Preparation method and application of lipidosome with good percutaneous permeability and suitable for entrapment of fat-soluble active matter

Abstract

The invention relates to a lipidosome with good percutaneous permeability and suitable for entrapment of fat-soluble active matters and a preparation method and application of the lipidosome, and the lipidosome is prepared from a phase A and a phase B. The preparation raw materials of the lipidosome comprise the following components in percentage by mass (100% by mass) of the total mass of the lipidosome, the phase A comprises the following components in percentage by weight: 0.2%-2% of fat-soluble active matter, 4%-8% of phospholipid, 0.1%-0.4% of cholesterol, 0.5%-2% of nonionic surfactant, 1%-8% of absolute ethyl alcohol and 0.02%-0.2% of antioxidant, and the phase B comprises the following components in percentage by weight: 5%-10.5% of polyhydric alcohol. The liposome is yellow or yellowish-brown transparent liquid, the particle size is 30-220nm, and the encapsulation efficiency is high and can reach 98%. After being placed for 60 days under an extreme condition, the sample character and the particle size are not obviously changed, the stability is good, the in-vitro and in-vivo percutaneous penetration performance and the slow release performance are good, active substances can be continuously released to deeper skin layers, and efficient effects are provided for fat-soluble active substances.

Description

Technical Field

[0001] This application belongs to the daily chemical field, and specifically relates to a liposome with good percutaneous penetration performance and suitable for encapsulating lipophilic active substances, its preparation method and application. Background Art

[0002] In recent years, with the popularization and promotion of efficacy requirements, consumers' demand for advanced efficacy skin care has shown an increasing trend, leading to the continuous expansion of the efficacy skin care market. In the development of skin care products, in order to achieve better effects, it is usually necessary to consider whether the efficacy active ingredients in the product can effectively exert their powerful efficacy on the human skin. For example, whether whitening and antioxidant ingredients can penetrate through the stratum corneum of the skin to reach the basal layer or related parts to play a role, and whether moisturizing and repairing ingredients can stay in the stratum corneum for a long time, that is, it is necessary to consider whether the active ingredients in the formula can have a high bioavailability.

[0003] However, the application of lipophilic active substances in cosmetics has the following three limitations: First, most lipophilic substances have very low solubility and poor water dispersibility. In conventional cosmetics, the lipophilic active ingredients are limited by the dosage form, resulting in a very low mass percentage, and unable to achieve the corresponding skin care efficacy when applied to the skin; Second, some lipophilic active substances are prone to oxidation, color change and inactivation, or are prone to crystallization and precipitation in traditional dosage forms, reducing the stability of cosmetics; Third, lipophilic substances have poor transdermal performance. Existing preparation technologies are usually traditional creams and emulsions with relatively large particle sizes, poor transdermal permeability and retention, affecting the skin care effect. Therefore, using nano-preparation technology to improve the solubility, water dispersibility, stability, transdermal permeability and retention of lipophilic active substances is the main direction for the application of lipophilic efficacy raw materials in efficacy cosmetics in recent years.

[0004] In order to improve the solubility of lipophilic active substances, improve water dispersibility and transdermal permeability, various nano-technologies have been used in the prior art to prepare nano-combinations of lipophilic substances, and their solubility and dispersibility have been greatly improved, but there are still obvious defects and deficiencies, such as:

[0005] A glabridin microcapsule and its preparation method. The prepared microcapsule has good stability and solubility, and the average particle size is 60-80 nm, which can achieve certain purposes of penetration, retention and slow release. However, a large amount of organic solvents such as dichloromethane and petroleum ether are used in the preparation method of this invention, and toxic substances will remain in the finished product, increasing the safety hazard of its application as a cosmetic raw material on the skin.

[0006] A ceramide nano-emulsion and its preparation method. Although the prepared nano-emulsion has good stability and water dispersibility, the average particle size of the prepared nano-emulsion is 100-500 nm, with a relatively large particle size, and the skin permeability and retention of ceramide are poor.

[0007] As a typical representative of new formulation technologies, liposomes have been widely used in the encapsulation and delivery of lipophilic active substances. Liposomes are closed vesicles composed of a bilayer formed by amphiphilic molecules. Their hydrophilic groups are curved arcs, connected to the lipophilic groups to form a structure similar to a "U" shape. When the hydrophobic chains in the two-sided "U" structures dock, a bilayer wall structure of the vesicle is formed. This structure of liposomes can encapsulate two types of substances, water-soluble and lipophilic. Lipophilic substances are dispersed in the phospholipid bilayer of liposomes, while water-soluble substances are encapsulated in the aqueous phase inside the vesicles. Compared with traditional formulations and other new formulation technologies, liposomes are more suitable for encapsulating and delivering lipophilic efficacy substances. First, liposomes can make the encapsulated substances water-based, effectively improving the dissolution of lipophilic active substances and expanding the application scenarios of active substances. Second, as a carrier, liposomes can improve the stability of efficacy substances. Third, the phospholipids and cholesterol in liposomes have a structure similar to the stratum corneum of the skin, with the characteristics of cell affinity and tissue compatibility, not only not irritating the skin but also playing a role in moisturizing, nourishing, and protecting the skin. Fourth, the small particle size effect of liposomes makes them easier to penetrate the stratum corneum, promoting the transdermal absorption of active substances and improving the bioavailability. Fifth, liposome-encapsulated drugs can target and retain in the skin, and the active substances are slowly released, reducing skin irritation and optimizing the dose of active substances.

[0008] Currently, many liposome technologies suitable for the encapsulation and delivery of lipophilic active substances have been developed, but there are still many obvious technical defects, such as:

[0009] A liposome solution containing licorice and glabridin and its preparation method. Although this invention uses liposome technology to solve the problems of poor dispersion ability and low solubility of glabridin in water, the preparation temperature described in this technology is 75°C, and the temperature exceeding 60°C will cause glabridin to decompose and inactivate, limiting the efficacy of the active substance in the skin.

[0010] A preparation method of liposomes with whitening and anti-aging effects. Although the low-temperature stability of the liposomes described in this invention is good, the encapsulation amount of its efficacy components is low. The encapsulation amount of glabridin in the preferred scheme is only 0.4%, and the particle size of the liposomes is greater than 150 nm.

[0011] A liposome and its preparation method. In this invention, two fat-soluble substances are encapsulated in liposomes. However, first, this technology uses two homogenization methods: vacuum homogenization and high-pressure homogenization, which makes the preparation process cumbersome and increases the production cost. Second, the invention does not conduct stability tests on the liposomes, so it is impossible to evaluate their storage stability during the shelf life. Third, the excipients in the invention's formula mainly include hydrogenated lecithin with hydrophilic and skin-friendly effects, triglyceride of caprylic / capric acid with stabilizing and antifreezing effects, and glycerol with moisturizing effects. The formula lacks components that promote transdermal penetration and absorption, which will slow down the transdermal absorption of fat-soluble active substances.

[0012] The prerequisite for the application of fat-soluble substance preparation technology in cosmetics is safety and non-irritation, and at the same time, a simple and low-cost preparation process is adopted; the prerequisite and material basis for fat-soluble skin care products to take effect is that the active substances can penetrate the skin to reach the relevant skin layers and can achieve high-concentration, long-term retention and slow release in the skin. Therefore, the preparation technology with small particle size effect and permeation-promoting effect is also one of the important prerequisite conditions for fat-soluble active substances to achieve efficient and multiple skin care effects. Summary of the Invention

[0013] Based on this, it is necessary for this application to provide a liposome with good transdermal penetration performance and suitable for encapsulating fat-soluble active substances and its preparation method. At the same time, the application of the above liposome in the preparation of cosmetics is provided. The stability, transdermal penetration performance and slow release performance of the above liposome or cosmetics containing liposomes are all better under extreme environments and in the formula.

[0014] The specific technical solutions are as follows:

[0015] A liposome with good transdermal penetration performance, and the preparation raw materials of the liposome include phase A and phase B, calculated as a percentage of the total mass of the liposome (calculated as 100% by mass).

[0016] Phase A contains the following components in the following contents:

[0017]

[0018] Phase B contains the following components in the following contents:

[0019] Polyhydric alcohol 5% - 10.5%.

[0020] In one embodiment, calculated as a percentage of the total mass of the liposome (calculated as 100% by mass).

[0021] Phase A contains the following components in the following contents:

[0022]

[0023] Phase B contains the following components in the following contents:

[0024] The polyol is 8% - 10.5%.

[0025] In one embodiment, the liposome further comprises a C phase, and the C phase contains p - hydroxyacetophenone and pentylene glycol; wherein, the mass percentage of p - hydroxyacetophenone in the liposome is 0.02% - 0.05%, and the mass percentage of pentylene glycol in the liposome is 0.05% - 2%. Optionally, the mass percentage of p - hydroxyacetophenone in the liposome is 0.03% - 0.05%, and the mass percentage of pentylene glycol in the liposome is 1% - 2%.

[0026] In one embodiment, the mass ratio of the phospholipid to cholesterol is (20:1) - (40:1). Optionally, the mass ratio of the phospholipid to cholesterol is (25:1) - (35:1).

[0027] In one embodiment, the phospholipid is one or more of 45% phosphatidylcholine, 75% phosphatidylcholine, and 90% phosphatidylcholine.

[0028] The method for preparing the above - mentioned liposome comprises the following steps:

[0029] At a certain temperature, add the phospholipid to the absolute ethanol and dissolve it completely, then add the lipophilic active substance, cholesterol, non - ionic surfactant, and antioxidant, and dissolve them completely to prepare phase A;

[0030] Dissolve and preheat the polyol and water at a certain temperature to prepare phase B;

[0031] Inject phase A into phase B at a constant speed and stir to prepare a coarse emulsion dispersion; and

[0032] Cool the coarse emulsion dispersion and then perform high - pressure homogenization to prepare liposomes.

[0033] In one embodiment, the preparation method further comprises the step of adding p - hydroxyacetophenone and pentylene glycol to the liposome and dispersing them evenly.

[0034] Use of the above - mentioned liposome or the liposome prepared by the above - mentioned preparation method in the preparation of cosmetics.

[0035] An essence with good percutaneous penetration performance, comprising the above - mentioned liposome or the liposome prepared by the above - mentioned preparation method.

[0036] Compared with the prior art, the present application has the following beneficial effects:

[0037] ① The liposome with good percutaneous penetration performance provided by this application is suitable for encapsulating various lipophilic active substances, effectively improving the application limitations of lipophilic active substances in skin care cosmetics. It can carry more active substances through the stratum corneum and penetrate deeper into the skin, continuously release the active substances to the corresponding positions in the skin layer, and more efficiently exert the skin care effects of lipophilic active substances.

[0038] ② The liposome with good percutaneous penetration performance provided by this application improves the skin permeability and skin retention of lipophilic active substances through a suitable formulation: the non-ionic surfactant in the formulation, as an edge activator of the liposome, endows the lipid membrane with elastic deformation ability, further reduces the interfacial tension together with phospholipids and cholesterol, and the liquidified liposome enhances the percutaneous penetration performance of the liposome. The non-ionic surfactant, as the penetration-enhancing component in the formulation, effectively improves the transdermal absorption of lipophilic active substances.

[0039] ③ The liposome with good percutaneous penetration performance provided by this application has good stability: the phospholipids and cholesterol in the formulation are compounded in a suitable ratio, which can reduce the influence of membrane phase transition caused by high temperature to a certain extent, thereby improving the encapsulation of the active substances by the liposome and enhancing the stability of the liposome in a high-temperature environment; at the same time, polyols, as cryoprotectants, can reduce the crystallinity of the aqueous phase and improve the stability of the liposome in low-temperature and freezing environments; therefore, the phospholipids, cholesterol, and polyols in the above formulation act synergistically to prepare a liposome with better stability in extreme environments.

[0040] ④ The liposome with good percutaneous penetration performance provided by this application has a suitable and small particle size, with a particle size of 30 - 220 nm. This particle size range can not only ensure the penetration of lipophilic active substances through the stratum corneum by the small particle size effect, but also ensure the effective retention and continuous release of the active substances in the skin, significantly increasing the skin penetration amount and skin retention amount of lipophilic substances.

[0041] ⑤ The liposome with good percutaneous penetration performance provided by this application has a high encapsulation rate. The encapsulation rate of the lipophilic active substance reaches 98%, and the liposome has good sustained-release performance, can continuously release the active substances, maintain an effective concentration for a long time, and better exert the skin care effects.

[0042] ⑥ The preparation method of the liposome with good percutaneous penetration performance provided by this application is simple, the process is easy to control, and it is suitable for large-scale industrial production. Description of the Drawings

[0043] Figure 1 It is the particle size distribution diagram of the liposome in Example 1;

[0044] Figure 2 It is the particle size distribution diagram of the liposome in Example 14;

[0045] Figure 3 Particle size distribution diagram of liposomes in Example 15;

[0046] Figure 4 Particle size distribution diagram of liposomes in Example 16;

[0047] Figure 5 Particle size distribution diagram of liposomes in Example 17;

[0048] Figure 6 Particle size distribution diagram of liposomes in Example 18;

[0049] Figure 7 Transmission electron micrograph of liposomes in Example 1;

[0050] Figure 8 Scanning comparison of the multiple light scattering stability analyzer for the samples of Example 19 and Comparative Example 17;

[0051] Figure 9 Comparison of in vitro cumulative permeation curves for the samples of Example 1 and Comparative Example 15;

[0052] Figure 10 Comparison of in vitro 24h cumulative permeation amount and skin retention amount for the samples of Example 1 and Comparative Example 15;

[0053] Figure 11 Comparison of relative in vitro percutaneous permeation curves by confocal Raman technology for the samples of Example 20 and Comparative Example 18;

[0054] Figure 12 Comparison of relative in vitro percutaneous permeation total amount by confocal Raman technology for the samples of Example 20 and Comparative Example 18;

[0055] Figure 13 Comparison of relative in vivo percutaneous permeation curves by confocal Raman technology for the samples of Example 19 and Comparative Example 17;

[0056] Figure 14 Comparison of relative in vivo percutaneous permeation total amount by confocal Raman technology for the samples of Example 19 and Comparative Example 17;

[0057] Figure 15 Comparison of in vitro release curves for the samples of Example 1 and Comparative Example 16;

[0058] Figure 16 Comparison of human melanin tests for the samples of Example 19 and Comparative Example 17. Detailed implementation manners

[0059] To make the above objects, features, and advantages of the present application more obvious and understandable, the following provides a detailed description of the specific embodiments of the present application with reference to the accompanying drawings. Many specific details are set forth in the following description to facilitate a full understanding of the present application. However, the present application can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without departing from the connotation of the present application. Therefore, the present application is not limited by the specific embodiments disclosed below.

[0060] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the technical field to which this application belongs. The terms used in the description of this application herein are for the purpose of describing specific embodiments only and are not intended to limit this application.

[0061] An embodiment of the present application provides a liposome with good percutaneous penetration performance and suitable for encapsulating lipophilic active substances. Calculated as a percentage of the total mass of the liposome (counted as 100% by mass), it comprises the following components:

[0062] Phase A comprises the following components in the following contents:

[0063]

[0064] Phase B comprises the following components in the following contents:

[0065] Polyol 5% - 10.5%.

[0066] In a specific example, the mass percentage of the lipophilic active substance in the liposome includes, but is not limited to, 0.2%, 0.4%, 0.6%, 0.8%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, or a range composed of any two values.

[0067] In a specific example, the mass percentage of phospholipid in the liposome includes, but is not limited to, 4%, 5%, 6%, 7%, 8%, or a range composed of any two values.

[0068] In a specific example, the mass percentage of cholesterol in the liposome includes, but is not limited to, 0.1%, 0.2%, 0.3%, 0.4%, or a range composed of any two values.

[0069] In a specific example, the mass percentage of the non-ionic surfactant in the liposome includes, but is not limited to, 0.5%, 0.6%, 0.8%, 1.0%, 1.2%, 1.5%, 1.8%, 2.0%, or a range composed of any two values.

[0070] In a specific example, the mass percentage of absolute ethanol in the liposome includes, but is not limited to, 1%, 2%, 3%, 4%, 5%, 6%, 8%, or a range composed of any two values.

[0071] In a specific example, the mass percentage of the antioxidant in the liposome includes 0.02%, 0.03%, 0.04%, 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.2%, or a range composed of any two values.

[0072] In a specific example, the mass percentage of the polyol in the liposome includes 5%, 6%, 7%, 7.5%, 8%, 9%, 10%, 10.5%, or a range composed of any two values.

[0073] In a specific example, the liposome further contains a C phase, where the C phase has an antiseptic effect. Optionally, the C phase contains p-hydroxyacetophenone and pentylene glycol; wherein, the mass percentage of p-hydroxyacetophenone in the liposome is 0.02% - 0.05%, and the mass percentage of pentylene glycol in the liposome is 0.05% - 2%. Optionally, the mass percentage of p-hydroxyacetophenone in the liposome is 0.03% - 0.05%, and the mass percentage of pentylene glycol in the liposome is 1% - 2%.

[0074] In a specific example, calculated as a percentage of the total mass of the liposome (counted as 100% by mass), it contains the following components:

[0075] Phase A contains the following components in the following amounts:

[0076]

[0077] Phase B contains the following components in the following amounts:

[0078] Polyol 5% - 10.5%,

[0079] Deionized water the balance;

[0080] Phase C contains the following components in the following amounts:

[0081] p-Hydroxyacetophenone 0.02% - 0.05%,

[0082] Pentylene glycol 0.05% - 2%.

[0083] In a specific example, calculated as a percentage of the total mass of the liposome (counted as 100% by mass), it contains the following components:

[0084] Phase A contains the following components in the following amounts:

[0085]

[0086] Phase B contains the following components in the following amounts:

[0087] Polyol: 7.5% - 10.5%,

[0088] Deionized water: the balance;

[0089] Phase C contains the following components in the following amounts:

[0090] p-Hydroxyacetophenone: 0.03% - 0.05%,

[0091] Pentanediol: 1% - 2%.

[0092] In a specific example, in terms of the percentage of the total mass of the liposome (counted as 100% by mass), it contains the following components:

[0093] Phase A contains the following components in the following amounts:

[0094]

[0095] Phase B contains the following components in the following amounts:

[0096] Polyol: 10.5%,

[0097] Deionized water: the balance;

[0098] Phase C contains the following components in the following amounts:

[0099] p-Hydroxyacetophenone: 0.05%,

[0100] Pentanediol: 2%.

[0101] In a specific example, in terms of the percentage of the total mass of the liposome (counted as 100% by mass), it contains the following components:

[0102] Phase A contains the following components in the following amounts:

[0103]

[0104] Phase B contains the following components in the following amounts:

[0105] Polyol: 10.5%,

[0106] Deionized water: the balance;

[0107] Phase C contains the following components in the following amounts:

[0108] p-Hydroxyacetophenone: 0.05%

[0109] Pentanediol: 2%.

[0110] In this application, cholesterol is an important supplementary component of the liposome wall material. When the unsaturated bonds of phospholipids reach the corresponding phase transition temperature, physical property changes will occur in the lipid membrane, resulting in leakage of liposomes. However, the compounding of cholesterol and phospholipids in an appropriate ratio can, to a certain extent, reduce the impact of membrane phase transition caused by high temperature, thereby improving the encapsulation of the active substance by liposomes and enhancing the stability of liposomes. In a specific example, the mass ratio of the phospholipid to cholesterol is (20:1) to (40:1). Optionally, the mass ratio of the phospholipid to cholesterol is (25:1) to (35:1). Optionally, the mass ratio of the phospholipid to cholesterol is (20:1) to (30:1).

[0111] In a specific example, the phospholipid is one or more of phosphatidylcholine containing 45%, 75%, and 90%. Among them, as an important component of the liposome wall material, phospholipids have good hydrophilicity and moisture retention, can nourish the skin, and at the same time have good biocompatibility with the skin cutin layer structure, can fuse with skin lipids, disrupt their bilayer arrangement structure, and promote the percutaneous penetration of active substances.

[0112] In a specific example, the non-ionic surfactant includes one or more of methyl glucoside sesquistearate, Tween-80, cetearyl glucoside, polyglyceryl-10 laurate, polyglyceryl-10 myristate, behenyl polyether-25, steareth-21, cetearyl alcohol polyether-25, ceteth-20, oleth-20, sucrose stearate.

[0113] Preferably, the non-ionic surfactant is a polyether non-ionic surfactant, including one or more of behenyl polyether-25, steareth-21, cetearyl alcohol polyether-25, ceteth-20, oleth-20. Non-ionic surfactants such as behenyl polyether-25, as the edge activator of liposomes, can enhance the fluidity of the membrane lipid membrane, endow the phospholipid membrane of liposomes with elastic deformation ability, reduce the interfacial tension, and liquefy liposomes, thereby enhancing the percutaneous penetration performance of liposomes and promoting the transdermal absorption of active ingredients. Further preferably, the non-ionic surfactant is one or more of behenyl polyether-25, steareth-21, cetearyl alcohol polyether-25. The three polyether non-ionic surfactants used synergistically maintain the shape and particle size stability of liposomes, and can still basically maintain the original particle size under the stability accelerated experiment.

[0114] In a specific example, the polyol includes one or more of glycerol and butanediol. As a humectant and cryoprotectant, the polyol has excellent moisture retention performance, provides a good skin feel, and at the same time can reduce the crystallinity of the aqueous phase, and synergistically with a certain proportion of phospholipids and cholesterol to improve the stability of liposomes in extreme environments.

[0115] In a specific example, the antioxidant includes one or more of pentaerythritol tetra(bis-tert-butylhydroxyhydrocinnamate), vitamin E, and vitamin E acetate. Among them, antioxidants such as pentaerythritol tetra(bis-tert-butylhydroxyhydrocinnamate) have antioxidant effects and delay the premature leakage of liposome contents caused by oxidation generated by unsaturated bonds.

[0116] In a specific example, the lipophilic active substances include one or more of glabridin, ceramide, bakuchiol, hydroxypinacolone retinoate, and resveratrol. Optionally, the ceramide can be ceramide III.

[0117] Among them, glabridin has the functions of inhibiting tyrosinase activity, inhibiting the generation of reactive oxygen species, and reducing inflammatory responses, and can provide multi-target and high-efficiency whitening, antioxidant, and anti-inflammatory effects. It is a natural plant whitening cosmetic raw material with quite high research and application value. Ceramide III has a long-lasting moisturizing effect, and at the same time can protect the skin barrier and repair skin sensitivity. It is a widely used moisturizing and repairing raw material. Bakuchiol has the functions of moisturizing, barrier repair ability, antioxidant, anti-inflammatory, and anti-aging. It is a mild and highly efficient cosmetic efficacy raw material. Hydroxypinacolone retinoate is a retinoid raw material with anti-aging and sebum secretion inhibitory effects. Resveratrol is a plant polyphenol substance with whitening, antioxidant, anti-inflammatory and other effects. Those skilled in the art can select components with corresponding effects as the lipophilic active substances of the liposomes of the present application according to actual needs.

[0118] The above liposomes are yellow or yellowish-brown transparent liquids with a particle size of 30 - 220 nm, a high encapsulation rate of up to 98%. After being placed under extreme conditions for 60 days, the sample properties and particle size did not change significantly, with good stability, and both in vitro and in vivo percutaneous penetration performance and sustained release performance are better. It can continuously release active substances to deeper layers of the skin, providing efficient efficacy for lipophilic active substances. At the same time, the raw material components of the liposomes do not contain a large amount of organic solvents, and are safe and non-irritating.

[0119] An embodiment of the present application provides a preparation method of the above liposomes, including the following steps 1 to 4:

[0120] Step 1, at a temperature of 45°C to 85°C, dissolve phospholipids in absolute ethanol, then add lipophilic active substances, cholesterol, non-ionic surfactants, and antioxidants, and mix and dissolve to prepare Phase A.

[0121] Specifically, at a temperature of 45°C to 85°C, phospholipids are added to absolute ethanol and stirred until completely dissolved. Then, other components in Phase A are added: lipophilic active substances, cholesterol, non-ionic surfactants, and antioxidants, and stirred until the solution is clear, transparent, and completely dissolved to obtain Phase A.

[0122] Specifically, the selectable temperature range is 45°C to 55°C, 55°C to 65°C, 65°C to 75°C, or 75°C to 85°C. Specifically, the selectable temperatures are 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, or 85°C.

[0123] Step 2, polyols and water are dissolved and preheated at a temperature of 45°C to 85°C to obtain Phase B.

[0124] Step 3, Phase A is uniformly injected into Phase B and stirred to prepare a coarse emulsion dispersion.

[0125] Specifically, the stirring rate is 200 to 10000 rpm, and the stirring time is 10 to 40 min; preferably, the stirring rate is 400 to 600 rpm, and the stirring time is 15 to 30 min.

[0126] Step 4, after the coarse emulsion dispersion is cooled, it is subjected to high-pressure homogenization to prepare liposomes.

[0127] In this application, high-pressure homogenization treatment makes the particle size of the liposome composition system smaller, more uniform, and more stable.

[0128] Specifically, after the coarse emulsion dispersion is cooled to room temperature, it is subjected to high-pressure homogenization to prepare liposomes.

[0129] Optionally, the homogenization pressure is 200 to 900 bar, and the number of homogenization times is 2 to 9 times. Preferably, the homogenization pressure is 300 to 500 bar, and the number of homogenization times is 3 to 5 times.

[0130] In a specific example, the preparation method further includes Step 5, adding p-hydroxyacetophenone and pentylene glycol to the liposomes prepared in Step 4 and dispersing them evenly.

[0131] Specifically, p-hydroxyacetophenone and pentylene glycol can be first mixed to prepare a Phase C solution, and the Phase C solution is added to the liposomes prepared in Step 4 and dispersed evenly.

[0132] In this application, a combination of ethanol injection and high-pressure homogenization is used to prepare liposomes with a smaller particle size, which can carry lipophilic active substances deeper into the skin and have good transdermal penetration performance, effectively promoting the transdermal absorption of lipophilic active substances and achieving long-acting slow-release and high-efficiency skin care effects.

[0133] One embodiment of the present application also provides the use of the above liposomes or liposomes prepared by the above preparation method in the preparation of cosmetics. The liposomes prepared in the present application have high transparency and can meet the requirements of all cosmetic formulations, especially aqueous formulations.

[0134] Optionally, the cosmetics include one or more of essence, skin care lotion, emulsion, gel, cream and facial mask liquid.

[0135] One embodiment of the present application also provides a liposome essence with good percutaneous penetration performance, including the above liposomes or liposomes prepared by the preparation method.

[0136] In a specific example, the mass percentage of liposomes in the essence is 10% - 50%, and the specific optional mass percentages are 20%, 25%, 30%, 35%, 40%, 45% or 50%. Optionally, the mass percentage of liposomes in the essence is 20% - 40%.

[0137] In a specific example, by mass percentage, the essence contains the following components: 20% - 40% liposomes, 3% - 5% glycerin, 5% - 8% propylene glycol, 5% - 8% butylene glycol, 1% - 3% pentylene glycol, 1% - 2% polyglyceryl-10 myristate, 0.03% - 0.05% EDTA-2Na, 0.4% - 0.6% AVC (acryloyldimethyltaurine ammonium / VP copolymer), 0.1% - 0.2% p-hydroxyacetophenone, and the balance is deionized water.

[0138] In a specific example, the preparation method of the above essence includes the following steps:

[0139] After dispersing AVC in 30% - 50% water, add glycerin, propylene glycol, butylene glycol, pentylene glycol, polyglyceryl-10 myristate, p-hydroxyacetophenone, and EDTA-2 sodium, heat to 80°C - 85°C, and keep warm (for example, keep warm for 20 min - 30 min); cool down (for example, cool down to 40°C - 45°C), then add the above liposomes and mix evenly, and make up water to 100% after cooling to room temperature.

[0140] The embodiments of the present application will be described in detail below in conjunction with examples. It should be understood that these examples are only used to illustrate the present application and not to limit the scope of the present application. For the experimental methods without specific conditions in the following examples, the guidance given in the present application should be preferentially referred to, and it can also be carried out according to the experimental manuals or conventional conditions in the art, or according to the conditions recommended by the manufacturers, or referring to the experimental methods known in the art.

[0141] In the following specific embodiments, regarding the measurement parameters of raw material components, without special instructions, there may be slight deviations within the weighing accuracy range. Regarding temperature and time parameters, acceptable deviations caused by instrument test accuracy or operation accuracy are allowed.

[0142] Example 1

[0143] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components by mass percentage: glabridin 1%, phospholipid 4%, cholesterol 0.13%, polyoxyethylene 25 behenyl ether 1.5%, absolute ethanol 4%, pentaerythritol tetra(3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate) 0.05%, glycerol 10.5%, p-hydroxyacetophenone 0.05%, pentylene glycol 2%, and the balance is deionized water, where the mass percentage ratio of phospholipid to cholesterol is 30:1.

[0144] This example also provides a preparation method for the above liposome, which specifically includes the following steps: Add phospholipid to absolute ethanol and stir at 45°C until completely dissolved; after the solution becomes clear and transparent, add glabridin, cholesterol, polyoxyethylene 25 behenyl ether, and pentaerythritol tetra(3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate), and stir until the solution becomes clear and transparent and completely dissolved to obtain Phase A; Dissolve and preheat glycerol and water at 45°C to obtain Phase B; Dissolve p-hydroxyacetophenone in pentylene glycol at room temperature to obtain Phase C; Inject Phase A into Phase B at a constant speed and mechanically stir at 500 rpm for 30 min to obtain a coarse emulsion dispersion; After the coarse emulsion dispersion is cooled to room temperature, perform high-pressure homogenization with a homogenization pressure of 500 bar and a homogenization times of 3 times to obtain the liposome; Finally, add Phase C after the liposome naturally cools and disperse evenly.

[0145] Example 2

[0146] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this example is polyoxyethylene 21 stearyl ether, and other components and preparation methods are the same as those in Example 1.

[0147] Example 3

[0148] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this example is cetostearyl alcohol polyoxyethylene ether-25, and other components and preparation methods are the same as those in Example 1.

[0149] Example 4

[0150] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of cholesterol and deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol. In this example, the mass percentage of cholesterol is 0.2%, that is, the ratio of the mass percentage of phospholipid to cholesterol is 20:1, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0151] Example 5

[0152] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of cholesterol and deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol. In this example, the mass percentage of cholesterol is 0.1%, that is, the ratio of the mass percentage of phospholipid to cholesterol is 40:1, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0153] Example 6

[0154] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glycerol and deionized water. In this example, the mass percentage of glycerol is 5%, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0155] Example 7

[0156] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glycerol and deionized water. In this example, the mass percentage of glycerol is 7.5%, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0157] Example 8

[0158] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the types and addition amounts of polyols and the addition amount of deionized water. In this example, the polyols used are glycerol and butanediol, and the mass percentages of glycerol and butanediol are 2% and 3% respectively, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0159] Example 9

[0160] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the types and addition amounts of polyols and the addition amount of deionized water. In this example, the polyols used are glycerol and butanediol, and the mass percentages of glycerol and butanediol are 3% and 2% respectively, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0161] Example 10

[0162] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glabridin and deionized water. In this example, the mass percentage of glabridin is 0.2%, and the balance is deionized water. Other components and the preparation method are the same as those in Example 1.

[0163] Example 11

[0164] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glabridin and deionized water. In this example, the mass percentage of glabridin is 0.4%, and the balance is deionized water. Other components and the preparation method are the same as those in Example 1.

[0165] Example 12

[0166] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glabridin and deionized water. In this example, the mass percentage of glabridin is 0.6%, and the balance is deionized water. Other components and the preparation method are the same as those in Example 1.

[0167] Example 13

[0168] This example provides a liposome with good percutaneous penetration performance. Compared with Example 1, the difference lies in the addition amounts of glabridin and deionized water. In this example, the mass percentage of glabridin is 0.8%, and the balance is deionized water. Other components and the preparation method are the same as those in Example 1.

[0169] Example 14

[0170] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components by mass percentage: ceramide III 1%, phospholipid 6%, cholesterol 0.2%, beheneth-25 1.5%, absolute ethanol 4%, pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate) 0.05%, glycerol 10.5%, p-hydroxyacetophenone 0.05%, pentylene glycol 2%, and the balance is deionized water, where the mass percentage ratio of phospholipid to cholesterol is 30:1.

[0171] This example also provides a method for preparing the above liposomes, which specifically includes the following steps: Add phospholipids to absolute ethanol and stir at 85 °C until completely dissolved; after the solution becomes clear and transparent, add ceramide, cholesterol, behenyl polyether-25, and pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate), and stir until the solution becomes clear and transparent and completely dissolved to obtain Phase A; Dissolve and preheat glycerol and water at 85 °C to obtain Phase B; Dissolve p-hydroxyacetophenone in pentylene glycol at room temperature to obtain Phase C; Slowly inject Phase A into Phase B and mechanically stir at 500 rpm for 30 min to obtain a coarse emulsion dispersion; After the coarse emulsion dispersion is cooled to room temperature, perform high-pressure homogenization with a homogenization pressure of 500 bar and a homogenization times of 3 times to obtain liposomes; Finally, add Phase C after the liposomes are naturally cooled and disperse evenly.

[0172] Example 15

[0173] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components by mass percentage: bakuchiol 1%, phospholipids 6%, cholesterol 0.2%, behenyl polyether-25 1.5%, absolute ethanol 4%, pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate) 0.05%, glycerol 10.5%, p-hydroxyacetophenone 0.05%, pentylene glycol 2%, and the balance is deionized water, where the mass percentage ratio of phospholipids to cholesterol is 30:1.

[0174] This example also provides a method for preparing the above liposomes, which specifically includes the following steps: Add phospholipids to absolute ethanol and stir at 45 °C until completely dissolved; after the solution becomes clear and transparent, add bakuchiol, cholesterol, behenyl polyether-25, and pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate), and stir until the solution becomes clear and transparent and completely dissolved to obtain Phase A; Dissolve and preheat glycerol and water at 45 °C to obtain Phase B; Dissolve p-hydroxyacetophenone in pentylene glycol at room temperature to obtain Phase C; Slowly inject Phase A into Phase B and mechanically stir at 500 rpm for 30 min to obtain a coarse emulsion dispersion; After the coarse emulsion dispersion is cooled to room temperature, perform high-pressure homogenization with a homogenization pressure of 500 bar and a homogenization times of 3 times to obtain liposomes; Finally, add Phase C after the liposomes are naturally cooled and disperse evenly.

[0175] Example 16

[0176] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components by mass percentage: hydroxypinacolone retinoate 1%, phospholipids 6%, cholesterol 0.2%, behenyl polyether-25 1.5%, absolute ethanol 4%, pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate) 0.05%, glycerol 10.5%, p-hydroxyacetophenone 0.05%, pentylene glycol 2%, and the balance is deionized water, where the mass percentage ratio of phospholipids to cholesterol is 30:1.

[0177] This example also provides a method for preparing the above liposome, which specifically includes the following steps: Add phospholipids to absolute ethanol and stir at 45 °C until completely dissolved; after the solution becomes clear and transparent, add hydroxypinacolone retinoate, cholesterol, beheneth-25, and pentaerythrityl tetra(bis-tert-butylhydroxyhydrocinnamate), and stir until the solution becomes clear and transparent and completely dissolved to obtain Phase A; Dissolve and preheat glycerol and water at 45 °C to obtain Phase B; Dissolve p-hydroxyacetophenone in pentylene glycol at room temperature to obtain Phase C; Slowly inject Phase A into Phase B and mechanically stir at 500 rpm for 30 min to obtain a coarse emulsion dispersion; After the coarse emulsion dispersion is cooled to room temperature, perform high-pressure homogenization with a homogenization pressure of 500 bar and a homogenization times of 3 times to obtain liposomes; Finally, add Phase C after the liposomes are naturally cooled and disperse evenly.

[0178] Example 17

[0179] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components in mass percentage: 2% of hydroxypinacolone retinoate, 8% of phospholipids, 0.27% of cholesterol, 1.5% of beheneth-25, 4% of absolute ethanol, 0.05% of pentaerythrityl tetra(bis-tert-butylhydroxyhydrocinnamate), 10.5% of glycerol, 0.05% of p-hydroxyacetophenone, 2% of pentylene glycol, and the balance is deionized water, where the ratio of the mass percentage of phospholipids to cholesterol is 30:1.

[0180] This example also provides a method for preparing the above liposome, which specifically includes the following steps: Add phospholipids to absolute ethanol and stir at 45 °C until completely dissolved; after the solution becomes clear and transparent, add hydroxypinacolone retinoate, cholesterol, beheneth-25, and pentaerythrityl tetra(bis-tert-butylhydroxyhydrocinnamate), and stir until the solution becomes clear and transparent and completely dissolved to obtain Phase A; Dissolve and preheat glycerol and water at 45 °C to obtain Phase B; Dissolve p-hydroxyacetophenone in pentylene glycol at room temperature to obtain Phase C; Slowly inject Phase A into Phase B and mechanically stir at 500 rpm for 30 min to obtain a coarse emulsion dispersion; After the coarse emulsion dispersion is cooled to room temperature, perform high-pressure homogenization with a homogenization pressure of 500 bar and a homogenization times of 3 times to obtain liposomes; Finally, add Phase C after the liposomes are naturally cooled and disperse evenly.

[0181] Example 18

[0182] This example provides a liposome with good percutaneous penetration performance, which is composed of the following raw material components by mass percentage: resveratrol 1%, phospholipid 6%, cholesterol 0.2%, behenyl alcohol polyether-25 1.5%, absolute ethanol 4%, pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate) 0.05%, glycerol 10.5%, p-hydroxyacetophenone 0.05%, pentylene glycol 2%, and the balance is deionized water, where the mass percentage ratio of phospholipid to cholesterol is 30:1.

[0183] This example also provides a preparation method of the above liposome, which specifically includes the following steps: adding phospholipid to absolute ethanol and stirring at 45°C until completely dissolved; after the solution is clear and transparent, adding resveratrol, cholesterol, behenyl alcohol polyether-25, and pentaerythrityl tetra(bis-tert-butyl hydroxyhydrocinnamate), and stirring until the solution is clear and transparent and completely dissolved to obtain phase A; dissolving and preheating glycerol and water at 45°C to obtain phase B; dissolving p-hydroxyacetophenone in pentylene glycol at room temperature to obtain phase C; injecting phase A into phase B at a constant speed and mechanically stirring at 500 rpm for 30 min to obtain a coarse emulsion dispersion; after the coarse emulsion dispersion is cooled to room temperature, performing high-pressure homogenization, with the homogenization pressure of 500 bar and the number of homogenization times of 3 times, to obtain the liposome; finally, adding phase C after the liposome is naturally cooled and dispersing evenly.

[0184] Example 19

[0185] This example provides a serum containing liposome, which contains the liposome prepared in Example 1 and is composed of the following raw material components by mass percentage: liposome in Example 1 20%, glycerol 3%, propylene glycol 5%, butylene glycol 5%, pentylene glycol 1%, polyglyceryl-10 myristate 1%, EDTA-2Na 0.03%, AVC (acryloyldimethyltauramide / VP copolymer) 0.4%, p-hydroxyacetophenone 0.2%, and the balance is deionized water.

[0186] This example also provides a preparation method of the above serum, which specifically includes the following steps: first dispersing AVC in 50% water, then adding glycerol, propylene glycol, butylene glycol, pentylene glycol, polyglyceryl-10 myristate, p-hydroxyacetophenone, and EDTA disodium, heating to 85°C, and holding for 30 min; cooling to 40°C, then adding the glabridin liposome in Example 1 and mixing evenly, and making up the water to 100% after cooling to room temperature.

[0187] Example 20

[0188] This example provides a serum containing liposome. Compared with Example 19, the difference is that 20% of the glabridin liposome in Example 1 in the serum is replaced by 40% of the ceramide liposome in Example 14, and the other components and preparation methods are the same as those in Example 19.

[0189] Comparative Example 1

[0190] Compared with Example 1, the difference lies in the preparation method of liposomes. In this comparative example, high-pressure homogenization is not used to treat the coarse emulsion dispersion, and other components and preparation methods are the same as those in Example 1.

[0191] Comparative Example 2

[0192] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is ceteth-20, and other components and preparation methods are the same as those in Example 1.

[0193] Comparative Example 3

[0194] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is oleth-20, and other components and preparation methods are the same as those in Example 1.

[0195] Comparative Example 4

[0196] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is methyl glucoside sesquistearate, and other components and preparation methods are the same as those in Example 1.

[0197] Comparative Example 5

[0198] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is Tween-80, and other components and preparation methods are the same as those in Example 1.

[0199] Comparative Example 6

[0200] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is cetearyl glucoside, and other components and preparation methods are the same as those in Example 1.

[0201] Comparative Example 7

[0202] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is polyglyceryl-10 laurate, and other components and preparation methods are the same as those in Example 1.

[0203] Comparative Example 8

[0204] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is polyglyceryl-10 myristate, and other components and preparation methods are the same as those in Example 1.

[0205] Comparative Example 9

[0206] Compared with Example 1, the difference lies in the type of non-ionic surfactant. The non-ionic surfactant used in this comparative example is sucrose stearate, and other components and preparation methods are the same as those in Example 1.

[0207] Comparative Example 10

[0208] Compared with Example 1, the difference lies in the addition amount of cholesterol and the addition amount of deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol. In this comparative example, the mass percentage of cholesterol is 0.67%, and the balance is deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol is 6:1. Other components and preparation methods are the same as those in Example 1.

[0209] Comparative Example 11

[0210] Compared with Example 1, the difference lies in the addition amount of cholesterol and the addition amount of deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol. In this comparative example, the mass percentage of cholesterol is 0.07%, and the balance is deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol is 60:1. Other components and preparation methods are the same as those in Example 1.

[0211] Comparative Example 12

[0212] Compared with Example 1, the difference lies in the addition amount of cholesterol and the addition amount of deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol. In this comparative example, the mass percentage of cholesterol is 0.05%, and the balance is deionized water, that is, the ratio of the mass percentage of phospholipid to cholesterol is 80:1. Other components and preparation methods are the same as those in Example 1.

[0213] Comparative Example 13

[0214] Compared with Example 1, the difference lies in the addition amount of glycerol and the addition amount of deionized water. In this comparative example, the mass percentage of glycerol is 2.5%, and the balance is deionized water. Other components and preparation methods are the same as those in Example 1.

[0215] Comparative Example 14

[0216] Compared with Example 1, the difference lies in not adding polyol. Other components and preparation methods are the same as those in Example 1.

[0217] Comparative Example 15

[0218] Prepare glabridin oil solution. 1% glabridin is dissolved in 99% caprylic / capric triglyceride by mass percentage to obtain 1% glabridin oil solution.

[0219] Comparative Example 16

[0220] Prepare a glabridin alcohol solution by dissolving 1% glabridin by mass in 99% ethanol by mass to obtain a 1% glabridin alcohol solution.

[0221] Comparative Example 17

[0222] This example provides a serum containing a fat-soluble active substance. Compared with Example 19, the difference is that 20% of the glabridin liposomes in Example 1 in the serum are replaced with 0.2% glabridin, and other components and preparation methods are the same as those in Example 19.

[0223] Comparative Example 18

[0224] This example provides a serum containing a fat-soluble active substance. Compared with Example 19, the difference is that 20% of the glabridin liposomes in Example 1 in the serum are replaced with 0.4% ceramide III, and other components and preparation methods are the same as those in Example 19.

[0225] Experimental Example 1

[0226] Compare the appearance properties and particle size of liposomes prepared with different contents of fat-soluble active substances and preparation methods.

[0227] Place the samples of Example 1, Examples 10 - 18, and Comparative Example 1 in an environment at room temperature without direct sunlight, visually observe their appearance properties, and measure their particle sizes using a Malvern laser particle size analyzer.

[0228] Table 1 Appearance properties and particle size measurement results of liposomes prepared with different fat-soluble active substances, contents, and preparation methods

[0229] Sample Appearance Particle size / nm PDI Example 1 Yellowish-brown transparent liquid, without stratification, leakage, aggregation, or flocculation 48.36 0.277 Example 10 Yellowish-brown transparent liquid, without stratification, leakage, aggregation, or flocculation 43.36 0.199 Example 11 Yellowish-brown transparent liquid, without stratification, leakage, aggregation, or flocculation 43.71 0.225 Example 12 Yellowish-brown transparent liquid, without stratification, leakage, aggregation, or flocculation 42.73 0.228 Example 13 Yellowish-brown transparent liquid, without stratification, leakage, aggregation, or flocculation 46.39 0.246 Example 14 Yellow transparent liquid, without stratification, leakage, aggregation, or flocculation 48.50 0.263 Example 15 Light yellow opaque liquid, without stratification, leakage, aggregation, or flocculation 114.1 0.322 Example 16 Yellow transparent liquid, without stratification, leakage, aggregation, or flocculation 44.56 0.256 Example 17 Yellow transparent liquid, without stratification, leakage, aggregation, or flocculation 49.89 0.265 Example 18 Yellow transparent liquid, without stratification, leakage, aggregation, or flocculation 64.50 0.085 Comparative Example 1 Yellowish-brown turbid liquid, without stratification, leakage, aggregation, or flocculation 443.35 0.283

[0230] The experimental results show (Table 1): The samples of Example 1, Examples 10 - 18, and Comparative Example 1 prepared in this application have no stratification, leakage, aggregation, or flocculation phenomena. The sample prepared in Comparative Example 1 was not subjected to high-pressure homogenization treatment, and its appearance was turbid and the particle size was too large, exceeding 400 nm. However, the samples prepared in Example 1 and Examples 10 - 18 have appearances, properties, and particle sizes that meet the actual application requirements at room temperature. Especially when the concentration of the fat-soluble active substance is high, they are still stable, and no crystallization precipitation or leakage phenomena are found. This indicates that the liposomes prepared according to the component contents and preparation methods used in this application have good properties. Among them, Figure 1 - Figure 6 They are respectively the particle size distribution diagrams of the liposomes of Example 1 and Examples 14 - 18. Figure 7 It is the transmission electron microscope image of the liposomes of Example 1.

[0231] Experimental Example 2

[0232] Compare the particle size and particle size stability of liposomes prepared with different types of non-ionic surfactants.

[0233] The samples of Examples 1 to 3 and Comparative Examples 2 to 9 were placed under conditions of room temperature, 45 °C, -15 °C, and freeze-thaw cycles (one freeze-thaw cycle was placing at -15 °C for 24 h and then at 45 °C for 24 h), and their particle sizes, PDI were characterized and the particle size stability was measured using a Malvern laser particle size analyzer.

[0234] Table 2 Results of particle size and particle size stability of liposomes prepared with different types of non-ionic surfactants

[0235]

[0236] The experimental results show (Table 2) that: for the samples of Examples 1 to 3 and Comparative Examples 2 to 3 prepared in this application, the particle sizes and PDI are relatively stable at room temperature, and the appearance is yellowish-brown and transparent. While for the samples of Comparative Examples 4 to 9, the particle sizes and PDI are larger at room temperature, and the dispersion stability of the liposomes is poor, the solution is turbid and shows a layering phenomenon; when Examples 1 to 3 and Comparative Examples 2 to 3 were placed under conditions of 45 °C, -15 °C and freeze-thaw cycles for 30 days, the particle sizes of Examples 1 to 3 hardly changed, while the particle sizes of Comparative Examples 2 to 3 increased to varying degrees. It shows that the three polyether-based non-ionic surfactants (behenyl alcohol polyether-25, stearyl alcohol polyether-21, cetearyl alcohol polyether-25) used in the liposomes prepared in this application have the effect of maintaining the liposome properties and particle size stability, and can still basically maintain the original particle size under the accelerated stability experiment, and together with the vesicle wall formed by the compounding of phospholipids and cholesterol, ensure the stability of the liposomes.

[0237] Experimental Example 3

[0238] Compare the properties and particle sizes of liposomes prepared with different mass percentage ratios of phospholipids and cholesterol.

[0239] The samples of Example 1, Examples 4 to 5, and Comparative Examples 10 to 12 were placed in an environment without direct sunlight at room temperature, and their appearance properties were visually observed, and their particle sizes were measured using a Malvern laser particle size analyzer.

[0240] Table 3 Appearance properties and particle size measurement results of liposomes prepared with different mass percentage ratios of phospholipids and cholesterol

[0241]

[0242]

[0243] The experimental results show (Table 3) that: the compounding ratios of phospholipids and cholesterol used in this application (20:1, 30:1, 40:1) have the effect of maintaining the stability of liposome properties. The liposomes prepared with this ratio do not show layering, leakage, aggregation, and flocculation phenomena, and their appearance, properties, and particle sizes meet the actual application requirements.

[0244] Experimental Example 4

[0245] Compare the appearance properties, particle size and particle size stability of liposomes prepared with different types and mass percentages of polyols.

[0246] Place the samples of Example 1, Examples 6 - 9, and Comparative Examples 13 - 14 at room temperature, -15°C, and freeze-thaw cycle (one freeze-thaw cycle is placing at -15°C for 24 h and then at 45°C for 24 h), visually observe the changes in appearance properties, and use a Malvern laser particle size analyzer to characterize the particle size, PDI, and measure the particle size stability.

[0247] Table 4 Results of appearance properties, particle size and particle size stability of liposomes prepared with different types and mass percentages of polyols

[0248]

[0249] The experimental results show (Table 4): For the samples of Example 1, Examples 6 - 9 prepared in this application, there are no phenomena of delamination, leakage, aggregation, and flocculation when placed at -15°C and under freeze-thaw cycle conditions, and the particle size hardly changes; while for the samples of Comparative Examples 13 - 14 with a small amount of polyol added and without polyol added, white flocculants and leakage and aggregation phenomena occur under -15°C and freeze-thaw cycle conditions. This shows that the polyol and its addition amount used in the liposomes prepared in this application improve the stability of liposomes in a frozen environment, and can still basically maintain the original particle size under the accelerated stability experiment, thus preparing liposomes with good stability in an extremely low temperature environment.

[0250] Experimental Example 5 Liposome Stability Test

[0251] Place the samples of Example 1 and Example 14 at room temperature, 45°C, 4°C, -15°C, freeze-thaw cycle (one freeze-thaw cycle is placing at -15°C for 24 h and then at 45°C for 24 h), light and dark conditions, and use a Malvern laser particle size analyzer to test the particle size stability for 7 days, 14 days, 30 days, 60 days, and 90 days.

[0252] Table 5 Results of stability investigation of liposomes of Example 1 and Example 14

[0253]

[0254] The experimental results show (Table 5): For the samples of Example 1 and Example 14 prepared in this application, there are no aggregation and leakage phenomena. After the prepared samples are placed under different extreme conditions for 60 days, the properties and particle size do not change significantly and still meet the actual application requirements, indicating that the liposome properties are relatively stable according to the components and contents used in this application.

[0255] Experimental Example 6 Stability Test of Liposomes in Essence

[0256] The samples of Example 19 and Comparative Example 17 were respectively placed in a Turbiscan Lab multiple light scattering stability analyzer to measure the dynamic changes of the essence within 24 h, scanning once every 30 min, so as to predict the stability of the fat-soluble active substance and its liposomes in the essence. The kinetic stability index (TSI) was used to analyze the stability and homogeneity of the samples. The smaller the TSI, the more homogeneous and stable the system is.

[0257] See the experimental results in Figure 8 : The TSI of Example 19 was significantly smaller than that of Comparative Example 17, indicating that the distribution of liquiritin after being encapsulated by the liposome technology was more homogeneous and stable in the essence, and the liposomes prepared in this application had good stability in the cosmetic formulation, improving the problem of the limited application of liquiritin in the cosmetic formulation.

[0258] Experimental Example 7 In Vitro Skin Permeability and Retention Test of Liposomes

[0259] The in vitro transdermal experiments were carried out on Example 1 and Comparative Example 15, Example 20 and Comparative Example 18 of this application respectively using Franz diffusion cells. The abdominal skin of guinea pigs weighing 200 - 250 g was used as the transdermal test barrier layer, and the intact and undamaged skin was fixed between the receiving pool and the supply pool. The effective diffusion area was 1.77 cm 2 , the volume of the receiving pool was about 12 ml, and the magnetic stirring speed was 300 r / min. The receiving pool was filled with the release medium (2% Tween 80 - 20% propylene glycol - normal saline was used as the release medium for Example 1 and Comparative Example 15; 2% SDS - 20% absolute ethanol - normal saline was used for Example 20 and Comparative Example 18), air bubbles were removed, stirring was started, and the temperature was kept constant at (37.0 ± 0.5)°C. Samples containing an equal amount of liquiritin were evenly coated on the skin surface. At the set times of 2 h, 4 h, 6 h, 8 h, 12 h, and 24 h, 1 ml of the receiving solution was aspirated with a long-headed sampling needle, and the sample solution was placed in an EP tube. First, the receiving pool was used to draw air bubbles with a non-porous puncture needle, and then 1 ml of the receiving solution was added to the receiving pool. The concentration of the fat-soluble active substance liquiritin in the receiving solution filtered through a 0.22 μm filter membrane was measured by high performance liquid chromatography respectively, and the cumulative permeation amount of the drug at different times was calculated. The cumulative permeation amount of the fat-soluble active substance was calculated according to the following formula:

[0260]

[0261] In the formula, Q n is the cumulative permeation amount at the nth time point per unit area (μg / cm 2 ); V ois the volume of the liquid in the receiving pool (ml); V is the sampling volume (ml); C n is the mass concentration of the drug in the receiving solution measured at the nth time point (μg / ml); C n-1 is the mass concentration of the drug in the receiving solution measured at the (n - 1)th sampling point (μg / ml); S is the effective area.

[0262] After 24 h, the mouse skin was cut into pieces and placed in a 10 mL EP tube. 3 mL of methanol was added and sonicated for 30 min. After sonication, it was centrifuged at 5000 r / min for 10 min, and the supernatant was collected into the EP tube. It was filtered through a 0.22 μm microporous filter membrane, and the filtrate was used to determine the concentrations of liposoluble active substances, glabridin and ceramide, in the receiving solution filtered through the 0.22 μm filter membrane by high performance liquid chromatography, and the skin retention amount of the drug was calculated. The skin retention amount of the liposoluble active substance was calculated according to the following formula: Q s = VC / A

[0263] In the formula, Qs is the skin retention amount of glabridin per unit area (μg / cm 2 ); V is the total volume of the skin extract (ml); A is the effective diffusion area (cm 2 ); C is the concentration of the drug in the skin extract (μg / ml).

[0264] See the experimental results in Figure 9 、 Figure 10 、 Figure 11 、 Figure 12 : It can be seen from the cumulative permeation curves of Figure 9 and Figure 11 that the cumulative permeation amounts of Example 1 and Example 20 are significantly higher than those of Comparative Example 15 and Comparative Example 18, respectively. It can be seen from the 24 h cumulative permeation amount and skin retention amount of Figure 10 that the 24 h cumulative permeation amounts of the glabridin liposome of Example 1 and the free glabridin solution of Comparative Example 15 are: 22.86 μg / cm 2 、11.09 μg / cm 2 , and their 24 h skin retention amounts are: 10.31 μg / cm 2 、5.89 μg / cm 2 , respectively. It can be seen that the permeation amount and retention amount of the glabridin liposome are higher than those of the free glabridin solution, and are increased by 2.06 times and 1.75 times respectively compared with the free glabridin. It can be seen from the 24 h cumulative permeation amount and skin retention amount of Figure 12 that the 24 h cumulative permeation amounts of the ceramide liposome essence of Example 20 and the free ceramide essence of Comparative Example 18 are: 2.01 μg / cm 2 、0.99 μg / cm 2, and their skin retention amounts at 24 h were respectively: 6.10 μg / cm 2 , 3.69 μg / cm 2 . It can be seen that both the permeation amount and the retention amount of the ceramide liposome essence are higher than those of the free ceramide essence, and are respectively increased by 2.03 times and 1.65 times compared with the free ceramide essence. The experimental results respectively conform to the action paths of the two fat-soluble active substances in the skin layer. Obviously, the liposome of the present application has the effect of improving the transdermal penetration and skin retention of fat-soluble active substances, thus making it have a more efficient skin care effect.

[0265] In-vivo skin transdermal penetration test of the liposome in Experimental Example 8

[0266] The relative transdermal penetration amounts of Example 19 of the present application and Comparative Example 17 were tested by confocal Raman spectroscopy. Six healthy volunteers aged 22-25 years were selected to participate in this test. Before the test, the subjects washed their bilateral forearms with clean water, exposed their forearms in a constant temperature and humidity environment (24°C ± 1°C, 55% ± 5%) and balanced for 30 min. During the balancing process, two 2 cm × 2 cm areas were respectively marked on the inner sides of the bilateral forearms of the subjects, 3-5 cm away from the wrist, as the areas to be tested. Before using the sample, the spectral signals of the skin background of the areas to be tested of the 6 subjects were detected by a confocal Raman spectrometer at a laser wavelength of 660 nm, and the acquisition range was 4000-400 cm -1 . After the background signal acquisition was completed, test samples containing the same concentration of active ingredients were respectively applied to the test areas on the inner sides of the left and right arms of the subjects. Half an hour after applying the sample, the remaining products were cleaned with soft filter paper. Six test sites were randomly selected for each skin area of each volunteer 0.5 h after using the sample for repeated testing. The scanning depth was from 0 μm on the skin surface to 30 μm in the skin layer, the scanning step size was 2 μm, and the exposure time for detection at each depth was 2 seconds. In this way, the Raman spectra of human skin before and after using the sample were collected. All the collected spectra were processed with the original Raman spectral data by LabSpec 6.0 software according to the same method, and the exported results were statistically analyzed. The relative transdermal penetration values of the active substances in different samples were judged by qualitative and semi-quantitative analysis methods using a mathematical model, and the skin depth-glabridin relative penetration amount curves at different time points were plotted. When the relative transdermal penetration amount > 0 at a certain depth, it means that the fat-soluble active substance glabridin has penetrated at this skin depth; the larger the value of the relative amount, the greater the transdermal penetration amount of glabridin.

[0267] See the experimental results in Figure 13 and Figure 14:After using the glabridin liposome essence sample of Example 19 for 0.5 h, the Raman signal of glabridin was detected in the skin, and the relative penetration amount showed an upward trend at 16-22 μm in the skin, indicating that the liposome prepared in this application can carry the lipophilic active substance glabridin to deeper layers of the skin; at the same active substance concentration, within the depth of 0-30 μm of human skin, the relative penetration amount of the active substance in the glabridin essence of Example 19 was higher than that of the free glabridin essence of Comparative Example 17, with a significant difference (P<0.001), and the total relative penetration amount of the former at the depth of 0-30 μm was 3.82 times that of the latter. It shows that the liposome of this application has excellent percutaneous penetration performance in humans.

[0268] Experimental Example 9 Test on the in vitro release behavior of liposomes

[0269] The in vitro release experiment was carried out on Example 1 and Comparative Example 16 of this application by the dynamic dialysis method. The cellulose semipermeable membrane was used as the dialysis membrane (cut-off molecular weight 3500 Da), and 35% ethanol-saline was used as the release medium. Take 1 ml of the samples of Example 1 and Comparative Example 16 each and place them in a dialysis bag. Seal both ends and place them in a beaker containing 50 mL of the release medium. Shake and release at (37±0.5)°C and 100 r / min on a shaker. Take 1 mL of samples at 2 h, 4 h, 6 h, 8 h, 12 h, 24 h, 48 h, and 72 h, and quickly supplement 1 mL of isothermal fresh release medium. The concentration of the lipophilic active substance glabridin in the obtained samples was determined by high performance liquid chromatography, and the cumulative release amount and cumulative release percentage of the drug at different times were calculated. The cumulative release amount and cumulative release percentage of the lipophilic active substance were calculated according to the following formula:

[0270]

[0271]

[0272] Q n is the cumulative release amount at the nth sampling point; C n is the release concentration of glabridin at the nth sampling point; V0 is the volume of the release medium, V is the volume of each sampling; Q(%) is the cumulative release percentage, Q n is the cumulative release amount at the nth sampling point, and M is the content of the lipophilic active substance glabridin in the system.

[0273] See the experimental results Figure 15, at the same active ingredient concentration, the release rate of glabridin in Comparative Example 16 is much faster than that in Example 1; in Comparative Example 16, the free glabridin is released to 50.85% after 6 hours and then tends to be stable, while the encapsulated glabridin liposome in Example 1 is only released 31.19% after 6 hours and continues to be released for 72 hours, and the release level is the same as that of free glabridin. This component can be slowly released from the liposome, probably due to the strong encapsulation effect of the liposome. It shows that the liposome prepared in this application can achieve a continuous, slow and linear release process, which helps to reduce the irritation caused by the sudden increase in local content after application and achieve the effect of long-term slow release.

[0274] Experimental Example 10 Testing the Skin Whitening Effect of Liposomes

[0275] The liposomes of Example 19 and Comparative Example 17 of this application were used for human melanin testing with a skin red and melanin tester Mexameter MX18. Thirteen subjects aged 22 to 25 were selected and the samples of Example 19 and Comparative Example 17 were used respectively to observe the skin whitening effect of the subjects after 4 weeks of use. The specific method is as follows: an equal amount of the sample was applied to two parts of each subject's forearm every day, and a skin red and melanin tester was used to measure the melanin MI value of the forearm skin of each subject before using the sample and after 1, 2, 3, and 4 weeks of using the sample (measured 3 times and the average value was taken, with an error of ±5). A significant decrease in the MI value indicates a significant skin whitening and brightening effect.

[0276] The experimental results are shown in Figure 16 , the melanin content decreased significantly after 1 week of applying the glabridin liposome essence of Example 19, and the melanin content decreased by 13.7% after 4 weeks of application; compared with the free glabridin essence of Comparative Example 17, there was a significant difference between the two after 2 weeks of application, and the melanin content decline rate of the essence of Example 19 was 2.8 times that of the essence of Comparative Example 17 after 4 weeks of application. It shows that the liposome prepared in this application has a significant whitening effect and excellent skin care effect in cosmetic formulations.

[0277] The technical features of the above-described embodiments can be combined arbitrarily. For the sake of brevity of description, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, it should be considered as the scope described in this specification.

[0278] The above-described embodiments only represent several implementation manners of this application, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of this application, several deformations and improvements can still be made, and these all belong to the protection scope of this application. Therefore, the protection scope of the patent of this application should be subject to the appended claims, and the specification and drawings can be used to explain the content of the claims.

Claims

1. A liposome with good percutaneous permeability and suitable for encapsulating lipophilic active substances, characterized in that, The raw materials for preparing the liposomes include Phase A and Phase B, and are expressed as percentages based on the total mass of the liposomes (counted as 100% by mass). Phase A contains the following components in the following contents: Phase B contains the following components in the following contents: Polyol: 5% - 10.5%.

2. The liposome according to claim 1, wherein Based on the percentage of the total mass of the liposomes (counted as 100% by mass), Phase A contains the following components in the following contents: Phase B contains the following components in the following contents: Polyol: 7.5% - 10.5%.

3. The liposome according to claim 1 or 2, characterized in that, The liposomes further contain Phase C, and Phase C contains p-hydroxyacetophenone and pentylene glycol; wherein, the mass percentage of p-hydroxyacetophenone in the liposomes is 0.02% - 0.05%, and the mass percentage of pentylene glycol in the liposomes is 0.05% - 2%; Optionally, the mass percentage of p-hydroxyacetophenone in the liposomes is 0.03% - 0.05%, and the mass percentage of pentylene glycol in the liposomes is 1% - 2%.

4. The liposome according to claim 1 or 2, characterized in that, The mass ratio of the phospholipid to cholesterol is (20:1) - (40:1), and optionally, the mass ratio of the phospholipid to cholesterol is (25:1) - (35:1).

5. The liposome according to claim 1 or 2, characterized in that, The phospholipid is one or more of those containing 45% phosphatidylcholine, 75% phosphatidylcholine, and 90% phosphatidylcholine.

6. The liposome according to claim 1 or 2, characterized in that, The non-ionic surfactant includes one or more of methyl glucoside sesquistearate, Tween-80, cetearyl glucoside, polyglyceryl-10 laurate, polyglyceryl-10 myristate, behenyl polyether-25, steareth-21, cetearyl alcohol polyether-25, ceteth-20, oleth-20, and sucrose stearate; Optionally, the non-ionic surfactant is a polyether non-ionic surfactant, including one or more of behenyl polyether-25, steareth-21, cetearyl alcohol polyether-25, ceteth-20, and oleth-20; More preferably, the non-ionic surfactant is one or more of behenyl polyether-25, steareth-21, and cetearyl alcohol polyether-25.

7. The liposome according to claim 1 or 2, characterized in that, The liposomes satisfy one or more of the following conditions: (1) The polyol includes one or more of glycerol and butanediol; (2) The antioxidant includes one or more of pentaerythrityl tetra(bis-tert-butylhydroxyhydrocinnamate), vitamin E, and vitamin E acetate; (3) The lipophilic active substance includes one or more of glabridin, ceramide, bakuchiol, hydroxypinacolone retinoate, and resveratrol.

8. The method for preparing the liposome according to any one of claims 1 to 7, characterized in that, It includes the following steps: At a certain temperature, add the phospholipid to anhydrous ethanol and dissolve it, then add the lipophilic active substance, cholesterol, non-ionic surfactant, and antioxidant, and mix and dissolve them to prepare Phase A; Dissolve and preheat the polyol and water at a certain temperature to prepare Phase B; Inject Phase A into Phase B at a constant speed and stir to prepare a coarse emulsion dispersion; and After the coarse emulsion dispersion is cooled, perform high-pressure homogenization to prepare liposomes; Optionally, the preparation method further includes the step of adding p-hydroxyacetophenone and pentylene glycol to the liposomes and dispersing them evenly. Optionally, the certain temperature is 45°C to 85°C. Optionally, the certain temperature is 45°C to 65°C; Optionally, the stirring rate is 200 to 10,000 rpm, and the stirring time is 10 to 40 min; Preferably, the stirring rate is 400 to 600 rpm, and the stirring time is 15 to 30 min; Optionally, the homogenization pressure is 200 to 900 bar, and the number of homogenization times is 2 to 9 times; Preferably, the homogenization pressure is 300 to 500 bar, and the number of homogenization times is 3 to 5 times.

9. Use of the liposome according to any one of claims 1 to 7 in the preparation of cosmetics. Optionally, the cosmetics include one or more of essence, skin care lotion, emulsion, gel, cream and facial mask liquid.

10. A liposome essence with good percutaneous penetration performance, characterized in that, Comprising the liposome according to any one of claims 1 to 7.

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