 200-300 nm when placed at -18°C and 45°C for 6 months; on the other hand, it can enhance the interaction with the skin, open the tight junctions between cells, improve the mucosal permeability, effectively strengthen the skin barrier function, and enhance the self-repair ability of the skin; thirdly, through the synergistic effect between ceramide, hydroxy-Centella glycoside and chitosan, it can effectively regulate the pathological microenvironment of the skin and prevent the occurrence of skin inflammation or skin sensitivity symptoms. At the same time, it can promote wound healing, is mild and non-irritating, has streamlined ingredients, reduces production costs, and has broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] To more clearly illustrate the technical solutions of the embodiments of this application, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some of the embodiments described in this application. For those skilled in the art, other drawings can be derived from these drawings without inventive effort.

[0024] Figure 1 A photo of the nanoliposomes A1 prepared in Example 1 of the present application for repairing the skin's natural barrier;

[0025] Figure 2 This is a graph showing the effects of the A1-A3 nanoliposomes and B1-B3 liposomes prepared in the examples of this application on the proliferation of HaCaT cells for repairing the skin's natural barrier;

[0026] Figure 3 This is a microscopic result showing the effects of A1-A3 nanoliposomes and B1-B3 liposomes prepared in the examples of this application on the proliferation of HaCaT cells;

[0027] Figure 4 This is a graph showing the effects of the A1-A3 nanoliposomes and B1-B3 liposomes prepared in the examples of this application on the moisturizing factor of HaCaT cells for repairing the skin's natural barrier;

[0028] Figure 5 This is a graph showing the cumulative skin penetration rate test results of the A1-A3 nanoliposomes and B1-B3 liposomes prepared in the examples of this application on the back skin of rats;

[0029] Figure 6 This is a graph showing the results of a VISIA red yeast rice chart test for evaluating the redness-removing efficacy of the A1-A3 nanoliposomes and B1-B3 liposomes for repairing the skin's natural barrier, prepared in the examples of this application. DETAILED DESCRIPTION

[0030] The following will be combined with the accompanying drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the described embodiments are part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0031] In the following description of this embodiment, the term "and / or" is used to describe the association relationship between associated objects, indicating that three possible relationships exist. For example, A and / or B can represent: A exists alone, B exists alone, and both A and B exist. A and B can be singular or plural. The character " / " generally indicates that the associated objects are in an "or" relationship.

[0032] In the following description of this embodiment, the term "at least one" refers to one or more, and "plurality" refers to two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, "at least one of a, b, or c", or "at least one of a, b, and c" can all represent: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, and c can be single or multiple.

[0033] Those skilled in the art should understand that in the following description of the embodiments of the present application, the order of serial numbers does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0034] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments and are not intended to limit the present application. The singular forms "a", "an" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms unless the context clearly indicates otherwise.

[0035] Unless otherwise specified, all raw materials and reagents used in the examples of this application were purchased from the market or prepared according to conventional methods well known to those skilled in the art.

[0036] In a first aspect, the present invention provides a nanoliposome for repairing the skin's natural barrier, wherein the components thereof include, by weight percentage of the total raw materials: 6-15% ceramide, 3-5% madecassoside, 5-15% solvent, 40-60% co-emulsifier, 3-10% emulsifier, 0.1-0.3% chitosan, and the balance being water to 100%;

[0037] The average particle size distribution of the nanoliposomes for repairing the natural barrier of the skin is 200-300 nm after being placed at a temperature of -18°C to 45°C for 6 months.

[0038] Among them, on the one hand, chitosan is compounded on the surface of nanoliposomes of hydroxy-madecassoside and ceramide by electrostatic attraction and hydrogen bonding, which can give the nanoliposomes excellent product stability, so that the particle size of the nanoliposomes that repair the skin's natural barrier remains at 200-300nm when placed at -18°C and 45°C for 6 months; on the other hand, it can enhance the interaction with the skin, open the tight junctions between cells, improve the mucosal permeability, effectively strengthen the skin barrier function, and enhance the skin's self-repair ability; thirdly, through the synergistic effect between ceramide, hydroxy-madecassoside and chitosan, it can effectively regulate the pathological microenvironment of the skin and prevent the occurrence of skin inflammation or skin sensitivity symptoms. At the same time, it can promote wound healing, is mild and non-irritating, has streamlined ingredients, reduces production costs, and has broad market application prospects.

[0039] In the embodiments of the present application, the solvent is preferably one or more of caprylyl glyceryl ether, butylene glycol laureth-23, PEG / PPG-dimethyl ether, trideceth-12, and n-octyl ether. The selection of these solvents allows ceramide and madecassoside to be completely dissolved in the solvents, forming a uniform and stable oil phase solution.

[0040] In the embodiments of the present application, the co-emulsifier is preferably one or more of glycerol, cholesterol, hexyldecanol, polyethylene glycol-400, 1,3-propanediol, polypropylene glycol, 1,2-pentanediol, phytosphingosine, and octyldodecanol. The selection of these co-emulsifiers can form an interfacial barrier on the droplet surface, slowing the migration of monomers from small droplets to larger droplets, thereby forming smaller droplets and improving the stability of the emulsion system.

[0041] In the examples of the present application, the emulsifier is preferably one or more of hydrogenated lecithin, soy lecithin, triglycerides, polysorbate, polyglyceryl-10 laurate, PEG-60 hydrogenated castor oil, and caprylic / capric triglyceride. The selection of these emulsifiers allows for effective mixing of the oil phase and the aqueous phase, allowing the components to form a stable and uniform emulsion, thereby maintaining product stability. Furthermore, these emulsifiers are mild, non-irritating, environmentally friendly, and suitable for large-scale applications.

[0042] The second aspect of the present application provides a method for preparing the nanoliposome for repairing the natural barrier of the skin according to the first aspect, the method comprising:

[0043] Dissolving ceramide and madecassoside in a solvent to obtain phase A;

[0044] Mix the emulsifier and the co-emulsifier to obtain phase B;

[0045] Add phase A to phase B, mix well, and homogenize under high pressure to obtain liposome emulsion;

[0046] Chitosan is added into the liposome emulsion for compounding to obtain the nano liposome for repairing the natural barrier of the skin.

[0047] It should be noted that the present application adopts an oil phase formed by ceramide and madecassoside and an aqueous phase formed by an emulsifier and a co-emulsifier, and the process of mixing the oil phase and the aqueous phase for pre-emulsification can increase the stability of the effective ingredients of the liposome and the soothing effect. After high-pressure homogenization treatment, the oil phase and the aqueous phase are mixed more fully through high-speed impact, shearing and cavitation, which can effectively encapsulate the active substance, so that ceramide and madecassoside are stably present in the lipid system, giving the liposome excellent comprehensive performance. At the same time, the particle size distribution of the prepared nanoliposomes can be made more uniform.

[0048] In the embodiments of the present application, when dissolving ceramide and madecassoside in a solvent, the temperature is preferably 20-30° C., for example, 20° C., 30° C., or any temperature within this range. By controlling the reaction temperature, a uniform and stable oil phase emulsion can be formed.

[0049] In the embodiments of the present application, the conditions for the high-pressure homogenization treatment are: the pressure is preferably 20-100 MPa, for example, 20 MPa, 50 MPa, 80 MPa, 90 MPa, 100 MPa or any pressure within this range; the temperature is preferably 40-80°C, for example, 40°C, 50°C, 60°C, 70°C, 80°C or any temperature within this range; the number of cycles is preferably 1-5 times, for example, 1 time, 3 times, 5 times or any number within this range. Among them, by controlling the pressure, temperature and time of the high-pressure homogenization treatment, the raw materials can be sheared, impacted and homogenized under high pressure, and the liposome components can be evenly dispersed. At the same time, the particle size distribution of the liposomes can be adjusted, thereby improving the stability of the liposomes and the product performance.

[0050] In the embodiment of the present application, when chitosan is added to the liposome emulsion for reaction, the reaction temperature is preferably 20-50°C, for example, 20°C, 30°C, 40°C, 50°C or any temperature within this range; the reaction time is preferably 10-20 min, for example, 10 min, 15 min, 20 min or any time within this range. Among them, by controlling the time and temperature of the reaction between chitosan and the liposome emulsion, chitosan can be uniformly and stably distributed on the surface of the nanoliposomes, giving the liposomes excellent product stability, and through the synergistic effect between ceramide, madecassoside and chitosan, it can effectively regulate the pathological microenvironment of the skin and prevent the occurrence of skin inflammation or skin sensitivity symptoms.

[0051] It should be noted that after the lipids in the skin's stratum corneum are damaged, the skin becomes chapped. Nanoliposomes that repair the skin's natural barrier play a leading role in restoring skin moisture. They can effectively associate water molecules and form a network structure to maintain skin moisture. At the same time, they can connect the lipid matrix and keratinocytes, enhance the skin's barrier function, and reduce harmful substances from entering the skin through pores and sweat glands, thereby having an anti-allergic effect.

[0052] The third aspect of this application provides the use of nanoliposomes for repairing the skin's natural barrier, prepared by the preparation method described in the second aspect, in the preparation of pharmaceuticals and cosmetics. The nanoliposomes for repairing the skin's natural barrier have excellent product stability, strengthen the skin barrier, enhance the skin's self-repair ability, regulate the skin's pathological microenvironment, and promote wound healing, thereby giving the product excellent product competitiveness.

[0053] The technical solution of the present application will be further described below in conjunction with specific embodiments.

[0054] Example 1

[0055] This embodiment provides a method for preparing A1-nanoliposomes for repairing the skin's natural barrier, which specifically includes:

[0056] S101: 8% ceramide, 4% madecassoside, 8% octylglyceryl ether, and 8% PEG / PPG-dimethyl ether were mixed and dispersed by ultrasonication at 30°C for 40 min to obtain phase A.

[0057] S102: 30% glycerol, 1% cholesterol, 2% 1,3-propylene glycol, 2% hexyldecanol, 5% hydrogenated lecithin, and 5% soy lecithin were heated at 60°C with magnetic stirring at 400 rpm for 40 min to obtain phase B;

[0058] S103: adding phase A to phase B, mixing, homogenizing, and shearing at a shear speed of 10,000 rpm, a shear temperature of 60°C, and a shear time of 5 min to obtain liposome colostrum, and subjecting the mixture to high-pressure homogenization at a pressure of 100 MPa and a temperature of 60°C for three cycles, and cooling to room temperature to obtain a liposome emulsion;

[0059] S104: Add the chitosan solution to the liposome emulsion, stir magnetically at 300 rpm at 45° C. for 20 min, and obtain A1-nanoliposomes that repair the skin's natural barrier.

[0060] according to Figure 1 As known, the prepared A1-nanoliposomes for repairing the skin's natural barrier are evenly distributed and have good dispersibility.

[0061] Example 2

[0062] This embodiment provides a method for preparing A2-nanoliposomes for repairing the skin's natural barrier, which specifically includes:

[0063] S201: 8% ceramide, 4% madecassoside, 5% butylene glycol lauryl polyether, 5% n-octyl ether, and 5% PEG / PGG-dimethyl ether were dispersed by ultrasonication at 20°C for 60 min to obtain phase A.

[0064] S202: 20% glycerol, 1% cholesterol, 3% polyethylene glycol-400, 3% 1,2-pentanediol, 10% hydrogenated lecithin, 5% PEG-60 hydrogenated castor oil, and 3% triglyceride were heated at 50°C with magnetic stirring at 400 rpm for 40 min to obtain phase B;

[0065] S203: adding phase A to phase B, mixing, homogenizing, and shearing at a shear speed of 8000 rpm, a shear temperature of 80°C, and a shear time of 10 min to obtain liposome colostrum, and subjecting the mixture to high-pressure homogenization at a pressure of 80 MPa and a temperature of 40°C for four cycles, and cooling to room temperature to obtain a liposome emulsion;

[0066] S204: adding the chitosan solution to the liposome emulsion, stirring with a magnetic force at 400 rpm for 10 min at 30° C. to obtain A2-nanoliposomes for repairing the skin's natural barrier.

[0067] Example 3

[0068] This embodiment provides a method for preparing A3-nanoliposomes for repairing the skin's natural barrier, which specifically includes:

[0069] S301: 8% ceramide, 4% madecassoside, 10% trideceth-12, and 5% n-octyl ether were dispersed by ultrasonication at 25°C for 60 minutes to obtain phase A.

[0070] S302: 25% glycerol, 1% cholesterol, 3% polypropylene glycol, 2% octyldodecanol, 5% caprylic / capric triglyceride, 3% polysorbate, and 3% polyglyceryl-10 laurate were heated at 50°C with magnetic stirring at 400 rpm for 40 min to obtain phase B;

[0071] S303: adding phase A to phase B, mixing, homogenizing, and shearing at a shear speed of 8000 rpm, a shear temperature of 80°C, and a shear time of 10 min to obtain liposome colostrum, and subjecting the mixture to high-pressure homogenization at a pressure of 90 MPa and a temperature of 50°C for 4 cycles, and cooling to room temperature to obtain a liposome emulsion;

[0072] S304: Add the chitosan solution to the liposome emulsion, and stir magnetically at 350 rpm for 15 minutes at 40° C. to obtain A3-nanoliposomes that repair the skin's natural barrier.

[0073] At the same time, in order to verify the comprehensive performance of the nanoliposomes for repairing the natural barrier of the skin prepared in the above examples, the present application provides the following comparative examples for detailed description.

[0074] Comparative Example 1

[0075] The component ratio, preparation operation and process parameters of the B1-liposome prepared in this comparative example are basically the same as those in Example 1, except that madecassoside is not added in this comparative example to prepare the B1-liposome.

[0076] Comparative Example 2

[0077] The component ratio, preparation operation and process parameters of the B2-liposome prepared in this comparative example are basically the same as those in Example 1, except that ceramide is not added in this comparative example to prepare the B2-liposome.

[0078] Comparative Example 3

[0079] The component ratio, preparation operation and process parameters of the B3-liposome emulsion prepared in this comparative example are basically the same as those in Example 1, except that chitosan is not added in this comparative example to prepare the B3-liposome emulsion.

[0080] In order to verify the comprehensive performance of the liposomes prepared in the examples of the present application, corresponding performance tests were performed. The liposomes prepared in the above examples 1-3 and comparative examples 1-3 were subjected to performance tests:

[0081] Test 1: Effect on HaCaT cell proliferation

[0082] Weigh 1 g of each of the liposomes prepared in Examples 1-3 and Comparative Examples 1-3, add them to 100 mL of DMEM medium and fully dissolve them; collect HaCaT cells in the logarithmic growth phase, inoculate them in 96-well plates, and adjust the HaCaT cell density to 1 × 104 cells / well; after adaptive culture for 1 day, remove the old culture medium, and add 100 μL of DMEM medium containing Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3, respectively. Use DMEM medium without sample as the control group, with 3 replicates per group; continue to culture for 48 hours, add 10 μL of CCK-8 solution to each well, incubate for 2 hours, measure the absorbance (A) of each well at 450 nm, and calculate the cell proliferation rate. The results are shown in Table 1. Figure 2 and Figure 3 .

[0083] Table 1 Effects on HaCaT cell proliferation

[0084]

[0085] The cell proliferation rate can reflect the repair effect of the sample to a certain extent. Cell proliferation is one of the important characteristics of cell life activities and is the basis of biological reproduction. The proliferation of single-celled organisms leads to an increase in the number of individual organisms. Multicellular organisms develop from a single cell, namely a fertilized egg, and cell proliferation is the basis of multicellular organism reproduction. Adult organisms still need cell proliferation, mainly to replace aging and dead cells, maintain the relative balance of individual cell numbers and the normal function of the body, and the body's wound healing, tissue regeneration, and pathological tissue repair all rely on cell proliferation. According to Table 1, Figure 2 and Figure 3 The test results show that all sample group experiments have a proliferation effect on HaCaT cells, and the proliferation rate of A1, A2 and A3 nanoliposomes that repair the skin's natural barrier is significantly better than that of B1, B2 and B3 liposomes.

[0086] Test 2: Effect on the moisturizing factor of HaCaT cells

[0087] Weigh 1 g of each liposome from Example 1-3 and Comparative Example 1-3, add them to 100 mL of DMEM medium and fully dissolve them; HaCaT cells are plated at 2 × 10 5 The cells were inoculated into a 24-well cell culture plate at a density of 500 μL per well and cultured for 24 h. The experiment was divided into a blank control group, Example 1, Example 2, Comparative Example 1, Comparative Example 2, and Comparative Example 3. The cells were incubated in an incubator for 24 h, and the supernatant was taken and the secretion of AQP-3 (aquaporin), FLG (filament aggregate protein), and Claudin-1 in HaCaT cells was tested using an Elisa kit. The results are shown in Table 2 and Figure 4 .

[0088] Table 2 Effects on HaCaT cell moisturizing factor

[0089]

[0090]

[0091] HaCaT cells can synthesize biological molecules related to skin moisturizing, such as hyaluronic acid (HA), aquaporin (AQP), tight junction protein (TJP), and filaggrin (FLG). Therefore, the moisturizing ability of liposomes can be reflected and their efficacy can be evaluated by detecting the effect of liposomes on the content of moisturizing-related proteins such as AQP, claudin-1 (claudin-1 is the main transmembrane protein that forms the epidermal cell barrier in TJP) and FLG in HaCaT cells. Figure 4 The test results show that the moisturizing abilities of A1, A2 and A3 are significantly better than those of B1, B2 and B3, and the moisturizing effect of A1 is the most significant.

[0092] Test 3: Human patch test

[0093] Weigh 3 g of the liposomes prepared in Examples 1-3 and Comparative Examples 1-3 and mix thoroughly with 97 g of a blank cream to obtain a ceramide cream containing madecassoside. Thirty subjects were selected for this experiment, and their reactions were recorded. The blank cream, the comparative example cream, and the example cream were applied to the curved side of the subject's forearm. Gently press the cream evenly onto the skin with the palm of your hand for 24 hours. After removing the spot tester, wait 30 minutes until the indentation disappears and observe the skin reaction. Skin reactions were observed again 48 hours after removing the spot tester. The results are shown in Table 3.

[0094] The classification of adverse skin reactions is determined according to the classification standards for adverse skin reactions in the "Safety Evaluation of Human Trial Tests" stipulated in the 2015 edition of the "Technical Specifications for Safety of Cosmetics" issued by the Ministry of Health:

[0095] Grade 0 (no skin reaction);

[0096] Grade 1 (light red spots on the skin);

[0097] Grade 2 (erythema, infiltration, and papules on the skin);

[0098] Grade 3 (erythema, edema, papules and blisters on the skin);

[0099] Grade 4 (erythema, edema and bullae on the skin).

[0100] Table 3 Human patch test results

[0101]

[0102] The principle of the patch test is to replicate the reaction process of allergic contact dermatitis on localized skin. This involves applying a small amount of allergen directly to the skin and observing whether a mild dermatitis is induced locally, thereby determining whether the skin is allergic to the tested allergen. The above results indicate that none of the samples in the seven experimental groups, A1 / A2 / A3 / B1 / B2 / B3, showed any adverse skin reactions, indicating that all seven experimental groups were non-irritating.

[0103] Test 4: Stability test

[0104] The nanoliposomes prepared in Examples 1-3 and Comparative Examples 1-3 for repairing the skin's natural barrier, along with pure water (control group), were placed in a sealed container and stored at -18°C, 4°C, room temperature, and 45°C for 180 days. The samples were then inspected for agglomeration or stratification at the initial stage, after 90 days, and after returning to room temperature after 180 days. The results were expressed as "yes / no" and the average particle size of the samples was measured. The results are shown in Table 4.

[0105] Table 4 Stability test results

[0106]

[0107]

[0108] The ability of a cosmetic to maintain its initial properties or properties unchanged over time under specified or reasonably foreseeable storage and use conditions. The control group in the above results is usually a pure aqueous solution, which has a particle size much smaller than that of the sample solution and has no practical reference value. The results show that the nanoliposomes A1, A2, and A3 that repair the skin's natural barrier, and the liposomes B1, B2, and B3, are all stable after dispersion in water, with no agglomeration or stratification. Furthermore, the nanoliposomes A1, A2, and A3 that repair the skin's natural barrier have smaller fluctuations than the B1, B2, and B3 liposomes, making the system more stable.

[0109] Test 5: Cumulative transdermal dose test

[0110] Take SD rats weighing 200-250g, shave the hair on their backs, kill them after 24 hours and cut the skin on their backs. After removing the subcutaneous fat, clean them with physiological saline, absorb the surface moisture and place them in the refrigerator for freezing. During the test, take out the rat skin from the refrigerator, place it in physiological saline to thaw at room temperature, and then clamp the rat skin between the receiving pool and the diffusion pool with the stratum corneum facing up. Measure 1mL of each sample prepared in Example 1 and Comparative Examples 1-3 and apply it evenly on the rat skin. Seal the top of the diffusion pool with plastic wrap and place it in a magnetic stirrer with a constant temperature water bath at 37°C and stirring at 200r / min. Take 0.5mL of samples after 2h, 4h, 8h, 12h and 24h. After each sampling, add 10% Triton solution to make the volume 1mL and filter it with a 0.45μm microporous filter membrane; at the same time, add 0.5mL of physiological saline to the receiving pool. The concentration of the active substance was tested by HPLC, and the cumulative transmittance was calculated. The results are shown in Table 6 and Figure 5 .

[0111] Table 6 Cumulative transdermal test results

[0112]

[0113] The above experimental results show that the permeability of each group of samples gradually increased within 24 hours, and the permeability of A1, A2 and A3 nanoliposomes that repair the skin's natural barrier is significantly better than that of B1, B2 and B3 liposomes.

[0114] Test 6: VISIA Red Yeast Chart Test

[0115] Weigh 3g of liposomes prepared in Examples 1-3 and Comparative Examples 1-3, mix them with 97g of blank cream to obtain a ceramide cream containing madecassoside. Seven subjects were selected in the experiment, and VISIA photos of the subjects' faces before and after use were recorded. Seven groups of samples of blank cream, comparative example component cream and example component cream corresponded to seven subjects respectively. The subjects signed an informed consent form before enrollment. Before each test, they used facial cleanser to clean the test area and sat quietly in a constant temperature and humidity environment (temperature 21.0℃±1.0℃; humidity: 50%±10%) for at least 20 minutes. Data collection of the subjects was arranged before using the samples and 28 days after use. Skin parameters and images were collected using VISIA each time. The results are shown in Tables 7 and Figure 6 .

[0116] Table 7 Hemoglobin test results

[0117]

[0118] According to the test results, hemoglobin can reflect the dilation of capillaries in the superficial dermis. Abnormally increased hemoglobin and capillary dilation can be used to measure the severity of skin inflammation and skin sensitivity. That is, reduced hemoglobin can support the soothing effect to a certain extent. According to Table 7 and Figure 6 The test results showed that A1, A2 and A3 nanoliposomes that repair the skin's natural barrier and B1, B2 and B3 liposomes all had a significant effect on reducing hemoglobin, and the total hemoglobin area reduction rate of A1, A2 and A3 nanoliposomes that repair the skin's natural barrier was better than that of B1, B2 and B3 liposomes.

[0119] The various embodiments in this specification are described in a progressive manner. The same or similar parts between the various embodiments can be referenced to each other, and each embodiment focuses on the differences from other embodiments.

[0120] The above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit the present application. Although the present application has been described in detail with reference to the aforementioned embodiments, a person skilled in the art should understand that the technical solutions described in the aforementioned embodiments can still be modified, or some or all of the technical features therein can be replaced by equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the present application.

Claims

1. A nanoliposome for repairing the skin's natural barrier, characterized in that: The composition thereof is calculated as follows according to the total mass percentage of the raw materials: ceramide 8%, madecassoside 4%, butylene glycol lauryl polyether 5%, n-octyl ether 5%, PEG / PGG-dimethyl ether 5%, glycerol 20%, cholesterol 1%, polyethylene glycol-400 3%, 1,2-pentanediol 3%, hydrogenated lecithin 10%, PEG-60 hydrogenated castor oil 5%, triglyceride 3%, chitosan 0.1-0.3%, and the balance is water to make up to 100%. The average particle size distribution of the nanoliposomes for repairing the natural barrier of the skin is 200-300 nm after being placed at a temperature of -18°C to 45°C for 6 months.

2. A method for preparing the nanoliposome for repairing the natural barrier of the skin according to claim 1, characterized in that: The method comprises: Dissolving ceramide and madecassoside in a solvent to obtain phase A; Mix the emulsifier and the co-emulsifier to obtain phase B; Add phase A to phase B, mix well, and homogenize under high pressure to obtain liposome emulsion; Chitosan is added into the liposome emulsion for compounding to obtain the nano liposome for repairing the natural barrier of the skin.

3. The method for preparing nanoliposomes for repairing the natural barrier of the skin according to claim 2, characterized in that: When dissolving ceramide and madecassoside in the solvent, the temperature is 20-30°C.

4. The method for preparing nanoliposomes for repairing the natural barrier of the skin according to claim 2, characterized in that: The conditions of the high-pressure homogenization treatment are: pressure of 20-100 MPa, temperature of 40-80° C., and number of cycles of 1-5 times.

5. The method for preparing nanoliposomes for repairing the natural barrier of the skin according to claim 2, characterized in that: When chitosan is added to the liposome emulsion for reaction, the reaction temperature is 20-50° C. and the reaction time is 10-20 min.

6. Use of nanoliposomes for repairing the natural barrier of the skin prepared by the preparation method according to any one of claims 2 to 5 in the preparation of medicines and cosmetics.

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