Multi-effect bionic liposome transdermal repair carrier as well as preparation and application thereof

Abstract

The invention relates to a multi-effect bionic liposome transdermal repair carrier and a preparation method and application thereof.The multi-effect bionic liposome transdermal repair carrier comprises active ingredients and a nano-carrier, and the active ingredients comprise sialic acid, saccharomycetes / rice fermentation product filtrate, arginine / lysine polypeptide and vitamin E in the mass ratio of (1-10): (2-8): (0.1-1): (0.5-4); the nano-carrier is prepared from the following raw materials: phospholipid, an emulsifier, polyhydric alcohol and water. Compared with the prior art, the anti-aging, whitening and moisturizing composition has the advantages of enhancing skin moisturizing and anti-oxidation effects, promoting collagen synthesis and inhibiting tyrosinase to achieve remarkable anti-aging, whitening and moisturizing effects and the like.

Description

Technical Field

[0001] This invention relates to the field of skincare products, and in particular to a multi-effect biomimetic liposome transdermal repair carrier and its preparation and application. Background Art

[0002] Currently, anti-aging and whitening ingredients in cosmetics mainly rely on the following categories of substances: Vitamins (such as retinol and vitamin C): While retinol can promote collagen production, it has issues with photosensitivity and high irritation, causing adverse reactions such as skin redness and peeling in some consumers. Vitamin C (ascorbic acid) has significant antioxidant effects, but its stability is poor and it is easily oxidized and inactivated, requiring special formulation technologies (such as microencapsulation or derivative forms) to maintain its activity. Peptides (such as palmitoyl pentapeptide and acetyl hexapeptide): These can stimulate collagen synthesis or inhibit neurotransmitter release (such as a botulinum toxin-like effect), but some peptides have poor permeability and limited long-term effects. Natural extracts (such as centella asiatica and resveratrol): Although they have high safety, some ingredients have a slow onset of action, and high concentrations may affect product stability. Existing technologies mostly use single active ingredients or simple combinations, making it difficult to simultaneously achieve multiple effects such as anti-wrinkle, moisturizing, and barrier repair, and some ingredients (such as retinol) have limited applicability to certain groups.

[0003] Currently, many cosmetics vary greatly in quality. Cosmetics containing active ingredients suffer from the following problems: Poor stability of active ingredients, easily degraded and inactivated. Many anti-aging and whitening skincare products on the market contain polyphenols, vitamins, or peptides as their active ingredients. These are sensitive to enzymes, light, heat, and pH levels. If added directly to the formula without nano-encapsulation, unstable active ingredients are easily inactivated. Because the stratum corneum of the skin has a strong barrier function, many active molecules, such as active peptides, have difficulty penetrating it, thus affecting their skincare effects on the target area, resulting in minimal anti-aging, whitening, and moisturizing repair effects. Single skincare mechanism: Due to the complexity of skin aging and whitening mechanisms, the synergistic effect of multiple active ingredients with different mechanisms of action is required to effectively exert anti-aging effects. However, current anti-aging and whitening skincare products on the market have single mechanisms of action, ultimately leading to unsatisfactory anti-aging and whitening effects. Summary of the Invention

[0004] The purpose of this invention is to provide a multi-effect biomimetic liposome transdermal repair carrier with anti-aging, whitening, moisturizing and repairing effects, as well as its preparation and application, to enhance skin hydration, moisturizing and antioxidant effects, promote collagen synthesis, and inhibit tyrosinase to achieve significant anti-aging, whitening and moisturizing effects.

[0005] The objective of this invention can be achieved through the following technical solution: a multi-effect biomimetic liposome transdermal repair carrier, comprising active ingredients and nanocarriers, wherein the active ingredients include sialic acid, yeast / rice fermentation product filtrate, arginine / lysine peptides, and vitamin E in a mass ratio of (1-10):(2-8):(0.1-1):(0.5-4). More preferably, the mass ratio of sialic acid to vitamin E in the composition is 2-4:1.

[0006] Furthermore, the raw materials for the nanocarrier include phospholipids, emulsifiers, polyols, and water.

[0007] Furthermore, based on the total mass of the multi-effect biomimetic liposome transdermal repair carrier as 100%, it includes 1-10% sialic acid, 2-8% yeast / rice fermentation product filtrate, 0.1-1% arginine / lysine peptides, 0.5-4% vitamin E, 0.1-2% phospholipids, 5-30% emulsifiers, 10-40% polyols, and the remainder water.

[0008] Further optimization, based on the total mass of the multi-effect biomimetic liposome transdermal repair carrier as 100%, includes 3-9% sialic acid, 5-8% yeast / rice fermentation product filtrate, 0.5-1% arginine / lysine peptide, 1-4% vitamin E, 0.1-2% phospholipid, 5-30% emulsifier, 10-40% polyol, and the balance being water.

[0009] Furthermore, the phospholipids include one or more of soybean lecithin, hydrogenated lecithin, egg yolk lecithin, hydrogenated soybean lecithin, and hydrogenated egg yolk lecithin;

[0010] The emulsifiers include one or more of the following: polyoxyethylene sorbitan fatty acid ester, polyglycerol fatty acid ester, polyoxyethylene fatty acid ester, polyoxyethylene castor oil emulsifier, polyoxyethylene hydrogenated castor oil emulsifier, PEG-8 caprylic / capric glyceride, polyglycerol emulsifier, poloxamer, cocoyl glucoside, triglycerides, pyrrolidones, polyglycerol esters, PEG-30 dihydroxystearate and cetearyl alcohol polyether, Tween 80, Tween 20, Tween 60, PEG-20 hydrogenated castor oil, PEG-40 hydrogenated castor oil, and PEG-60 hydrogenated castor oil.

[0011] The polyols include one or more of glycerol, propylene glycol, 1,3-butanediol, 1,3-propanediol, 1,2-pentanediol, ethoxydiethylene glycol, 1,2-hexanediol, dipropylene glycol, isopropanol, polyethylene glycol-200, PPG-10 sorbitol, octyldodecyl alcohol, and dipropylene glycol.

[0012] Furthermore, the raw materials for the nanocarrier also include a pH adjuster at a mass percentage of 1-2%.

[0013] Furthermore, the pH adjuster includes tromethamine.

[0014] This invention also provides a method for preparing a multi-effect biomimetic liposome transdermal repair carrier, comprising the following steps:

[0015] (1) Mix vitamin E with emulsifier, polyol and phospholipid to obtain mixture A;

[0016] (2) Mix sialic acid, yeast / rice fermentation product filtrate, arginine / lysine peptide, pH adjuster, polyol and water to obtain mixture B;

[0017] (3) Pour mixture A into mixture B and mix well to obtain mixture C;

[0018] (4) The mixture C is nano-processed to obtain a liposome composition with anti-aging, whitening, moisturizing and repairing effects.

[0019] Furthermore, in step (1), the mixing conditions are: stirring at 30-65°C until homogeneous;

[0020] In step (2), the mixing conditions are: stirring at 30-65℃ until homogeneous;

[0021] In step (3), the mixing conditions are: stirring at 30-65℃ until homogeneous;

[0022] In step (4), the nano-sizing process is homogenization using a high-pressure homogenizer at a pressure of 500-1200 bar.

[0023] This invention also provides a skincare product containing the aforementioned multi-effect biomimetic liposome transdermal repair carrier. Skincare products include lotions, creams, freeze-dried powders, and toners.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] (1) This invention selects four specific ratios of active ingredients: sialic acid (N-acetylneuraminic acid), yeast / rice ferment filtrate, conospirol (arginine / lysine polypeptide), and vitamin E (tocopherol) to achieve multi-target synergistic effects. Sialic acid makes the skin fair and beautiful; yeast / rice ferment filtrate reduces oxidative stress caused by ultraviolet rays and environmental pollution, protects the skin from free radical damage, promotes skin hydration and barrier function, softens the stratum corneum and replenishes moisture, keeping the skin moisturized, crystal clear and smooth; conospirol (arginine / lysine polypeptide) regulates skin nerve signals, reduces wrinkle formation, promotes collagen production, and delays cell aging. Vitamin E (tocopherol) scavenges free radicals, prevents lipid peroxidation, and delays photoaging, ultimately achieving anti-aging, whitening, moisturizing and repairing effects.

[0026] (2) This invention rationally combines multiple active ingredients with nanocarrier technology to form a liposome that acts on anti-aging, whitening, moisturizing, and repairing. This invention utilizes novel nanotechnology to encapsulate and deliver four active ingredients—sialic acid, yeast / rice ferment filtrate (maifune), arginine / lysine polypeptide (conopeptide), and vitamin E (tocopherol)—using a nanocarrier. Based on the structure and characteristics of the nanocarrier, the composition obtained through nano-encapsulation exhibits good stability and water dispersibility, increasing the solubility of the active ingredients while also reducing their irritation. This allows the active ingredients to reach sufficient concentrations in the product to exert their corresponding functional effects. While effectively improving the solubility of each active ingredient, it also provides a stable storage space, preventing unnecessary degradation or inactivation of the active ingredients before storage and use, which is beneficial for increasing the concentration of active substances in anti-aging and whitening products. Attached Figure Description

[0027] Figure 1 The figures represent cumulative permeation and retention in the skin; in the figure, compared with free components, &&P<0.01;

[0028] Figure 2 For observing skin penetration using laser confocal microscopy;

[0029] Figure 3 The uptake behavior of HaCaT and HDF cells was compared; among them, compared with free RhoB, ∠P < 0.05, ∠P < 0.01, Figure 3 A and 3C are HDF cells. Figure 3 B and 3D are HaCaT cells;

[0030] Figure 4 The results are from the cell proliferation experiment; among them: compared with the normal group, #P<0.05, ##P<0.01; compared with the free component, &P<0.05, &&P<0.01;

[0031] In each figure, compared with the model group, **P<0.01; compared with Comparative Examples 1, 2, 3, 4, ##P<0.01; compared with Comparative Example 5, %%P<0.01; compared with Comparative Example 6, @@P<0.01; compared with Examples 4 and 5, &&P<0.01. Detailed Implementation

[0032] The present invention will now be described in detail with reference to the accompanying drawings and specific embodiments. These embodiments are based on the technical solution of the present invention and provide detailed implementation methods and specific operating procedures. However, the scope of protection of the present invention is not limited to the following embodiments.

[0033] This invention utilizes a specific ratio of sialic acid (N-acetylneuraminic acid), yeast / rice fermentation product filtrate, conospirin (arginine / lysine polypeptide), and vitamin E (tocopherol) to achieve a multi-target synergistic effect. Through various skincare mechanisms, it enhances skin hydration, moisturization, and antioxidant effects, promotes collagen synthesis, and inhibits tyrosinase, achieving significant anti-aging, whitening, and moisturizing effects. Specific details are as follows:

[0034] Sialic acid (N-acetylneuraminic acid) acts on epidermal barrier-related proteins, promotes keratinocyte differentiation, enhances skin barrier function, regulates epidermal water transport, improves skin hydration, relieves redness in sensitive skin, and can also effectively inhibit tyrosinase, reduce melanin production, and adjust the skin's melanin metabolism balance, thereby achieving the effect of making the skin white and beautiful.

[0035] The sialic acid used in this invention is a commercially available product.

[0036] Yeast / rice ferment filtrate (McFoun); as a core ingredient of the Mageline brand, it possesses anti-inflammatory and antioxidant properties. It can reduce oxidative stress caused by ultraviolet radiation and environmental pollution, protecting the skin from free radical damage. The key ingredient in McFoun, Alpha-KG, can activate cell proliferation, promote skin hydration and barrier function, soften the stratum corneum, and replenish moisture, leaving the skin moisturized, radiant, and smooth.

[0037] The yeast / rice fermentation product filtrate of this invention was obtained using a method reported in the published literature by Mageline, as shown in the following examples where the yeast / rice fermentation product filtrate was prepared by the following method:

[0038] (1) Add water to rice powder and heat and stir to make 30% rice slurry. Add β-glucanase to the rice slurry. The amount of enzyme added is 0.005% of the mass of rice slurry and the enzyme activity is 200U / g. Perform enzymatic hydrolysis to obtain rice slurry hydrolysate.

[0039] (2) Inoculate the yeast strain Saccharomyces veronae into the rice slurry hydrolysate. The fermentation conditions are: fermentation temperature 35℃, fermentation time 36 hours, and pH controlled between 6 and 6.5 during the fermentation process. After fermentation, rice fermentation liquid is obtained.

[0040] (3) The fermented rice fermentation liquid is concentrated 10 times by reverse osmosis technology to efficiently enrich active substances. After heating and sterilization, it is cooled and filtered to obtain concentrated rice fermentation liquid, namely yeast / rice fermentation product filtrate.

[0041] Conospiropeptides (arginine / lysine peptides) can specifically act on neurotransmitter receptors, such as acetylcholine receptors, to regulate nerve signals in the skin, potentially helping to reduce muscle contraction and thus wrinkle formation. By blocking specific ion channels, conospiropeptides may affect skin cell function, including cell proliferation and apoptosis, promote collagen production, and delay cell aging.

[0042] The conopod peptide used in this invention is a commercially available product.

[0043] Vitamin E (tocopherol) is a classic fat-soluble antioxidant that can scavenge free radicals, prevent lipid peroxidation, and delay photoaging.

[0044] The vitamin E used in this invention is a commercially available product.

[0045] Nanoliposomes generally refer to small, single-chambered liposomes with a particle size of less than 100 nm. They can encapsulate water-soluble or lipid-soluble active ingredients and have advantages such as sustained release, biodegradability, and non-immunogenicity and toxicity. Their nanoscale size and good deformability give them a strong ability to penetrate physiological tissue barriers. Nanocarriers used in cosmetics / pharmaceuticals can significantly improve the skin penetration of active ingredients. Encapsulating active ingredients with different mechanisms of action within the same nanoliposome and delivering them through the skin can achieve sustained release and long-lasting effects, while also realizing the synergistic effect of multiple active and multi-target components, significantly enhancing efficacy.

[0046] This invention uses nano-encapsulation technology to form liposomes from four active ingredients.

[0047] The following detailed description is provided through specific embodiments. Unless otherwise specified, the raw materials and testing methods involved in this invention are commonly used in the field.

[0048] Example 1

[0049] Preparation of a multi-effect biomimetic liposome transdermal repair carrier:

[0050] (1) Mix 2% vitamin E, 2% lecithin, 10% 1,2-pentanediol, 5% poloxamer, and 25% Tween-60 at 45°C to form a homogeneous and clear liquid A.

[0051] (2) Mix 8% sialic acid, 6% yeast / rice fermentation product filtrate (mafuyin), 1% arginine / lysine polypeptide (conotazopeptide), 10% glycerol, 20% 1,3-propanediol, 2% tromethamine, and the balance of water at 45°C to form a homogeneous and clear liquid B.

[0052] (3) Pour liquid A into liquid B and mix, then stir at 45°C to form a homogeneous liquid C.

[0053] (4) After homogenizing liquid C at 1200 bar in a high-pressure homogenizer, a multi-effect biomimetic liposome transdermal repair carrier was obtained.

[0054] Example 2

[0055] Preparation of a multi-effect biomimetic liposome transdermal repair carrier:

[0056] (1) Mix 1% vitamin E, 1% lecithin, 10% 1,2-hexanediol, 5% cocoyl glucoside, and 10% PEG-20 hydrogenated castor oil at 45°C to form a homogeneous and clear liquid A.

[0057] (2) Mix 3% sialic acid, 5% yeast / rice fermentation product filtrate (mafuyin), 0.5% arginine / lysine polypeptide (conotazolide), 10% glycerol, 5% 1,2-propanediol, 1.4% tromethamine, and the balance of water at 45°C to form a homogeneous and clear liquid B.

[0058] (3) Pour liquid A into liquid B and mix, then stir at 45°C to form a homogeneous liquid C.

[0059] (4) After homogenizing liquid C at 1200 bar in a high-pressure homogenizer, a multi-effect biomimetic liposome transdermal repair carrier was obtained.

[0060] Example 3

[0061] Preparation of a multi-effect biomimetic liposome transdermal repair carrier:

[0062] (1) Mix 0.5% vitamin E, 0.1% lecithin, 2% ethoxydiethylene glycol, 3% dipropylene glycol, and 5% polyglycerol at 45°C to form a homogeneous and clear liquid A.

[0063] (2) Mix 1% sialic acid, 2% yeast / rice fermentation product filtrate (mafuyin), 0.1% arginine / lysine polypeptide (conopeptide), 5% 1,2-propanediol, 1% tromethamine, and the remainder water at 45°C to form a homogeneous and clear liquid B.

[0064] (3) Pour liquid A into liquid B and mix, then stir at 45°C to form a homogeneous liquid C.

[0065] (4) After homogenizing liquid C at 1200 bar in a high-pressure homogenizer, a multi-effect biomimetic liposome transdermal repair carrier was obtained.

[0066] Using Example 1 as a control, examples with different ratios of sialic acid and vitamin E were set up.

[0067] Example 4

[0068] Preparation of a multi-effect biomimetic liposome transdermal repair carrier:

[0069] (1) Mix 1.6% vitamin E, 2% lecithin, 10% 1,2-pentanediol, 5% poloxamer, and 25% Tween-60 at 45°C to form a homogeneous and clear liquid A.

[0070] (2) Mix 8.4% sialic acid, 8% yeast / rice fermentation product filtrate (mafuyin), 1% arginine / lysine polypeptide (conopeptide), 10% glycerol, 20% 1,3-propanediol, 2% tromethamine, and the balance of water at 45°C to form a homogeneous and clear liquid B.

[0071] (3) Pour liquid A into liquid B and mix, then stir at 45°C to form a homogeneous liquid C.

[0072] (4) After homogenizing liquid C at 1200 bar in a high-pressure homogenizer, a multi-effect biomimetic liposome transdermal repair carrier is obtained.

[0073] Example 5

[0074] Preparation of a multi-effect biomimetic liposome transdermal repair carrier

[0075] (1) Mix 4% vitamin E, 2% lecithin, 10% 1,2-pentanediol, 5% poloxamer, and 25% Tween-60 at 45°C to form a homogeneous and clear liquid A.

[0076] (2) Mix 6% sialic acid, 8% yeast / rice fermentation product filtrate (mafuyin), 1% arginine / lysine polypeptide (conopeptide), 10% glycerol, 20% 1,3-propanediol, 2% tromethamine, and the balance of water at 45°C to form a homogeneous and clear liquid B.

[0077] (3) Pour liquid A into liquid B and mix, then stir at 45°C to form a homogeneous liquid C.

[0078] (4) After homogenizing liquid C at 1200 bar in a high-pressure homogenizer, a multi-effect biomimetic liposome transdermal repair carrier is obtained.

[0079] Comparative Example

[0080] Using Example 1 as a control, the active ingredient composition was replaced with an equal amount of active ingredient, and nanocomposites containing one or more different active ingredients were prepared using the same method.

[0081] Comparative Example 1

[0082] It contains only the active ingredient sialic acid (17%), and the other ingredients and preparation method are the same as in Example 1.

[0083] Comparative Example 2

[0084] It contains only 17% of the active ingredient yeast / rice fermentation product filtrate (Maifuyin), and the other ingredients and preparation method are the same as in Example 1.

[0085] Comparative Example 3

[0086] It contains only 17% of the active ingredient arginine / lysine polypeptide (cono peptide), and the other ingredients and preparation method are the same as in Example 1.

[0087] Comparative Example 4

[0088] It contains only the active ingredient, vitamin E 17%, and the other ingredients and preparation method are the same as in Example 1.

[0089] Comparative Example 5

[0090] It contains 8% sialic acid, 6% yeast / rice fermentation product filtrate (mafuyin), 3% arginine / lysine polypeptide (conopeptide), and 0% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0091] Comparative Example 6

[0092] It contains 8% sialic acid, 6% yeast / rice fermentation product filtrate (mafuyin), 0% arginine / lysine polypeptide (conopeptide), and 3% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0093] Comparative Example 7

[0094] It contains 14% sialic acid, 0% yeast / rice fermentation product filtrate (mafuyin), 1% arginine / lysine polypeptide (conopeptide), and 2% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0095] Comparative Example 8

[0096] It contains 0% sialic acid, 14% yeast / rice fermentation product filtrate (mafuyin), 1% arginine / lysine polypeptide (conopeptide), and 2% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0097] Comparative Example 9

[0098] It contains 11% sialic acid, 6% yeast / rice fermentation product filtrate (mafuyin), 0% arginine / lysine polypeptide (conopeptide), and 0% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0099] Comparative Example 10

[0100] It contains 15% sialic acid, 0% yeast / rice fermentation product filtrate (mafuyin), 0% arginine / lysine polypeptide (conopeptide), and 2% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0101] Comparative Example 11

[0102] It contains 0% sialic acid, 15% yeast / rice fermentation product filtrate (mafuyin), 0% arginine / lysine polypeptide (conopeptide), and 2% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0103] Comparative Example 12

[0104] It contains 0% sialic acid, 0% yeast / rice fermentation product filtrate (mafuyin), 15% arginine / lysine polypeptide (conopeptide), and 2% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0105] Comparative Example 13

[0106] It contains 11% sialic acid, 0% yeast / rice fermentation product filtrate (mafuyin), 0% arginine / lysine polypeptide (conopeptide), and 6% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0107] Comparative Example 14

[0108] It contains 11% sialic acid, 4% yeast / rice fermentation product filtrate (mafuyin), 1.5% arginine / lysine polypeptide (conopeptide), and 0.5% vitamin E. Other ingredients and preparation methods are the same as in Example 1.

[0109] Comparative Example 15

[0110] Sialic acid 8%, yeast / rice fermentation product filtrate (Maifuyin) 6%, arginine / lysine polypeptide (conopeptide) 1%, vitamin E (tocopherol) 2% were mixed in a solvent, namely dimethyl sulfoxide (DMSO):water (volume ratio of 8:2), to obtain a free composition with the same content as in Example 1.

[0111] The average particle size of the products obtained in Examples 1-5 and Comparative Examples 1-14 above is less than 100 nm, and the PDI is 0.05-0.4. The products obtained in each example and comparative example were placed in sealed containers and stored at -20℃, room temperature, 4℃, and 45℃ for 3 months respectively. No stratification or precipitation occurred, and the particle size did not change significantly, indicating that the products obtained in each example and comparative example have good stability.

[0112] Performance testing:

[0113] Performance Testing 1: Evaluation of Irritation of Chicken Embryo Allantoic Membrane

[0114] Chicken embryos: Purchase chickens and incubate them in an incubator for 9 days, ensuring the air cell faces upwards during incubation.

[0115] Experimental Method: Take 9-day-old chicken embryos and evenly spread 0.2 ml (or g) of the test substance onto the chorioallantoic membrane (CAM) of the embryo, contacting it with the CAM for 30 seconds. Then gently rinse with physiological saline for about 30 seconds to remove the test substance until the CAM surface is clean. Three chicken embryos are prepared for each test substance in parallel. Negative and positive controls are set up simultaneously, with physiological saline as the negative control and NaOH as the positive control.

[0116] Observation of indicators: Observe the changes in CAM vessels within 5 minutes and record the initial time of CAM vessel congestion, hemorrhage and coagulation.

[0117] The multifunctional biomimetic liposome transdermal repair carriers obtained in Examples 1-5 were diluted 10 times. 0.2 mL of the sample was taken and dropped onto the surface of the chorioallantoic membrane (CAM) of chicken embryos. The changes in CAM blood vessels were observed within 5 min, and the initial time of CAM blood vessel congestion, hemorrhage and coagulation was recorded. The stimulation score IS was calculated.

[0118] IS=[(301-secH)×5+(301-secL)×7+(301-secC)×9] / 300

[0119] In the formula, secH is the initial time of surface congestion, s; secL is the initial time of surface bleeding, s; and secC is the initial time of surface clotting, s.

[0120] Calculate the mean of repeated trials and classify the irritation of the test substance according to the magnitude of the mean. The mean values ​​of 0–0.9, 1.0–4.9, 5.0–8.9, and 9–21.0 are respectively classified as no irritation, mild irritation, moderate irritation, and severe irritation.

[0121] Test results showed that when the liposome composition diluted 10 times was in contact with the chicken embryo allantoic membrane for 300 seconds, there was no bleeding, vascular dissolution, or coagulation in the capillaries, and the reaction scores were 0.07, 0.06, 0.08, 0.07, and 0.06, indicating that the multi-effect biomimetic liposome transdermal repair carrier described in Examples 1-5 after 10-fold dilution had good safety and was non-irritating.

[0122] Performance testing 2 patch test

[0123] Test method: The safety of the product was assessed using a human skin patch test.

[0124] Test substance: The compositions described in Examples 1-5.

[0125] Subjects: A total of 50 people, 25 males and 25 females, aged 25 to 35 years, who met the criteria for voluntary inclusion, and were divided into 5 groups of 10 people each;

[0126] Patch test method: Using a qualified patch tester, the liposome composition obtained in Examples 1-5 was diluted 10 times and a blank control (i.e., containing no substances) was applied to the inside of the patch tester at a concentration of approximately 0.02–0.025 g. The patch was then applied to the flexor side of the subject's forearm with special adhesive tape (the blank control was applied next to the experimental group) for 24 hours. After removing the patch tester, the skin reaction was observed after a 30-minute interval until the indentation disappeared. The skin reaction was graded according to the skin reaction grading standards in the "Cosmetic Safety Technical Regulations 2015". Skin reactions were observed again 24 hours and 48 hours after removing the patch tester.

[0127] The results showed that none of the 50 subjects developed light red spots, erythema, edematous erythema, significant redness and swelling, infiltration or papules, or papules or vesicles, indicating that the multi-effect biomimetic liposome transdermal repair carrier described in Examples 1-5 is non-irritating to human skin.

[0128] Performance testing 3 In vitro skin penetration test

[0129] Comparative Example 15 (Free RhoB), Carrier Prepared in Example 1 (RhoB Nanocarrier): Sialic acid contained in the multi-effect biomimetic transdermal liposome repair carrier (RhoB nanocarrier) prepared in Example 1 was used as a fluorescent label, and a free RhoB solution of equal concentration was prepared as a control sample (free RhoB).

[0130] Skin permeation experiments on ex vivo porcine skin were conducted using the vertical Franz diffusion cell method. The skin was fixed between the receiving chamber and the supply chamber. The experiment was divided into two groups: a free RhoB group and a RhoB nanocarrier group. 0.5 g each of a 5% RhoB nanocarrier complex (i.e., the product prepared in Example 1 was added to the receiving solution as the RhoB nanocarrier, with an addition amount of 5 wt%) and a free RhoB complex (with the same free RhoB concentration as the RhoB nanocarrier) were placed in the supply chamber. PBS was used as the receiving solution, and the mixture was stirred and diffused at 37°C. After 2 and 4 hours, residual sample on the skin was gently wiped away, and the skin from the target area was removed. The skin was rinsed again, thoroughly cleaned, and dried. The sample was frozen and sectioned, and the sections were observed using a laser confocal microscope. Representative areas were selected and photographed.

[0131] Depend on Figure 1 As can be seen, the cumulative skin permeation of sialic acid per unit area in the free RhoB group and the RhoB nanocarrier group was 5.36 μg / cm² over 24 hours. 2 and 18.23 μg / cm 2 The skin retention amount was 35.29 μg / cm³. 2 and 87.16 μg / cm 2 When delivered through the skin via a carrier, compared to the free RhoB group containing sialic acid, the cumulative skin penetration of sialic acid in the RhoB nanocarrier multi-effect biomimetic liposome transdermal repair carrier increased by 240.1%, and the skin retention increased by 147.0%. This indicates that encapsulation by a nanocarrier can effectively promote the skin penetration and retention of active ingredients, thereby improving their skin bioavailability.

[0132] Depend on Figure 2 As can be seen, the free RhoB group was mostly concentrated in the stratum corneum after 2 hours and failed to penetrate the stratum corneum barrier. The RhoB nanocarrier group had already penetrated the stratum corneum barrier after 2 hours. With the extension of time, the fluorescence penetration depth of the RhoB nanocarrier group in the skin increased at 6 hours, reaching a skin depth of 430.2 μm. The results show that, within the same time period, the fluorescence intensity and penetration depth of the RhoB nanocarrier group in the skin were significantly stronger than those of the free RhoB group, indicating that the nanocarrier technology used in the multi-effect biomimetic liposome transdermal repair carrier obtained in Example 1 can promote the rapid and effective delivery of encapsulated active ingredients to the deep skin tissues, reaching the dermis.

[0133] Performance testing 4: Observation of cell uptake behavior using laser confocal microscopy

[0134] HDF and HaCaT cells in logarithmic growth phase were cultured at 3.0 × 10⁶ cells per dish. 5Cells were seeded in 35 mm confocal dishes and cultured for 24 h. Then, DMEM medium containing free RhoB (the product obtained in Comparative Example 15) and RhoB nanocarrier (the product obtained in Example 1) was added and incubated for 2 h and 4 h, respectively. After incubation, the medium was discarded, and the cells were washed three times with PBS solution. The cells were then fixed with 4% paraformaldehyde and stained with DAPI solution for 15 min each. The cells were observed and photographed under a 60x objective lens using a laser confocal microscope.

[0135] The results are as follows Figure 3 As shown, where, Figure 3 A and 3C are HDF cells. Figure 3 B and 3D are HaCaT cells, composed of... Figure 3 It was observed that after 2 hours of co-incubation, the intracellular fluorescence intensity of the free RhoB group was weak, while the fluorescence intensity of the RhoB-encapsulated nanocarrier had entered the cells and was significantly stronger than that of the free RhoB. After 4 hours of incubation, the fluorescence intensity of the RhoB nanocarrier further increased. These results indicate that compared to free RhoB, the RhoB nanocarrier can be taken up by more HaCaT and HDF cells, effectively delivering the encapsulated active ingredients into skin target cells to exert their effects, thus enhancing the cell entry efficiency and intracellular accumulation of the active ingredients.

[0136] Performance testing 5. Cell proliferation experiment

[0137] HDF and HaCaT cells in logarithmic growth phase were respectively treated with 4 × 10⁻⁶ 3 pcs / hole, 8×10 3 The culture medium was seeded at a density of 100 μL / well in 96-well plates and incubated at 5% CO2 and 37°C for 24 h. 100 μL of DMEM complete medium containing the multi-functional biomimetic liposome transdermal repair carrier prepared in Example 1 (the amount of multi-functional biomimetic liposome transdermal repair carrier added was such that the concentration of sialic acid in the medium was 20, 40, and 80 μg / mL, i.e., the liposome group) and DMEM complete medium containing the composition obtained in Comparative Example 15 (where the concentration of free sialic acid in the medium was the same as that of the liposome group, i.e., the free component) were added to each well. The control group was treated with 100 μL of DMEM complete medium (i.e., the free component). Figure 4 The cells were cultured in three replicates (within the normal control group) for 48 hours, and cell proliferation was measured using the CCK-8 assay.

[0138] Depend on Figure 4 It can be seen that, compared with the normal group, a certain concentration of free components and the multi-effect biomimetic liposome transdermal repair carrier of Example 1 can promote the proliferation of HaCaT and HDF cells (P<0.05 or P<0.01). Compared with the free components, the multi-effect biomimetic liposome transdermal repair carrier can significantly promote the proliferation of HDF cells (P<0.05 or P<0.01).

[0139] Performance testing 6ROS fluorescence intensity detection

[0140] HDF cells were loaded at 4 × 10 4 Cells were seeded at a density of 500 μL per well in 24-well plates. After 24 h of culture, the supernatant was discarded. The model group was added to DMEM medium containing 0.6 mmol / L H2O2, while the experimental groups were added to DMEM medium containing 0.6 mmol / L H2O2 and 400 mg / L of the multi-effect biomimetic liposome transdermal repair vector obtained in Examples 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 4, 5, and 1. A blank control group without H2O2 was also included. After 24 h of culture, DMEM medium containing 20 μM DCFH-DA was added and incubated for another 20 min. Cells were washed three times with PBS, and fluorescence intensity was observed under a fluorescence microscope. After collecting the cells, fluorescence intensity was detected by flow cytometry. Using 40 μg / mL as the experimental concentration, the same concentration of free sialic acid was prepared.

[0141] As shown in Table 1, compared with the model group, Comparative Examples 1-15, Example 4, Example 5 and Example 1 all significantly reduced the ROS fluorescence intensity (P<0.01).

[0142] Compared with Comparative Examples 1-4, Example 1 significantly suppressed ROS fluorescence intensity (P<0.01), indicating that the antioxidant effect of four components encapsulated in the nanocarrier—sialic acid, yeast / rice fermentation product filtrate (mafuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—is better than that of single components encapsulated in the nanocarrier.

[0143] Compared with Comparative Examples 5-13, Example 1 significantly suppressed ROS fluorescence intensity (P<0.01), indicating that the multi-effect biomimetic liposome transdermal repair carrier prepared by combining four active ingredients in Example 1—sialic acid, yeast / rice fermentation product filtrate (mafuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—had better suppression of ROS fluorescence intensity than any combination of three or two active ingredients (p<0.01), demonstrating that the combined use of active ingredients has a synergistic effect.

[0144] Compared with Comparative Example 14, the weight ratio of sialic acid, vitamin E (tocopherol), yeast / rice fermentation product filtrate (mafuyin), and arginine / lysine polypeptide (conopeptide) in Example 1 is not within the scope of this invention, resulting in a decrease in ROS inhibition rate and failure to achieve the best effect.

[0145] Compared with Examples 4 and 5, when the mass ratio of sialic acid to vitamin E is (2-4):1, the intracellular ROS inhibition effect of liposomes prepared by sialic acid and vitamin E at other mass ratios is better than that of liposomes prepared by other mass ratios of sialic acid and vitamin E.

[0146] Compared with the free composition (Comparative Example 15), the inhibition rate of ROS by the free component was 13.74% (i.e., (model group AU - free composition AU) / model group AU * 100%), while the inhibition rate of ROS by the multi-effect biomimetic liposome transdermal repair carrier of Example 1 was 36.76%. Compared with the free component, the multi-effect biomimetic liposome transdermal repair carrier of Example 1 can significantly reduce the intracellular ROS level of HDF cells (P < 0.01), resist damage caused by oxidative stress, and its effect is significantly better than that of the same dose of free component. Example 1 can significantly reduce ROS fluorescence intensity (P < 0.01).

[0147] Table 1. Average fluorescence intensity (AU)

[0148]

[0149]

[0150] Note: ** indicates comparison with the model group, p<0.01; ## indicates comparison with Comparative Examples 1-4, p<0.01; %% indicates comparison with Comparative Examples 5-13, p<0.01; && indicates comparison with Comparative Example 14, p<0.01; @@ indicates comparison with Comparative Example 15, p<0.01; ^^ indicates comparison with Examples 4 and 5, p<0.01.

[0151] Performance testing 7 Moisturizing and repairing factor testing

[0152] HaCaT cells were used at a rate of 2 × 10 4Cells were seeded at a density of 500 μL per well into multiple 24-well cell culture plates and incubated at 37°C with 5% CO2 for 24 h. 100 μL of DMEM complete medium containing the multi-effect biomimetic liposome transdermal repair carrier from each example (the amount of multi-effect biomimetic liposome transdermal repair carrier added ensures that the concentration of sialic acid in the medium is 40 μg / mL) and DMEM complete medium containing the comparative component solutions (where the concentration of the comparative components in the medium is consistent with the concentration of the active ingredient in the multi-effect biomimetic liposome transdermal repair carrier prepared in the examples) were added to each well. The blank control group received only 100 μL of DMEM complete medium, while the positive control group received 100 μL of DMEM complete medium containing vitamin C (vitamin C concentration 40 μg / mL). Each group was divided into three replicates. The plates were incubated in a CO2 incubator for 24 h. The supernatant was collected, and the contents of AQP3, FLG, HA, and Claudin1 were measured using an ELISA kit.

[0153] As shown in Table 2, compared with the blank control group and the positive group, Comparative Examples 1-15, Example 4, Example 5 and Example 1 significantly promoted the secretion of FLG, AQP3, HA and Claudin1 by cells (P<0.05 or P<0.01).

[0154] Compared with Comparative Examples 1-4, each example significantly promoted the secretion of FLG, AQP3, HA and Claudin1 by cells (P<0.05 or P<0.01), indicating that the moisturizing and repairing effect of four components encapsulated in the nanocarrier—sialic acid, yeast / rice ferment filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—is better than that of single components encapsulated in the nanocarrier.

[0155] Compared with Comparative Examples 5-13, Example 1 significantly promoted the secretion of FLG, AQP3, HA and Claudin1 by cells, indicating that the moisturizing and repairing ability of the liposomes prepared by combining the four active ingredients of sialic acid, yeast / rice fermentation product filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol) in each example is better than that of any combination of three or two active ingredients (p<0.01), indicating that the combined use of active ingredients has a synergistic effect.

[0156] Compared with Comparative Example 14, the weight ratio of sialic acid, vitamin E (tocopherol), yeast / rice fermentation product filtrate (maifune), and arginine / lysine polypeptide (conopeptide) in Example 1 is not within the scope of this invention, resulting in a decrease in the effect of promoting cell secretion of FLG, AQP3, HA, and Claudin1, and failing to achieve the best moisturizing and repairing effect.

[0157] Compared with Examples 4 and 5, Example 1 showed that when the mass ratio of sialic acid to vitamin E was (2-4):1, the liposomes prepared with sialic acid to vitamin E at other mass ratios were better at promoting the secretion of FLG, AQP3, HA, and Claudin1. This indicates that when the mass ratio of sialic acid to vitamin E was (2-4):1, the prepared liposome composition had a more significant moisturizing and repairing effect.

[0158] Compared with the free composition (Comparative Example 15), the examples showed a significant increase in the contents of FLG, AQP3, HA and Claudin1 (P < 0.01), indicating that the encapsulated liposome composition with anti-aging, moisturizing and repairing effects has a more significant moisturizing and repairing effect than the same dose of free component.

[0159] Table 2. Content of FLG, AQP3, HA, and Claudin1 secreted by cells.

[0160]

[0161]

[0162] Note: ** indicates comparison with blank control and positive group, p<0.01; ## indicates comparison with comparative examples 1-4, p<0.01; %% indicates comparison with comparative examples 5-11, p<0.01; && indicates comparison with comparative example 12, p<0.01; @@ indicates comparison with comparative example 13, p<0.01; ^^ indicates comparison with examples 4 and 5, p<0.01.

[0163] Performance testing: Detection of anti-aging factors

[0164] HDF cells were loaded at 4 × 10 4 Cells were seeded at a density of 500 μL per well into multiple 24-well cell culture plates and incubated at 37°C with 5% CO2 for 24 h. 100 μL of DMEM complete medium containing the products obtained in Examples 1, 4, 5, and Comparative Examples 1-15 was added to each well (the amount added in each example and comparative example ensured that the concentration of the active ingredient in the medium was 40 μg / mL). The blank control group received only 100 μL of DMEM complete medium. Each group had three replicates. The model group received 100 μL of DMEM complete medium containing vitamin C (vitamin C concentration 40 μg / mL). The plates were incubated in a CO2 incubator for 24 h. The supernatant was collected, and the contents of MMP-1, MMP-3, Col I, and Col III were measured using an ELISA kit.

[0165] As shown in Table 3, compared with the model group, Comparative Examples 1-15, Example 4, Example 5 and Example 1 significantly reduced the levels of MMP-1 and MMP-3 secreted by cells and significantly promoted the levels of COL I and COL III secreted by cells.

[0166] Compared with Comparative Examples 1-4, Example 1 significantly reduced the levels of MMP-1 and MMP-3 secreted by cells and significantly promoted the levels of COL I and COL III secreted by cells. This indicates that the antioxidant effect of the four components encapsulated in the nanocarrier—sialic acid, yeast / rice fermentation product filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—is better than that of the single components encapsulated in the nanocarrier—sialic acid, yeast / rice fermentation product filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol).

[0167] Compared with Comparative Examples 5-13, Example 1 significantly inhibited the secretion of MMP-1 and MMP-3 by cells, as well as the secretion of COL I and COL III by cells. This indicates that the liposomes prepared by combining four active ingredients in Example 1—sialic acid, yeast / rice fermentation product filtrate (mafuyin), arginine / lysine polypeptide (conopeptide), and vitamin E (tocopherol)—have better collagen-promoting and anti-aging effects than any combination of three or two active ingredients (p<0.01). This demonstrates that the combined use of active ingredients has a synergistic effect.

[0168] Compared with Comparative Example 14, the weight ratio of sialic acid, vitamin E (tocopherol), yeast / rice fermentation product filtrate (mafuyin), and arginine / lysine polypeptide (conopeptide) in Example 1 is not within the scope of this invention, and the effects of inhibiting cell secretion of MMP-1 and MMP-3 and promoting cell secretion of COL I and COL III are not as good as in Example 1.

[0169] Compared with Examples 4 and 5, when the mass ratio of sialic acid to vitamin E is (2-4):1, the liposomes prepared by sialic acid to vitamin E have better effects on inhibiting the secretion of MMP-1 and MMP-3 and promoting the secretion of COL I and COL III than those prepared by other mass ratios of sialic acid to vitamin E.

[0170] Compared with the free component (Comparative Example 15), Example 1 significantly increased the content of COLⅠ and COLⅢ and decreased the content of MMP-1 and MMP-3. This indicates that the anti-aging effect of the encapsulated multi-effect biomimetic liposome transdermal repair carrier is more significant and significantly better than that of the same dose of free component.

[0171] Table 3. Content of MMP-1, MMP-3, COL I, and COL III secreted by cells.

[0172]

[0173]

[0174] Note: ** indicates comparison with the model group, p<0.01; ## indicates comparison with Comparative Examples 1-4, p<0.01; %% indicates comparison with Comparative Examples 5-11, p<0.01; && indicates comparison with Comparative Example 12, p<0.01; @@ indicates comparison with Comparative Example 13, p<0.01; ^^ indicates comparison with Examples 4 and 5, p<0.01.

[0175] Performance testing of tyrosinase activity and melanin production content.

[0176] Take B16F10 cells in the logarithmic growth phase and distribute them at a density of 5 × 10⁶ cells per well. 4 Cells were seeded at a density of 100 μL in multiple 24-well plates and cultured for 24 h. The cells were then divided into a blank control group, a model group, and an experimental group, with three replicates in each group. The model group received DMEM complete medium containing only 100 nmol / L α-MSH. The experimental groups received DMEM complete medium containing 100 nmol / L α-MSH, the products obtained from each comparative example, and the products obtained from each embodiment (the amount added to each comparative example and embodiment was such that the effective ingredient was 40 μg / mL in the medium). The blank control group received only 100 μL of DMEM complete medium. After culturing for another 48 hours, 200 μL of cell lysis buffer containing 1% (V%) Triton X-100 was added to each well, and the cells were frozen at -80℃ for 30 min. The cell lysis buffer was collected and centrifuged, and 100 μL of the supernatant was transferred to a 96-well plate. 100 μL of 0.1% (W / V) L-DOPA solution was added, and the plate was incubated at 37℃ for 2 h. The absorbance (A) of each well was measured at a wavelength of 495 nm using a microplate reader.

[0177] The intracellular melanin content was then determined using the NaOH lysis method. B16F10 cells were lysed at 5 × 10⁻⁶. 4Cells were seeded at a density of 2 mL / mL in multiple 6-well plates and cultured for 24 h. 100 nM α-MSH was added to each well to induce melanin expression (except for the blank control group) to construct an α-MSH-induced melanin overexpression model. DMEM complete medium containing 100 nmol / L α-MSH, the products obtained from each comparative example, and the products obtained from each embodiment was added (the amount added to each comparative example and embodiment was 40 μg / mL of the active ingredient in the medium). The blank control group received only 100 μL of DMEM complete medium, while the model group received 100 μL of DMEM complete medium containing vitamin C (vitamin C concentration 40 μg / mL). Each group had 3 replicates. After 48 h of culture, the supernatant was discarded, and the cells were washed three times with PBS. 300 μL of 1.0 mmol / L NaOH solution (containing 10% DMSO) was added to each well, and the cells were lysed at 80 °C for 1 h. The absorbance (A) of each well was measured at 405 nm.

[0178] As shown in Table 4, compared with the model group, Comparative Examples 1-15, Example 4, Example 5 and Example 1 can significantly reduce the tyrosinase activity and melanin content in melanocytes.

[0179] Compared with Comparative Examples 1-4, Example 1 significantly reduced tyrosinase activity and melanin content in melanocytes, indicating that the whitening effect of nanocarrier encapsulating four components—sialic acid, yeast / rice ferment filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—is superior to that of nanocarrier encapsulating single components—sialic acid, yeast / rice ferment filtrate (Maifuyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol).

[0180] Compared with Comparative Examples 5-13, Example 1 significantly reduced tyrosinase activity and melanin content in melanocytes, indicating that the whitening effect of the liposomes prepared by combining four active ingredients in Example 1—sialic acid, yeast / rice fermentation product filtrate (maifune), arginine / lysine polypeptide (conopeptide), and vitamin E (tocopherol)—was better than any combination of three or two active ingredients (p<0.01). This demonstrates that the combined use of active ingredients has a synergistic effect.

[0181] Compared with Comparative Example 14, the weight ratio of sialic acid, vitamin E (tocopherol), yeast / rice fermentation product filtrate (mafuyin), and arginine / lysine polypeptide (conopeptide) in Example 1 is not within the scope of this invention, resulting in a decrease in the inhibitory effect on tyrosinase activity and melanin content in melanocytes, and thus failing to achieve the best effect.

[0182] Compared with Examples 4 and 5, Example 1 showed that when the mass ratio of sialic acid to vitamin E was (2-4):1, the inhibitory effect on tyrosinase activity and melanin content in melanocytes was better than that on liposomes prepared with other mass ratios of sialic acid and vitamin E. This further demonstrates that when the mass ratio of sialic acid to vitamin E is (2-4):1, it can significantly inhibit tyrosinase activity and melanin content in melanocytes, resulting in a better whitening effect.

[0183] Compared with the free component (Comparative Example 15), the whitening effect of the encapsulated multi-effect biomimetic liposome transdermal repair carrier is more significant and is significantly better than that of the same dose of the free component.

[0184] Table 4 Tyrosinase activity and melanin production content

[0185] Group Tyrosinase activity (%) Melanin production content (%) Blank control group 47.55±3.21 67.82±3.21 Model group 99.90±4.23 100.00±4.23 Example 1 <![CDATA[80.62±2.66 **##％％&&@@^^ ]]> <![CDATA[87.09±2.26 **##％％&&@@^^ ]]> Example 4 <![CDATA[86.62±5.21 ** ]]> <![CDATA[90.02±1.93 ** ]]> Example 5 <![CDATA[86.03±3.46 ** ]]> <![CDATA[90.21±1.96 ** ]]> Comparative Example 1 <![CDATA[92.79±3.54 ** ]]> <![CDATA[93.12±1.56 ** ]]> Comparative Example 2 <![CDATA[92.15±2.18 ** ]]> <![CDATA[93.23±1.37 ** ]]> Comparative Example 3 <![CDATA[93.29±4.52 ** ]]> <![CDATA[93.78±2.08 ** ]]> Comparative Example 4 <![CDATA[91.53±2.08 ** ]]> <![CDATA[93.56±1.84 ** ]]> Comparative Example 5 <![CDATA[91.02±2.54 ** ]]> <![CDATA[92.28±1.59 ** ]]> Comparative Example 6 <![CDATA[90.98±4.13 ** ]]> <![CDATA[92.17±1.55 ** ]]> Comparative Example 7 <![CDATA[91.12±3.62 ** ]]> <![CDATA[92.32±1.82 ** ]]> Comparative Example 8 <![CDATA[91.54±3.11 ** ]]> <![CDATA[92.08±2.05 ** ]]> Comparative Example 9 <![CDATA[92.03±2.05 ** ]]> <![CDATA[93.02±2.54 ** ]]> Comparative Example 10 <![CDATA[92.12±4.07 ** ]]> <![CDATA[92.98±1.77 ** ]]> Comparative Example 11 <![CDATA[91.73±2.29 ** ]]> <![CDATA[92.65±2.13 ** ]]> Comparative Example 12 <![CDATA[91.82±1.56 ** ]]> <![CDATA[92.39±1.82 ** ]]> Comparative Example 13 <![CDATA[91.08±2.05 ** ]]> <![CDATA[92.84±1.63 ** ]]> Comparative Example 14 <![CDATA[89.94±1.65 ** ]]> <![CDATA[91.68±1.02 ** ]]> Comparative Example 15 <![CDATA[95.45±1.53 ** ]]> <![CDATA[98.06±2.93 ** ]]>

[0186] Note: ** indicates comparison with the model group, p<0.01; ## indicates comparison with Comparative Examples 1-4, p<0.01; %% indicates comparison with Comparative Examples 5-11, p<0.01; && indicates comparison with Comparative Example 12, p<0.01; @@ indicates comparison with Comparative Example 13, p<0.01; ^^ indicates comparison with Examples 4 and 5, p<0.01.

[0187] Performance testing 10 Anti-aging and wrinkle reduction effect evaluation

[0188] Test Methods: Two hundred healthy women (45-55 years old) were selected and divided into 20 groups of 10 each. Each group used the products obtained in Examples 1-5 and Comparative Examples 1-15, respectively. Participants used the samples twice daily, morning and evening, for 56 days. Participants were not allowed to use other products during the study period. Test results were evaluated after 56 days of sample use. Before the trial, skin wrinkles and skin elasticity were measured. Skin wrinkles and skin elasticity were measured again after 7, 14, 28, and 56 days of product use. Facial skin wrinkles were measured using a Skin Visiometer SV600; skin elasticity was measured using a skin elasticity meter. The dual MPA580 was used for measurement, and the average value of each group of 10 people was calculated.

[0189] Wrinkle change rate (%) = (average wrinkle value of each group after using the product in the example / comparative example - average wrinkle value of each group before use) / average wrinkle value of each group before use * 100%. The wrinkle improvement rate in Table 5 is calculated by taking the positive value of the change rate. The results are shown in Table 5.

[0190] Table 5. Wrinkle improvement rate after different periods of product use in Examples 1-5 and Comparative Examples 1-15.

[0191]

[0192]

[0193] Table 5 shows that the products in Examples 1-5 all exhibited significant wrinkle-reducing effects after use, with better results observed over longer usage. The wrinkle-reducing and anti-aging performance of Example 1 was significantly superior to that of Comparative Examples 1-14, indicating that the synergistic effect of the four components encapsulated in the nanocarrier described in this invention—sialic acid, yeast / rice fermentation product filtrate (maifune), arginine / lysine polypeptide (conopeptide), and vitamin E (tocopherol)—is crucial for maximizing the wrinkle-reducing and anti-aging effects of the active ingredients. Compared to the free component (Comparative Example 15), this demonstrates that the wrinkle-reducing and anti-aging performance of the encapsulated multi-effect biomimetic liposome transdermal repair carrier is significantly superior to that of the same dose of free component.

[0194] Performance Testing 11 Moisturizing Efficacy Test

[0195] Test samples: Products obtained from Examples 1-5 and Comparative Examples 1-15

[0196] Test subjects: 200 healthy women (45-55 years old) were selected and divided into 20 groups of 10 each. Before each test, the subjects rested for 30 minutes in a laboratory environment with a temperature of 21±1℃ and a relative humidity of 50±5%RH.

[0197] The subject wiped their forearm with a dry tissue. The experimenter marked a 3cm x 3cm test area on the inside of both forearms of the subject as the area for the test sample. The experimenter measured the transepidermal water loss rate of the skin on the inside of the forearm. The experimenter applied ordinary transparent tape to the test area, then peeled off the tape, and repeated this operation 15 times before applying the sample.

[0198] Test method: Transdermal water loss after tape peeling was measured using a Tewameter™ 300 (Courage & Khazak, Germany) to measure the recovery of skin barrier function and evaluate the soothing and repairing effects of the test samples.

[0199] The results were tested at the following four time periods: immediately after the tape peeling operation (t=0), 10 minutes after the tape peeling operation, and again on day 1 and day 3.

[0200] Test results: Change rate of transepidermal water loss in skin

[0201] Table 6. Changes in transepidermal water loss of the stratum corneum (%)

[0202]

[0203]

[0204] Transepidermal water loss rate of skin stratum corneum = (Transepidermal water loss rate measurement after use - Transepidermal water loss rate measurement before use) ÷ Transepidermal water loss rate measurement before use × 100%

[0205] The pre-use transdermal water loss measurement refers to the transdermal water loss rate measured immediately after peeling (t=0).

[0206] The "measured value" in this formula refers to the transepidermal water loss (TEWL) value of the stratum corneum measured by the Tewameter™ 210. The higher the TEWL value, the more water is lost through the skin, and the worse the barrier function of the stratum corneum. Therefore, the greater the decrease in the TEWL value (i.e., the greater the absolute value of the TEWL change rate), the stronger the moisturizing effect on the skin.

[0207] As can be seen from the results in Table 6 above, the change rate of TEWL in Example 1 is greater than that in Comparative Examples 1-14, proving that when all four components—sialic acid, yeast / rice fermentation product filtrate (Maifoyin), arginine / lysine peptide (conopeptide), and vitamin E (tocopherol)—are added to the nanocarrier described in this invention, the TEWL is lower than when only one or two of them are added, resulting in better skin barrier repair and water-locking moisturizing effects. Compared with the free component (Comparative Example 15), this demonstrates that the multi-effect biomimetic liposome transdermal repair carrier after encapsulation has significantly better skin barrier repair and water-locking moisturizing effects than the same dose of free component.

[0208] In summary, it can be seen that this invention, through the rational combination of four functional ingredients and combined with nanocarrier technology, forms a multi-effect biomimetic transdermal repair carrier with good anti-aging, whitening, moisturizing and repairing effects.

Claims

1. A multi-effect biomimetic liposome transdermal repair carrier, characterized in that, This multi-effect biomimetic liposome transdermal repair carrier includes active ingredients and nanocarriers. The active ingredients include sialic acid, yeast / rice fermentation product filtrate, arginine / lysine peptides, and vitamin E in a mass ratio of (1-10):(2-8):(0.1-1):(0.5-4).

2. The multi-effect biomimetic liposome transdermal repair carrier according to claim 1, characterized in that, The raw materials for the nanocarrier include phospholipids, emulsifiers, polyols, and water.

3. The multi-effect biomimetic liposome transdermal repair carrier according to claim 2, characterized in that, Based on the total mass of the multi-effect biomimetic liposome transdermal repair carrier as 100%, it includes 1-10% sialic acid, 2-8% yeast / rice fermentation product filtrate, 0.1-1% arginine / lysine peptides, 0.5-4% vitamin E, 0.1-2% phospholipids, 5-30% emulsifiers, 10-40% polyols, and the balance being water.

4. The multi-effect biomimetic liposome transdermal repair carrier according to claim 2, characterized in that, The phospholipids include one or more of the following: soybean lecithin, hydrogenated lecithin, egg yolk lecithin, hydrogenated soybean lecithin, and hydrogenated egg yolk lecithin. The emulsifiers include one or more of the following: polyoxyethylene sorbitan fatty acid esters, polyglycerol fatty acid esters, polyoxyethylene fatty acid esters, polyoxyethylene castor oil emulsifiers, polyoxyethylene hydrogenated castor oil emulsifiers, PEG-8 caprylic / capric glycerides, polyglycerol emulsifiers, poloxamer, cocoyl glucoside, triglycerides, pyrrolidones, polyglycerol esters, PEG-30 dihydroxystearate and cetearyl alcohol polyethers, Tween 80, Tween 20, Tween 60, PEG-20 hydrogenated castor oil, PEG-40 hydrogenated castor oil, and PEG-60 hydrogenated castor oil.

5. The multi-effect biomimetic liposome transdermal repair carrier according to claim 2, characterized in that, The polyols include one or more of glycerol, propylene glycol, 1,3-butanediol, 1,3-propanediol, 1,2-pentanediol, ethoxydiethylene glycol, 1,2-hexanediol, dipropylene glycol, isopropanol, polyethylene glycol-200, PPG-10 sorbitol, octyldodecyl alcohol, and dipropylene glycol.

6. The multi-effect biomimetic liposome transdermal repair carrier according to claim 3, characterized in that, The raw materials for the nanocarrier also include a pH adjuster at a mass percentage of 1-2%.

7. The multi-effect biomimetic liposome transdermal repair carrier according to claim 6, characterized in that, The pH adjuster mentioned includes tromethamine.

8. A method for preparing the multi-effect biomimetic liposome transdermal repair carrier as described in claim 1, characterized in that, Includes the following steps: (1) Mix vitamin E with emulsifier, polyol and phospholipid to obtain mixture A; (2) Mix sialic acid, yeast / rice fermentation product filtrate, arginine / lysine peptide, pH adjuster, polyol and water to obtain mixture B; (3) Pour mixture A into mixture B and mix well to obtain mixture C; (4) The mixture C was nano-sized to obtain a multi-effect biomimetic liposome transdermal repair carrier.

9. The method for preparing the multi-effect biomimetic liposome transdermal repair carrier according to claim 8, characterized in that, In step (1), the mixing conditions are: stirring at 30-65℃ until homogeneous; In step (2), the mixing conditions are: stirring at 30-65℃ until homogeneous; In step (3), the mixing conditions are: stirring at 30-65℃ until homogeneous; In step (4), the nano-sizing process is homogenization using a high-pressure homogenizer at a pressure of 500-1200 bar.

10. A skincare product, characterized in that, Contains the multi-effect biomimetic liposome transdermal repair carrier as described in any one of claims 1-7.

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