Ginsenoside Rg3 nano-liposome easier to transdermally absorb as well as preparation method and application of ginsenoside Rg3 nano-liposome

Abstract

The invention discloses a ginsenoside Rg3 nano-liposome easier for transdermal absorption as well as a preparation method and application thereof, and belongs to the technical field of liposome preparation. The ginsenoside Rg3 nano-liposome easier to transdermally absorb, provided by the invention, is prepared from the following raw materials in parts by mass: 5 to 20 parts of polyhydric alcohol, 2 to 10 parts of ginsenoside Rg3, 1 to 8 parts of phospholipid, 1 to 5 parts of cholesterol, 1 to 2 parts of resveratrol, 0 to 2 parts of a compound functional substance and 1 to 5 parts of water. The ginsenoside Rg3 nano-liposome easier to transdermally absorb, provided by the invention, has excellent anti-oxidation and anti-inflammatory characteristics through a compound synergistic effect of all the components, can regulate the skin state, comprehensively protect the skin from being damaged by free radicals and inflammation, solves the skin problem caused by external stimulation, and has a good application prospect. The skin care composition has the effects of protecting skin, maintaining skin barrier function, improving skin redness and improving skin stability, can be compounded with any skin care preparation for use, and has wide market application prospect.

Description

Technical Field

[0001] The present application belongs to the technical field of liposome preparation, and in particular relates to a ginsenoside Rg3 nanoliposome that is easier to be absorbed through the skin, and a preparation method and application thereof. Background Art

[0002] Ginsenoside Rg3 can restore the structural and functional characteristics of damaged skin, inhibit inflammatory reactions at the site of skin lesions, regulate the balance of oxidation and anti-oxidation of skin tissue, and improve fibroblast proliferation by increasing the thickness of the epidermis and dermis, promote the wound healing process, and inhibit the early formation and proliferation of scars. Ginsenoside Rg3 liposomes are usually prepared using liposome technology, but they still have the characteristics of low solubility and short half-life, which limits their bioavailability in the human body. Therefore, it is necessary to develop a ginsenoside Rg3 liposome with excellent comprehensive performance.

[0003] The prior art with publication number CA 103417479 A discloses a method for preparing ginsenoside Rg3 liposomes, comprising completely dissolving phospholipids, cholesterol, and ginsenoside Rg3 in n-butanol, subjecting the mixture to reduced pressure rotary evaporation, adding a phosphate buffer solution with a pH value of 6.0-7.5, and subjecting the mixture to high-pressure homogenization circulation treatment to obtain a liposome suspension with an average particle size of 100 nm, and subjecting the mixture to spray drying and freeze drying to obtain ginsenoside Rg3 liposomes.

[0004] However, the liposomes prepared as above have the following problems: first, the single ginsenoside Rg3 active component leads to a large amount of liposomes used and high cost; second, the prepared ginsenoside Rg3 liposomes have low solubility and poor permeability, resulting in poor bioavailability. Summary of the invention

[0005] The present application discloses a ginsenoside Rg3 nanoliposome that is easier to be absorbed through the skin, and a preparation method and application thereof, aiming to solve the technical problems of existing liposomes such as large usage, high cost, low solubility, poor permeability and poor bioavailability.

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

[0007] The first aspect of the present application provides a ginsenoside Rg3 nanoliposome that is easier to be absorbed through the skin, and its components, calculated by weight of raw materials, include:

[0008] Polyol 5-20 parts;

[0009] Ginsenoside Rg3 2-10 parts;

[0010] 1-8 parts of phospholipids;

[0011] Cholesterol 1-5 parts;

[0012] 1-2 parts of resveratrol;

[0013] 0-2 parts of compound efficacy ingredients;

[0014] 1-5 parts water;

[0015] Wherein, the compounded effective substance is one or more of VC ethyl ether, folic acid and bosine.

[0016] In combination with the first aspect, preferably, the components thereof include, by weight of raw materials:

[0017] Polyol 10-15 parts;

[0018] Ginsenoside Rg3 8-10 parts;

[0019] 1-5 parts of phospholipids;

[0020] 1-2 parts cholesterol;

[0021] Resveratrol 0.5-2 parts;

[0022] 0-2 parts of compound efficacy ingredients;

[0023] 1-5 parts water;

[0024] Wherein, the compounded effective substance is one or more of VC ethyl ether, folic acid and bosine.

[0025] In combination with the first aspect, preferably, the particle size of the ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin is 100-150 nm, and the encapsulation rate is ≥90%.

[0026] In combination with the first aspect, preferably, the polyol is one or more of glycerol, butanediol, 1,2-hexanediol, 1,3-propylene glycol, polyethylene glycol, and sorbitol.

[0027] In combination with the first aspect, preferably, the phospholipid is one or more of lecithin, hydrogenated lecithin, soybean lecithin, egg yolk lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine.

[0028] The second aspect of the present application provides a method for preparing the ginsenoside Rg3 nanoliposomes that are easier to be absorbed through the skin as described in the first aspect, the method comprising:

[0029] Phospholipids, cholesterol, resveratrol and ginsenoside Rg3 were dissolved in ethanol and then subjected to rotary evaporation to form a lipid film;

[0030] After dissolving ginsenoside Rg3, resveratrol, polyols and compounded functional substances in water, the lipid film is added and subjected to ultrasonic and high-pressure homogenization treatment to obtain the ginsenoside Rg3 nanoliposomes which are more easily absorbed through the skin.

[0031] In combination with the second aspect, preferably, when the phospholipids, cholesterol, resveratrol and ginsenoside Rg3 are dissolved in ethanol and then subjected to rotary evaporation, the temperature is 40-60°C.

[0032] In combination with the second aspect, preferably, the temperature of the ultrasonic treatment is 30-50°C, the time is 30-60min, and the ultrasonic power is 50-200W.

[0033] In combination with the second aspect, preferably, the conditions of the high-pressure homogenization treatment are: pressure of 100-800 bar, and number of cycles of 1-5 times.

[0034] The third aspect of the present application provides the use of ginsenoside Rg3 nanoliposomes which are more easily absorbed through the skin and are prepared by the preparation method of the second aspect in the preparation of skin care products.

[0035] Compared with the prior art, the advantages or beneficial effects of the embodiments of the present application include at least:

[0036] The ginsenoside Rg3 nanoliposomes provided in the present application, which are more easily transdermally absorbed, are made according to raw materials including ginsenoside Rg3, polyols, resveratrol, phospholipids, cholesterol, compounded functional substances and water. On the one hand, the permeability of the liposomes can be greatly improved, and the active components can be delivered to the deep layers of the skin, thereby fundamentally improving the skin condition. On the second hand, through the synergistic effect of the compounding of the components, the liposomes have excellent antioxidant and anti-inflammatory properties, can regulate the skin condition, and comprehensively protect the skin from free radicals and inflammation, while solving skin problems caused by external stimuli, maintaining the skin barrier function, improving skin redness, and improving skin stability. On the third hand, the solubility of the active components can be greatly improved, the residence time on the skin can be prolonged, and its bioavailability can be increased, so that the usage amount is greatly reduced, and the usage cost is reduced. In addition, the liposome has a high encapsulation rate, which can reach up to 92.4%. On the fourth hand, the liposomes can be endowed with excellent skin affinity and are suitable for a variety of skins. At the same time, they can be compounded with any skin care preparations, so that the usage amount is greatly reduced and excellent effects are achieved, reducing costs, and having broad market application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following briefly introduces the drawings required for use in the description of the embodiments. Obviously, the drawings described below are only some embodiments recorded in the present application, and for ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work.

[0038] Figure 1IC of positive control VC in ABTS free radical scavenging experiment of ginsenoside Rg3 nanoliposomes which are easier to be absorbed through skin prepared in the present application example 50 picture;

[0039] Figure 2 A statistical graph showing the results of ABTS free radical scavenging rate of ginsenoside Rg3 nanoliposomes which are easier to be absorbed through the skin and prepared in the examples of the present application;

[0040] Figure 3 This is a statistical chart of the cell viability results of the ginsenoside Rg3 nanoliposomes that are easier to be absorbed through the skin prepared in the examples of the present application;

[0041] Among them, the comparison between UV group and blank group####P<0.0001; compared with UV group*P<0.05,**P<0.01,***P<0.005,****P<0.0001;

[0042] Figure 4 This is a statistical chart of the results of the cell anti-inflammatory experiment of the ginsenoside Rg3 nanoliposomes which are easier to be absorbed through the skin and prepared in the examples of the present application;

[0043] Among them, the comparison between LPS group and blank group ####P<0.0001; compared with LPS group *P<0.05, ****P<0.0001. DETAILED DESCRIPTION

[0044] The technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the drawings in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0045] In the following description of this embodiment, the term "and / or" is used to describe the association relationship of associated objects, indicating that there may be three relationships. For example, A and / or B can represent: A exists alone, B exists alone, and A and B exist at the same time. A and B can be singular or plural. The character " / " generally indicates that the associated objects before and after are in an "or" relationship.

[0046] In the following description of this embodiment, the term "at least one" refers to one or more, and "plurality" refers to two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single items or plural items. For example, "at least one of a, b or c", or "at least one of a, b and c", can all represent: a, b, c, ab (i.e., a and b), ac, bc, or abc, where a, b, c can be single or multiple, respectively.

[0047] Those skilled in the art should understand that in the following description of the embodiments of the present application, the order of serial numbers does not mean the order of execution, some or all of the steps can be executed in parallel or sequentially, and the execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0048] The terms used in the embodiments of the present application are only for the purpose of describing specific embodiments, and are not intended to limit the present application. The singular forms "a", "an" and "the" used in the embodiments of the present application and the appended claims are also intended to include plural forms, unless the context clearly indicates other meanings.

[0049] Unless otherwise specified, all raw materials and reagents used in the examples of the present application are purchased on the market or prepared according to conventional methods well known to those skilled in the art.

[0050] In the first aspect, the present invention provides a ginsenoside Rg3 nanoliposome that is easier to be absorbed through the skin, and its components, calculated by weight of raw materials, include:

[0051] Polyol 5-20 parts;

[0052] Ginsenoside Rg3 2-10 parts;

[0053] 1-8 parts of phospholipids;

[0054] Cholesterol 1-5 parts;

[0055] 1-2 parts of resveratrol;

[0056] 0-2 parts of compound efficacy ingredients;

[0057] 1-5 parts water;

[0058] Wherein, the compounded effective substance is one or more of VC ethyl ether, folic acid and bosine.

[0059] Among them, on the one hand, it can greatly improve the permeability of liposomes, and can deliver active ingredients to the deep layers of the skin, fundamentally improving the skin condition; on the second hand, through the synergistic effect of the compounding of various components, the liposomes have excellent antioxidant and anti-inflammatory properties, can regulate the skin condition, and comprehensively protect the skin from free radicals and inflammation, while solving skin problems caused by external stimuli, maintaining the skin barrier function, improving skin redness, and improving skin stability; on the third hand, it can greatly improve the solubility of active ingredients, prolong the residence time on the skin, increase its bioavailability, and greatly reduce the usage amount, reduce the cost of use, and the liposome encapsulation rate is high, up to 92.4%; on the fourth hand, it can give liposomes excellent skin affinity, suitable for a variety of skins, and at the same time, it can be compounded with any skin care preparations, so that the dosage is greatly reduced and excellent effects are achieved, reducing costs, and has broad market application prospects.

[0060] In the embodiment of the present application, the components include, by weight of raw materials: 10-15 parts of polyols; 8-10 parts of ginsenoside Rg3; 1-5 parts of phospholipids; 1-2 parts of cholesterol; 0.5-2 parts of resveratrol; 0-2 parts of compound efficacy; 1-5 parts of water; wherein the compound efficacy is preferably one or more of VC ethyl ether, folic acid, and bosine. Among them, by selecting these raw materials and controlling the content of each component, the liposome can be endowed with excellent product stability, antioxidant and anti-inflammatory properties, permeability, solubility and skin affinity.

[0061] The second aspect of the present application provides a method for preparing the ginsenoside Rg3 nanoliposomes that are easier to be absorbed through the skin as described in the first aspect, the method comprising:

[0062] Phospholipids, cholesterol, resveratrol and ginsenoside Rg3 were dissolved in ethanol and then subjected to rotary evaporation to form a lipid film;

[0063] After dissolving ginsenoside Rg3, resveratrol, polyols and compounded functional substances in water, the lipid film is added and subjected to ultrasonic and high-pressure homogenization treatment to obtain the ginsenoside Rg3 nanoliposomes which are more easily absorbed through the skin.

[0064] It should be noted that the preparation process adopted in this application is to prepare liposomes by a two-step method, which can make the two main active components (ginsenoside Rg3 and resveratrol) not only exist in the core material of the liposome, but also exist on the wall material of the liposome, so that the comprehensive performance of the liposome is greatly improved. In addition, the particle size of the liposomes treated by this preparation process is uniform, and the preparation process is simple and the operating conditions are mild.

[0065] It should be noted that through the selection of each component and the control of the proportion, it can be endowed with excellent antioxidant properties. Through the synergistic effect of each component, when liposomes are used on the skin, they can smoothly supply nutrients to the skin by dilating capillaries and promoting blood circulation, thereby inhibiting skin aging.

[0066] It should be noted that after the lipids in the stratum corneum of the skin are damaged, the skin becomes chapped. Ginsenoside Rg3 nanoliposomes play a leading role in restoring skin moisture. They can effectively associate water molecules and form a network structure to maintain skin moisture. At the same time, they can connect the lipid matrix and keratinized cells to enhance the barrier function of the skin and reduce the entry of harmful substances into the skin through the pores and sweat glands, thus having an anti-inflammatory effect.

[0067] The third aspect of the present application provides the use of the ginsenoside Rg3 nanoliposomes that are easier to be absorbed through the skin prepared by the preparation method described in the second aspect in the preparation of skin care products. Among them, based on the excellent stability, antioxidant and anti-inflammatory properties, permeability, solubility and skin affinity of the above-mentioned ginsenoside Rg3 nanoliposomes, the product is endowed with excellent product competitiveness.

[0068] The technical solution of the present application will be further described below in conjunction with specific embodiments.

[0069] Example 1

[0070] This embodiment provides a method for preparing A1-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin, specifically comprising:

[0071] S101: 5 parts of lecithin, 1 part of cholesterol, 3 parts of ginsenoside Rg3 and 1 part of resveratrol were added into a dry single-mouth round flask, and anhydrous ethanol was added to dissolve them at 60°C and 500rpm magnetic stirring to obtain a lipid mixture, which was subjected to reduced pressure rotary evaporation at 60°C to remove the anhydrous ethanol in the bottle and form a uniform A1-lipid film on the inner wall of the flask;

[0072] S102: 5 parts of ginsenoside Rg3, 1 part of resveratrol, 20 parts of glycerol, 1 part of bosine and 5 parts of water are stirred magnetically at 50°C and 500rpm until the solution becomes clear, and then A1-lipid film is added. Under constant temperature conditions of 50°C, a high-speed shearing instrument is used at 10,000rpm for 3 minutes, and ultrasonic treatment is performed at 100W power and 50°C for 10 minutes. After high-pressure homogenization treatment at a pressure of 600 bar and a temperature of 50°C, the mixture is cycled 4 times and cooled to room temperature to obtain A1-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin.

[0073] Example 2

[0074] This embodiment provides a method for preparing A2-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin, specifically comprising:

[0075] S201: 5 parts of lecithin, 1 part of cholesterol, 3 parts of ginsenoside Rg3 and 1 part of resveratrol were added into a dry single-necked round flask, and anhydrous ethanol was added to dissolve them at 60°C and 500rpm magnetic stirring to obtain a lipid mixture, which was subjected to reduced pressure rotary evaporation at 60°C to remove the anhydrous ethanol in the bottle and form a uniform A2-lipid film on the inner wall of the flask;

[0076] S202: 10 parts of ginsenoside Rg3, 0.5 parts of resveratrol, 5 parts of glycerol, 5 parts of butylene glycol, 5 parts of 1,2-hexanediol, 1 part of VC ethyl ether, 0.5 parts of bosine and 5 parts of water are stirred under magnetic stirring at 50°C and 500rpm until the solution becomes clear, and then A2-lipid film is added. Under constant temperature conditions of 50°C, a high-speed shearing instrument is used at 10,000rpm for 3 minutes, and ultrasonic treatment is performed at 100W power and 50°C for 10 minutes. After high-pressure homogenization treatment at a pressure of 600 bar and a temperature of 50°C, the mixture is cycled 4 times and cooled to room temperature to obtain A2-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin.

[0077] Example 3

[0078] This embodiment provides a method for preparing A3-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin, specifically comprising:

[0079] S301: 1 part of soybean lecithin, 2 parts of cholesterol, 3 parts of ginsenoside Rg3 and 0.3 parts of resveratrol were added into a dry single-mouth round flask, and anhydrous ethanol was added to dissolve them at 60°C and 500rpm magnetic stirring to obtain a lipid mixture, which was subjected to reduced pressure rotary evaporation at 60°C to remove the anhydrous ethanol in the bottle and form a uniform A3-lipid film on the inner wall of the flask;

[0080] S302: 5 parts of ginsenoside Rg3, 5 parts of glycerol, 3 parts of butylene glycol, 2 parts of 1,2-hexanediol, 0.2 parts of resveratrol, 2 parts of folic acid and 5 parts of water are stirred under magnetic stirring at 50°C and 500rpm until the solution is clear, and then A3-lipid film is added. Under constant temperature conditions of 50°C, a high-speed shearing instrument is used at 10000rpm for 3 minutes, and ultrasonic treatment is performed at 100W power and 50°C for 10 minutes. After high-pressure homogenization treatment at a pressure of 600 bar and a temperature of 50°C, the mixture is cycled 4 times and cooled to room temperature to obtain A3-ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin.

[0081] At the same time, in order to verify the comprehensive performance of the ginsenoside Rg3 nanoliposomes prepared in the above examples, the present application provides the following comparative examples for detailed description.

[0082] Comparative Example 1

[0083] The component ratio, preparation operation and process parameters of the B1-liposome prepared in this comparative example are basically the same as those in Example 1, except that ginsenoside Rg3 is not added to the active component in this comparative example to prepare B1-liposome.

[0084] Comparative Example 2

[0085] The component ratio, preparation operation and process parameters of the B2-liposome prepared in this comparative example are basically the same as those in Example 1, except that resveratrol is not added as the active component in this comparative example to prepare B2-liposome.

[0086] Comparative Example 3

[0087] The component ratio, preparation operation and process parameters of the B3-liposome prepared in this comparative example are basically the same as those in Example 1, except that in this comparative example, ginsenoside Rg3 and resveratrol are not added as active components to prepare B3-liposome.

[0088] Comparative Example 4

[0089] The component ratio, preparation operation and process parameters of the B4-liposome prepared in this comparative example are basically the same as those in Example 1, except that in this comparative example, only ginsenoside Rg3 and resveratrol are added as active components, and no compound functional substances are added to prepare B4-liposome.

[0090] Comparative Example 5

[0091] The component ratio, preparation operation and process parameters of the B5-liposome prepared in this comparative example are basically the same as those in Example 1, except that in this comparative example, lecithin, cholesterol, ginsenoside Rg3, resveratrol, glycerol and bosine are directly dissolved in water and then treated with ultrasound and high-pressure homogenization, which are recorded as B5-liposomes.

[0092] In order to verify the comprehensive performance of the liposomes prepared in the examples of the present application, corresponding performance tests were performed. The liposomes prepared in the above examples 1-3 and comparative examples 1-5 were subjected to performance tests:

[0093] Test 1: Stability test

[0094] The liposomes prepared in the above Examples 1-3 and Comparative Examples 1-5 were placed in a sealed container and placed at -18°C, 4°C, room temperature and 45°C for 180 days. After returning to room temperature, the samples were checked for agglomeration or stratification at the initial state, 90 days and 180 days respectively. The results can be expressed as "yes / no" and the average particle size of the samples was tested. The results are shown in Table 1.

[0095] Table 1 Stability test results

[0096]

[0097] As can be seen from Table 1, the liposomes prepared in Examples 1-3 of the present application all showed good stability, and no agglomeration and stratification occurred after being placed at -18°C, 4°C, room temperature and 45°C for 180 days. The particle size and particle size variation of the A2-ginsenoside Rg3 nanoliposomes, which are more easily transdermally absorbed, prepared in Example 2 were the smallest, and the system was the most stable. It can be seen that the preparation method provided by the present invention is of great significance for promoting the stability of the product.

[0098] Test 2: Encapsulation efficiency test

[0099] The encapsulation efficiency was determined by high performance liquid chromatography (HPLC), and the encapsulation efficiency was calculated after liquid phase analysis. The results are shown in Table 2.

[0100] Table 2 Encapsulation efficiency test results

[0101] Group A1 A2 A3 B1 B2 B3 B4 B5 Encapsulation efficiency 92.0 92.4 91.9 91.4 91.5 90.6 90.1 89.8

[0102] As shown in Table 2, the liposomes prepared in Examples 1-3 and Comparative Examples 1-5 have high encapsulation efficiency and good stability when measured from the perspective of physicochemistry.

[0103] Test 3: Free Radical Scavenging Test

[0104] Dilute the sample to a final concentration of VC ethyl ether of 10 mg / ml, take 10 μl of the sample to be tested and mix it with 190 μl of 0.25 mmol / LABTS working solution / PBS, shake well, incubate at room temperature for 6 minutes, and measure the absorbance at 405 nm. The absorbance of the sample to be tested and the ABTS working solution is recorded as A1, the absorbance of the sample to be tested and PBS is recorded as A2, and the absorbance of the ABTS working solution and PBS is recorded as A3. Then the ABTS clearance rate = [1-(A1-A2) / A3]×100%. The positive control is vitamin C with sample concentrations of 200, 150, 125, 100, 75, 50, and 25 μg / mL. The IC of the positive control VC 50 At 0.040g / L-0.070g / L, the test system was considered effective and repeated three times.

[0105] The present invention uses the liposome emulsions prepared in Examples 1-3 and Comparative Examples 1-5 as test samples to conduct ABTS free radical scavenging experiments to obtain the free radical scavenging rates of each liposome emulsion. The results are shown in Table 3. Figure 1 and Figure 2 shown.

[0106] Table 3 Test results of ABTS free radical scavenging

[0107]

[0108]

[0109] Depend on Figure 1 The IC of the positive control VC is 50 The test system is considered to be effective within the range of 40 μg / mL-70 μg / mL. Figure 2 It can be seen that the ABTS free radical scavenging rate of the liposome emulsion prepared in Example 2 is the highest, with an average value of 70.23%, followed by Example 3 and Example 1, and the ABTS free radical scavenging rate of the liposome emulsions prepared in Comparative Examples 1 to 5 is lower than that in the Examples. This shows that the liposome emulsion provided in the present application can effectively scavenge free radicals and has antioxidant properties.

[0110] Test 4: Anti-aging test

[0111] The prepared liposomes were tested for anti-UV oxidation efficacy. Examples 1-3 and Comparative Examples 1-5 were diluted with DMEM medium at a final concentration of 10 μg / ml of active ingredients. HDF cells in logarithmic growth phase were cultured at 8×10 3 Each well was inoculated in a 96-well plate, with 100 μL of the system in each well. The outermost circle of the 96-well plate was sealed with 100 μL of PBS. After the cells were completely attached to the wall, they were replaced with complete culture medium containing the sample to be tested. After continuing to culture the cells for 24 hours, the CCK-8 method was used to detect cell activity. The cells cultured with the sample dilution for 24 hours were irradiated with UV for 5 minutes, and the cell activity was detected by CCK8 method after 24 hours of culture in the incubator. The results are shown in Figure 3 .

[0112] Depend on Figure 3 The results show that the cell viability of the group without drug administration after UVB damage was 42.47%, and the cell viability level of the UV group was significantly lower than that of the normal group, indicating that the UV damage HDF model was successfully established. Compared with the blank control, the nanoliposomes prepared in Examples 1-3 and Comparative Examples 1-5 can significantly improve the viability of UV-damaged human epidermal cells, and the difference is statistically significant (P < 0.01). Among them, Example 1 has the strongest anti-UV damage performance, and the effects of Examples 1-3 are better than those of the comparative examples, indicating that the liposomes provided by the present application have excellent anti-UV oxidation efficacy.

[0113] Test 5: Anti-inflammatory test

[0114] RAW 264.7 cells (mouse mononuclear macrophage leukemia cells) in the logarithmic growth phase were inoculated in a 24-well plate at a cell density of (2-5)×10 5 / mL, 0.5mL per well, incubate for 24h. After 24h, remove the supernatant and add 0.5mL of the following samples respectively:

[0115] ①Solvent control group: DMEM medium (2% FBS; 1% double antibody) was used to act on cells for 24 hours;

[0116] ②LPS (lipopolysaccharide) inflammation model group: 1 μg / mL LPS continuously stimulated RAW 264.7 cells for 24 h;

[0117] ③ Experimental group: The diluted samples (Examples 1-3 and Comparative Examples 1-5) + LPS (1 μg / ml) were used to act on the cells for 24 hours. After the cells were treated with the samples for 24 hours, the supernatant was aspirated into a centrifuge tube with a pipette, centrifuged at 1000 r / min for 5 minutes, the precipitate was discarded and the supernatant was aspirated, and the change in the amount of inflammatory factors in the supernatant was detected using a TNF-αIL-6 inflammatory factor detection kit. The detection method was strictly in accordance with the instructions on the detection kit.

[0118] TNF-α (tumor necrosis factor-α) and IL-6 (interleukin-6) are key cytokines in the inflammatory process. Their expression levels can reflect the degree of inflammation activity. By measuring the levels of these cytokines, the anti-inflammatory potential of the test substance can be evaluated. Figure 3 As shown, the levels of TNF-α and IL-6 in the LPS group were significantly higher than those in the normal group, indicating that the inflammatory induction model was successfully established. Both the examples and the comparative examples can significantly inhibit the increase in the levels of inflammatory factors induced by LPS, and the difference is statistically significant (P < 0.01). The effects of different samples show certain differences. The inhibitory effects of Examples 1-3 on inflammatory factors TNF-α and IL-6 are better than those of Comparative Examples 1-5, indicating that the liposomes provided by the present application are more conducive to the active substances to exert the inhibitory effect on the inflammatory response of body cells.

[0119] Test 6: Human patch test

[0120] Weigh 3g of liposomes prepared in Examples 1-3 and Comparative Examples 1-5, mix with 97g of blank cream, and obtain ginsenoside Rg3 nanoliposome cream. 30 subjects were selected in the experiment, and the subjects' reactions were recorded. The blank cream, the comparative example component cream, and the example component cream were all applied to the curved side of the subject's forearm, and gently pressed with the palm of the hand to evenly apply it on the skin for 24 hours. After removing the spot tester, the interval was 30min, and the skin reaction was observed after the indentation disappeared. The skin reaction was observed again 48 hours after the spot tester was removed. The results are shown in Table 3.

[0121] The classification of skin adverse reactions is determined according to the skin adverse reaction classification standards in the "Safety Evaluation of Human Trial Tests" stipulated in the 2015 edition of the "Technical Specifications for Safety of Cosmetics":

[0122] Grade 0 (no skin reaction);

[0123] Grade 1 (light red spots on the skin);

[0124] Grade 2 (erythema, infiltration and papules on the skin);

[0125] Grade 3 (erythema, edema, papules and blisters on the skin);

[0126] Grade 4 (skin erythema, edema and bullae).

[0127] Table 4 Human patch test results

[0128]

[0129] The principle of the patch test is to reproduce the reaction process of allergic contact dermatitis on the local skin, that is, after a small amount of allergen directly contacts the skin, observe whether mild dermatitis is induced locally, so as to judge whether the skin is allergic to the tested allergen. The above results show that Examples 1-3 and Comparative Examples 1-5 did not have adverse skin reactions, which means that the samples in the 8 experimental groups were non-irritating.

[0130] Test 7: Cumulative transdermal penetration test

[0131] Take SD rats weighing 200-250g, shave the hair on their backs, kill them after 24 hours, cut the skin on their backs, remove the subcutaneous fat, clean them with physiological saline, absorb the surface moisture and place them in the refrigerator for later use. During the test, take out the mouse skin from the refrigerator, place it in physiological saline to thaw at room temperature, and then clamp the mouse skin between the receiving pool and the diffusion pool with the stratum corneum facing up. Measure 1mL of each sample prepared in Example 1 and Comparative Examples 1-4 and apply it evenly on the mouse skin. Seal the top of the diffusion pool with plastic wrap and place it in a magnetic stirrer, keep it in a constant temperature water bath at 37°C, and stir at 200r / min. Take 0.5mL of samples after 2h, 4h, 8h, 12h, and 24h. After each sampling, add 10% Triton solution to make the volume to 1mL and filter it with a 0.45μm microporous filter membrane; at the same time, add 0.5mL of physiological saline to the receiving pool. The concentration of the active substance was tested by HPLC, and the cumulative permeability was calculated. The results are shown in Table 5 and Figure 4 .

[0132] Table 5 Cumulative transdermal penetration test results

[0133]

[0134] The above experimental results show that the permeability of each group of samples gradually increased within 24 hours, and the permeability of ginsenoside Rg3 nanoliposomes A1, A2 and A3, which are easier to permeate through the skin, is significantly better than that of samples B1, B2, B3, B4 and B5, indicating that the liposomes provided in the examples of the present application have excellent permeability.

[0135] The various embodiments in this specification are described in a progressive manner, and the same or similar parts between the various embodiments can be referenced to each other, and each embodiment focuses on the differences from other embodiments.

[0136] The above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit the present application. Although the present application has been described in detail with reference to the aforementioned embodiments, a person of ordinary skill in the art should understand that the technical solutions recorded in the aforementioned embodiments may still be modified, or some or all of the technical features therein may be replaced by equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the present application.

Claims

1. A ginsenoside Rg3 nanoliposome that is easier to absorb through the skin. It is characterized in that Its components include, by weight of raw materials: Polyol 5-20 parts; Ginsenoside Rg3 2-10 parts; 1-8 parts of phospholipids; Cholesterol 1-5 parts; 1-2 parts of resveratrol; 0-2 parts of compound efficacy ingredients; 1-5 parts water; Wherein, the compounded effective substance is one or more of VC ethyl ether, folic acid and bosine.

2. The ginsenoside Rg3 nanoliposome that is easier to be absorbed through the skin according to claim 1, It is characterized in that Its components include, by weight of raw materials: Polyol 10-15 parts; Ginsenoside Rg3 8-10 parts; 1-5 parts of phospholipids; 1-2 parts cholesterol; Resveratrol 0.5-2 parts; 0-2 parts of compound efficacy ingredients; 1-5 parts water; Wherein, the compounded effective substance is one or more of VC ethyl ether, folic acid and bosine.

3. The ginsenoside Rg3 nanoliposome that is more easily absorbed through the skin according to claim 1 or 2, It is characterized in that The particle size of the ginsenoside Rg3 nanoliposome is 100-150nm, and the encapsulation rate is ≥90%.

4. The ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin according to claim 1 or 2, It is characterized in that The polyol is one or more of glycerol, butanediol, 1,2-hexanediol, 1,3-propylene glycol, polyethylene glycol, and sorbitol.

5. The ginsenoside Rg3 nanoliposome that is more easily absorbed through the skin according to claim 1 or 2, It is characterized in that The phospholipid is one or more of lecithin, hydrogenated lecithin, soybean lecithin, egg yolk lecithin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidylserine.

6. A method for preparing ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin according to any one of claims 1 to 5, It is characterized in that The method comprises: Phospholipids, cholesterol, resveratrol and ginsenoside Rg3 were dissolved in ethanol and then subjected to rotary evaporation to form a lipid film; After dissolving ginsenoside Rg3, resveratrol, polyols and compounded functional substances in water, the lipid film is added and subjected to ultrasonic and high-pressure homogenization treatment to obtain the ginsenoside Rg3 nanoliposomes which are more easily absorbed through the skin.

7. The method for preparing ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin according to claim 6, It is characterized in that When the phospholipids, cholesterol, resveratrol and ginsenoside Rg3 are dissolved in ethanol and then subjected to rotary evaporation, the temperature is 40-60°C.

8. The method for preparing ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin according to claim 6, It is characterized in that The temperature of the ultrasonic treatment is 30-50°C, the time is 30-60min, and the ultrasonic power is 50-200W.

9. The method for preparing ginsenoside Rg3 nanoliposomes that are more easily absorbed through the skin according to claim 6, It is characterized in that The conditions of the high-pressure homogenization treatment are: pressure of 100-800 bar, and number of cycles of 1-5 times.

10. Use of ginsenoside Rg3 nanoliposomes which are easier to be absorbed through the skin, prepared according to the preparation method according to any one of claims 6 to 9, in the preparation of skin care products.

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