Preparation method and application of super-hydrophilic high-transdermal ceramide liposome

Abstract

The invention provides a super-hydrophilic high-transdermal ceramide liposome and a preparation method thereof. The invention further provides a preparation method of the super-hydrophilic high-transdermal ceramide liposome. The invention further provides an application of the super-hydrophilic high-transdermal ceramide liposome in cosmetics. The invention also provides a cosmetic. According to the preparation method and application of the super-hydrophilic high-transdermal ceramide lipidosome, the super-hydrophilic high-transdermal ceramide lipidosome is obtained, industrial mass production is easy, use of a surfactant is avoided, and the super-hydrophilic high-transdermal ceramide lipidosome has the advantages of being good in water solubility, high in transparency, good in stability, high in transdermal efficiency, remarkable in effect and the like.

Description

Technical Field

[0001] The present invention belongs to the technical field of cosmetics, and relates to a preparation method and application of a super-hydrophilic and highly transdermal ceramide liposome, specifically to a super-hydrophilic and highly transdermal ceramide liposome, its preparation method, and its application in cosmetics. Background Art

[0002] Ceramide is a class of complex lipids formed by connecting sphingosine fatty acids through amide bonds. The ceramide family has a very wide application in cosmetics and has recognized effects such as strengthening and repairing the skin barrier, moisturizing and locking water, anti-aging, and improving skin texture. Although ceramide has the above functions, due to its oil-soluble nature, it is difficult to directly add it to solubilizing formulations and is extremely prone to crystallization and precipitation. Developing stability technology for ceramide is the key to solving its application pain points.

[0003] US20240225987A9 provides a liquid crystal technology for stabilizing ceramide, including an aqueous phase and an oil phase. Among them, the oil phase components are: skin lipids, emulsifiers, and oils, and the skin lipids include ceramide, cholesterol or its derivatives, and fatty acids. This technology still maintains liquid crystal state stability at high temperature for four weeks. CN114848533A (a preparation method for efficiently encapsulating composite ceramide by nano-liposome) efficiently encapsulates multiple composite ceramides through liposome solid particle technology. It is mentioned that this technology, as a delivery technology for lipophilic molecules, mainly utilizes the characteristics of solid oils cooling and solidifying to form a solid structure, restricting the fluidity and diffusivity of the embedded active ingredients, thereby improving the chemical stability of the active ingredients.

[0004] Although the above technologies can improve the stability of ceramide in formulations, they are not suitable for most water-based transparent formulations on the market. The liposome technology can well solve the stability problem of ceramide in transparent aqueous agents. The liposome technology in cosmetics is an advanced carrier system that uses tiny spherical vesicles (liposomes) to encapsulate and deliver active ingredients. Liposomes are composed of a phospholipid bilayer membrane. On the one hand, its structure is highly similar to the cell membrane, and this biocompatibility enables it to better fuse with skin tissues, promoting the more precise delivery of active ingredients in cosmetics to the deep layer of the skin and improving skin care efficiency; on the other hand, the bilayer membrane encapsulation can also reduce the irritation of active ingredients to the skin. Since the active ingredients encapsulated by liposomes are gradually released, it avoids the allergy or discomfort that may be caused by direct contact of high-concentration ingredients with the skin, and is especially suitable for people with sensitive skin. In addition, liposomes can effectively protect active ingredients from damage by external environments such as light, heat, and oxygen, significantly improving the stability of the ingredients.

[0005] Traditional liposomes are mostly composed of natural lecithin from soybeans or egg yolks as the basic framework. The carbon chain length of their fatty acids is usually 16 - 20 carbon atoms. The ceramide liposomes formed by such natural lecithin face the challenges of unstable self - structure and inability to be stored for a long time. Research has found that when using fatty acids with longer carbon chains to form liposomes, the stability of the liposome membrane can be enhanced, and drug leakage can be reduced; in addition, long - chain fatty acids and ceramide can jointly construct a highly ordered lipid structure, which is very beneficial for the repair of the skin barrier. The uniformly and tightly arranged lipid layer helps to enhance the skin barrier function, promote its repair process, and improve the overall health status of the skin. Such a lipid combination not only supports the skin in retaining moisture but also effectively resists the invasion of the external environment. However, the presence of long carbon chains will lead to a decrease in the hydrophilicity of liposomes and a poor compatibility with aqueous formulations. Therefore, obtaining a liposome model with both stability and hydrophilicity is the key to solving the application difficulties of ceramide - type liposomes. Summary of the Invention

[0006] In view of the above - mentioned disadvantages of the prior art, the purpose of the present invention is to provide a preparation method and application of super - hydrophilic and highly transdermal ceramide liposomes, which are used to solve the problems that the prior art cannot simultaneously improve the water - solubility and stability of liposomes; the surface modification technology of liposomes with macromolecules such as chitosan can improve the aqueous solution, but is not conducive to stability; optimizing the stability of liposomes with a coagulation thickening system is not conducive to the skin feel of raw materials, application range, and active substance release efficiency; the operation steps of using a combination of lecithin and hydrogenated phosphatidylcholine as the liposome shell structure are cumbersome and the production efficiency is low.

[0007] To achieve the above - mentioned purpose and other related purposes, in the first aspect, the present invention provides a super - hydrophilic and highly transdermal ceramide liposome, which includes a ternary biomimetic phospholipid polymer, ceramide, hydrogenated lecithin, cholesterol, co - solvent, and water.

[0008] In the second aspect, the present invention provides a preparation method of a super - hydrophilic and highly transdermal ceramide liposome, including: mixing and reacting the components in the super - hydrophilic and highly transdermal ceramide liposome provided in the first aspect of the present invention according to the ratio to provide the required super - hydrophilic and highly transdermal ceramide liposome.

[0009] In the third aspect, the present invention provides a super - hydrophilic and highly transdermal ceramide liposome, which is obtained by the method provided in the second aspect of the present invention.

[0010] In the fourth aspect, the present invention provides the use of the super - hydrophilic and highly transdermal ceramide liposome provided in the first aspect of the present invention or the super - hydrophilic and highly transdermal ceramide liposome provided in the third aspect of the present invention in cosmetics.

[0011] The fifth aspect of the present invention provides a cosmetic, which includes the superhydrophilic and highly transdermal ceramide liposome provided by the first aspect of the present invention or the superhydrophilic and highly transdermal ceramide liposome provided by the third aspect of the present invention.

[0012] As described above, the present invention provides a preparation method and application of a superhydrophilic and highly transdermal ceramide liposome. By providing a novel phospholipid-based polymer material that can adjust the ratio and structure of the hydrophobic and hydrophilic ends and participating in the encapsulation of ceramide under specific process parameters, a superhydrophilic and highly stable ceramide liposome is obtained. This ceramide liposome is easy to be mass-produced industrially, avoids the use of surfactants, and the obtained liposome has the advantages of good water solubility, high transparency, good stability, high transdermal efficiency, and remarkable efficacy. In practical applications, it is found that adding 0.1% - 10% of the liposome to the aqueous formulation can achieve complete dissolution and has superhydrophilicity; adding 1% of the liposome has a clinical repair effect and high transdermal permeability. Description of the Drawings

[0013] Figure 1 Shown is the product appearance diagram of liposome samples 1# - 3# prepared in Examples 1 - 3 of the present invention.

[0014] Figure 2 Shown is the scanning electron microscope image of the freeze-dried powder in Example 1 of the present invention.

[0015] Figure 3 Shown are the microscopic structure diagrams of the optical microscopes of liposome samples 1# - 3# prepared in Examples 1 - 3 of the present invention. Among them, a is the microscopic structure diagram of the optical microscope of liposome sample 1#, b is the microscopic structure diagram of the optical microscope of liposome sample 2#, and c is the microscopic structure diagram of the optical microscope of liposome sample 3#.

[0016] Figure 4 Shown is the particle size diagram of the liposome in Example 1 of the present invention.

[0017] Figure 5 Shown is the comparison diagram of the chick embryo chorioallantoic membrane test in Example 1 of the present invention. Among them, 0S is the test diagram at 0 seconds of the test, and 300S is the test diagram at 300 seconds of the test.

[0018] Figure 6 Shown is the column comparison diagram of the changes in the content and promotion rate of filaggrin (FLG) in Example 1 of the present invention.

[0019] Figure 7 Shown is the column comparison diagram of the changes in the skin water content of different samples in Example 1 of the present invention.

[0020] Figure 8Shown is a comparison graph of the appearances of liposome samples prepared in Example 1, Example 4, and Example 5 in Test Example 10 of the present invention and commercially available ceramide NP liposomes.

[0021] Figure 9 Shown is a graph of the dispersion of liposome samples prepared in Example 1, Example 4, and Example 5 in Test Example 10 of the present invention and commercially available ceramide NP liposomes in an aqueous solution. Detailed implementation manners

[0022] The inventors of the present application have developed a super-hydrophilic and highly transdermal ceramide liposome. A preparation method of the super-hydrophilic and highly transdermal ceramide liposome is also provided. It is proposed that by using a ternary biomimetic phospholipid polymer with a penetration-enhancing effect shown in the following formula (I), and by adjusting the amounts of x, y, z, m and the type of R group, different specific ternary polymers can be obtained, including but not limited to ternary polymer A, ternary polymer B, ternary polymer C, etc. Under specific process conditions, they participate in the traditional liposome encapsulation process, thereby enhancing the structural rigidity and water solubility of the liposome, making the prepared ceramide liposome have the advantages of good water solubility, high transparency in the formulation, good stability, high transdermal efficiency, and remarkable efficacy, and can be applied to cosmetics. Thus, the present invention is completed, and the specific elaboration is as follows.

[0023] The first aspect of the present invention provides a super-hydrophilic and highly transdermal ceramide liposome, which includes a ternary biomimetic phospholipid polymer, ceramide, hydrogenated lecithin, cholesterol, a co-solvent, and water.

[0024] In a specific implementation manner, the super-hydrophilic and highly transdermal ceramide liposome includes the following components by weight percentage:

[0025] Ternary biomimetic phospholipid polymer 0.01 - 5%, specifically such as 0.01 - 0.05%, 0.05 - 0.1%, 0.1 - 2%, 2 - 5%;

[0026] Ceramide 0.01 - 6%, specifically such as 0.01 - 0.1%, 0.1 - 5%, 5 - 6%;

[0027] Hydrogenated lecithin 1 - 10%; specifically such as 1 - 2%, 2 - 7%, 7 - 10%;

[0028] Cholesterol 0.01 - 5%, specifically such as 0.01 - 0.1%, 0.1 - 2%, 2 - 3%, 3 - 5%;

[0029] Co-solvent 1 - 50%; specifically such as 1 - 10%, 10 - 40%, 40 - 50%;

[0030] The balance is water.

[0031] In a preferred embodiment, the superhydrophilic high transdermal ceramide liposome, by weight percentage, comprises the following components:

[0032] Ternary biomimetic phospholipid polymer 0.1 - 2%; specifically such as 0.1 - 0.5%, 0.5 - 1%, 1 - 2%;

[0033] Ceramide 0.1 - 5%; specifically such as 0.1 - 0.5%, 0.5 - 1%, 1 - 3%, 3 - 5%;

[0034] Hydrogenated lecithin 2 - 7%; specifically such as 2 - 4%, 4 - 6%, 6 - 7%;

[0035] Cholesterol 0.1 - 2%; specifically such as 0.1 - 0.5%, 0.5 - 1%, 1 - 2%;

[0036] Cosolvent 10 - 40%; specifically such as 10 - 20%, 20 - 30%, 30 - 40%;

[0037] The balance is water.

[0038] In a specific embodiment, the structure of the ternary biomimetic phospholipid polymer is as shown in formula (I):

[0039]

[0040] In formula (1), x > 0; z > 0; y ≥ 0; 0 ≤ m ≤ 25; R is selected from H, -OH, -COOH, -NH2, -O - Na + , -OCH2CH2OH or epoxy group.

[0041] In a preferred embodiment, the ternary biomimetic phospholipid polymer is selected from at least one of ternary polymer A, ternary polymer B, and ternary polymer C; the structures of the ternary polymer A, ternary polymer B, and ternary polymer C are as shown in formula (I), wherein, in the ternary polymer A, x = 10, y = z = 5, m = 3, R is -OCH2CH2OH, in the ternary polymer B, x = 5, y = 6, z = 11, m = 11, R is -OH, and in the ternary polymer C, x = 13, y = z = 6, m = 3, R is -O - Na + .

[0042] The above ternary polymers A, B, and C are all produced by Ori. For example, ternary polymer A obtained a new INCI name: BUTYL METHACRYLATE / HEMA / METHACRYLOXYETHYL PHOSPHORYLCHOLINE COPOLYMER through PCPC in 2023 by Ori, and obtained a new Chinese INCI name for the new raw material: Phosphorylcholine Polymer - MBH through the record-filing of the drug regulatory agency. The INCI name of the terpolymer C in cosmetics is: POLYQUATERNIUM-65, and the standard Chinese name is: Polyquaternium-65.

[0043] The above ternary biomimetic phospholipid polymer is a biomimetic material polymerized from bioactive 2-methacryloyloxyethyl phosphorylcholine and other monomers containing acrylic double bonds such as fatty acid methacrylate. It is a type of amphiphilic polymer material similar to the phospholipid structure in cell membranes, with adjustable hydrophobic and hydrophilic ends. It usually contains amphoteric end groups and alkane non-polar molecular chains. The amphoteric end groups are hydrophilic, and the alkane molecular chains are hydrophobic. Due to the controllability of the length of the non-polar molecular chain (i.e., carbon chain) of this biomimetic phospholipid polymer, and the controllability of the active groups such as carboxyl and hydroxyl groups contained in the amphoteric end groups, it can be regarded as an ideal material for preparing liposomes. By controlling the length of the hydrophobic end chain, when the biomimetic phospholipid polymer participates in liposome encapsulation, it can intersperse in the oil phase of traditional liposomes and jointly build the liposome bilayer structure, effectively making up for the defect that liposomes are easily ruptured. The raw material itself has better stability than natural phospholipids and is not easily oxidized. At the same time, the hydrophilic groups (including but not limited to hydroxyl (-OH), carboxyl (-COOH), amino (-NH2) or epoxy groups, etc.) that can be selectively incorporated into the hydrophilic end of the biomimetic phospholipid polymer make the outer layer of the liposome bilayer membrane more likely to combine with water, enhancing the performance of liposomes in aqueous formulations and improving the utilization efficiency of ceramides.

[0044] In a specific embodiment, the ceramide (general CAS: 100403-19-8) is selected from at least one of ceramide EOP (INCI: CERAMIDE EOP), ceramide NG / ceramide NS (INCI: CERAMIDE NG / CERAMIDE NS), ceramide NP (INCI: CERAMIDE NP), ceramide AS (INCI: CERAMIDE AS), ceramide AP (INCI: CERAMIDE AP), or ceramide AH (INCI: CERAMIDE AH).

[0045] In a specific embodiment, the CAS number of the hydrogenated lecithin is 92128-87-5.

[0046] In a specific embodiment, the CAS number of cholesterol is 57-88-5.

[0047] In a specific embodiment, the co-solvent is a common organic solvent in cosmetics. Specifically, the co-solvent is selected from at least one of 1,3-butanediol, 1,2-propanediol, glycerol, pentanediol, hexanediol, or ethoxydiglycol.

[0048] In a specific embodiment, the mass ratio of the ternary biomimetic phospholipid polymer to hydrogenated lecithin is 4:4-60, specifically such as 4:4-20, 4:20-22, 4:22-60, preferably 4:20-22, and more preferably 4:21.

[0049] The second aspect of the present invention provides a method for preparing a super-hydrophilic and highly transdermal ceramide liposome, including: mixing the components in the super-hydrophilic and highly transdermal ceramide liposome provided in the first aspect of the present invention according to the ratio to provide the required super-hydrophilic and highly transdermal ceramide liposome.

[0050] In a specific embodiment, the mixing reaction includes the following steps:

[0051] 1) Mix and grind the ternary biomimetic phospholipid polymer and hydrogenated lecithin to obtain a mixture.

[0052] 2) Divide the co-solvent into a first part of the co-solvent and a second part of the co-solvent. After heating the first part of the co-solvent in the oil phase, add the mixture and cholesterol and perform the first stirring and mixing to obtain a primary liposome, and then add ceramide and perform the second stirring and mixing to obtain a first mixed solution.

[0053] 3) Add the second part of the co-solvent and water, heat in the water phase, and then perform the third stirring and mixing to obtain a second mixed solution.

[0054] 4) Perform the fourth stirring and mixing of the second mixed solution and the first mixed solution, and then perform a homogenization treatment to obtain the required super-hydrophilic and highly transdermal ceramide liposome.

[0055] In step 1), the grinding is carried out in a mortar. Sufficient grinding causes intermolecular interactions to occur.

[0056] In step 1), the grinding time is 10-15 min. Specifically, when grinding, when the volume of the grinding tool such as a mortar remains unchanged and is calculated based on accommodating 100 g of the mixture, the grinding time is 10-15 min.

[0057] In step 2), the first part of the co-solvent is selected from at least one of 1,3-butanediol, 1,2-propanediol, glycerol, pentanediol, hexanediol, or ethoxydiglycol, and is preferably ethoxydiglycol.

[0058] In step 2) or 3), the second part of the cosolvent is selected from at least one of 1,3 - butanediol, 1,2 - propanediol, glycerol, pentanediol, hexanediol or ethoxydiglycol, and is preferably glycerol.

[0059] In step 2), the mass ratio of the first part of the cosolvent to the second part of the cosolvent added is 2 - 5:20 - 35, specifically such as 2 - 5:20 - 25, 2 - 5:25 - 33, 2 - 5:33 - 35, preferably 2 - 5:25 - 33, and more preferably 3 - 4.5:25 - 33.

[0060] In step 2), the heating temperature of the oil phase is 75 - 85 °C, specifically such as 75 - 78 °C, 78 - 82 °C, 82 - 85 °C, and is preferably 80 °C.

[0061] In step 2), the first stirring and mixing and the second stirring and mixing are carried out in an emulsifying tank.

[0062] In step 2), the stirring speed of the first stirring and mixing is 30 - 40 r / min, specifically such as 30 - 33 r / min, 33 - 37 r / min, 37 - 40 r / min, and is preferably 35 r / min.

[0063] In step 2), the time of the first stirring and mixing is 5 - 15 min, specifically such as 5 - 6 min, 6 - 14 min, 14 - 15 min, and is preferably 6 - 14 min.

[0064] In step 2), the stirring speed of the second stirring and mixing is 15 - 25 r / min, specifically such as 15 - 18 r / min, 18 - 22 r / min, 22 - 25 r / min, and is preferably 20 r / min.

[0065] In step 2), the time of the second stirring and mixing is 2 - 10 min, specifically such as 2 - 3 min, 3 - 9 min, 9 - 10 min, and is preferably 3 - 9 min. The ceramide is completely dissolved.

[0066] In step 3), the heating temperature of the aqueous phase is 75 - 85 °C, specifically such as 75 - 78 °C, 78 - 82 °C, 82 - 85 °C, and is preferably 80 °C.

[0067] In step 3), the third stirring and mixing is carried out in an aqueous phase tank.

[0068] In step 3), the stirring speed of the third stirring and mixing is 450 - 550 r / min, specifically such as 450 - 480 r / min, 480 - 520 r / min, 520 - 550 r / min, and is preferably 500 r / min.

[0069] In step 3), the time of the third stirring and mixing is 8 - 22 min, specifically such as 8 - 10 min, 10 - 20 min, 20 - 22 min, and preferably 10 - 20 min. This enables the raw materials of the third stirring and mixing to be fully mixed.

[0070] In step 4), the fourth stirring and mixing is carried out in an emulsifying tank.

[0071] In step 4), the time of the fourth stirring and mixing is 25 - 65 min, specifically such as 25 - 30 min, 30 - 60 min, 60 - 65 min, and preferably 30 - 60 min.

[0072] In step 4), the fourth stirring and mixing is carried out after cooling, and the temperature is cooled to 15 - 25 °C, specifically such as 15 - 18 °C, 18 - 22 °C, 22 - 25 °C, and preferably 20 °C.

[0073] In a preferred embodiment, the cooling rate is 3 - 5 °C / min, and preferably 4 °C / min.

[0074] In step 4), the stirring speed of the fourth stirring and mixing is 55 - 65 r / min, specifically such as 55 - 58 r / min, 58 - 62 r / min, 62 - 65 r / min, and preferably 60 r / min.

[0075] In step 4), the homogenization treatment includes a three - stage pressure - boosting procedure carried out in sequence; the homogenization pressure for the first - stage pressure - boosting is 5500 - 6500 psi, specifically such as 5500 - 5800 psi, 5800 - 6200 psi, 6200 - 6500 psi, and preferably 6000 psi; the number of cycles for the first - stage pressure - boosting is 2 - 3 times, and preferably 2 times; the homogenization pressure for the second - stage pressure - boosting is 14500 - 15500 psi, specifically such as 14500 - 14800 psi, 14800 - 15200 psi, 15200 - 15500 psi, and preferably 15000 psi; the number of cycles for the second - stage pressure - boosting is 3 - 4 times, and preferably 3 times; the homogenization pressure for the third - stage pressure - boosting is 19500 - 20500 psi, specifically such as 19500 - 19800 psi, 19800 - 20200 psi, 20200 - 20500 psi, and preferably 20000 psi; the number of cycles for the third - stage pressure - boosting is 3 - 4 times, and preferably 3 times.

[0076] The third aspect of the present invention provides a super - hydrophilic and highly transdermal ceramide liposome, which is obtained by the method provided in the second aspect of the present invention.

[0077] The fourth aspect of the present invention provides the use of the superhydrophilic and highly transdermal ceramide liposome provided in the first aspect of the present invention or the superhydrophilic and highly transdermal ceramide liposome provided in the third aspect of the present invention in cosmetics.

[0078] The fifth aspect of the present invention provides a cosmetic, comprising the superhydrophilic and highly transdermal ceramide liposome provided in the first aspect of the present invention or the superhydrophilic and highly transdermal ceramide liposome provided in the third aspect of the present invention.

[0079] In a specific embodiment, the addition amount of the superhydrophilic and highly transdermal ceramide liposome in the cosmetic is 0.1-10 wt%.

[0080] In a specific embodiment, the cosmetic can be formulated into various product forms, including but not limited to basic cosmetics, facial makeup cosmetics, body skin care products, body washing products, etc. By formulating the cosmetic into various products, the corresponding effects of the active substances contained in the cosmetic can be better exerted, such as moisturizing, anti-aging, repair, soothing, whitening, anti-dandruff, oil control, antibacterial, fragrance retention, etc.

[0081] In a specific embodiment, there is no special limitation on the dosage form of the cosmetic, and it can be reasonably selected according to different purposes.

[0082] In a specific embodiment, the cosmetic further comprises one or more conventional other active ingredients. The content of the other active ingredients is a conventional content in the art.

[0083] When the embodiments give numerical ranges, it should be understood that unless otherwise specified in the present invention, both endpoints of each numerical range and any value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as commonly understood by those skilled in the art of this technology. Except for the specific methods, equipment, and materials used in the examples, according to the knowledge of those skilled in the art of this technology and the description of the present invention, any methods, equipment, and materials similar or equivalent to the methods, equipment, and materials described in the embodiments of the present invention can also be used to implement the present invention.

[0084] Unless otherwise specified, the experimental methods, detection methods, and preparation methods disclosed in the present invention all adopt the conventional techniques in the fields of molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields in this technical field.

[0085] As used herein, the terms "individual" or "patient" refer to humans, wild animals, and domestic animals suffering from diseases, preferably mammals, especially humans.

[0086] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the protection scope of the present invention.

[0087] The following specific examples illustrate the implementation manners of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention from the content disclosed in this specification. The present invention can also be implemented or applied through other different specific implementation manners. Various details in this specification can also be modified or changed based on different viewpoints and applications without departing from the spirit of the present invention.

[0088] Example 1

[0089] 4.2 parts of hydrogenated lecithin and 0.8 part of terpolymer A were placed in a mortar and ground for 12 min. After sufficient grinding, intermolecular interactions occurred to obtain a mixture. The temperature of the emulsifying tank was set at 80 °C and the rotation speed was 35 r / min. 3 parts of ethoxydiglycol were added to the emulsifying tank to reach the reaction temperature. Then the mixture and 0.45 part of cholesterol were added for the first stirring and mixing reaction for 10 min. After the primary liposomes were formed, the rotation speed was set at 20 r / min, and 1 part of ceramide NP was added for the second stirring and mixing reaction for 6 min until the ceramide was completely dissolved to obtain the first mixed solution.

[0090] The temperature of the aqueous phase tank was set at 80 °C and the rotation speed was 500 r / min. 25 parts of glycerol and 65.55 parts of sterilized water were successively added to the aqueous phase tank, and the third stirring and mixing reaction was carried out for 15 min until the above raw materials were fully mixed to obtain the second mixed solution.

[0091] The rotation speed of the emulsifying tank was set at 60 r / min. The second mixed solution in the aqueous phase tank was slowly pumped into the emulsifying tank, and immediately cooled at a cooling rate of 4 °C / min to 20 °C. The second mixed solution and the first mixed solution were subjected to the fourth stirring and mixing reaction for 45 min, and then high-pressure homogenization treatment was carried out. The homogenization treatment included a three-stage pressure increase procedure carried out in sequence. The homogenization pressure for the first-stage pressure increase was 6000 psi; the number of cycles for the first-stage pressure increase was 2 times; the homogenization pressure for the second-stage pressure increase was 15000 psi; the number of cycles for the second-stage pressure increase was 3 times; the homogenization pressure for the third-stage pressure increase was 20000 psi; the number of cycles for the third-stage pressure increase was 3 times. The product was discharged to obtain liposome sample 1#. The specific components and their contents added in liposome sample 1# are shown in Table 1 below.

[0092] Example 2

[0093] The specific preparation steps and their conditions are the same as those in Example 1, and the specific contents of the selected components of the liposome sample are the same as those in Example 1. The difference lies in the specific types of the selected components of the liposome sample, specifically the types of ceramides are different, and liposome sample 2# is obtained. The specific components added and their contents in liposome sample 2# are shown in Table 1 below.

[0094] Example 3

[0095] The specific preparation steps and their conditions are the same as those in Example 1, and the specific types of the selected components of the liposome sample are the same as those in Example 1. The difference lies in the specific contents of the selected components of the liposome sample, and liposome sample 3# is obtained. The specific components added and their contents in liposome sample 3# are shown in Table 1 below.

[0096] Example 4

[0097] The specific preparation steps and their conditions are the same as those in Example 1, and the specific contents of the selected components of the liposome sample are the same as those in Example 1. The difference lies in the specific types of the selected components of the liposome sample, specifically the types of ternary biomimetic phospholipid polymers are different, and liposome sample 4# is obtained. The specific components added and their contents in liposome sample 4# are shown in Table 1 below.

[0098] Example 5

[0099] The specific preparation steps and their conditions are the same as those in Example 1, and the specific contents of the selected components of the liposome sample are the same as those in Example 1. The difference lies in the specific types of the selected components of the liposome sample, specifically the types of ternary biomimetic phospholipid polymers are different, and liposome sample 5# is obtained. The specific components added and their contents in liposome sample 5# are shown in Table 1 below.

[0100] Comparative Example 1

[0101] The specific preparation steps and their conditions are the same as those in Example 1. The difference between the liposome sample and that in Example 1 is that no ternary biomimetic phospholipid polymer is added, and liposome comparative sample 1* is obtained. The specific components added and their contents in liposome comparative sample 1* are shown in Table 1 below.

[0102] Comparative Example 2

[0103] The specific components added and their contents are the same as those of the liposome sample in Example 1. The difference between the liposome sample and that in Example 1 is that the homogenization treatment of the three-stage pressure boost procedure is not carried out in the preparation method, and only the homogenization treatment is carried out 3 times at 20000 ps, and liposome comparative sample 2* is obtained. The specific components added and their contents in liposome comparative sample 2* are shown in Table 1 below.

[0104] Comparative Example 3

[0105] The specific preparation steps and their conditions are the same as those in Example 1. The difference between the liposome sample of this example and that of Example 1 is that chitosan is used instead of the ternary biomimetic phospholipid polymer to obtain the liposome comparative sample 3*. The specific components and their contents added in the liposome comparative sample 3* are shown in Table 1 below.

[0106] Comparative Example 4

[0107] The specific components and their contents added are the same as those in the liposome sample of Example 1. The difference between the liposome sample of this example and that of Example 1 is that the cooling rate before the fourth stirring and mixing in the preparation method is 1°C / min to obtain the liposome comparative sample 4*. The specific components and their contents added in the liposome comparative sample 4* are shown in Table 1 below.

[0108] Table 1

[0109]

[0110] Examples 6 - 7

[0111] The specific components and their contents added are the same as those in the liposome sample of Example 1. The difference between the liposome samples of this example and that of Example 1 is the different preparation conditions in the preparation method to obtain the liposome samples 6# and 7#. The different preparation conditions from Example 1 are shown in Table 2 below.

[0112] Table 2

[0113]

[0114]

[0115] Test Example 1

[0116] The liposome samples 1# - 3# prepared in Examples 1 - 3 were observed for their appearance. The specific details are shown in Figure 1 . From Figure 1 it can be seen that the product appearances of the liposome samples 1# - 3# are characteristic - colored transparent aqueous solutions. The colors of the ceramide liposomes with different types and contents are slightly different, probably due to the refractive index differences of different ceramides. There are no solid particles precipitated in the product, indicating that the ceramide in the system has been completely encapsulated, realizing the possibility of applying ceramide in a transparent aqueous agent; the product appearance is a characteristic - colored transparent liquid state, indicating that the liposome particles are evenly dispersed in the aqueous agent system, the hydrophilic property of the particle surface is extremely good, and the particles have reached the nanometer level, which is more conducive to transdermal absorption.

[0117] Test Example 2

[0118] The liposome sample 1# prepared in Example 1 was freeze - dried. The microscopic image of its freeze - dried powder under a scanning electron microscope is shown in Figure 2 . From Figure 2It can be seen that the liposomes are in complete form without rupture, presenting regular spherical shapes, with uniform sizes, and the particle sizes are in the range of about 100 - 200 nm, and the particle size dispersity is good.

[0119] The dilutions of liposome samples 1# - 3# prepared in Examples 1 - 3 were observed under a microscope, and the microscopic images thereof under the microscope are shown in Figure 3 . From Figure 3 it can be seen that different ceramide liposomes of liposome samples 1# - 3# are all distributed in a particulate form with consistent structures and uniform sizes. This morphology is conducive to increasing the stability of the liposomes. In addition, no insoluble substances were seen in the visual images, indicating that the ceramide has been completely encapsulated and completely dissolved in the water-based medium.

[0120] Test Example 3

[0121] The liposome sample 1# prepared in Example 1 was measured by dynamic light scattering (DLS) technology, and the particle size distribution of liposome sample 1# is shown in Figure 4 . Among them, the horizontal axis represents the particle diameter (in nanometers), and the vertical axis represents the proportion or intensity of particles of each size.

[0122] From Figure 4 it can be seen that the curve of liposome sample 1# presents a relatively sharp and symmetric peak, centered at about 100 nanometers, which is consistent with the liposome particle size under the scanning electron microscope. This indicates that most liposome particles are concentrated near this target size. The width of the peak is narrow, meaning that the liposome preparation under this condition has good monodispersity, that is, a low coefficient of variation of particle size and relatively consistent particle sizes. In addition, there are almost no small peaks or tails significantly deviating from the main peak in the figure, further confirming that liposome sample 1# has ideal uniformity and a small size distribution range. Such a particle size distribution is crucial for ensuring the effectiveness and safety of liposomes as drug carriers.

[0123] Test Example 4

[0124] The appearance stability of liposome samples 1# - 3# prepared in Examples 1 - 3 and liposome comparative samples 1* - 4* prepared in Comparative Examples 1 - 4 was observed and tested, and the specific results are shown in Table 3.

[0125] As can be seen from Table 3, under appropriate material ratios and operating processes, the liposome samples 1#-3# prepared by the present invention have excellent storage stability for different ceramide liposomes, while the liposome control samples 1*-4* have defects. Specifically, the liposome control sample 1* showed a pink semi-transparent state immediately at day 0, indicating that the liposome particles in this system could not be well dissolved in the external aqueous phase, confirming that liposomes without ternary biomimetic phospholipid polymers have poor solubility, and demulsification and stratification occurred after 60 days at room temperature, demonstrating the promoting effect of the ternary biomimetic phospholipid polymer on the structural stability of liposomes. The liposome control sample 2* had precipitates during storage at room temperature and 5°C, indicating that liposomes without the three-stage stepwise pressure homogenization process are more likely to aggregate and rupture. This is because of the poor particle size uniformity, resulting in easier mutual attraction and sedimentation between particles, forming large particles until demulsification. The liposome control sample 3* showed flocculent precipitation after 90 days at room temperature, indicating that under the same conditions, although chitosan coating can provide better water solubility for liposomes, since it does not participate in the formation of the phospholipid bilayer, it cannot help improve the stability of liposomes. The liposome control sample 4* reduced the cooling rate at the end of the emulsification stage, resulting in demulsification of liposomes during storage, probably because the structure of lecithin was damaged by long-term high temperature.

[0126] Table 3

[0127]

[0128] Examples 8-10

[0129] The specific preparation steps and their conditions are the same as those in Example 1. The difference from the liposome samples in Example 1 lies in the different addition amounts of the ternary biomimetic phospholipid polymer, and liposome samples 8#-10# are obtained. The specific components and their contents added in liposome samples 8#-10# are shown in Table 4 below.

[0130] Table 4

[0131] Example 1 Comparative Example 1 Example 8 Example 9 Example 10 Ceramide NP 1 1 1 1 1 Ternary Polymer A 0.8 0 0.3 0.5 1.5 Hydrogenated Lecithin 4.2 5 4.7 4.5 3.5 Cholesterol 0.45 0.45 0.45 0.45 0.45 Ethoxydiglycol 3 3 3 3 3 Glycerol 25 25 25 25 25 Water 65.55 65.55 65.55 65.55 65.55

[0132] Test Example 5

[0133] The liposome sample 1# prepared in Example 1, the liposome control sample 1* prepared in Comparative Example 1, and the liposome samples 8#-10# prepared in Examples 8-10 were respectively tested by the fluorescence labeling method. The specific components and their contents added in the corresponding samples are shown in Table 4 above. The test results of the ceramide encapsulation efficiency of the samples obtained by the fluorescence labeling method are shown in Table 5 below.

[0134] The test conditions of the fluorescence labeling method are as follows:

[0135] Principle: Label ceramide with a fluorescent dye, and determine the encapsulation rate through fluorescence intensity.

[0136] Steps: Combine the fluorescent dye (NBD-Ceramide) with ceramide. After preparing liposomes, measure the total fluorescence intensity using a fluorescence spectrophotometer. Add a quenching agent (Triton X-100) to disrupt the liposomes and measure the fluorescence intensity of free ceramide.

[0137] The calculation formula for the encapsulation efficiency of ceramide is as follows: Encapsulation efficiency = (Total fluorescence intensity - Free fluorescence intensity) / Total fluorescence intensity × 100%

[0138] Table 5

[0139] Entrapment Efficiency / % Example 1 99.6 Comparative Example 1 85.2 Example 8 92.8 Example 9 96.5 Example 10 98.2

[0140] As can be seen from Table 5, by measuring the encapsulation efficiency of ceramide at different addition amounts of the ternary biomimetic phospholipid polymer, analyze the effects of the ternary biomimetic phospholipid polymer on the encapsulation efficiency and encapsulation stability. At the optimal addition amount (Example 1), the measured encapsulation efficiency of ceramide is 99.6%. At this time, the addition amount of the ternary biomimetic phospholipid polymer is 0.8%, and the mass ratio to hydrogenated lecithin is 4:21; as the addition amount of the ternary biomimetic phospholipid polymer decreases and the addition amount of hydrogenated lecithin increases, the encapsulation efficiency of ceramide gradually decreases. When the addition amount of the ternary biomimetic phospholipid polymer is 0 (Comparative Example 1), the encapsulation efficiency of ceramide is only 85.2%, and the effect is poor; while when the addition amount of the ternary biomimetic phospholipid polymer increases to 1.5% (Example 10), the encapsulation efficiency of ceramide is still lower than the optimal condition, indicating that only when the ternary biomimetic phospholipid polymer and hydrogenated lecithin are in an appropriate ratio, there is the optimal encapsulation efficiency for ceramide, proving that the bilayer membrane structure is the densest at this time, the leakage amount of ceramide is the lowest, and the liposome stability is the best.

[0141] Test Example 6

[0142] Perform the chicken embryo chorioallantoic membrane test on the liposome sample 1# prepared in Example 1 according to the "Chicken Embryo Chorioallantoic Membrane Test for Eye Irritation and Corrosion of Cosmetics - SN / T 2329-2009". The results of the chicken embryo experiment are shown in Figure 5 . From Figure 5 it can be seen that the liposome sample 1# prepared in Example 1 is safe and non-irritating.

[0143] Test Example 7

[0144] The liposome sample 1# prepared in Example 1 is added at a content of 0.1% - 0.8% in cosmetics for in vitro repair testing. The test method is to use a UVB-irradiated HaCaT cell model to cause cell damage, and then detect the expression of FLG to evaluate and verify the repair effect of the sample on the skin barrier. The specific test results are shown in Figure 6 .

[0145] From Figure 6It is known that filaggrin (FLG) is an important molecule that connects keratin fibers in the stratum corneum. Increasing the FLG content plays an important role in maintaining the integrity of the skin barrier function. The test results of the FLG content show that adding liposome sample 1# with a content of 0.1%-0.8% can significantly increase the filaggrin content. The promotion rate of liposome sample 1# with a content of 0.8% for FLG is 37.89%, indicating that samples with low addition content have strong repair effects and contribute to improving the skin barrier function.

[0146] Test Example 8

[0147] Subjects aged 20-45 years were selected, with 15 males and 15 females. In the same emulsion matrix formula, liposome sample 1# prepared in Example 1 was added to the aqueous phase, with addition amounts of 1% and 3%; ceramide NP powder was added to the oil phase, with addition amounts of 0.01% and 0.03%. To make the total amount of active substances the same, after the subjects used the samples for 0, 1, 2, and 4 hours, the skin water content was measured with a CM825 probe, and the data was compared with that before use. The specific results are shown in Figure 7 .

[0148] It can be Figure 7 seen that clinical trials show that using liposome sample 1# prepared in Example 1, that is, a product with ceramide NP encapsulated in liposomes, can significantly improve the skin water content in the short term and make the skin softer and smoother. In contrast, adding ceramide NP powder to the oil phase, that is, using unencapsulated ceramide, although it also has a certain moisturizing effect, its effect is usually not as significant as that of the liposome-encapsulated form. This shows that liposome encapsulation is beneficial to the delivery of ceramide, improves the utilization rate of ceramide, and enables it to exert better effects.

[0149] Test Example 9

[0150] To explore the solubility of ceramide liposomes with different ternary biomimetic phospholipid polymers under various pH conditions, aqueous solutions with a mass fraction of 50% containing liposome samples prepared in Example 1, Example 4, Example 5, and Comparative Example 1 were respectively prepared, and the pH of the solution was adjusted to 3.0, 7.0, and 11.0 with citric acid and sodium citrate, and the precipitation of ceramide was observed. The results are shown in Table 6.

[0151] Table 6

[0152]

[0153] As can be seen from Table 6, the surface charge of liposomes is usually determined by the dissociation state of hydrophilic groups. When the environmental pH value changes, this may affect the ionization state of the head groups, thereby affecting the solubility of liposomes. For example, at a lower pH value, some groups may be less ionized, reducing the electrostatic repulsion, which may lead to the coalescence or precipitation of liposomes, thus reducing the solubility of liposomes. According to the test results in Table 6, the liposomes without the ternary biomimetic phospholipid polymer precipitated at pH values of 3.0 and 11.0; the liposomes added with ternary biomimetic phospholipid polymers such as terpolymer A and terpolymer C did not precipitate under various pH conditions; the liposomes added with ternary biomimetic phospholipid polymers such as terpolymer B precipitated at a pH value of 11.0. From the above results, it can be seen that the ternary biomimetic phospholipid polymer can improve the solubility of liposomes at different pH values by adjusting the types and proportions of hydrophilic groups, but the effects of different ternary biomimetic phospholipid polymers on solubility are slightly different, which may be due to the different effects of different hydrophilic groups on the surface charge density of liposomes.

[0154] Test Example 10

[0155] Select the liposome samples prepared in Example 1, Example 4, and Example 5 and compare them with the commercially available ceramide NP liposomes. Observe the appearance as shown in Figure 8 and observe the dispersion in the aqueous solution as shown in Figure 9 .

[0156] From Figure 8 it can be seen that the liposome samples prepared in Example 1, Example 4, and Example 5 are transparent liquids in appearance, while the commercially available same products are semi-transparent liquids. From Figure 9 it can be seen that after dispersing the liposome samples prepared in Example 1, Example 4, and Example 5 in the aqueous phase, they present a blue-light-emitting clear and transparent liquid, while after dispersing the commercially available products with the same ceramide content in the aqueous phase, they present a milky and turbid liquid. The above results show that through the highly hydrophilic modification of the surface of the ceramide liposomes in the present invention, it helps ceramide to form nano-scale dispersion in the aqueous phase (blue light is the characteristic manifestation of nano-scale particles dispersed in water), greatly improving the poor solubility of commercially available traditional ceramide liposomes in water, which is beneficial to the application and promotion of ceramide.

[0157] As described above, it is only the preferred embodiment of the present invention and does not impose any formal or substantial limitations on the present invention. It should be noted that for those of ordinary skill in the art of this technology, without departing from the method of the present invention, several improvements and supplements can still be made, and these improvements and supplements should also be regarded as the protection scope of the present invention. Those skilled in the art, without departing from the spirit and scope of the present invention, when making some equivalent changes such as minor modifications, decorations and evolutions using the technical content disclosed above, are all equivalent embodiments of the present invention; at the same time, any equivalent changes, modifications and evolutions made to the above embodiments based on the essential technology of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A ceramide liposome, comprising a ternary biomimetic phospholipid polymer, ceramide, hydrogenated lecithin, cholesterol, a co-solvent, and water.

2. The ceramide liposome according to claim 1, wherein The ceramide liposome, by weight percentage, comprises the following components: Ternary biomimetic phospholipid polymer 0.01 - 5%; Ceramide 0.01 - 6%; Hydrogenated lecithin 1 - 10%; Cholesterol 0.01 - 5%; Co-solvent 1 - 50%; The balance is water.

3. The ceramide liposome according to any one of claims 1-2, characterized in that, Comprising any one or more of the following conditions: A1) The structure of the ternary biomimetic phospholipid polymer is shown in formula (I): In formula (1), x > 0; z > 0; y ≥ 0; 0 ≤ m ≤ 25; R is selected from -H, -OH, -COOH, -NH2, -O - Na + , -OCH2CH2OH or epoxy group; Preferably, the ternary bionic phospholipid polymer is selected from at least one of ternary polymer A, ternary polymer B, and ternary polymer C; the structures of the ternary polymer A, ternary polymer B, and ternary polymer C are shown in formula (I), wherein in the ternary polymer A, x = 10, y = z = 5, m = 3, and R is -OCH2CH2OH; in the ternary polymer B, x = 5, y = 6, z = 11, m = 11, and R is -OH; in the ternary polymer C, x = 13, y = z = 6, m = 3, and R is -O - Na + ; A2) The ceramide is selected from at least one of ceramide EOP, ceramide NG / ceramide NS, ceramide NP, ceramide AS, ceramide AP, or ceramide AH; A3) The co-solvent is selected from at least one of 1,3 - butanediol, 1,2 - propanediol, glycerol, pentanediol, hexanediol, or ethoxydiglycol; A4) The mass ratio of the ternary biomimetic phospholipid polymer to hydrogenated lecithin is 4:4 - 60.

4. A method for preparing a ceramide liposome, comprising: Mix the components in the ceramide liposome according to any one of claims 1 - 3 in proportion to provide the required ceramide liposome.

5. The preparation method of the ceramide liposome according to claim 4, characterized in that, The mixing reaction comprises the following steps: 1) Mix and grind the ternary biomimetic phospholipid polymer and hydrogenated lecithin to obtain a mixture; 2) Divide the co-solvent into a first part of the co-solvent and a second part of the co-solvent. After heating the first part of the co-solvent in the oil phase, add the mixture and cholesterol and perform the first stirring and mixing to obtain primary liposomes, and then add ceramide and perform the second stirring and mixing to obtain a first mixed solution; 3) Add the second part of the co-solvent and water, heat in the water phase, and then perform the third stirring and mixing to obtain a second mixed solution; 4) Perform the fourth stirring and mixing of the second mixed solution and the first mixed solution, and then perform a homogenization treatment to obtain the required ceramide liposome.

6. The preparation method of the ceramide liposome according to claim 5, wherein Comprising any one or more of the following conditions: B1) In step 1), the grinding time is 10 - 15 min; B2) In step 2), the first part of the co-solvent is selected from at least one of 1,3 - butanediol, 1,2 - propanediol, glycerol, pentanediol, hexanediol, or ethoxydiglycol; preferably, the first part of the co-solvent is ethoxydiglycol; B3) In step 2) or 3), the second part of the co-solvent is selected from at least one of 1,3 - butanediol, 1,2 - propanediol, glycerol, pentanediol, hexanediol, or ethoxydiglycol; preferably, the second part of the co-solvent is glycerol; B4) In step 2), the mass ratio of the first part of the co-solvent to the second part of the co-solvent added is 2 - 5:20 - 35; B5) In step 2), the oil phase heating temperature is 75 - 85 °C; B6) In step 2), the stirring speed of the first stirring and mixing is 30 - 40 r / min; B7) In step 2), the time of the first stirring and mixing is 5 - 15 min; B8) In step 2), the stirring speed of the second stirring and mixing is 15 - 25 r / min; B9) In step 2), the time of the second stirring and mixing is 2 - 10 min; B10) In step 2), the water phase heating temperature is 75 - 85 °C; B11) In step 3), the stirring speed of the third stirring and mixing is 450 - 550 r / min; B12) In step 3), the time for the third stirring and mixing is 8 - 22 min; B13) In step 4), the time for the fourth stirring and mixing is 25 - 65 min; B14) In step 4), the fourth stirring and mixing is carried out after cooling, and the temperature is cooled to 15 - 25 °C; preferably, the cooling rate is 3 - 5 °C / min; B15) In step 4), the stirring speed of the fourth stirring and mixing is 55 - 65 r / min; B16) In step 4), the homogenization treatment includes a three - stage pressure - increasing procedure carried out in sequence; the homogenization pressure for the first - stage pressure increase is 5500 - 6500 psi; the number of cycles for the first - stage pressure increase is 2 - 3 times; the homogenization pressure for the second - stage pressure increase is 14500 - 15500 psi; the number of cycles for the second - stage pressure increase is 3 - 4 times; the homogenization pressure for the third - stage pressure increase is 19500 - 20500 psi; the number of cycles for the third - stage pressure increase is 3 - 4 times.

7. A ceramide liposome obtained by the method according to any one of claims 4 - 6.

8. Use of the ceramide liposome according to any one of claims 1 - 3 or the ceramide liposome according to claim 7 in cosmetics.

9. A cosmetic comprising the ceramide liposome according to any one of claims 1 - 3 or the ceramide liposome according to claim 7.

10. The cosmetic according to claim 9, characterized in that, The addition amount of the ceramide liposome in the cosmetic is 0.1 - 10 wt%.

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