Active composition based on ginseng vesicle wrapping and application thereof

Abstract

The invention discloses an active composition based on ginseng vesicle wrapping and application of the active composition. The active composition is prepared from ginseng extracellular vesicles, asiaticoside, ergothioneine and sodium hyaluronate, the ginseng vesicles are extracted through a gradient centrifugation method, and the active ingredients are efficiently entrapped in the vesicles by adopting an ultrasonic loading technology. The invention relates to the technical field of cosmetics, ginseng vesicles are synergistically compatible with asiaticoside, ergothioneine and sodium hyaluronate for the first time, the stability, the skin permeability and the targeting property of active ingredients are remarkably improved by utilizing a nano-scale delivery system of the vesicles, and meanwhile, the irritation is reduced. Experiments show that the composition can significantly improve the activity of fibroblasts, inhibit the generation of active oxygen and promote the synthesis of collagen. Human body tests prove that the transdermal rate is increased by more than two times compared with that of free components, and no sensitization is caused. The active composition can be applied to essence, masks, emulsions and other dosage forms, and has efficient anti-aging, repairing and anti-oxidation functions.

Description

Technical Field

[0001] The present invention relates to the technical field of skin care products, and more particularly to an active composition encapsulated by ginseng vesicles and an application thereof. Background Art

[0002] Current anti-aging cosmetics generally have the following problems:

[0003] (1) Poor stability of active ingredients: Traditional antioxidants (such as vitamin C and retinol) are easily degraded by light and heat, resulting in reduced efficacy.

[0004] (2) Insufficient skin permeability: Large molecular active ingredients (such as polysaccharides and peptides) have difficulty penetrating the stratum corneum and have low bioavailability.

[0005] (3) Irritation risk: High concentrations of active ingredients (such as acid and niacinamide) may cause skin sensitivity.

[0006] (4) Insufficient synergistic effect: The target of a single ingredient is limited, making it difficult to achieve multi-pathway anti-aging.

[0007] Therefore, there is an urgent need for an anti-aging system that combines efficient penetration, stable release and multi-target synergy. Summary of the Invention

[0008] In response to the problems of poor permeability, insufficient stability and high irritation of existing anti-aging ingredients, the purpose of the present invention is to provide an active composition encapsulated by ginseng vesicles, which improves the targeting, stability and safety of the active ingredients through a nano-delivery system.

[0009] To achieve the above object, the present invention provides the following technical solutions:

[0010] A ginseng vesicle-encapsulated active composition comprises the following components in parts by mass: 50-80 parts of ginseng extracellular vesicles, 5-15 parts of asiaticoside, 3-10 parts of ergothioneine, and 2-8 parts of sodium hyaluronate.

[0011] Preferably, the ginseng extracellular vesicles are extracted by gradient centrifugation, which specifically comprises the following steps:

[0012] (1) Wash fresh ginseng tubers and squeeze the juice. Centrifuge the filtrate at 800-1500g for 10-30 minutes to remove cell debris.

[0013] (2) The supernatant was centrifuged at 2500-4000 g for 10-30 min to remove large particles;

[0014] (3) further centrifugation at 8000-12000 g for 40-70 min to remove free protein;

[0015] (4) Finally, the precipitate was collected by centrifugation at 120,000-180,000 g for 80-100 min and resuspended in phosphate buffer.

[0016] Preferably, the gradient centrifugation is carried out at 4°C throughout the entire process, and the precipitate after centrifugation is sterilized by filtration through a 0.22 μm filter membrane.

[0017] Preferably, the active ingredient is loaded into the ginseng vesicles by ultrasonic loading method, with ultrasonic power of 100-200W, frequency of 20-40kHz, time of 10-30min, and loading temperature of 25-37°C.

[0018] Preferably, the molar ratio of asiaticoside to ergothioneine is 1:0.5-1:2, and the encapsulation efficiency is ≥85%.

[0019] Preferably, the molecular weight of the sodium hyaluronate is 50-200 kDa, and the concentration is 0.1-0.5% (w / v).

[0020] The present invention also provides a use of the active composition in the preparation of anti-aging cosmetics, wherein the dosage form of the cosmetics includes essence, facial mask, emulsion or microneedle patch.

[0021] The present invention has the following beneficial effects:

[0022] (1) Highly efficient penetration: The vesicle particle size is 100-150nm. According to the Franz diffusion cell test, the transdermal rate is more than 2 times higher than that of the free component.

[0023] (2) Synergistic anti-aging: Centella asiatica promotes collagen synthesis, ergothioneine scavenges free radicals, and ginseng vesicles inhibit the inflammatory factor IL-6.

[0024] (3) Low irritation: The risk of direct contact of ingredients with the skin is reduced through vesicle encapsulation, and the patch test shows a sensitization rate of 0%. DETAILED DESCRIPTION

[0025] The technical solutions of the present invention are further described below in conjunction with the embodiments. Obviously, the embodiments described are only some embodiments of the present invention, rather than all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention. The methods or operations used in the embodiments, unless otherwise specified, are conventional methods or routine operations in the art.

[0026] This application provides general and / or specific descriptions of the materials and experimental methods used in the experiments. In the following examples, unless otherwise specified, % represents wt%, i.e., percentage by weight. All reagents or instruments used without manufacturer indication are commercially available conventional reagents.

[0027] Example 1: Gradient Centrifugation Extraction of Ginseng Vesicles

[0028] Raw material processing: Take 500g of fresh ginseng tubers, wash and chop them, add pre-cooled PBS (4℃, 1:1w / v) to squeeze the juice, coarsely filter through 4 layers of gauze, and then finely filter through a 0.45μm filter membrane to obtain clarified ginseng juice.

[0029] Differential centrifugation: First centrifugation: clarify the ginseng juice by centrifugation at 4°C and 1200g for 20 minutes, discard the precipitate, and collect supernatant I; second centrifugation: supernatant I is centrifuged at 4°C and 3000g for 20 minutes, discard the precipitate, and collect supernatant II; third centrifugation: supernatant II is centrifuged at 4°C and 10000g for 60 minutes, discard the precipitate, and collect supernatant III; ultracentrifugation: supernatant III is centrifuged at 4°C and 150000g for 90 minutes, collect the precipitate, resuspend it in 10 mL PBS at a ratio of 1 g of precipitate, sterilize it with a 0.22 μm filter membrane, and obtain the ginseng vesicle solution.

[0030] Example 2: Ultrasonic loading of active composition

[0031] Formula: ginseng vesicle solution (containing 60 parts of vesicles), 10 parts of asiaticaside, 5 parts of ergothioneine, and 5 parts of sodium hyaluronate (100kDa).

[0032] Loading process: asiaticoside and thioneine are dissolved in PBS (pH 6.0), mixed with the ginseng vesicle solution of Example 1, and ultrasonically processed under ice bath condition (power 150W, frequency 28kHz, pulse mode: working 2s / interval 1s, total duration 20min); Sodium hyaluronate is added and stirred to complete dissolution; Unencapsulated components are removed by centrifugation at 10000g for 30min, and the supernatant is concentrated by ultrafiltration (molecular weight cut-off 100kDa) to obtain a loaded active composition. The encapsulation efficiency of asiaticoside and the encapsulation efficiency of thioneine are determined to be 88.3%, and 91.5%.

[0033] Example 3: High-Centella Asiaticoside Formula

[0034] Formula: ginseng vesicle solution (containing 60 parts of vesicles), 15 parts of asiaticoside, 3 parts of ergothioneine, and 5 parts of sodium hyaluronate (100kDa).

[0035] Loading process: Same as Example 2, the encapsulation efficiency of asiaticoside was determined to be 82.1%, and the encapsulation efficiency of ergothioneine was 88.7%.

[0036] Embodiment 4: high ergothioneine formula

[0037] Formula: ginseng vesicle solution (containing 60 parts of vesicles), 5 parts of asiaticaside, 10 parts of ergothioneine, and 5 parts of sodium hyaluronate (100kDa).

[0038] Loading process: Same as Example 2, the encapsulation efficiency of asiaticoside was determined to be 75.6%, and the encapsulation efficiency of ergothioneine was 93.4%.

[0039] Example 5: Low Sodium Hyaluronate Formula

[0040] Formula: ginseng vesicle solution (containing 80 parts of vesicles), 10 parts of asiaticoside, 5 parts of ergothioneine, and 2 parts of sodium hyaluronate (100kDa).

[0041] Loading process: Same as Example 2, the encapsulation efficiency of asiaticoside was determined to be 85.2%, and the encapsulation efficiency of ergothioneine was 89.7%.

[0042] Example 6: Composite Antioxidant Formula

[0043] Formula: ginseng vesicle solution (containing 60 parts of vesicles), 8 parts of asiaticaside, 2 parts of coenzyme Q10, 6 parts of ergothioneine, and 5 parts of sodium hyaluronate (100kDa).

[0044] Loading process: Same as Example 2, the encapsulation efficiency of asiaticoside was 80.5%, the encapsulation efficiency of ergothioneine was 90.2%, and the encapsulation efficiency of coenzyme Q10 was 78.3%.

[0045] Comparative Example 1: Free active ingredient

[0046] The formulation was the same as that of Example 2, except that the ultrasonic loading step was omitted, and the ginseng vesicles and the active ingredients were directly mixed without centrifugation to remove free components.

[0047] Test 1: Transdermal absorption test (Franz diffusion cell method)

[0048] Adopt ex vivo pigskin (thickness 0.5mm, remove subcutaneous fat) as transdermal barrier, donor pool adds the test sample that 0.5mL contains thioneine 1mg / mL, receptor pool is the PBS solution (containing 0.01%NaN3) of pH7.4, maintain 37 DEG C of constant temperature and 600rpm magnetic agitation, in 2h, 4h, 8h, 12h, 24h time point sampling and supplement equal-volume fresh receptor liquid, by high performance liquid chromatography (HPLC, C18 chromatographic column, mobile phase acetonitrile-water (20:80), flow velocity 1mL / min, detection wavelength 254nm) quantitative analysis thioneine cumulative penetration amount, calculate transdermal rate and steady-state flow rate.The results are shown in Table 1.

[0049] Table 1 Transdermal absorption test

[0050]

[0051] As shown in Table 1, the transdermal penetration rates of all Example groups were significantly higher than those of Comparative Example 1 (p<0.001), indicating that the ginseng vesicle delivery system effectively enhanced the penetration of active ingredients. Among them, Example 6 (Coenzyme Q10) had the highest transdermal penetration rate (44.1%), which may be related to the lipid solubility of Coenzyme Q10, which enhances the fluidity of the vesicle membrane. Example 3 (High-Asianthoside) had a lower transdermal penetration rate (38.5%) due to a decreased encapsulation efficiency (82.1% vs. 88.3% in the baseline group). Example 5 (Low-Sodium Hyaluronate) had a significantly lower steady-state flux due to a decrease in system viscosity (p<0.01).

[0052] Test 2: Cellular anti-aging efficacy test (HFF-1 fibroblast model)

[0053] The samples of Examples 2-6 and Comparative Example 1 (0.1 mg / mL) were incubated with human skin fibroblasts (HFF-1, 5×10 3 Cells were co-cultured at 450 nm for 48 hours. Cell viability was determined using the CCK-8 assay (450 nm absorbance); ROS clearance was determined using a DCFH-DA probe (Ex / Em = 488 / 525 nm fluorescence intensity); and collagen synthesis was measured using a hydroxyproline detection kit (colorimetric, 558 nm). Data were statistically analyzed using ANOVA and Tukey's test, with the blank control group as the reference (100%). Results are shown in Table 2.

[0054] Table 2 Anti-aging efficacy test

[0055]

[0056] As shown in Table 2, embodiment 2-6 group cell viability, ROS inhibition rate and collagen synthesis are significantly better than comparative example 1 group (p<0.001), confirming that encapsulation system promotes bioavailability. Wherein, embodiment 4 (high ergothioneine) ROS inhibition rate is the highest (75%), showing that ergothioneine concentration improves antioxidant capacity; Embodiment 6 (coenzyme Q10) is due to multiple antioxidant mechanisms (coenzyme Q10+ ergothioneine), and collagen synthesis amount is the highest (25.6 μ g / mL); Embodiment 5 (low sodium hyaluronate) is due to moisturizing deficiency, and cell viability and collagen synthesis significantly decline (p<0.01).

[0057] Experimental Example 3: Human Patch Test (Skin Safety Assessment)

[0058] 330 healthy subjects applied samples of Examples 2-6 and Comparative Example 1 to their backs for 48 hours. Reactions, such as erythema and edema, were observed and scored on a scale of 0-3. The sensitization rate was the proportion of subjects experiencing a reaction (≥ 1 point), and the irritation index was the average score for each group. The results are shown in Table 3.

[0059] Table 3 Human patch test

[0060] Group Sensitization rate (%) Average irritation index Example 2 0％ 0.12±0.05 Example 3 3.3％ 0.25±0.08* Example 4 0％ 0.15±0.06 Example 5 6.7％ 0.38±0.12** Example 6 0％ 0.13±0.05 Comparative Example 1 13.3％ 0.87±0.21***

[0061] As shown in Table 3, all Example groups showed significantly lower sensitization rates and irritation indices than Comparative Example 1 (p<0.001), confirming that ginseng vesicle encapsulation reduces the risk of direct skin contact with the ingredients. Example 5 (low sodium hyaluronate) showed an elevated sensitization rate of 1.7% (p<0.01) due to insufficient moisturizing. Example 3 (high asiaticoside) also showed mild irritation, with a sensitization rate of 3.3% (p<0.05), likely due to the high concentration of asiaticoside. Examples 4 and 6 demonstrated the best safety, with irritation indices showing no significant differences from the baseline group (p>0.05).

[0062] In summary, the present invention synergizes ginseng vesicles with asiaticoside, ergothioneine, and sodium hyaluronate for the first time, and utilizes the nanoscale delivery system of the vesicles to significantly improve the stability, skin permeability and targeting of the active ingredients, while reducing irritation. Experiments have shown that the composition can significantly enhance fibroblast viability (increased by 65% compared to the blank group), inhibit the generation of reactive oxygen species (reduced by 65%), and promote collagen synthesis (increased by 30%). Human trials have verified that its transdermal rate is more than twice that of the free ingredients, and it is non-sensitizing. The active composition of the present invention can be applied to dosage forms such as essences, facial masks, and lotions, and has efficient anti-aging, repair, and antioxidant functions.

[0063] The above description is merely a preferred embodiment of the present invention. The scope of protection of the present invention is not limited to the above embodiment. All technical solutions based on the concept of the present invention are within the scope of protection of the present invention. It should be noted that for those skilled in the art, various improvements and modifications that do not depart from the original invention should also be considered within the scope of protection of the present invention.

Claims

1. An active composition based on ginseng vesicles, characterized in that: The invention comprises the following components in parts by weight: 50-80 parts of ginseng extracellular vesicles, 5-15 parts of asiaticoside, 3-10 parts of ergothioneine and 2-8 parts of sodium hyaluronate.

2. The active composition according to claim 1, characterized in that The ginseng extracellular vesicles are extracted by gradient centrifugation, which specifically includes the following steps: (1) Wash fresh ginseng tubers and squeeze the juice. Centrifuge the filtrate at 800-1500g for 10-30 minutes to remove cell debris. (2) The supernatant was centrifuged at 2500-4000 g for 10-30 min to remove large particles; (3) further centrifugation at 8000-12000 g for 40-70 min to remove free protein; (4) Finally, the precipitate was collected by centrifugation at 120,000-180,000 g for 80-100 min and resuspended in phosphate buffer.

3. The active composition according to claim 2, characterized in that The gradient centrifugation was carried out at 4° C. throughout the process, and the precipitate was sterilized by filtration through a 0.22 μm filter membrane after centrifugation.

4. The active composition according to any one of claims 1 to 3, characterized in that The active ingredient is loaded into the ginseng vesicles through an ultrasonic loading method, with an ultrasonic power of 100-200W, a frequency of 20-40kHz, a time of 10-30min, and a loading temperature of 25-37°C.

5. The active composition according to claim 4, characterized in that The molar ratio of asiaticoside to ergothioneine is 1:0.5-1:2, and the encapsulation efficiency is ≥85%.

6. The active composition according to claim 1, characterized in that The molecular weight of the sodium hyaluronate is 50-200 kDa, and the concentration is 0.1-0.5% (w / v).

7. Use of the active composition according to any one of claims 1 to 6 in the preparation of anti-aging cosmetics, wherein the dosage form of the cosmetics comprises an essence, a facial mask, an emulsion or a microneedle patch.

Images