A black ginseng biomimetic delivery body, a preparation method and application thereof

Abstract

The present application relates to the field of cosmetic technology, and particularly relates to a black ginseng biomimetic delivery body, a preparation method and application thereof.The black ginseng biomimetic delivery body comprises: 40 wt%-60 wt% of glycerol, 1 wt%-3 wt% of hydrogenated lecithin, 0.3 wt%-1 wt% of polyglyceryl-10 myristate, 0.2 wt%-1 wt% of black ginseng extract, 0.05 wt%-0.15 wt% of arginine, 0.01 wt%-0.1 wt% of carboxymethyl chitosan, 0.01 wt%-0.02 wt% of collagen and the rest of water.The black ginseng biomimetic delivery body has better cell repair efficacy and soothing efficacy, can effectively improve the sedimentation problem of the black ginseng solution, enhance the stability of the black ginseng solution and the biomimetic delivery body, and effectively improve the penetration and absorption of the black ginseng and collagen.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, and in particular to a biomimetic delivery system for black ginseng, its preparation method, and its application. Background Technology

[0002] Black ginseng is a processed ginseng product, following red ginseng, made through a nine-steaming and nine-drying process. It contains various rare ginsenosides, especially ginsenoside Rg3, which is its main component. Rg3 possesses pharmacological activities such as antioxidant, anti-inflammatory, anti-melanoma proliferation, and anti-photoaging effects. Therefore, black ginseng is rarer, more valuable, and more suitable for cosmetic use than ordinary white ginseng. However, in black ginseng solutions, especially at higher concentrations, the total and reversible precipitation levels gradually increase with prolonged storage, limiting its addition in formulations. Therefore, developing a stable skincare ingredient that leverages the advantages of Rg3 has become a hot topic in current skincare research and development.

[0003] Cationic liposomes are novel nanocarriers composed of cationic lipids, neutral phospholipids, and cholesterol. Their positively charged surfaces allow for efficient binding through electrostatic interactions, forming stable lipid complexes with excellent biocompatibility and encapsulation capacity. Their positively charged surfaces can interact with the negatively charged cell membranes, significantly enhancing the adsorption and penetration of the delivery system into cells, thus facilitating targeted delivery and efficient release of active ingredients. Furthermore, cationic liposomes can encapsulate various drugs or biomolecules, protecting their stability and reducing the impact of external environmental factors, making them widely applicable in drug, gene, and protein delivery.

[0004] Ginsenosides have poor solubility in water and oil, resulting in very low absorption efficiency when directly added to cosmetics. In the pharmaceutical field, liposome encapsulation is commonly used to enhance the absorption of ginsenosides. For example, patent application CN120114392A describes phospholipid-type liposomes constructed using ginsenosides, soybean lecithin, and Tween-80; patent application CN117752612A describes phospholipid-type liposomes constructed using ginsenosides, phospholipids, and cholesterol; patent application CN111228219A only describes blank liposome carriers prepared using ginsenoside Rg3 and its analogues; patent application CN119074581A describes ginsenoside Rg3 nanoliposomes prepared using ginsenosides, phospholipids, cholesterol, resveratrol, etc.; and patent application CN114053225A uses phytosterols or ginsenoside compounds to replace cholesterol as membrane stabilizers, resulting in a positively charged membrane surface and high delivery efficiency.

[0005] However, patent applications CN120114392A, CN117752612A, and CN111228219A all employ a method of dissolving in organic solvents followed by rotary evaporation, which easily leads to organic solvent residue problems, thus irritating the skin. Patent application CN119074581A contains cholesterol to enhance the membrane structure; however, since cholesterol is mostly derived from animals, its application has certain limitations. Patent application CN114053225A's cationic liposomes are prepared using thin-film dispersion, vacuum drying, reverse-phase evaporation, or freeze-thaw methods, which are relatively cumbersome preparation methods.

[0006] Therefore, existing technologies mention using ginsenosides as membrane materials or having a positively charged membrane surface to form cationic liposomes; however, some patents in the pharmaceutical field often involve toxic organic solvents such as chloroform and acetone in their preparation methods, which cannot be used in the cosmetics field; or the liposome system contains cholesterol, which is mostly of animal origin. In cosmetics, especially export products, the review of products of animal origin is more complicated, thus limiting its application. Summary of the Invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a bipolar, pH-responsive charge-reversing biomimetic delivery system for black ginseng, its preparation method and application. The bipolar biomimetic delivery system for black ginseng has superior cell repair and soothing effects, can effectively improve the sedimentation problem of black ginseng solution, enhance the stability of black ginseng solution and biomimetic delivery system, and can effectively improve the penetration and absorption of black ginseng and collagen.

[0008] This invention provides a biomimetic delivery system for black ginseng, comprising:

[0009] Glycerin 40 wt%~60 wt%, hydrogenated lecithin 1 wt%~3 wt%, polyglycerol-10 myristate 0.3 wt%~1 wt%, black ginseng extract 0.2 wt%~1 wt%, arginine 0.05 wt%~0.15 wt%, carboxymethyl deacetylated chitosan 0.01 wt%~0.1 wt%, collagen 0.01 wt%~0.02 wt%, and the balance being water.

[0010] This invention also provides a method for preparing the above-described biomimetic black ginseng delivery body, comprising the following steps:

[0011] S1. After dispersing hydrogenated lecithin with glycerol, it is mixed with polyglycerol-10 myristate at 65~75℃ to obtain mixed solution I;

[0012] The black ginseng extract was partially dissolved in water and then neutralized with arginine to obtain a black ginseng solution with pH=7. The black ginseng solution, carboxymethyl deacetylated chitosan, collagen and the remaining water were mixed at 65~70℃ to obtain mixed solution II.

[0013] S2. Mix the mixed solution I and mixed solution II thoroughly and then hydrate them;

[0014] S3. The solution after hydration in step S2 is passed through a high-pressure microjet to obtain a biomimetic delivery body for black ginseng.

[0015] Preferably, in step S1, mixing at 65~75℃ means stirring at 65~75℃, and the stirring rate is 300~800 rpm.

[0016] Preferably, in step S1, mixing at 65~70℃ means stirring at 65~70℃, and the stirring rate is 300~800 rpm.

[0017] Preferably, in step S2, mixing the mixed solution I and mixed solution II includes:

[0018] Add the mixed solution II to the stirred mixed solution I, and stir until well mixed;

[0019] The stirring and mixing rate is 300~800 rpm.

[0020] Preferably, in step S2, the hydration time is 1 to 3 minutes.

[0021] Preferably, in step S3, the pressure of the high-pressure microjet is 800~1200 bar; and the number of times is 3~5.

[0022] The present invention also provides the application of the above-described biomimetic delivery body of black ginseng, or the biomimetic delivery body of black ginseng prepared by the above-described preparation method, in the preparation of pharmaceuticals, cosmetics or skin care products.

[0023] The present invention also provides the application of the above-described biomimetic delivery body of black ginseng, or the biomimetic delivery body of black ginseng prepared by the above-described preparation method, in the preparation of aqueous essence.

[0024] The present invention also provides an aqueous essence comprising phase A, phase B and phase C;

[0025] Phase A includes deionized water, butanediol, 1,2-hexanediol, xanthan gum, p-hydroxyacetophenone, polyacrylate cross-linked polymer-6, acrylate / C10~30 alkanol acrylate cross-linked polymer and allantoin;

[0026] Phase B includes arginine;

[0027] Phase C includes dipotassium glycyrrhizate, a biomimetic black ginseng delivery system, and black ginseng extract;

[0028] The biomimetic delivery body for black ginseng is the biomimetic delivery body for black ginseng described above, or the biomimetic delivery body for black ginseng prepared by the preparation method described above.

[0029] The biomimetic delivery system for black ginseng provided by this invention has superior cell repair and soothing effects, can effectively improve the sedimentation problem of black ginseng solution, enhance the stability of black ginseng solution and biomimetic delivery system, and can effectively improve the penetration and absorption of black ginseng and collagen. Attached Figure Description

[0030] Figure 1 The fluorescence penetration depth maps of the samples from Example 1 and Comparative Example 8 of this invention are shown.

[0031] Figure 2 Appearance diagrams of the aqueous essence of Application Example 1 and Comparative Application Example 1;

[0032] Figure 3 These are test images of the appearance and skin feel dimensions of the aqueous essence in Application Example 1 and Comparative Application Example 1 of the present invention. Detailed Implementation

[0033] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0034] This invention provides a biomimetic delivery system for black ginseng, comprising:

[0035] Glycerin 40 wt%~60 wt%, hydrogenated lecithin 1 wt%~3 wt%, polyglycerol-10 myristate 0.3 wt%~1 wt%, black ginseng extract 0.2 wt%~1 wt%, arginine 0.05 wt%~0.15 wt%, carboxymethyl deacetylated chitosan 0.01 wt%~0.1 wt%, collagen 0.01 wt%~0.02 wt%, and the balance being water.

[0036] The glycerol content can be 45 wt%, 50 wt%, 55 wt%, or 60 wt%.

[0037] The content of the hydrogenated lecithin can be 1 wt%, 1.5 wt%, 2 wt%, 2.5 wt%, or 3 wt%.

[0038] The content of the polyglycerol-10 myristate can be 0.3 wt%, 0.5 wt%, 0.7 wt%, or 1 wt%.

[0039] The content of the black ginseng extract can be 0.2 wt%, 0.5 wt%, 0.7 wt%, or 1 wt%.

[0040] The arginine content can be 0.05 wt%, 0.07 wt%, 0.085 wt%, 0.1 wt%, 0.12 wt%, or 0.15 wt%.

[0041] The content of the carboxymethyl deacetylated chitosan can be 0.01 wt%, 0.02 wt%, 0.05 wt%, 0.07 wt%, or 0.1 wt%.

[0042] The collagen content can specifically be 0.01 wt%, 0.012 wt%, 0.015 wt%, 0.017 wt%, or 0.02 wt%.

[0043] The water can be deionized water.

[0044] The pH of the biomimetic delivery system for black ginseng is 6.5 to 7.5, for example, 7.

[0045] The collagen mentioned is commercially available.

[0046] The black ginseng extract contains ginsenoside Rg3. This invention does not limit the content of ginsenoside Rg3. This invention does not have any special restrictions on the source of the black ginseng extract; it can be prepared according to the method described in patent application CN116159011A.

[0047] The biomimetic black ginseng delivery device provided by this invention is a double-charge-layer, pH-responsive charge-reversal biomimetic black ginseng delivery device. The biomimetic black ginseng delivery device includes: a continuous phase and spheres dispersed in the continuous phase; the spheres include a core, a first shell layer formed on the outer surface of the core, and a second shell layer formed on the outer surface of the first shell layer;

[0048] The continuous phase consists of deionized water, glycerol, and arginine.

[0049] The core is composed of collagen;

[0050] The first shell layer is composed of hydrogenated lecithin, polyglycerol-10 myristate, and black ginseng extract. Specifically, hydrogenated lecithin forms a lipid bilayer as a spherical basic framework; polyglycerol-10 myristate is embedded to regulate the hydrophobic chain and enhance the stability of the basic framework; at the same time, ginsenoside Rg3 from the black ginseng extract is inserted into the lipid membrane (spherical basic framework), and its hydrophilic glycosyl groups are exposed on the surface to form a biomimetic modification layer (liposome).

[0051] The second shell is composed of carboxymethyl deacetylated chitosan. Under pH ≈ 7 conditions, the negatively charged carboxymethyl deacetylated chitosan wraps around the entire liposome through electrostatic interaction, forming a stable outer shell.

[0052] The biomimetic delivery system for black ginseng provided by this invention does not contain cholesterol and uses a surfactant (polyglycerol-10 myristate ester) to enhance the membrane structure.

[0053] The biomimetic delivery system for black ginseng provided by this invention does not contain organic solvents, such as toxic chloroform and acetone; it fundamentally solves the problem of organic solvent residue and is gentler on the skin.

[0054] This invention also provides a method for preparing the above-described biomimetic black ginseng delivery body, comprising the following steps:

[0055] S1. After dispersing hydrogenated lecithin with glycerol, it is mixed with polyglycerol-10 myristate and heated to 65~75℃ to obtain mixed solution I;

[0056] Dissolve the black ginseng extract in some water, then neutralize with arginine to obtain a black ginseng solution with pH=7; mix the black ginseng solution, carboxymethyl deacetylated chitosan, collagen and the remaining water, and heat to 65~70℃ to obtain mixed solution II;

[0057] S2. Mix the mixed solution I and mixed solution II thoroughly and then hydrate them;

[0058] S3. The solution after hydration in step S2 is passed through a high-pressure microjet to obtain a biomimetic delivery body for black ginseng.

[0059] Regarding step S1:

[0060] Hydrogenated lecithin was dispersed with glycerol and then mixed with polyglycerol-10 myristate and heated to 65-75°C (e.g., 70°C) to obtain mixed solution I.

[0061] Dissolve the black ginseng extract in some water, then neutralize with arginine to obtain a black ginseng solution with pH=7; mix the black ginseng solution, carboxymethyl deacetylated chitosan, collagen and the remaining water, and heat to 65~70℃ (e.g., 68℃) to obtain mixed solution II.

[0062] The present invention does not impose any special restrictions on the ratio of the partial water to the remaining water, as long as the partial water is sufficient to dissolve the black ginseng extract.

[0063] In some embodiments of the present invention, mixing at 65~75°C is equivalent to stirring and mixing at 65~75°C, and the stirring and mixing rate is 300~800 rpm; for example, 500 rpm.

[0064] In some embodiments of the present invention, mixing at 65~70°C is equivalent to stirring and mixing at 65~70°C, and the stirring and mixing rate is 300~800 rpm; for example, 500 rpm.

[0065] Regarding step S2:

[0066] Mix the mixed solution I and mixed solution II thoroughly and then hydrate them.

[0067] In some embodiments of the present invention, mixing the mixed solution I and the mixed solution II includes:

[0068] Add the mixed solution II to the stirred mixed solution I, and stir until well mixed.

[0069] The stirring speed is 300~800 rpm; for example, 500 rpm.

[0070] In some embodiments of the present invention, the hydration time is 1 to 3 minutes, for example, 2 minutes.

[0071] Regarding step S3:

[0072] The solution after hydration in step S2 was passed through a high-pressure microjet to obtain a biomimetic delivery body for black ginseng.

[0073] In some embodiments of the present invention, the pressure of the high-pressure microjet is 800~1200 bar, for example 1000 bar; and the number of times is 3~5 times, for example 4 times.

[0074] The preparation method of the biomimetic delivery body for black ginseng provided by this invention is simpler.

[0075] The preparation method of the black ginseng biomimetic delivery body provided by the present invention does not require the use of cholesterol or toxic organic solvents.

[0076] The present invention also provides the application of the above-described biomimetic delivery body of black ginseng, or the biomimetic delivery body of black ginseng prepared by the above-described preparation method, in the preparation of pharmaceuticals, cosmetics or skin care products.

[0077] The present invention also provides the application of the above-described biomimetic delivery body of black ginseng, or the biomimetic delivery body of black ginseng prepared by the above-described preparation method, in the preparation of aqueous essence.

[0078] The present invention also provides an aqueous essence composed of phase A, phase B and phase C;

[0079] Phase A is composed of deionized water, butanediol, 1,2-hexanediol, xanthan gum, p-hydroxyacetophenone, polyacrylate cross-linked polymer-6, acrylate / C10~30 alkanol acrylate cross-linked polymer and allantoin;

[0080] Phase B is arginine;

[0081] Phase C is composed of dipotassium glycyrrhizate, a biomimetic black ginseng delivery system, and black ginseng extract;

[0082] The black ginseng biomimetic delivery body is the black ginseng biomimetic delivery body described above, or the black ginseng biomimetic delivery body prepared by the preparation method described above.

[0083] In some embodiments of the present invention, the aqueous essence is composed of the following components:

[0084] Deionized water 80 wt%~85 wt%;

[0085] Butanediol 3 wt%~7 wt%;

[0086] 1,2-Hexanediol 0.4 wt%~0.8 wt%;

[0087] Xanthan gum 0.05 wt%~0.15 wt%;

[0088] p-Hydroxyacetophenone 0.2 wt%~0.8 wt%;

[0089] Polyacrylate crosspolymer-6 0.05 wt%~0.15 wt%;

[0090] Acrylic (ester) crosspolymer / C10~30 alkanol acrylate crosspolymer 0.1 wt%~0.5 wt%;

[0091] Allantoin 0.05 wt%~0.15 wt%;

[0092] Arginine 0.1 wt%~0.4 wt%;

[0093] Dipotassium glycyrrhizate 0.2 wt%~0.6 wt%;

[0094] Black ginseng biomimetic delivery system 8 wt%~12 wt%;

[0095] The sum of the amounts of the components is 100%.

[0096] Specifically, the content of the deionized water is 80 wt%, 82 wt%, 82.76 wt%, 83 wt%, and 85 wt%.

[0097] The content of butanediol is 3 wt%, 4 wt%, 5 wt%, 6 wt%, and 7 wt%.

[0098] The xanthan gum content is 0.05 wt%, 0.08 wt%, 0.1 wt%, 0.12 wt%, and 0.15 wt%.

[0099] The content of p-hydroxyacetophenone is 0.2 wt%, 0.4 wt%, 0.5 wt%, 0.6 wt%, and 0.8 wt%.

[0100] The content of the polyacrylate crosspolymer-6 is 0.05 wt%, 0.07 wt%, 0.1 wt%, 0.12 wt%, and 0.15 wt%.

[0101] The content of the acrylate / C10~30 alkanol acrylate crosspolymer is 0.1 wt%, 0.2 wt%, 0.3 wt%, 0.4 wt%, and 0.5 wt%.

[0102] The allantoin content is 0.05 wt%, 0.08 wt%, 0.1 wt%, 0.12 wt%, and 0.15 wt%.

[0103] The content of arginine is 0.1 wt%, 0.2 wt%, 0.22 wt%, 0.3 wt%, and 0.4 wt%.

[0104] The content of dipotassium glycyrrhizate is 0.2 wt%, 0.3 wt%, 0.4 wt%, 0.5 wt%, and 0.6 wt%.

[0105] The content of the black ginseng biomimetic delivery body is 8 wt%, 9 wt%, 10 wt%, 11 wt%, and 12 wt%.

[0106] In some embodiments of the present invention, the preparation method of the aqueous essence includes the following steps:

[0107] a) After stirring and mixing the A phase material, cool it to 50~60℃;

[0108] b) Add phase B, stir and mix well, and cool to 25~35℃;

[0109] c) Add phase C and mix thoroughly at 500 rpm to obtain the aqueous essence.

[0110] In step a), the mixing temperature is 80~90℃, for example 85℃; the stirring speed is 300~800 rpm, for example 500 rpm. The temperature after cooling is 55℃.

[0111] In step b), the stirring speed is 800~1000 rpm; for example, 900 rpm. The temperature after cooling is 30℃.

[0112] In step c), the stirring speed is 300~800 rpm; for example, 500 rpm.

[0113] In this invention, when the biomimetic delivery system of black ginseng with a double charge layer comes into contact with human skin (normal skin pH≈5, inflamed areas pH<4.5), the carboxymethyl deacetylated chitosan is protonated, changing from a negatively charged state to a positively charged state. At this time, the larger molecular weight carboxymethyl deacetylated chitosan separates from the cationic core due to electrostatic repulsion; simultaneously, because the skin cell membrane is negatively charged, it can promote the penetration of the positively charged cationic core, thereby enhancing the targeted and local release effect of Rg3 and collagen carried by the cationic core, effectively improving the penetration and absorption of black ginseng and collagen.

[0114] This invention also provides a green and environmentally friendly method for preparing a pH-responsive charge-reversing biomimetic black ginseng delivery system with a double charge layer, the preparation process of which avoids the use of toxic organic solvents.

[0115] To further illustrate the present invention, the following detailed description of a biomimetic delivery system for black ginseng, its preparation method, and its application, in conjunction with embodiments, is provided by the present invention, but should not be construed as limiting the scope of protection of the present invention.

[0116] 1. Synergistic Efficiency Implementation Plan

[0117] Table 1. Sample Formulation Table for Type III Collagen Test (mass percentage, %)

[0118]

[0119] In Table 1, the black ginseng extract was prepared according to the method of step 1 in Example 1 of patent application CN116159011A.

[0120] (1) Synergistic effect of type III collagen test

[0121] Human fibroblasts (HFF-1 cells, Hunan Fenghui Biotechnology Co., Ltd.) were seeded at 12,000 cells / well in 96-well plates and incubated at 37 ℃ with 5% CO2 for 24 h. Five test groups were set up: a blank control group, a positive control group, Experimental Example 1, Comparative Example 1, and Comparative Example 2. According to the test protocol, drug was administered after the cell growth rate reached 80%–90%. After drug administration, the 96-well plates were incubated at 37 ℃ with 5% CO2 for 24 h. After incubation, the cells were observed and photographed under an inverted fluorescence microscope, and the average fluorescence intensity was calculated using ImageJ software. GraphPad Prism 8.0 software was used for statistical analysis and graph plotting. Quantitative data are expressed as x±s. One-way ANOVA was used to analyze differences between groups. P<0.05 was considered statistically significant, P<0.05 was considered statistically significant, and P<0.01 was considered highly significant. The asterisk (*, **, ***, ****) is a common method for indicating significance, representing the degree of significance of differences in means. One asterisk (*) indicates a significance level of 0.05, two asterisks (**) indicate a significance level of 0.01, three asterisks (***) indicate a significance level of 0.001, and four asterisks (****) indicate a significance level of 0.0001. The results are shown in Table 2.

[0122] Table 2 Summary of COLⅢ results in HFF-1 samples

[0123]

[0124] As shown in Table 2, compared with the blank control group, Experimental Example 1 and Comparative Examples 1 and 2 all showed significant upregulation effects on type III collagen in human fibroblasts, with upregulation rates of 20.49%, 7.18%, and 8.75%, respectively. This indicates that the combined use of black ginseng extract and collagen is significantly better than the promoting effect of using either substance alone.

[0125] The Combination Index (CI) is calculated using the Bliss independent model method, and the calculation formula is shown in equation (1):

[0126] E(ABC) > E(theoretical) = 1 - (1 - E) A (1-E) B (1-E) C Formula (1);

[0127] Among them, E A =Net effect of drug A alone at the "combined concentration";

[0128] E B=Net effect of drug B acting alone at the "combined concentration";

[0129] E C =Net effect of drug C acting alone at the "combined concentration";

[0130] When CI>1, it indicates that the two drugs have a synergistic effect. E(theoretical) = "theoretical effect of two drugs combined" based on the Bliss model. E(ABC) = the effect when drugs A, B, and C are used together. When E(ABC) > E(theoretical), it indicates that there is a synergistic effect.

[0131] Based on the results in Table 2, the synergistic index was calculated using the Bliss independent model method. E(AB) = 0.205 > E(theoretical) = 0.153, further indicating that the sample of 0.5 wt% black ginseng extract and 0.015 wt% collagen has a synergistic effect on promoting type III collagen.

[0132] (2) Synergistic effect cell repair efficacy test

[0133] HaCat cells (Wuhan Pronosei Life Sciences Co., Ltd.) were seeded at 60,000 cells / well in two wells of a culture-insert 2 well and cultured for 24 h in a 37 ℃ CO2 incubator with a volume content of 5%. Five test samples were set up: negative-0h (BC), negative-24h (NC), experimental example 1, comparative example 1, and comparative example 2. According to the test protocol, the scratch insertion plug was removed and photographed, and the drugs were administered to each group, with two replicates for each group. After drug administration, the 6-well plate was placed in an incubator (37 ℃, CO2 with a volume content of 5%) and cultured for 24 h. Cell morphology was observed under a microscope and photographed. Image J software was used for image area analysis, and the scratch healing percentage was calculated using formula (2):

[0134] (Area of ​​scratched blank area - 0H — Area of ​​scratched blank area - 24H) / (Area of ​​scratched blank area - 0H) × 100 Equation (2);

[0135] GraphPad Prism 8 software was used for statistical analysis and chart generation. Quantitative data are expressed as mean ± standard deviation (x±s). One-way ANOVA was used to analyze differences between groups. A p-value < 0.05 was considered statistically significant, p < 0.05 was considered highly significant, and p < 0.01 was considered extremely significant. Asterisks (*, **, ***, ****) are a common method for indicating significance, representing the degree of significance of differences in means. One asterisk (*) indicates a significance level of 0.05, two asterisks (**) indicate a significance level of 0.01, three asterisks (***) indicate a significance level of 0.001, and four asterisks (****) indicate a significance level of 0.0001. The results are shown in Table 3.

[0136] Table 3 Summary of HaCat cell scratch test results for samples

[0137]

[0138] Table 3 shows that, compared with the blank control group, Experimental Example 1 and Comparative Example 1 both showed significant healing effects on HaCat cell scratch wound healing, with upregulation rates of 191.10% and 72.06%, respectively; Comparative Example 2 showed no significant healing effect on HaCat cell scratch wound healing. This indicates that the combined use of black ginseng extract and collagen, which does not normally have cell repair function, significantly enhances the repair effect compared to using black ginseng extract alone.

[0139] The Combination Index (CI) was calculated using the Bliss independent model method, and the calculation formula is shown in Equation (1).

[0140] When CI>1, it indicates that the two drugs have a synergistic effect. E(theoretical) = "theoretical effect of two drugs combined" based on the Bliss model. E(ABC) = the effect when drugs A, B, and C are used together. When E(ABC) > E(theoretical), it indicates that there is a synergistic effect.

[0141] Based on the results in Table 3, the synergistic index was calculated using the Bliss independent model method. E(AB) = 1.91 > E(theoretical) = 0.74, further indicating that the sample composed of 0.5 wt% black ginseng extract and 0.015 wt% collagen has a synergistic effect on the repair efficacy.

[0142] (3) Synergistic soothing efficacy test

[0143] Ana-1 cells (Hunan Fenghui Biotechnology Co., Ltd.) were seeded at 4500 cells / well in 96-well plates and cultured for 24 h in a 37℃, 5% CO2 incubator. Six samples were set up: blank (BC), negative control (NC), positive control (PC), Experimental Example 1, Comparative Example 1, and Comparative Example 2. According to the test protocol, 1 mg / mL lipopolysaccharide aqueous solution (LPS solution) was diluted to 2 μg / mL and added to each well at 50 μL. An equal volume of culture medium was added to the negative control group. The plates were then cultured at 37℃, 5% CO2 incubator for 4 h. Working solutions were prepared using complete culture medium as a diluent according to the sample test concentration. An equal volume of complete culture medium was added to both the negative and positive control groups. The plates were then cultured at 37℃, 5% CO2 incubator for 24 h, and cell morphology was observed under a microscope. The supernatant from the plates was collected, and the TNF-α concentration was detected by ELISA. GraphPad Prism 8 software was used for statistical analysis and graph generation. Quantitative data are expressed as mean ± standard deviation (x±s). One-way ANOVA was used to analyze differences between groups. A p-value < 0.05 was considered statistically significant, p < 0.05 was considered highly significant, and p < 0.01 was considered extremely significant. Asterisks (*, **, ***, ****) are a common method for indicating significance, representing the degree of significance of differences in means. One asterisk (*) indicates a significance level of 0.05, two asterisks (**) indicate a significance level of 0.01, three asterisks (***) indicate a significance level of 0.001, and four asterisks (****) indicate a significance level of 0.0001. The results are shown in Table 4.

[0144] Table 4 Summary of TNF-α release results from Ana-1 cells by samples

[0145]

[0146] Table 4 shows that, compared with the negative control group, Experimental Example 1, Comparative Example 1, and Comparative Example 2 all showed significant inhibitory effects on the release of TNF-α from Ana-1 cells, with inhibition rates of 45.87%, 42.47%, and 22.37%, respectively, demonstrating a soothing effect. This indicates that the combined use of black ginseng extract and collagen has a significantly better soothing effect than using black ginseng extract alone.

[0147] The analysis was performed using the highest single agent (HSA) model, and the calculation formula is shown in equation (3):

[0148] Synergistic effect = measured combined inhibition rate - max(inhibition rate of single drug A, inhibition rate of single drug B) Equation (3);

[0149] When the measured inhibition rate of the combination is greater than that of the strongest single drug, it indicates a synergistic effect; when the measured inhibition rate of the combination is less than or equal to that of the strongest single drug, it indicates an additive / no synergistic effect.

[0150] Based on the results in Table 4, combined with the highest single-drug (HSA) model, the measured combined inhibition rate of 45.87% was greater than the inhibition rate of the strongest single drug of 42.47%, further indicating that the sample of 0.5% black ginseng extract and 0.015% collagen has a synergistic effect on the soothing effect.

[0151] 2. Encapsulating synergistic components to form a pH-responsive charge-reversal biomimetic delivery system for black ginseng.

[0152] The components and mass content of Examples 1-2 and Comparative Examples 3-5 are shown in Table 5.

[0153] Table 5. Components and mass content (%) of Examples 1-2 and Comparative Examples 3-5

[0154]

[0155] In Table 5, the black ginseng extract was prepared according to the method of step 1 in Example 1 of patent application CN116159011A.

[0156] Example 1

[0157] Preparation of biomimetic delivery systems for black ginseng:

[0158] (1) After dispersing hydrogenated lecithin with glycerol, it was mixed with polyglycerol-10 myristate at 70°C (stirring speed of 500 rpm) to obtain mixed solution I;

[0159] (2) Dissolve the black ginseng extract (the black ginseng extract was prepared according to the method of step 1 in Example 1 of patent application CN116159011A) in a portion of water, then neutralize it with arginine to obtain a black ginseng solution with pH=7; mix the black ginseng solution, carboxymethyl deacetylated chitosan, collagen and the remaining water at 68°C (stirring speed is 500 rpm) to obtain mixed solution II;

[0160] (3) Add the mixed solution II to the mixed solution I which is being stirred (stirring speed is 500 rpm), stir at 500 rpm to mix well, and hydrate for 2 min;

[0161] (4) The hydrated solution was passed through a high-pressure microjet four times at a pressure of 1000 bar to obtain a black ginseng biomimetic delivery body with pH=7.

[0162] The particle size was detected using a NanoBrook 90Plus PALS high-sensitivity zeta potential and particle size analyzer. The particle size was 59 nm and the zeta potential was -9.6 mV. The delivery body was negatively charged on the outside, which enhanced the stability of the delivery body.

[0163] Example 2

[0164] The difference from Example 1 is as follows:

[0165] The pH of the biomimetic delivery body of black ginseng obtained in Example 1 was adjusted to 5 using HCl solution to simulate the state when the sample was applied to the skin. Due to the change in pH, the carboxymethyl deacetylated chitosan responded spontaneously, and the amino group was protonated, changing from the original negative charge to a positive charge. At this time, the Zeta potential was 12.5 mV.

[0166] Comparative Example 3

[0167] The difference from Example 1 is as follows:

[0168] Without the addition of hydroxymethyl deacetylated chitosan, after the simulated sample was applied to the skin, the outer layer of hydroxymethyl deacetylated chitosan separated from the core biomimetic delivery body due to electrostatic interaction. At this point, the particle size was 54 nm and the potential was 3.89 mV. Due to the reduced particle size and the presence of a positive charge, the biomimetic delivery body would be more conducive to penetration. The separated outer layer also carried a positive charge, which would help to exert the anti-inflammatory and soothing effects of hydroxymethyl deacetylated chitosan itself.

[0169] Comparative Example 4

[0170] The difference from Example 1 is as follows:

[0171] Increasing the amount of hydroxymethyl deacetylated chitosan to 0.5% caused the overall system to become unstable, with the particle size becoming 96 nm and the appearance becoming less transparent. This indicates that the concentration and dispersibility of hydroxymethyl deacetylated chitosan can also affect the stability of the system, and excessively high concentrations may lead to particle aggregation.

[0172] Comparative Example 5

[0173] The difference from Example 1 is as follows:

[0174] Increasing the amount of hydroxymethyl deacetylated chitosan to 1% caused the overall system to become unstable, with the particle size increasing to 114 nm and the appearance becoming less transparent. This indicates that the concentration and dispersibility of hydroxymethyl deacetylated chitosan can affect the stability of the system, and excessively high concentrations may lead to particle aggregation.

[0175] 3. Stability test of pH-responsive charge reversal biomimetic delivery system for black ginseng

[0176] Example 1 was placed at 4°C, -15°C, 25°C, and 48°C for 30 days, 60 days, and 90 days, respectively. The results are shown in Table 6.

[0177] Table 6 Statistical table of high and low temperature stability test results in Example 1

[0178]

[0179] As can be seen from Table 6, the biomimetic delivery system for black ginseng did not exhibit agglomeration or stratification, and the particle size and variation range were small. The system was stable and reliable, indicating that the biomimetic delivery system for black ginseng provided by this invention is of great significance for promoting product stability.

[0180] Comparative examples 3 to 5 were placed at 4°C, -15°C, 25°C, and 48°C respectively for 30 days. The results are shown in Table 7.

[0181] Table 7. Statistical table of high and low temperature stability test results for Comparative Examples 3-5

[0182]

[0183] 4. Osmosis test

[0184] Samples from Example 1 and Comparative Example 6 (formulations shown in Table 8) were taken and sulfonyl rhodamine salt was added to each sample to achieve a sulfonyl rhodamine salt content of 0.02% by mass. The resulting samples were then prepared into a 20% (w / w) fluorescent solution using deionized water as test samples. Ex vivo Bama miniature pig skin (0.8–1 cm) was used as the in vitro transdermal test material, with PBS as the receiving medium and a Franz diffusion cell used to measure the permeation absorption. The parameters were set at 37.5℃ and 600 rpm. After permeation for 30 min, the pig skin surface was cleaned with PBS and sectioned. The distribution of fluorescence signals in the pig skin was observed under a fluorescence microscope, and the fluorescence penetration depth was analyzed using ImageJ software. The transdermal absorption effect of the double-charged, pH-responsive charge-reversed black ginseng biomimetic delivery system and the unencapsulated black ginseng collagen complex was evaluated. The test results are shown below. Figure 1 As shown. Figure 1 The images show the fluorescence penetration depth of the samples from Example 1 and Comparative Example 6 of this invention.

[0185] Table 8. Permeability test formulations for Example 1 and Comparative Example 6 (mass content, %)

[0186]

[0187] Figure 1In the comparison of fluorescence penetration depth between Example 1 and Comparative Example 6, it can be seen that the penetration depth of Example 1 is 219 μm, while that of Comparative Example 6 is 177 μm. The penetration effect of Example 1 is 23.73% higher than that of Comparative Example 6, which is a significant difference, indicating that the encapsulated liposomes have a better penetration effect.

[0188] Application examples

[0189] The black ginseng biomimetic delivery system obtained in Example 1 was used to prepare an aqueous essence, and the steps are as follows:

[0190] (1) After mixing the A phase material at 85°C and 500 rpm, cool it down to 55°C;

[0191] (2) Add phase B material, stir at 900 rpm to mix well, and cool to 30°C;

[0192] (3) Add phase C material and stir at 500 rpm to obtain aqueous essence sample;

[0193] That is, Application Example 1; the difference between Application Example 1 and Application Example 2 is that the black ginseng delivery medium is replaced with black ginseng extract (the mass concentration of black ginseng extract is the same in both).

[0194] The specific component formulation is shown in Table 9.

[0195] Table 9. Component formulation of aqueous essence (mass content, %)

[0196]

[0197] Figure 2 The images show the appearance of the aqueous extracts of Application Example 1 and Comparative Application Example 1, where (a) is the appearance of the aqueous extract of Application Example 1 and (b) is the appearance of the aqueous extract of Comparative Application Example 1. Comparing the colors of the two, it can be seen that the aqueous extract of Application Example 1 is lighter in color, reducing the appearance of black ginseng and making it easier to apply in products.

[0198] A questionnaire survey was conducted after product use, comparing before and after results. The overall appearance, smoothness, moisturizing effect, hydration, softness, smoothness, and skin radiance of Application Example 1 were evaluated. Evaluation indicators were scaled from 1 to 10, ranging from unsatisfactory to satisfactory. Data analysis was performed. Results are as follows: Figure 3 As shown, Figure 3 These are test images of the appearance and skin feel of the aqueous essence in Application Example 1 and Comparative Application Example 1 of the present invention. Figure 3 It can be seen that Application Example 1 is superior to Comparative Application Example 2 in all dimensions.

[0199] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A biomimetic delivery system for black ginseng, characterized in that, include: Glycerin 40 wt%~60 wt%, hydrogenated lecithin 1 wt%~3 wt%, polyglycerol-10 myristate 0.3 wt%~1 wt%, black ginseng extract 0.2 wt%~1 wt%, arginine 0.05 wt%~0.15 wt%, carboxymethyl deacetylated chitosan 0.01 wt%~0.1 wt%, collagen 0.01 wt%~0.02 wt%, and the balance being water.

2. A method for preparing the black ginseng biomimetic delivery body according to claim 1, characterized in that, Includes the following steps: S1. After dispersing hydrogenated lecithin with glycerol, it is mixed with polyglycerol-10 myristate at 65~75℃ to obtain mixed solution I; The black ginseng extract was partially dissolved in water and then neutralized with arginine to obtain a black ginseng solution with a pH of 7. The black ginseng solution, carboxymethyl deacetylated chitosan, collagen and the remaining water were mixed at 65-70°C to obtain mixed solution II; S2. Mix the mixed solution I and mixed solution II thoroughly and then hydrate them; S3. The solution after hydration in step S2 is passed through a high-pressure microjet to obtain a biomimetic delivery body for black ginseng.

3. The preparation method according to claim 2, characterized in that, In step S1, mixing at 65~75℃ means stirring and mixing at 65~75℃, and the stirring and mixing rate is 300~800 rpm.

4. The preparation method according to claim 2, characterized in that, In step S1, mixing at 65~70℃ means stirring and mixing at 65~70℃, and the stirring and mixing rate is 300~800 rpm.

5. The preparation method according to claim 2, characterized in that, In step S2, mixing the mixed solution I and mixed solution II includes: Add the mixed solution II to the stirred mixed solution I, and stir until well mixed; The stirring and mixing rate is 300~800 rpm.

6. The preparation method according to claim 2, characterized in that, In step S2, the hydration time is 1 to 3 minutes.

7. The preparation method according to claim 2, characterized in that, In step S3, the pressure of the high-pressure microjet is 800~1200 bar; the number of times is 3~5.

8. The use of the black ginseng biomimetic delivery body according to claim 1, or the black ginseng biomimetic delivery body prepared by any one of claims 2 to 7, in the preparation of pharmaceuticals, cosmetics, or skin care products.

9. The application of the black ginseng biomimetic delivery body according to claim 1, or the black ginseng biomimetic delivery body prepared by any one of claims 2 to 7, in the preparation of aqueous essence.

10. A water-based essence, characterized in that, Including phase A, phase B, and phase C; Phase A includes deionized water, butanediol, 1,2-hexanediol, xanthan gum, p-hydroxyacetophenone, polyacrylate cross-linked polymer-6, acrylate / C10~30 alkanol acrylate cross-linked polymer and allantoin; Phase B includes arginine; Phase C includes dipotassium glycyrrhizate, a biomimetic black ginseng delivery system, and black ginseng extract; The black ginseng biomimetic delivery body is the black ginseng biomimetic delivery body according to claim 1, or the black ginseng biomimetic delivery body prepared by any one of claims 2 to 7.

Images