A lees extract liposome, and a preparation method and use thereof

Abstract

The application discloses a kind of lees extract liposome and its preparation method and purposes.The lees extract liposome, including liposome membrane and the lees extract solution and polysaccharide enclosed in the liposome membrane, the raw material of the liposome membrane includes phospholipid, cholesterol and active protein;The active protein is selected from one or two of lees alcohol-soluble protein and rice protein;The polysaccharide is selected from one or two of arabinoxylan and beta-glucan.The lees extract liposome of the application can effectively load active ingredients in lees extract solution by constructing stable phospholipid bilayer structure, not only can protect active substances from oxidative degradation, but also can improve its transdermal diffusion rate, so as to enhance skin repair and function improvement effect.In addition, the lees extract liposome of the application can significantly regulate skin tactile friction behavior, so that the application process is more smooth, reduces adhesion, significantly improves experience.

Description

Technical Field

[0001] This invention relates to the field of fine chemicals, and in particular to a liposome derived from distillers' grains, its preparation method, and its uses. Background Technology

[0002] In recent years, with the continuous expansion of the global skincare market and the increasing health awareness of consumers, consumers' requirements for skincare products are no longer limited to basic functionality and safety, but also place greater emphasis on the tactile experience and skin feel quality during use. Tactile perception, as a core dimension of consumers' contact experience with skincare products, has received increasing attention. The spreading, application, and absorption of skincare products on the skin surface trigger different tactile sensations, such as stickiness, smoothness, or roughness. This type of tactile experience has become one of the important factors affecting product comfort and acceptance. There is a good correlation between skin tactile friction behavior and the skin feel of skincare products, such as spreadability, stickiness, and smoothness. Therefore, measuring the skin tactile friction coefficient can provide an objective and quantitative technical means for evaluating the skin feel of skincare products. However, current methods for controlling the tactile friction of skincare raw materials on the skin surface are still relatively limited, and there is a lack of material systems that can simultaneously address both active ingredient delivery and tactile optimization.

[0003] Distillers' grains are a byproduct of the fermentation and distillation of cereal crops. Their extracts or extracts are rich in various components such as protein, cellulose, organic acids, amino acids, vitamins, and phenolic compounds, and have been reported for use in the food and cosmetic fields. For example, Chinese patent CN109805393B discloses a distillers' grains extract and its preparation method. The method involves heating and stirring the distillers' grains in a mixed solvent of ethanol / water to extract and collect the extract. The extract is then loaded onto a solid-phase extraction column containing packing material, eluted, and dried to obtain a product capable of processing large quantities of distillers' grains and efficiently extracting antioxidant components, suitable for use as a food ingredient. Chinese patent CN117064824A discloses a method for preparing cosmetic raw materials using baijiu (Chinese liquor) lees extract. This method involves enzymatic hydrolysis, solid-liquid separation, and membrane separation to obtain a concentrated extract of lees with a molecular weight less than 10,000 Da. This concentrated extract is then formulated with glycerin and other substances to create a toner for repairing cell damage.

[0004] Polyphenols, as the main active components in sake lees extracts or extracts, have the effects of improving dry skin, maintaining skin elasticity, and enhancing skin's moisturizing ability. However, the unsaturated bonds in the molecular structure of polyphenols are easily degraded under light, heat, and oxidative conditions, leading to a decrease in activity. Furthermore, while directly adding sake lees extracts or extracts to skincare products can improve skin condition to some extent, it has limitations such as low transdermal absorption efficiency and poor stability, thus restricting its further application in skincare.

[0005] Liposomes are vesicle structures composed of a phospholipid bilayer, similar in structure to biological membranes. They possess advantages such as good biocompatibility, biodegradability, and low toxicity, thus finding wide application in pharmaceuticals, food, and skincare products. Liposomes exhibit natural biocompatibility, strong cell regulation capabilities, and targeted delivery characteristics, making them considered excellent carriers of active ingredients. By loading active ingredients into liposomes, the stability of active substances can be improved to a certain extent, and their delivery efficiency in the skin can be enhanced. Chinese patent CN120923351 discloses a polyphenol-modified lipid material for encapsulating small molecule drugs and achieving targeted delivery; Chinese patent CN121265810A discloses a milk thistle nanoliposome with excellent long-lasting sustained-release effects. These technologies mainly focus on the protective and delivery functions of liposomes for active ingredients.

[0006] However, directly mixing sake lees extract with substances like glycerin for use in skincare products presents challenges such as poor stability, low transdermal efficiency, and insufficient skin feel regulation. Current liposome technology primarily targets purified substances with relatively simple components. When unpurified sake lees extract is directly introduced into liposome preparation systems, large molecular impurities (such as proteins and polysaccharides) may interact with the liposome surface or phospholipids through adsorption, affecting the ordered structure of the phospholipid bilayer. Simultaneously, electrolytes and organic acids in the extract may alter the ionic strength and pH of the medium, weakening the hydration layer and electrostatic repulsion on the phospholipid membrane surface, leading to liposome aggregation, fusion, or even lysis during preparation or storage, resulting in precipitation and aggregation. Furthermore, due to the complex composition of the extract, the surface properties of liposomes (such as charge and hydration layer thickness) are difficult to design specifically, potentially causing unpleasant skin feel such as stickiness, blockage, or dryness when spread on the skin. Existing research largely focuses on single active ingredient systems, with limited studies on the compatibility with complex natural extract systems.

[0007] How to proactively optimize the spreadability, smoothness, and post-absorption dryness of formulations through material structure design while ensuring the stability and delivery function of liposome membranes is a problem that has not yet been solved by existing technologies. Summary of the Invention

[0008] To address the problems of poor stability, low transdermal efficiency, and insufficient skin feel regulation of sake lees extract in existing skin care technologies, this invention provides a sake lees extract liposome, its preparation method, and its uses to overcome the shortcomings of existing technologies. The liposomes of this invention effectively load sake lees extract by constructing a stable phospholipid bilayer structure. This not only protects the active substances in the sake lees extract from oxidative degradation but also effectively improves its transdermal diffusion rate, thereby enhancing skin repair and functional improvement effects. Simultaneously, the sake lees extract liposomes of this invention can significantly regulate skin tactile friction behavior, making the application process smoother, reducing adhesion, and significantly improving the user experience.

[0009] To achieve this objective, the present invention adopts the following technical solution:

[0010] A first aspect of the present invention provides a liposome of a distiller's grains extract, comprising a liposome membrane and a distiller's grains extract and polysaccharides encapsulated within the liposome membrane, wherein the raw materials of the liposome membrane include phospholipids, cholesterol and active proteins;

[0011] The active protein is selected from one or two of distillers' grains prolysin and rice protein;

[0012] The polysaccharide is selected from one or both of arabinoxylan and β-glucan.

[0013] Another aspect of the present invention provides a method for preparing liposomes from distillers' grains extract as described above, comprising the following steps:

[0014] a) Lecithin, cholesterol, active protein, and organic solvent are mixed for the first time to obtain the oil phase; the distiller's grains extract and polysaccharide are mixed for the second time to obtain the aqueous phase;

[0015] b) The oil phase and the aqueous phase are mixed for a third time to obtain liposomes of the distillers' grains extract.

[0016] Another aspect of the present invention provides the use of the lees extract liposomes as described above in the preparation of cosmetics.

[0017] Another aspect of the present invention provides a cosmetic comprising liposomes of sake lees extract as described above.

[0018] Another aspect of the present invention provides the use of the lees extract liposomes as described above or the cosmetics as described above in at least one of the following:

[0019] C1) Enhances the skin's antioxidant capacity;

[0020] C2) Improves the skin's moisturizing ability;

[0021] C3) Improves skin tactile sensation.

[0022] Another aspect of the present invention provides the use of active proteins and polysaccharides in improving the stability of liposomes from distillers' grains extract, wherein the active protein is selected from one or two of distillers' grains prolysin and rice protein; and the polysaccharide is selected from one or two of arabinoxylan and β-glucan.

[0023] Compared with the prior art, the present invention has the following advantages:

[0024] (1) The present invention significantly improves the stability and transdermal efficiency of the distillers' grains extract by liposome encapsulation;

[0025] (2) The liposomes of the wine lees extract of the present invention form a biomimetic lubrication system that can regulate the skin's tactile friction, which significantly improves the skin feel when applying skin care products;

[0026] (3) This invention reveals the mechanism of action between liposome structural parameters and skin friction behavior, providing a theoretical basis for skin tactile friction regulation technology;

[0027] (4) The raw materials for constructing liposomes from the lees extract of the present invention are green and the process is highly feasible. It is suitable for industrial production and has good market application prospects.

[0028] (5) The present invention encapsulates the active ingredients of the distillers' grains extract into the liposome membrane by using polysaccharides and active proteins, thereby improving the stability of the liposome membrane. No stratification or deposition aggregation occurred within 7 days of standing at room temperature. Attached Figure Description

[0029] Figure 1 This is a standard curve for determining the polyphenol content in the lees extract in the examples.

[0030] Figure 2a These are polarized light microscope images of the liposomes of the distillers' grains extract prepared in Examples 1-4 and Comparative Examples 1-3.

[0031] Figure 2b This is a TEM image of the liposomes of the distillers' grains extract prepared in Example 2.

[0032] Figure 3 The rheological properties of the liposomes prepared from the distillers' grains extract in Examples 1-4 and Comparative Examples 1-3 are shown in the diagram.

[0033] Figure 4 The graph shows the DPPH free radical scavenging rate of liposomes from the distillers' grains extract and the distillers' grains extracts of Examples 1-4 and Comparative Examples 1-3.

[0034] Figure 5 The graph shows the hydration results of the liposomes of the distillers' grains extract and the distillers' grains extracts of Examples 1-4 and Comparative Examples 1-3.

[0035] Figure 6 This is a schematic diagram of the Franz diffusion cell in Application Example 1.

[0036] Figure 7a The graph shows the cumulative permeation of the distiller's grains extract in Application Example 1.

[0037] Figure 7b The graph shows the cumulative permeation of liposomes from the distillers' grains extract in Examples 1-4 and Comparative Examples 1-3 of Application Example 1.

[0038] Figure 8 This is a schematic diagram of the three-dimensional force sensor used in Application Example 2.

[0039] Figure 9a The graph shows the friction coefficient results between the liposomes of the distillers' grains extract in Examples 1-4 and Comparative Examples 1-3 and the skin.

[0040] Figure 9b This is a graph showing the change in the coefficient of friction between the liposomes of the lees extract in Example 2 and the skin over time.

[0041] Figure 10 These are actual photographs taken after the liposomes of the distillers' grains extract prepared in Examples 1-4 and Comparative Examples 1-3 have been placed at room temperature for 7 days. Detailed Implementation

[0042] The active substances in distillers' grains extract include polyphenols and polysaccharides. Polyphenols have a large molecular weight (up to several thousand Daltons) and contain multiple hydroxyl groups, making them prone to hydrogen bonding, resulting in strong hydrophilicity and poor lipid solubility. The stratum corneum of the skin is mainly composed of lipids, forming a hydrophobic barrier that hinders the penetration of hydrophilic molecules. Therefore, polyphenols have difficulty directly penetrating the stratum corneum to enter the dermis. This paper addresses the problems of low transdermal efficiency and poor oxidative stability of polyphenols in distillers' grains extract in existing technologies.

[0043] This invention provides a liposome of distillers' grains extract, its preparation method, and its uses. Using distillers' grains extract as the active core, solid nanoparticles (rice protein peptides, distillers' grains prolyl protein) are used to modify lecithin and cholesterol to construct liposome carriers, resulting in liposomes of distillers' grains extract with uniform morphology, high encapsulation efficiency, and good skin compatibility. This solves the technical problems of poor stability and low transdermal efficiency of polyphenolic active ingredients in distillers' grains extract, as well as the lack of targeted regulation of skin tactile friction and poor skin feel in existing liposomes.

[0044] A first aspect of the present invention provides a liposome of a distiller's grains extract, comprising a liposome membrane and a distiller's grains extract and polysaccharides encapsulated within the liposome membrane, wherein the raw materials of the liposome membrane include phospholipids, cholesterol and active proteins;

[0045] The active protein is selected from one or two of distillers' grains prolysin and rice protein;

[0046] The polysaccharide is selected from one or both of arabinoxylan and β-glucan.

[0047] This invention innovatively introduces active proteins into the membrane structure of liposomes, constructing a composite interface membrane that combines high mechanical strength with good lubricity. This composite interface membrane not only effectively encapsulates and protects the polyphenolic active substances in the internal distillers' grains extract, but also interacts directly with the stratum corneum of the skin, optimizing the tribological behavior during the application process from the source.

[0048] Furthermore, the liposome membrane of the present invention is encapsulated with polysaccharides. On the one hand, the polysaccharides prevent the sedimentation and aggregation of liposomes; on the other hand, the unique rheological properties of polysaccharides and active proteins endow the liposomes of sake lees extract with a smooth, light and non-sticky excellent skin feel, achieving a perfect unity of efficacy and experience.

[0049] Therefore, the liposomes of the sake lees extract of the present invention can form a controllable lubricating layer similar to a liquid crystal state during the application process, thereby improving the skin feel during the use of skin care products, including smoothness, spreadability and low stickiness.

[0050] In some embodiments, the liposomes from the distillers' grains extract are nanoemulsions with a particle size of <65 nm, preferably <50 nm. They are water-in-oil suspension liposome emulsions.

[0051] In some embodiments, the liposomes of the distillers' grains extract, when observed under a polarizing microscope, exhibit a Maltese cross pattern, an optical feature confirming the formation of a laminar liquid crystal structure. According to prior art reports, skincare products with a laminar liquid crystal structure offer significant advantages over ordinary skincare products: First, the laminar liquid crystal structure forms an ordered protective layer at the oil-water interface, effectively inhibiting droplet aggregation and delaying phase separation, thus exhibiting excellent storage stability; second, the stacking pattern of the laminar liquid crystal is highly similar to the laminar structure of intercellular lipids in the stratum corneum of human skin, therefore this structure has good skin compatibility and can promote the fusion and penetration of active ingredients at the skin interface; third, the laminar liquid crystal structure can encapsulate active ingredients, thereby improving their stability and slowing their release, maximizing their effectiveness; fourth, the laminar liquid crystal structure provides good spreadability and lubrication during application, while avoiding the sticky, heavy, and other unpleasant sensations common in traditional thickening systems, achieving a refreshing and non-sticky user experience. This invention utilizes polysaccharides and active proteins to successfully construct liposomes of distillers' grains extract with a layered liquid crystal structure without the need for additional thickeners. This system exhibits high viscosity, good spreadability, and excellent stability. Under a polarizing microscope, it displays a distinct Maltese cross structure, indicating its complete ordered liquid crystal structure and potential as a functional cosmetic matrix.

[0052] In some embodiments, the phospholipid is selected from one or more of soybean phospholipids, lecithin, hydrogenated soybean phospholipids, hydrogenated lecithin, phosphatidylethanolamine, distearate phosphatidylcholine, and polyethylene glycol-derived phospholipids. In one specific embodiment, it is soybean phospholipid.

[0053] In some embodiments, the relative molecular weight of the rice protein peptide is 500-1000 Da. The rice protein peptide of the present invention is produced from rice as a single raw material through processes such as extraction, enzymatic hydrolysis and / or microbial fermentation, filtration, sterilization, and drying. It is a product with peptides having a relative molecular mass of 500-1000 Da as its main component, and it conforms to the provisions of GB31611 "National Food Safety Standard for Plant Protein Peptides for Food Processing".

[0054] In some embodiments, the relative molecular weight of the arabinoxylan is 5-65 kDa. The arabinoxylan of the present invention is produced from sugarcane bagasse through processes such as washing, pressing, sodium hydroxide extraction, precipitation, purification, and drying.

[0055] In some embodiments, the relative molecular weight of the β-glucan is 10-70 kDa. Preferably, it is oat β-glucan.

[0056] In some embodiments, the raw materials of the distillers' grains extract liposomes comprise the following components in weight percentages: 10-20 wt% lecithin and cholesterol, 0.05-0.2 wt% active protein, 0.01-0.1 wt% polysaccharide, with the balance being distillers' grains extract; the mass ratio of lecithin to cholesterol is 30:(3-7).

[0057] In some embodiments, the raw materials for the distillers' grains extract liposomes contain 10 wt%, 12 wt%, 14 wt%, 16 wt%, 18 wt%, and 20 wt% lecithin and cholesterol.

[0058] In some embodiments, the raw materials for the distillers' grains extract liposomes comprise 0.05 wt%, 0.08 wt%, 0.1 wt%, 0.12 wt%, 0.14 wt%, 0.16 wt%, and 0.2 wt% of active proteins.

[0059] In some embodiments, the raw materials for the distillers' grains extract liposomes contain 0.01 wt%, 0.02 wt%, 0.03 wt%, 0.06 wt%, 0.08 wt%, and 0.1 wt% polysaccharides.

[0060] In some embodiments, the mass ratio of phospholipids to cholesterol is 30:(3-7), or it can be 30:(3-5.5), or it can be 30:(5-7), or it can be 30:3, 30:4, 30:5, 30:6, or 30:7.

[0061] In some embodiments, the method for preparing the distiller's grains extract is as follows:

[0062] A1) The distillers' grains powder was degreased using n-hexane to obtain degreased distillers' grains powder;

[0063] A2) Mix water with the defatted distiller's grains powder and sonicate to separate the solid and liquid, and take the supernatant to obtain the distiller's grains extract.

[0064] The applicant screened the solvents used for degreasing. When ethanol was used as the solvent, the ethanol and the lees powder were mixed together and difficult to separate, resulting in a significant loss of the effective components of the lees powder. When petroleum ether was used as the solvent, the petroleum ether was not easy to remove and would remain in the lees extract. However, when hexane was used for degreasing, it had high selectivity for polyphenols, and the degreasing process resulted in clear stratification. In addition, hexane was easy to recover.

[0065] In some implementations, the degreasing process is performed more than once, typically three times.

[0066] In some embodiments, the defatting process further includes the removal of n-hexane. Methods for removing n-hexane include, but are not limited to, vacuum drying; those skilled in the art can select the most suitable method based on specific experimental conditions and requirements. In one specific embodiment, n-hexane is removed under a vacuum of -0.09 MPa at room temperature.

[0067] In some embodiments, in A1), the particle size of the lees powder is about 150-180 μm.

[0068] In some embodiments, in A1), the mass-to-volume ratio of the distiller's grains powder to n-hexane can be 1g:(1-10)mL, 1g:(1-5)mL, 1g:(4-6)mL, 1g:(5-10)mL, 1g:1mL, 1g:2mL, 1g:3mL, 1g:4mL, 1g:5mL, 1g:6mL, 1g:7mL, 1g:7mL, 1g:8mL, 1g:9mL, or 1g:10mL.

[0069] In some embodiments, in A1), the method for preparing the lees powder is as follows: drying and pulverizing the original lees liquid to obtain the lees powder.

[0070] In some specific embodiments, the drying temperature is 20-60°C, or it can be 20-35°C, or it can be 28-48°C, or it can be 45-60°C, or it can be 45°C.

[0071] In some specific embodiments, the process includes rinsing twice with deionized water before drying to remove surface impurities.

[0072] In some embodiments, in A2), the volume-to-mass ratio of water to defatted distiller's grains powder is (10-30) mL: 1 g, or it can be (10-21) mL: 1 g, or it can be (18-22) mL: 1 g, or it can be (21-30) mL: 1 g, or it can be 10 mL: 1 g, 15 mL: 1 g, 18 mL: 1 g, 20 mL: 1 g, 22 mL: 1 g, 25 mL: 1 g, or 30 mL: 1 g.

[0073] In some embodiments, in A2), the ultrasonic treatment temperature is 30-60℃, or 30-55℃, or 50-60℃, or 30℃, 40℃, 45℃, 50℃, 55℃, or 60℃. In this application, the ultrasonic treatment temperature cannot be too high or too low. Ultrasonic treatment temperatures above 60℃ will damage the polyphenol and polysaccharide content in the distiller's grains extract, while temperatures below 40℃ will cause polyphenols and polysaccharides to degrade and transform into other forms, such as quinones, melanoidins (brown substances), aglycones, and oligosaccharides.

[0074] In some embodiments, in A2), the ultrasonic treatment power is 400-600 W, and the extraction time is 40-80 min. Preferably, it is 500 W and 60 min.

[0075] In some embodiments, in A2), the solid-liquid separation is performed by centrifugation at 4°C and 10,000 g.

[0076] In some embodiments, in A2), the supernatant is filtered, and the microporous membrane used for filtration has a pore size of 0.22 μm.

[0077] In some embodiments, the relative molecular weight of the alcohol-soluble protein is 10-30 kDa.

[0078] In some embodiments, the method for preparing the prolysin from distiller's grains is as follows:

[0079] B1) The lees powder and water are subjected to a first ultrasonic treatment to separate the solid and liquid, and the first filter residue is obtained.

[0080] B2) The first filter residue and sodium chloride aqueous solution are subjected to a second ultrasonic treatment to separate the solid and liquid, and a second filter residue is obtained.

[0081] B3) The second filter residue and alcohol solution are subjected to a third ultrasonic treatment, solid-liquid separation, concentration, and drying to obtain the alcohol-soluble protein from the wine lees.

[0082] In one specific implementation method, the preparation method of alcohol-soluble protein from distiller's grains is as follows:

[0083] A certain amount of distiller's grains raw material was pretreated. The pretreatment included drying the distiller's grains in an oven, grinding the completely dried distiller's grains into powder using a grinder, passing the powder through a 60-mesh sieve, and storing the fine powder in a sealed bag for later use, thus obtaining distiller's grains powder. Deionized water was added at a material-to-liquid ratio of 1:15 (g / mL), and the ultrasonic power was set to 250 W, the ultrasonic temperature to 50℃, and the ultrasonic time to 30 min. The mixture was then centrifuged at 4000 r / min for 20 min, and the first filter residue was obtained. 5% NaCl solution was added at a material-to-liquid ratio of 1:14 (g / mL) to remove globulins. The ultrasonic power was set to 300 W, the ultrasonic temperature to 30℃, and the ultrasonic time to 40 min. The mixture was then centrifuged, and the supernatant obtained was the globulin solution. The residue was collected by filtering through gauze. Add 65% ethanol solution to the residue, set the material-to-liquid ratio to 1:16 (g / mL), the ultrasonic power to 240 W, the ultrasonic temperature to 35℃, and the ultrasonic time to 60 min, extract the alcohol-soluble protein, concentrate it under reduced pressure in a rotary evaporator at 42℃ water bath, place the concentrate in a 90 mm petri dish, and freeze-dry it in a vacuum freeze dryer at -53℃ for 24 h to obtain the alcohol-soluble protein from the distillers' grains.

[0084] Another aspect of the present invention provides a method for preparing liposomes from distillers' grains extract as described above, comprising the following steps:

[0085] a) Lecithin, cholesterol, active protein, and organic solvent are mixed for the first time to obtain the oil phase; the distiller's grains extract and polysaccharide are mixed for the second time to obtain the aqueous phase;

[0086] b) The oil phase and aqueous phase are mixed for a third time and homogenized to obtain the liposomes of the distillers' grains extract.

[0087] In some embodiments, the mass-to-volume ratio of the active protein to the organic solvent is 1g:(1000-3000)mL, or it can be 1g:1000mL, 1g:2000mL, or 1g:3000mL.

[0088] In some embodiments, the volume ratio of the oil phase to the water phase is 1:(0.01-0.5), or it can be 1:0.01, 1:0.1, 1:0.2, 1:0.3, 1:0.4, or 1:0.5.

[0089] In some embodiments, the organic solvent is selected from one or both of anhydrous ethanol and chloroform.

[0090] In some embodiments, the temperature of the first mixing is 40-60°C, or it can be 40°C, 45°C, 50°C, 55°C, or 60°C. The temperature of the first mixing should not be too high or too low; if it is too high, the liposomes will degrade and the antioxidant properties of the polyphenols will be compromised, while if it is too low, the dissolution will be slow.

[0091] In some embodiments, the temperature of the second mixing is 40-60°C, or it can be 40°C, 45°C, 50°C, 55°C, or 60°C.

[0092] In some embodiments, the temperature of the third mixing is 40-60°C, or it can be 40°C, 45°C, 50°C, 55°C, or 60°C.

[0093] In some embodiments, the first, second, and third mixing processes are performed by stirring. In some specific embodiments, the magnetic stirring speed is 400 rpm / min.

[0094] In some specific embodiments, the homogenization speed is 4000-6000 rpm / min, and the homogenization time is 1-10 min. The homogenization speed can also be 4000 rpm / min, 4500 rpm / min, 5000 rpm / min, 5500 rpm / min, or 6000 rpm / min; and the homogenization time can be 1 min, 2 min, 4 min, 6 min, 8 min, or 10 min.

[0095] In some embodiments, homogenization further includes a post-processing step, which is vacuum evaporation.

[0096] In some specific embodiments, the temperature of the vacuum evaporation is 60~65°C, or it can be 60°C, 61°C, 62°C, 63°C, 64°C, or 65°C. The temperature of vacuum evaporation should not be too high, otherwise it will cause the solvent (especially ethanol) to boil violently and the liposomes to be lost.

[0097] In some specific embodiments, the time for vacuum evaporation is 30-50 min, or it can be 30 min, 40 min, or 50 min.

[0098] In some specific embodiments, the vacuum degree of the reduced pressure evaporation is 0.01-0.2 MPa, or it can be 0.01 MPa, 0.05 MPa, 0.07 MPa, 0.09 MPa, 0.1 MPa, 0.15 MPa, 0.17 MPa, or 0.2 MPa.

[0099] Another aspect of the present invention provides the use of the lees extract liposomes as described above in the preparation of cosmetics.

[0100] In some embodiments, the cosmetic is a skin care preparation, which is one or more of the following: solution, suspension, emulsion, cream, ointment, gel, powder, and spray. Preferably, it is an emulsion. The cosmetic can be further prepared into product forms such as face masks, sprays, and powders from the skin care preparation.

[0101] Another aspect of the present invention provides a cosmetic comprising liposomes of sake lees extract as described above.

[0102] Another aspect of the present invention provides the use of the lees extract liposomes as described above or the cosmetics as described above in at least one of the following:

[0103] C1) Enhances the skin's antioxidant capacity;

[0104] C2) Improves the skin's moisturizing ability;

[0105] C3) Improves skin tactile sensation.

[0106] In some embodiments, the antioxidant effect refers to enhancing the skin's ability to scavenge DPPH free radicals. This invention found that, compared to sake lees extract, sake lees extract liposomes exhibited approximately 1.74 times higher DPPH free radical scavenging rate at 12 hours.

[0107] In some embodiments, skin moisturizing refers to increasing skin hydration. This invention has found that applying sake lees extract liposomes to the skin can increase skin hydration by 36.59% after 1 hour and still increase it by 33.41% after 24 hours.

[0108] In some embodiments, improving skin feel refers to reducing the coefficient of friction and enhancing skin smoothness; improving extensibility and enhancing the spreading experience. This invention has discovered that applying sake lees extract liposomes to the skin can reduce the coefficient of friction and improve skin feel.

[0109] Another aspect of the present invention provides the use of active proteins and polysaccharides in improving the stability of distillers' grains extract, wherein the active protein is selected from one or two of distillers' grains prolysin and rice protein; and the polysaccharide is selected from one or two of arabinoxylan and β-glucan.

[0110] Compared to adding active proteins or polysaccharides alone, the liposomes formed by simultaneously adding active proteins and polysaccharides exhibit significantly enhanced antioxidant and moisturizing properties, as well as superior skin penetration. After standing at room temperature for 7 days, no sedimentation, stratification, or aggregation was observed, demonstrating excellent storage stability. Furthermore, the liposomes of this invention effectively reduce the coefficient of friction on the skin surface during use, providing a silky, lightweight skin feel. The liposome system of this invention can be used as a functional carrier in various skin care preparations such as serums, lotions, and creams.

[0111] Before further describing specific embodiments of the present invention, it should be understood that the scope of protection of the present invention is not limited to the specific embodiments described below; it should also be understood that the terminology used in the embodiments of the present invention is for describing specific embodiments and not for limiting the scope of protection of the present invention. Test methods in the following embodiments that do not specify specific conditions are generally performed under conventional conditions or as recommended by the respective manufacturers.

[0112] When numerical ranges are given in the embodiments, it should be understood that, unless otherwise stated in the present invention, both endpoints of each numerical range and any value between the two endpoints may be selected. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art. In addition to the specific methods, apparatus, and materials used in the embodiments, based on the knowledge of the prior art possessed by one of ordinary skill in the art and the description of this invention, any prior art methods, apparatus, and materials similar to or equivalent to those described, apparatus, and materials in the embodiments of this invention may be used to implement the present invention.

[0113] Unless otherwise stated, the experimental methods, detection methods, and preparation methods disclosed in this invention all employ conventional techniques in molecular biology, biochemistry, chromatin structure and analysis, analytical chemistry, cell culture, recombinant DNA technology, and related fields.

[0114] The preparation method of distiller's grains extract includes the following steps:

[0115] 1) Take fresh rice wine lees (purchased from Bright Food (Shanghai) Co., Ltd.), rinse twice with deionized water, and dry to constant weight to obtain dried lees; crush the dried lees with a pulverizer and pass through an 80-mesh sieve (particle size of 200μm) to obtain lees powder.

[0116] 2) The lees powder and n-hexane (HPLC grade) were mixed at a mass-volume ratio of 1g:5mL and defatted (repeated 3 times) to remove the n-hexane and obtain defatted lees powder; the n-hexane was removed under a vacuum of -0.09Mpa at room temperature.

[0117] 3) Mix deionized water and defatted distiller's grains powder at a volume-to-mass ratio of 20 mL / g, and extract by ultrasonication at 50℃ and 500W for 60 min. Then centrifuge at 4℃ and 10000 g, and filter the supernatant through a 0.22 μm microporous membrane to obtain the distiller's grains extract.

[0118] The determination of polyphenol content in distiller's grains extract includes the following steps:

[0119] Take 0.2 mL of gallic acid standard solution with a concentration range of 30-300 μg / mL, add 1.0 mL of 10 v / v% Folin-Ciocalteu solution, mix well, and react in the dark for 10 min. Then add 0.5 mL of 7.5% sodium carbonate solution, shake to mix, and incubate at room temperature in the dark for 45 min. Measure the absorbance at 756 nm, and perform three replicates. Plot a standard curve with gallic acid standard solution concentration on the x-axis and absorbance on the y-axis. Take 0.2 mL of distiller's grains extract and measure the OD following the same steps. 756Substitute the values ​​into the standard curve (y = 0.00374x + 0.00414, R0). 2 =0.99899, ​​where x represents the concentration in μg / mL and y represents the absorbance. See [reference needed]. Figure 1 Calculate the polyphenol content, and express the result as gallic acid equivalent per gram of dry matter.

[0120] The polyphenol concentration (x, μg / mL) of the test solution was calculated by substituting the measured absorbance (y) into the standard curve equation. The measured polyphenol content in the distiller's grains extract was 64.03 μg / mL.

[0121] Rice protein peptides (Jiangxi Hengding Food Co., Ltd.), distillers' grains prolyl protein (self-made), arabinoxylan (Xi'an Laina Biotechnology Co., Ltd.), and oat β-glucan (Guangzhou Zhongkang Food Co., Ltd.) are white to light yellow powders, with β-glucan content ≥80%.

[0122] Example 1: Liposomes from Distillers' Grains Extract and Their Preparation Method

[0123] Example 1 provides liposomes from distillers' grains extract and their preparation method. The preparation method includes the following steps:

[0124] 1) Dissolve 0.1 g of rice protein peptide, 15 g of soybean lecithin and 2.5 g of cholesterol in 200 mL of anhydrous ethanol, heat in a water bath at 55 °C and stir until completely dissolved, and use as the oil phase.

[0125] Take 100 mL of distiller's grains extract and 0.05 g of arabinoxylan, stir to dissolve, and preheat to 55°C to prepare the aqueous phase.

[0126] 2) The aqueous phase was added dropwise to the oil phase using a syringe, and the mixture was stirred at 600 rpm / min for 30 min. Then, it was homogenized at 5000 rpm / min for 5 min, followed by stirring with a magnetic stirrer at 400 rpm / min and cooling to room temperature. Finally, ethanol was removed by vacuum evaporation at 0.1 MPa and 60℃ to obtain liposomes from the distillers' grains extract.

[0127] The materials used in its preparation process are shown in Table 1.

[0128] Example 2: Liposomes from Distillers' Grains Extract and Their Preparation Method

[0129] Example 2 provides liposomes from distillers' grains extract and their preparation method. The preparation method includes the following steps:

[0130] 1) Dissolve 0.1 g of distillers' grains protein, 15 g of soybean lecithin and 2.5 g of cholesterol in 200 mL of anhydrous ethanol, heat in a water bath at 55 °C and stir until completely dissolved, and use as the oil phase.

[0131] Take 100 mL of distiller's grains extract and 0.05 g of arabinoxylan, stir to dissolve, and preheat to 55°C to prepare the aqueous phase.

[0132] 2) The aqueous phase was added dropwise to the oil phase using a syringe, and the mixture was stirred at 600 rpm / min for 30 min. Then, it was homogenized at 5000 rpm / min for 5 min, followed by stirring with a magnetic stirrer at 400 rpm / min and cooling to room temperature. Finally, ethanol was removed by vacuum evaporation at 0.1 MPa and 60℃ to obtain liposomes from the distillers' grains extract.

[0133] The materials used in its preparation process are shown in Table 1.

[0134] Example 3: Liposomes from Distillers' Grains Extract and Their Preparation Method

[0135] Example 3 provides liposomes from distillers' grains extract and their preparation method. The preparation method includes the following steps:

[0136] 1) Dissolve 0.1 g of rice protein peptide, 15 g of soybean lecithin and 2.5 g of cholesterol in 200 mL of anhydrous ethanol, heat in a water bath at 55 °C and stir until completely dissolved, and use as the oil phase.

[0137] Take 100 mL of distiller's grains extract and 0.05 g of β-glucan, stir to dissolve, and preheat to 55°C to prepare the aqueous phase.

[0138] 2) The aqueous phase was added dropwise to the oil phase using a syringe, and the mixture was stirred at 600 rpm / min for 30 min. Then, it was homogenized at 5000 rpm / min for 5 min, followed by stirring with a magnetic stirrer at 400 rpm / min and cooling to room temperature. Finally, ethanol was removed by vacuum evaporation at 0.1 MPa and 60℃ to obtain liposomes from the distillers' grains extract.

[0139] The materials used in its preparation process are shown in Table 1.

[0140] Example 4: Liposomes from Distillers' Grains Extract and Their Preparation Method

[0141] Example 4 provides liposomes from distillers' grains extract and their preparation method. The preparation method includes the following steps:

[0142] 1) Dissolve 0.1 g of rice protein peptide, 15 g of soybean lecithin and 2.5 g of cholesterol in 200 mL of anhydrous ethanol, heat in a water bath at 55 °C and stir until completely dissolved, and use as the oil phase.

[0143] Take 100 mL of distiller's grains extract and 0.05 g of β-glucan, stir to dissolve, and preheat to 55°C to prepare the aqueous phase.

[0144] 2) The aqueous phase was added dropwise to the oil phase using a syringe, and the mixture was stirred at 600 rpm / min for 30 min. Then, it was homogenized at 5000 rpm / min for 5 min, followed by stirring with a magnetic stirrer at 400 rpm / min and cooling to room temperature. Finally, the ethanol was removed by vacuum evaporation at 0.1 MPa and 65℃ to obtain the liposomes from the distillers' grains extract.

[0145] The materials used in its preparation process are shown in Table 1.

[0146] Table 1 Raw materials for preparation

[0147]

[0148] Comparative Example 1

[0149] The difference between Comparative Example 1 and Example 2 is that no arabinoxylan was added to the aqueous phase, and no distillers' grains-soluble protein was added to the oil phase; all other aspects were the same as in Example 1. The materials used in the preparation process are shown in Table 1.

[0150] Comparative Example 2

[0151] The difference between Comparative Example 2 and Example 2 is that arabinoxylan was not added to the aqueous phase; otherwise, they are the same as in Example 1. The materials used in the preparation process are shown in Table 1.

[0152] Comparative Example 3

[0153] The difference between Comparative Example 3 and Example 2 is that no alcohol-soluble protein from distillers' grains was added to the oil phase; all other aspects are the same as in Example 1. The materials used in its preparation process are shown in Table 1.

[0154] Example 5 Characterization of liposomes from distillers' grains extract

[0155] 5.1 Encapsulation efficiency determination

[0156] The liposome emulsions of distillers' grains extract obtained in Examples 1-4 and Comparative Examples 1-3 were transferred to ultrafiltration centrifuge tubes and centrifuged at 7500×g for 10 min at 4°C. The lower filtrate of the ultrafiltration centrifuge tubes was collected, and the content of gallic acid (GA) was determined by spectrophotometry (see the determination of polyphenol content in distillers' grains extract). The encapsulation efficiency was calculated according to the following equation:

[0157]

[0158] In the formula, C 总 The concentration of gallic acid in the liposomes of distillers' grains extract, C 游离 This represents the concentration of free gallic acid after ultrafiltration.

[0159] The encapsulation rates of Examples 1-4 and Comparative Examples 1-3 are shown in Table 2.

[0160] Table 2 Encapsulation efficiency

[0161]

[0162] As shown in Table 2, the encapsulation efficiency of Comparative Examples 1-3 was less than 58%, while the encapsulation efficiency of the Examples was greater than 60%, with Example 2 having the highest encapsulation efficiency of 62.80%. Compared with Comparative Examples 1-3, the encapsulation efficiency of Examples 1-4 was significantly improved.

[0163] 5.2 Particle size and zeta potential determination

[0164] The particle size and zeta potential of the liposome emulsions of the distillers' grains extracts in Examples 1-4 and Comparative Examples 1-3 were determined using a dynamic light scattering instrument (Nano-ZS90, Malvern).

[0165] The lipid concentration of the liposome emulsions of the distillers' grains extract in each example and comparative example was diluted to 1 mg / mL with PBS (0.01M, pH 7.0) to avoid multiple scattering. The emulsions were equilibrated in the instrument for 180 s and tested at 25°C.

[0166] The particle size and zeta potential of Examples 1-4 and Comparative Examples 1-3 are shown in Table 3.

[0167] Table 3 Particle size and Zeta potential

[0168]

[0169] As shown in Table 3, compared with Comparative Examples 1-3, the liposomes from the distillers' grains extract in Examples 1-4 had a smaller particle size, below 65 nm, while the particle size of Comparative Examples 2-3 was greater than 65 nm. The zeta potential also met the requirements for liposome stability.

[0170] 5.3 Polarized light microscopy analysis

[0171] The liposome emulsion of distillers' grains extract was coated onto a glass slide and observed using a SCOPE53 polarizing microscope (Oplin Optoelectronic Equipment Co., Ltd., Hangzhou, China).

[0172] The liposomes of the distillers' grains extract were observed under polarized light and TEM. Figure 2a , 2b As shown.

[0173] As shown in Figure 2, the liposomes of the distillers' grains extracts in Examples 1-4 and Comparative Examples 1-3 exhibit the typical characteristics of a layered structure, with an ordered structure resembling Maltese crucifera.

[0174] The liquid crystal content was calculated according to the method in reference 1 (Wang Y, Li J, Shang Y, Zeng X. Study on the development of wax emulsion with liquid crystal structure and its moisturizing and frictional interactions with skin. Colloids and Surfaces B:Biointerfaces 171:335-42 (2018).), and the results are shown in Table 4.

[0175] Table 4

[0176]

[0177] As shown in Table 4, the liquid crystal content of the liposome emulsions from sake lees extract in Examples 1-4 is significantly higher than that in Comparative Examples 1-3; and the liquid crystal content of the liposome emulsion from sake lees extract in Example 2 is the highest at 1.91%, with a more uniform and complete structure and a higher encapsulation rate. Higher liquid crystal content indicates better moisturizing properties and a skin barrier-like effect, signifying successful liposome preparation.

[0178] 5.4 Rheological property analysis

[0179] Rheological tests were performed at room temperature using a rotational rheometer (MARS 60, HAAKE, Vreden, Germany). A 1° conical plate rotor with a diameter of 60 mm was used, and the test gap was set to 52 μm. Before testing, the Peltier plate temperature control was set to 25°C, and the rotor was zero-point calibrated. Then, 1 mL of the liposome emulsion of the distillers' grains extract from each example and comparative example was loaded, and rheological experiments were conducted according to the preset test procedure: first, a continuous flow test was performed, with the shear rate gradually increased from 0 to 1000 s⁻¹. −1 The duration is 60 seconds.

[0180] like Figure 3 As shown, the liposome emulsions of sake lees extract in Examples 1-4 and Comparative Examples 1-3 all exhibit shear-thinning properties, demonstrating good spreadability during application. Furthermore, Example 2 showed the lowest viscosity when unapplied, indicating a smoother skin feel.

[0181] Application Example 1: Analysis of the antioxidant properties of liposomes

[0182] To evaluate the stabilizing effect of liposomes on encapsulated active ingredients, this application example 1 conducted accelerated stability testing experiments on the distillers' grains extract and the liposome emulsions obtained from each example and comparative example, while also examining changes in their antioxidant properties.

[0183] The final concentration of gallic acid in the liposome emulsions of the distillers' grains extracts of Examples 1-4 and Comparative Examples 1-3 was controlled to be 50 μg / mL.

[0184] Subsequently, the liposome emulsions of the distillers' grains extracts from the above examples and comparative examples were placed in a UV incubator at 50°C, and samples were taken at 0 h, 2 h, 4 h, 6 h, 10 h and 12 h. The antioxidant activity was determined using a DPPH kit (Shanghai Yuanye Biotechnology Co., Ltd., Shanghai, China).

[0185] Mix 1.5 mL of the liposome emulsions obtained in Examples 1-4 and Comparative Examples 1-3 with 1.5 mL of DPPH solution (0.2 mmol in 95% ethanol) and keep in the dark for 30 min.

[0186] A control group was set up, consisting of 1.5 mL of liposome emulsions obtained in Examples 1-4 and Comparative Examples 1-3 mixed with 1.5 mL of 95% ethanol and kept in the dark for 30 min.

[0187] A blank control group was also set up, consisting of 1.5 mL of DPPH solution mixed with 1.5 mL of 95% ethanol and kept in the dark for 30 min.

[0188] In addition, a pure distiller's grains extract group was established, which was not encapsulated by liposomes.

[0189] The absorbance of the three groups was measured at 517 nm using a microplate reader. The scavenging activity was calculated using the following formula:

[0190]

[0191] In the formula, A1 is the absorbance value of 1.5 mL of liposome emulsion or distillers' grains extract mixed with 1.5 mL of DPPH solution; A2 is the control group, i.e., the absorbance value of 1.5 mL of liposome emulsion or distillers' grains extract mixed with 1.5 mL of 95% ethanol; A0 is the blank group, i.e., the absorbance value of 1.5 mL of DPPH solution mixed with 1.5 mL of 95% ethanol. Results are shown below. Figure 4 .

[0192] from Figure 4 It can be seen that, as time goes on, the DPPH free radical scavenging rate of the liposome emulsions of sake lees extract in Examples 1-4 and Comparative Examples 1-3 shows a linear decrease; the DPPH free radical scavenging rate of the liposome emulsions of sake lees extract in Examples 1-4 and Comparative Examples 1-3 after encapsulation with liposomes tends to stabilize by day 12, and has a significantly improved DPPH free radical scavenging ability compared with pure sake lees extract.

[0193] At 0h, the DPPH radical scavenging rate of the distiller's grains extract was 85.32%, and the DPPH radical scavenging rate of Example 2 was 88.37%.

[0194] At 12 hours, the DPPH free radical scavenging rates of Examples 1-4 were 57.80%, 60.60%, 52.00%, and 55.30%, respectively, while the DPPH free radical scavenging rates of Comparative Examples 1-3 were 34.30%, 35.20%, and 35.00%, respectively. The DPPH free radical scavenging rate of the distillers' grains extract was only 22.07%. Notably, the DPPH free radical scavenging rate of the liposome emulsion of the distillers' grains extract in Example 2 was approximately 1.74 times higher than that of the distillers' grains extract, indicating that the liposomes can protect the distillers' grains extract and prevent the oxidation of polyphenols in it.

[0195] Application Example 2: Skin Hydration Study

[0196] The hydration levels of the sake lees extract liposome emulsions from Examples 1-4 and Comparative Examples 1-3 were measured after application to the skin using a Derma Unit SSC 3 skin oil-moisture-pH meter from CK (Germany). The measurement protocol is as follows:

[0197] Three areas were selected on the inner sides of each of the test subjects' arms, numbered 1, 2, 3, 4, 5, and 6. Each area was a 4×5cm rectangle. The application areas on the arms of Examples 1-4, Comparative Examples 1-3, and the blank control were randomly arranged. Before the experiment, the test subjects signed an informed consent form, agreeing not to use any lotions, oils, creams, topical alcohol, moisturizing soaps, shower gels, or any other products that affect the skin's moisture content on the inner sides of the forearms within 12 hours before and during the test. During the test, the test subjects were not allowed to wash, wet, or wipe their forearms, nor apply any other products to them. The test subjects were not allowed to drink hot beverages or caffeinated drinks within 1 hour before any test. For each test, the test subjects arrived at the laboratory 10 minutes before the test to rest. All tests were conducted under conditions of 20–24°C and 40–60% relative humidity.

[0198] Before the first test, the subject cleaned and dried the inside of their forearm with water, then rested for 15 minutes before the test began. The skin moisture content was recorded without any product applied (time: 0). Afterwards, the product was applied to each test area. The dosage for each product was 2 μL / cm². 2 Apply evenly with your fingers for 30 seconds. Test the skin hydration level in the applied area at 0 h, 1 h, 3 h, 6 h, 12 h, and 24 h after application, taking five test points in each area. The obtained skin hydration value is the average of the data from the five test points.

[0199] Meanwhile, a pure distiller's grains extract group was established, which was not encapsulated by liposomes.

[0200] The moisturizing performance curves of the liposome emulsions and pure sago extracts of Examples 1-4 and Comparative Examples 1-3 are shown below. Figure 5 As shown.

[0201] from Figure 5 It can be seen that, compared with pure sake lees extract, the sake lees extract liposome emulsions of Examples 1-4 and Comparative Examples 1-3 significantly improved skin hydration and had a good long-lasting moisturizing effect after being applied to the skin. The sake lees extract liposomes of Example 2 increased skin hydration by 36.59% ((1h hydration - 0h hydration) / 0h hydration) after 1 hour of application, and still increased skin hydration by 33.41% ((24h hydration - 0h hydration) / 0h hydration) after 24 hours. At 24 hours, the skin hydration of the sake lees extract liposomes of Comparative Examples 1-3 was between 35-40 au.

[0202] Application Example 3: Study on cumulative skin penetration

[0203] A Franz diffusion cell equipped with a Strat-M membrane (Transdermal Diffusion Test Model, 25 mm, Millipore, Germany) was used (effective diffusion area: 1.766 cm²). 2 The transdermal permeability of pure distiller's grains extract, liposomes of distiller's grains extracts from Examples 1-4 and Comparative Examples 1-3 was investigated.

[0204] like Figure 6 As shown, the Franz diffusion cell consists of a supply chamber and a receiving chamber, with the Strat-M membrane fixed between them using horseshoe clamps. 8 mL of PBS buffer (0.01 M, pH 7.4) was added to the receiving chamber, and the mixture was incubated in a 37°C circulating water bath for 30 min to equilibrate, initiating the permeation experiment. Each example was performed in triplicate, with each replicate conducted in an independent Franz diffusion cell.

[0205] Then, 2 mL of pure distillers' grains extract, 2 mL of liposome solutions of distillers' grains extract obtained in Examples 1-4 and Comparative Examples 1-3, were respectively placed into the supply chamber of the corresponding Franz diffusion cell. The supply and receiving chambers were covered with sealing caps to reduce solution evaporation. The magnetic stirrer was set to a speed of 250 rpm, and the bottom of the receiving chamber was maintained at a constant temperature of 37°C through a circulating water bath. At different time points (0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h, and 24 h), 1 mL of sample was taken from the sampling port through a sampling needle, and an equal volume of PBS buffer (0.01 M, pH 7.4) was added through the sealing cap to maintain a constant volume in the receiving chamber. The absorbance of the 1 mL sample taken at each time point was measured spectrophotometrically at a wavelength of 756 nm. The absorbance was substituted into the standard curve (y = 0.00374x + 0.00414, R0). 2 =0.99899) The content of GA was determined by calculation, and the cumulative permeability Q (mg / cm³) was calculated using the following formula. 3 Each experiment was repeated 6 times.

[0206]

[0207] Where C i Let C be the GA concentration of the i-th (i=n-1) sampling needle. n This refers to the GA concentration in the sampling needle at the nth time (n is 1-9, corresponding to 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h and 24 h respectively), V and V i These are the volumes of the receiving chamber PBS (8 mL) and the sample volume (1 mL), respectively, and S is the osmotic area (1.766 cm²). 2 ).

[0208] Cumulative permeability results are shown in Figure 7a and 7b .in, Figure 7a The cumulative permeation of pure distiller's grains extract at 30 min and 60 min, respectively; Figure 7b The cumulative penetration amount of the liposome emulsions of the distillers' grains extract in Examples 1-4 and Comparative Examples 1-3 at 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 10 h, 12 h and 24 h, respectively.

[0209] from Figure 7a It was found that the pure distiller's grains extract without liposome encapsulation released rapidly, with a cumulative permeation of 21.12 mg / cm³ at 30 min. 2It is almost completely released within 1 hour. This indicates that it is almost completely released and penetrates the skin within 1 hour, but the rapid release means that most of the active ingredients remain on the skin surface and cannot stay and continue to penetrate into deeper layers of the skin.

[0210] from Figure 7b It can be seen that the cumulative permeation of the liposome emulsions of distillers' grains extract in Examples 1-4 and Comparative Examples 1-3 gradually increased over time, eventually reaching a release plateau, indicating that the liposome encapsulation of the distillers' grains extract can achieve a sustained-release effect. At 24 h, the cumulative permeation of Examples 1-4 reached 33.10 mg / cm³, respectively. 2 34.20 mg / cm 2 32.50 mg / cm 2 33.60 mg / cm 2 This is far higher than the amount of pure distiller's grains extract that permeates in 30 minutes (approximately 21.12 mg / cm³). 2 The cumulative release amounts of Comparative Examples 1-3 reached 26.52 mg / cm³. 2 28.59 mg / cm 2 and 27.50 mg / cm 2 Among them, Example 2 showed the highest cumulative penetration, indicating that the liposomes formed by combining distillers' grains prolysin with arabinoxylan in Example 2 had a significant cumulative penetration effect.

[0211] Application Example 4

[0212] The tribological behavior of liposomes from distillers' grains extract was investigated using a finger ruber based on a three-dimensional force sensor (ATI), see reference 2 (Zeng X, Yang H, Han D, Liu C, Chang X, Zhang M, et al. An invivo methodology for studying the interfacial interactions between skin and hair with respect to tactile friction and sensory properties. Measurement179:109499 (2021)). Figure 8 As shown, a three-dimensional force sensor is used to record the coefficient of friction in real time during the finger sliding process.

[0213] The test was performed using the right index finger of a healthy 25-year-old adult woman, and the procedure is as follows:

[0214] First, the artificial skin (for the preparation of the artificial skin, see reference 3, Morales-Hurtado M, Zeng X, Gonzalez-Rodriguez P, Ten Elshof JE, van der Heide E. A new water absorbable mechanical epidermal skin equivalent: The combination of hydrophobic PDMS and hydrophilic PVA hydrogel. Journal of the Mechanical Behavior of BiomedicalMaterials 46:305-17 (2015)) was fixed to the top of the friction tester with double-sided tape. The artificial skin used for each test was rectangular, with a long side of 5 cm and a short side of 1.2 cm.

[0215] Then, 30 mg of the sake lees extract liposome emulsion from Examples 1-4 and Comparative Examples 1-3 was placed on the left end of the artificial skin. The tester used their index finger to slide the sake lees extract liposome emulsion from one end of the long side of the artificial skin to the opposite end, making six back-and-forth motions for each test. During the sliding process, the sliding speed and normal load were kept essentially constant. A normal force of 0.2 ± 0.05 N was used for the test, and the coefficient of friction (COF) (obtained by a three-dimensional force sensor) at 0 min, 1 min, 2 min, and 5 min was recorded and statistically analyzed.

[0216] Figure 9a The changes in the coefficient of friction of the liposome emulsions of sake lees extract from Examples 1-4 and Comparative Examples 1-3 after application to the skin surface for different durations are shown. Figure 9b As shown in Example 2, the frictional behavior of the liposomes from the distillers' grains extract exhibits a typical Stribeck curve lubrication pattern, which gradually transitions from hydrodynamic lubrication to mixed lubrication and then to boundary lubrication.

[0217] The Stribeck curve is divided into three regions: 1) Boundary lubrication zone: the leftmost part of the curve, where the Hersey number is very low, making adhesive wear and abrasive wear likely. This indicates high friction between the liposomes and the skin, resulting in a feeling of tightness, roughness, and skin pulling; 2) Mixed lubrication zone: the concave transition area in the middle of the curve, where wear is much less than in the boundary lubrication zone. This indicates a sharp drop in friction between the liposomes and the skin to its lowest point, making the product feel instantly melt, burst with moisture, and become exceptionally smooth. This is the golden moment for the skin feel and one of the key experiences pursued by formulators; 3) Hydrodynamic lubrication zone: the rightmost part of the curve, where the Hersey number is high, theoretically resulting in zero wear (no contact wear).

[0218] In the initial application phase (0 min, i.e., after the first application of the sake lees extract liposome emulsion to the left end, followed by 6 back-and-forth motions, also known as the first cycle), the liposomes form a relatively thick liposome film composed of liposome emulsion droplets at the interface during friction with the skin. The contact between the finger and skin is primarily hydrodynamic lubrication, resulting in a low coefficient of friction. After 1 min of application (1 minute after the first friction cycle, continuing with 6 back-and-forth motions, also known as the second cycle), boundary lubrication occurs. This is likely because the liposome droplets gradually break down and reassemble on the skin surface into a thin film formed by liposomes. The film thickness decreases, the contact surfaces partially separate through the film, and the lubrication state transitions to mixed lubrication. Over time, moisture gradually evaporates, further reducing the film thickness. After 5 min of application (5 minutes after the first friction cycle, continuing with 6 back-and-forth motions, also known as the third cycle), the coefficient of friction (COF) of the sake lees extract liposome emulsion shows a relatively flat trend. At this point, the COF mainly depends on the residual sensation of the sake lees extract liposome emulsion on the skin.

[0219] In summary, during the application process (1-5 min), Example 2 showed the lowest coefficient of friction; Examples 1-4 showed a significant reduction compared to Comparative Examples 1-3, indicating that the liposomes prepared by combining arabinoxylan and prolysin can reduce the coefficient of friction and improve skin feel.

[0220] Application Example 5: Stability Test

[0221] The liposomes prepared from the lees extract in Examples 1-4 and Comparative Examples 1-3 were placed in clean, transparent containers, sealed, and stored at room temperature for 7 days. The appearance, layering, flocculation, and precipitation of the liposome system were observed and recorded, along with any turbidity, discoloration, or phase separation. The results are shown below. Figure 10 .

[0222] from Figure 10It can be seen that after being placed at room temperature for 7 days, Comparative Example 1, which did not add arabinoxylan to the aqueous phase and did not add prolysin to the oil phase, began to show obvious sedimentation and stratification; Comparative Example 2, which did not add arabinoxylan to the aqueous phase and added prolysin to the oil phase, showed increased precipitation and severe aggregation.

[0223] In Comparative Example 3, where arabinoxylan was added to the aqueous phase but no prolysin from distillers' grains was added to the oil phase, suspended matter appeared, but no obvious precipitation or stratification occurred. This indicates that adding polysaccharides to the aqueous phase of the distillers' grains extract and adding active proteins to the liposome membrane can improve the stability of the liposomes from the distillers' grains extract.

[0224] The results above demonstrate that polysaccharides significantly improve the kinetic stability of the system by utilizing their thickening effect, thus preventing liposome sedimentation and aggregation.

[0225] In summary, the liposomes constructed from distillers' grains extract in this invention not only exhibit excellent physicochemical properties in terms of structural characterization, particle size, potential, encapsulation efficiency, and liquid crystal phase characteristics, but also demonstrate significant functional advantages in rheological properties and tactile friction behavior. Tribological tests show that the liposomes undergo a dynamic transition from hydrodynamic lubrication and mixed lubrication to boundary lubrication during application. The liposome system prepared in this invention exhibits good spreadability, stability, and tactile sensation regulation on the skin surface, significantly improving the user experience and application performance of the active ingredients from distillers' grains. Therefore, this invention not only achieves efficient loading and delivery of distillers' grains extract, but also provides a novel and scalable material system for the regulation of tactile friction on the skin, possessing the potential for wide application in skin care products.

[0226] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any changes or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in the present invention should be included within the scope of protection of the present invention.

Claims

1. A lees extract liposome, characterized by, It includes a liposome membrane and a distiller's grains extract and polysaccharides encapsulated within the liposome membrane, wherein the raw materials of the liposome membrane include phospholipids, cholesterol and active proteins; The active protein is selected from one or two of distillers' grains prolyl protein and rice protein peptides; The polysaccharide is selected from one or both of arabinoxylan and β-glucan.

2. The liposomes of the distillers' grains extract as described in claim 1, characterized in that, The phospholipid is selected from one or more of soybean phospholipids, lecithin, hydrogenated soybean phospholipids, hydrogenated lecithin, phosphatidylethanolamine, distearate phosphatidylcholine, and polyethylene glycol-derived phospholipids; And / or, the relative molecular weight of the prolysin is 10-30 kDa; And / or, the relative molecular weight of the rice protein peptide is 500-1000 Da; And / or, the relative molecular weight of the arabinoxylan is 5-65 kDa; And / or, the relative molecular weight of the β-glucan is 10-70 kDa; And / or, the liposomes from the distillers' grains extract, when observed under a polarizing microscope, exhibit a Maltese cross pattern.

3. The liposomes of the distillers' grains extract as described in claim 2, characterized in that, The raw materials of the liposomes extracted from distillers' grains contain the following components in the following mass percentages: 10-20 wt% lecithin and cholesterol, 0.05-0.2 wt% active protein, 0.01-0.1 wt% polysaccharide, and the balance being distillers' grains extract; the mass ratio of lecithin to cholesterol is 30:(3-7).

4. The liposomes of the distillers' grains extract as described in claim 3, characterized in that, Includes at least one of 1) or 2): 1) The preparation method of the distillers' grains extract is as follows: A1) The distillers' grains powder was degreased using n-hexane to obtain degreased distillers' grains powder; A2) Mix water with the defatted distiller's grains powder and sonicate to separate the solid and liquid, and take the supernatant to obtain the distiller's grains extract; 2) The preparation method of the alcohol-soluble protein from distillers' grains is as follows: B1) The lees powder and water are subjected to a first ultrasonic treatment to separate the solid and liquid, and the first filter residue is obtained. B2) The first filter residue and sodium chloride aqueous solution are subjected to a second ultrasonic treatment to separate the solid and liquid components and obtain the second filter residue; B3) The second filter residue and alcohol solution are subjected to a third ultrasonic treatment, followed by solid-liquid separation, concentration, and drying to obtain the alcohol-soluble protein from the distillers' grains. Preferably, the solid-liquid separation is centrifugation; Preferably, in B1), the mass-to-volume ratio of the distiller's grains powder to water is 1g:(10-20)L; Preferably, in B2), the mass-to-volume ratio of the first filter residue to the sodium chloride aqueous solution is 1 g: (10-20) L; Preferably, in B3), the mass-to-volume ratio of the second filter residue to the alcohol solution is 1 g: (10-20) L; Preferably, the temperature of the ultrasonic treatment is 30-60℃.

5. The method for preparing liposomes from distillers' grains extract according to any one of claims 1-4, characterized in that, Includes the following steps: a) Lecithin, cholesterol, active protein, and organic solvent are mixed for the first time to obtain the oil phase; the distiller's grains extract and polysaccharide are mixed for the second time to obtain the aqueous phase; b) The oil phase and aqueous phase are mixed for a third time and homogenized to obtain the liposomes of the distillers' grains extract.

6. The preparation method according to claim 5, characterized in that, The ratio of the active protein to the organic solvent is 1:(1000-3000). And / or, the ratio of the oil phase to the water phase is 1:(0.01-0.5); And / or, the organic solvent is selected from one or both of anhydrous ethanol and chloroform; And / or, the temperature of the first mixing is 40-60°C; And / or, the temperature of the second mixing is 40-60°C; And / or, the temperature of the third mixing is 40-60°C; And / or, the homogenization further includes a post-treatment, which is vacuum evaporation to remove organic solvents; And / or, the homogenization rotation speed is 4000-6000 rpm / min, and the homogenization time is 1-10 min; Preferably, the temperature for the reduced-pressure evaporation is 60~65℃; Preferably, the vacuum degree of the reduced pressure evaporation is 0.01-0.2 MPa.

7. Use of the liposomes extracted from distillers' grains as described in any one of claims 1-4 in the preparation of cosmetics.

8. A cosmetic product, characterized in that, Liposomes containing the distillers' grains extract as described in any one of claims 1-4.

9. The use of the lees extract liposomes as described in any one of claims 1-4 or the cosmetic as described in claim 8 in at least one of the following: C1) Enhances the skin's antioxidant capacity; C2) Improves the skin's moisturizing ability; C3) Improves skin tactile sensation.

10. Use of active proteins and polysaccharides in improving the stability of liposomes from distillers' grains extract, wherein the active protein is selected from one or two of distillers' grains prolysin and rice protein; and the polysaccharide is selected from one or two of arabinoxylan and β-glucan.

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