Whitening and acne-removing plant extract composition, preparation method and application in medical cosmetology

Abstract

The invention discloses a whitening and anti-acne plant extract composition, a preparation method and its application in medical cosmetology. Most natural Chinese herbal extracts have relatively poor water solubility and fat solubility, which is not conducive to skin transdermal absorption and affects the overall effect of cosmetics. The present invention uses lipid nanoparticles as a matrix, and encapsulates extracts such as Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa in the inner core to form a new drug carrier system, which can improve the permeability of Chinese herbal extracts and prolong the action time. In addition, Scutellaria baicalensis and Forsythia suspensa extracts have a good therapeutic effect on skin acne, and the addition of Angelica dahurica makes it have good antioxidant and whitening effects. The whitening and anti-acne composition prepared by the lipid nanoparticles can not only effectively inhibit the growth of acne, but also play a certain role in whitening and inhibiting melanin, and also has the effect of keeping the skin shiny.

Description

Technical Field

[0001] The invention belongs to the field of natural plant extraction, and specifically relates to a whitening and anti-acne composition, in particular to a whitening and anti-acne composition and a preparation process thereof. Background Art

[0002] Angelica dahurica is fragrant, white, and slightly warm. It was first recorded as a medicine in Shennong's Herbal Classic. Because of its special aromatic smell, it is also called fragrant Angelica dahurica. It is used externally as a beauty medicine and is often used in ancient beauty recipes. The Rihuazi Materia Medica records that it can "remove facial skin and smooth blemishes and tumors." The Compendium of Materia Medica says that it has the effect of "growing skin, adjusting color, and can be used as facial fat." Facial fat is the popular facial mask today.

[0003] Lianqiao is a Chinese medicine, which is mentioned in Shennong's Herbal Classic. In autumn, the fruit is harvested when it is just ripe and still green, and then impurities are removed, steamed, and dried. It is commonly called Qingqiao; when the fruit is fully ripe, it is harvested, dried, and impurities are removed. It is commonly called Laoqiao (Huangqiao). Qingqiao is the best and is used raw. It is mainly produced in Shanxi, Henan, Shaanxi, Shandong and other places, with the largest production in Shanxi and Henan. Lianqiao has the effects of clearing away heat and detoxifying, reducing swelling and dispersing knots, and evacuating wind and heat. It is used for carbuncle, scrofula, breast carbuncle, erysipelas, wind-heat cold, the beginning of febrile disease, warm and heat entering the camp, high fever and thirst, coma and spots, and hot stranguria and astringent pain.

[0004] Scutellaria baicalensis is bitter and cold. It enters the lung, gallbladder, spleen, large intestine and small intestine meridians. It can reduce the heat in the human body after taking it. It is a cool Chinese herbal medicine that can clear away heat and dampness, purge fire and detoxify, stop bleeding and stabilize the fetus. In clinical practice, Scutellaria baicalensis is often used to treat damp-heat summer heat, chest tightness and nausea, damp-heat fullness, diarrhea and jaundice, lung heat cough, high fever and thirst, blood heat vomiting and bleeding, carbuncle, sore and toxicity, and fetal movement disorder. Baicalin in Scutellaria baicalensis extract also has the effect of anti-oxidation and inhibiting melanin.

[0005] However, the water solubility and fat solubility of the extracts of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa are very poor, which is not conducive to skin absorption and even affects the overall effect of the drug. In view of this, the present application uses lipid nanoparticles to encapsulate the extracts of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa, greatly enhancing their skin permeability and also prolonging the action time of the Chinese herbal extracts. Summary of the invention

[0006] In view of the shortcomings of the prior art, the purpose of the present invention is to prepare a whitening and anti-acne composition, which has the characteristics of anti-oxidation, wrinkle removal, and skin glossiness. To achieve the above purpose, the present invention is implemented by the following technical solutions:

[0007] S1. Preparation of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts: weigh 50g of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa respectively, grind them into 70-300 mesh using a high-speed multifunctional grinder at 34000r / min, soak them in 70% ethanol with a mass fraction of 1:10 for overnight, reflux at above 80℃ for 1-6h, repeat twice, filter with diatomaceous earth until clear, and collect the filtrate; combine the two filtrates, distill under reduced pressure at 60℃, concentrate into an extract, and store in a refrigerator.

[0008] S2. Preparation of lipid nanoparticles: Weigh the prescribed amount of solid lipids and soybean lecithin, add them to 5 mL of ethanol, and dissolve them by ultrasonic. Then weigh a certain amount of Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa extracts in proportion, add them to the lipid solution, stir until the extracts are completely dissolved, and heat in a water bath to 60-90°C for insulation; weigh another 50 mL of purified water, heat in a water bath to 6-15°C for insulation; under magnetic stirring at 3000 r / min, quickly add the organic phase containing the extracts into the purified water to disperse to form a white emulsion, maintain a water bath temperature of 60-90°C, continue high-speed stirring for 30 minutes, evaporate the ethanol, and add purified water at the same temperature to a total volume of 50 mL; then put the solution into a high-speed dispersing homogenizer for homogenization at a speed of 20000 r / min for 10 minutes. After the homogenization, quickly put the emulsion into an ice water bath to obtain solid lipid nanoparticles.

[0009] S3. Preparation of lipid nanoparticle gel: Weigh 0.5 g of Carbomer 940 and add it to 50 mL of purified water, and continue stirring until it is evenly dispersed; take another 50 mL of solid lipid nanoparticle solution and add it to the Carbomer 940 solution, and stir to mix evenly; during the stirring process, add triethylamine to adjust the pH of the solution to between 6.5 and 7.5 to form a gel solution, thereby obtaining a solid lipid nanoparticle gel.

[0010] Preferably, the solid lipid used in step S2 is glyceryl distearate.

[0011] Compared with the prior art, the present invention has the following beneficial effects:

[0012] 1. The lipid nanoparticle gel of the present invention contains no hormones, no pigments, no synthetic fragrances, no preservatives or other harmful chemical additives.

[0013] 2. The extracts of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa used in the present invention can not only protect the skin, but also play a role in maintaining the body.

[0014] 3. The lipid nanoparticle gel prepared by the present invention can adhere to human skin for a long time, prolonging the action time of the whitening composition, and the skin permeability is also greatly enhanced.

[0015] 4. The present invention is simple to operate, has low production cost, and is conducive to mass production.

[0016] 5. The lipid nanoparticle gel prepared by the present invention has a stable two-dimensional network structure, which ensures the stability of the extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 This is a TEM image of the solid lipid nanoparticles prepared in the present invention.

[0018] Figure 2 This is a TEM image of the solid lipid nanoparticle gel prepared in the present invention.

[0019] Figure 3 A bar graph of DPPH clearance rates of VC control, Examples 1-5 and Comparative Examples 1-7 were prepared for the present invention.

[0020] Figure 4 The melanin content detection bar graph of Examples 1-5 and Comparative Examples 1-7 prepared by the present invention is shown.

[0021] Figure 5 This is the transdermal Qi-t relationship diagram of Comparative Example 8 and Example 6. DETAILED DESCRIPTION

[0022] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0023] Preparation of lipid nanoparticle gel

[0024] 1. Materials and Methods

[0025] 1. Preparation of Lipid Nanoparticle Gels

[0026] The preparation process of the lipid nanoparticle gel of a specific embodiment 1 is as follows:

[0027] S1. Preparation of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts: weigh 50g of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa respectively, grind them into 200 mesh using a high-speed multifunctional grinder at 34000r / min, soak them in 70% ethanol with a mass fraction of 1:10 overnight, reflux at above 80°C for 1h, repeat twice, filter with diatomaceous earth until clear, and collect the filtrate; combine the two filtrates, distill under reduced pressure at 60°C in vacuum, concentrate into an extract, and store in a refrigerator.

[0028] S2. Preparation of lipid nanoparticles: Weigh the prescribed amount of distearin and soybean lecithin, add to 5 mL of ethanol, and dissolve by ultrasonication. Then weigh 10 mg of Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa extracts respectively, add to the lipid solution, stir until the extracts are completely dissolved, and heat to 60°C in a water bath to keep warm. Weigh another 50 mL of purified water, heat to 6°C in a water bath to keep warm. Under magnetic stirring at 3000 r / min, quickly add the organic phase containing the extracts into the purified water to disperse to form a white emulsion. Maintain the water bath temperature at 60°C and continue high-speed stirring for 30 min. Volatilize the ethanol, and add purified water at the same temperature to a total volume of 50 mL. Place the solution in a high-speed dispersing homogenizer for homogenization at a speed of 20,000 r / min for 10 min. After homogenization, quickly place the emulsion in an ice water bath to obtain solid lipid nanoparticles.

[0029] S3. Preparation of lipid nanoparticle gel: Weigh 0.5 g of Carbomer 940 and add it to 50 mL of purified water, and continue stirring until it is evenly dispersed; take another 50 mL of solid lipid nanoparticle solution and add it to the Carbomer 940 solution, and stir to mix evenly; during the stirring process, add triethylamine to adjust the pH of the solution to between 6.5 to form a gel solution, thereby obtaining a solid lipid nanoparticle gel.

[0030] The preparation process of the lipid nanoparticle gel of a specific embodiment 2 is substantially the same as that of embodiment 1, except that 15 mg of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts are weighed in step S2.

[0031] The preparation process of the lipid nanoparticle gel of a specific embodiment 3 is substantially the same as that of embodiment 1, except that 20 mg of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts are weighed in step S2.

[0032] The preparation process of the lipid nanoparticle gel of a specific embodiment 4 is substantially the same as that of embodiment 1, except that 25 mg of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts are weighed in step S2.

[0033] The preparation process of the lipid nanoparticle gel of a specific embodiment 5 is substantially the same as that of embodiment 1, except that 30 mg of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts are weighed in step S2.

[0034] The preparation process of a specific lipid nanoparticle gel of Comparative Example 1 is as follows:

[0035] S1. Preparation of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts: weigh 50g of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa respectively, grind them into 200 mesh using a high-speed multifunctional grinder at 34000r / min, soak them in 70% ethanol by mass at a solid-liquid ratio of 1:10 overnight, reflux at above 80°C for 1h, repeat twice, filter with diatomaceous earth until clear, and collect the filtrate; combine the two filtrates, distill under reduced pressure at 60°C in vacuum, concentrate into an extract, and store in a refrigerator.

[0036] S2. Preparation of lipid nanoparticles: Weigh the prescribed amount of distearin and soybean lecithin, add to 5 mL of ethanol, and dissolve by ultrasonication. Then weigh 30 mg of Scutellaria baicalensis and Forsythia suspensa extracts respectively, add to the lipid solution, stir until the extracts are completely dissolved, and heat to 60°C in a water bath to keep warm. Weigh another 50 mL of purified water, heat to 6°C in a water bath to keep warm. Under magnetic stirring at 3000 r / min, quickly add the organic phase containing the extracts into the purified water to disperse to form a white emulsion. Maintain the water bath temperature at 60°C and continue high-speed stirring for 30 min. Volatilize the ethanol, and add purified water at the same temperature to a total volume of 50 mL. Place the solution in a high-speed dispersing homogenizer for homogenization at a speed of 20,000 r / min for 10 min. After homogenization, quickly place the emulsion in an ice-water bath to obtain solid lipid nanoparticles.

[0037] S3. Preparation of lipid nanoparticle gel: Weigh 0.5 g of Carbomer 940 and add it to 50 mL of purified water, and continue stirring until it is evenly dispersed; take another 50 mL of solid lipid nanoparticle solution and add it to the Carbomer 940 solution, and stir to mix evenly; during the stirring process, add triethylamine to adjust the pH of the solution to between 6.5 to form a gel solution, thereby obtaining a solid lipid nanoparticle gel.

[0038] The preparation process of the lipid nanoparticle gel of a specific comparative example 2 is substantially the same as that of comparative example 1, except that only Angelica dahurica and Scutellaria baicalensis are added in step S2.

[0039] The preparation process of the lipid nanoparticle gel of a specific comparative example 3 is substantially the same as that of comparative example 1, except that only Angelica dahurica and Forsythia suspensa are added in step S2.

[0040] The preparation process of the lipid nanoparticle gel of a specific comparative example 4 is substantially the same as that of comparative example 1, except that only Angelica dahurica is added in step S2.

[0041] The preparation process of the lipid nanoparticle gel of a specific comparative example 5 is substantially the same as that of comparative example 1, except that only Scutellaria baicalensis is added in step S2.

[0042] The preparation process of the lipid nanoparticle gel of a specific comparative example 6 is substantially the same as that of comparative example 1, except that only Forsythia suspensa is added in step S2.

[0043] The preparation process of the lipid nanoparticle gel of a specific comparative example 7 is substantially the same as that of comparative example 1, except that Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa are not added in step S2.

[0044] 2. In vitro antioxidant activity assay

[0045] DPPH free radical scavenging ability assay

[0046] Dissolve 40 mg DPPH in 1000 mL methanol to obtain a 0.04 mg / mL DPPH methanol solution, and prepare a sample solution with a concentration of 1 mg / mL. Take 0.2 mL of each sample solution and add 7.8 mL DPPH methanol solution. React in the dark for 30 minutes, measure the absorbance at 517 nm, use distilled water instead of the sample as a blank control, use methanol instead of DPPH methanol solution as a sample blank, and use vitamin C as a positive control.

[0047] DPPH free radical scavenging capacity / %=(A 对照组 -A 样品组 ) / A 对照组 ×100

[0048] 3. Tyrosinase Inhibitory Activity Assay

[0049] In the biosynthesis of skin melanin, tyrosinase is mostly used for DOPA to form DOPAquinone, which then forms melanin after a series of spontaneous reactions.

[0050] The inhibition rate of hyaluronidase activity was determined by the modified Elson-Morgan method. During the experiment, hyaluronidase can indiscriminately hydrolyze the 1,4-bond between β-N-acetylglucosamine and D-glucuronic acid in hyaluronic acid to obtain β-N-acetylglucosamine, which can condense with acetylacetone under alkaline conditions to generate a chromogen 2-methyl-3-diacetylpyrrole derivative, and the chromogen reacts with p-dimethylaminobenzaldehyde in concentrated hydrochloric acid and ethanol to produce color.

[0051] Prepare a sample solution with a concentration of 1 mg / mL for standby use. Take 0.1 mL of 0.25 mmol / L CaCl2 solution, add 0.5 mL of hyaluronidase solution (1250 U / mL), and keep it at 37°C for 20 min; add 0.5 mL of sample solution and continue to keep it at 37°C for 20 min; add 0.5 mL of 0.5 g / L sodium hyaluronate solution, keep it at 37°C for 30 min, and then place it at room temperature for 5 min; then add 0.1 mL of 0.4 mol / L NaOH solution and 0.5 mL of acetylacetone solution, place it in a boiling water bath and heat it for 15 min, then immediately cool it with cold water for 5 min; add 1 mL of Ehrlich reagent, dilute it with 3 mL of anhydrous ethanol, mix it well, and place it for 20 min for color development. Use dipotassium glycyrrhizinate as a positive control and measure the absorbance value with a UV-visible spectrophotometer.

[0052] Tyrosinase activity inhibition rate = [1-(CD) / (AB)] × 100%

[0053] Among them, A: absorbance value of control solution (acetate buffer is used instead of sample solution); B: absorbance value of control blank solution (acetate buffer is used instead of sample solution and enzyme solution); C: absorbance value of sample solution; D: absorbance value of sample blank solution (acetate buffer is used instead of enzyme solution).

[0054] First scan liquid A in the range of 450nm-700nm to determine the maximum absorption wavelength.

[0055] 4. In vitro antibacterial test

[0056] Acne is also called facial acne. The main cause is the infection of the face by Propionibacterium acnes and Staphylococcus aureus, and the secretion of oil leads to clogged pores. Therefore, we use the antibacterial ring experiment to test the antibacterial effect of the sample, and the selected bacteria are mainly Propionibacterium acnes and Staphylococcus aureus.

[0057] 2. Results

[0058] Figure 1 , Figure 2 The TEM images of lipid nanoparticles and solid lipid nanoparticle gels are shown in Figure 1. Through transmission electron microscopy, it was observed that the particle size of solid lipid nanoparticles before and after being prepared into gels was almost unchanged, with a particle size of about 200 nm, all of which were spherical or quasi-spherical, with a smooth and round surface, and no drug crystals were observed to precipitate.

[0059] Table 1

[0060]

[0061] Figure 3It is a bar graph of DPPH clearance rate of VC control, embodiment 1-5, and comparative example 1-7. As can be seen from the figure, with the increase of the content of Angelica dahurica, Scutellaria baicalensis, and Fructus Forsythiae, the DPPH clearance rate of embodiment 1-5 also gradually increases, so the antioxidant performance is also continuously enhanced. As can be seen from embodiment 1 and comparative examples 4-7, the DPPH clearance rate of the sample with Angelica dahurica added is higher than that of the sample with Angelica dahurica embedded, indicating that Angelica dahurica mainly plays the role of antioxidant and whitening. In addition, the antioxidant performance of the mixed addition of Angelica dahurica, Scutellaria baicalensis, and Fructus Forsythiae is better than that of the single Angelica dahurica, and the reason may be that the active substances of Angelica dahurica, Scutellaria baicalensis, and Fructus Forsythiae undergo certain chemical reactions in the solid lipid nanoparticles, which greatly enhances their antioxidant performance.

[0062] Whitening is closely related to melanin, so the whitening effect of the product is judged by the inhibition rate of melanin. As can be seen from Table 1, as the content of herbal plant extracts continues to increase, the tyrosinase inhibition rate is also increasing. From Examples 1-5 and the positive comparative example, it can be seen that the whitening effect of this product is almost the same as that of the products on the market.

[0063] The inhibitory effect of the sample on acne was determined by the size of the inhibition ring. It was stipulated that the extremely active sample (diameter>20mm) was scored as 4; the active sample (diameter 15-20mm) was scored as 3; the antibacterial active sample (diameter 9-14mm) was scored as 2; the antibacterial active sample (diameter 8-4mm) was scored as 1; and the sample with no antibacterial activity (diameter <4mm) was scored as 0, thus obtaining Table 2.

[0064] Table 2 Inhibition ring test results

[0065] Propionibacterium acnes Staphylococcus aureus Example 5 4 4 Comparative Example 1 3 3 Comparative Example 2 1 2 Comparative Example 3 1 1 Comparative Example 4 0 0 Comparative Example 5 2 1 Comparative Example 6 2 2 Comparative Example 7 0 0

[0066] As can be seen from Table 2, Example 5 has a good antibacterial effect on Propionibacterium acnes and Staphylococcus aureus. By comparing Example 5 with Comparative Examples 1-7, it can be seen that Scutellaria baicalensis and Forsythia suspensa play an antibacterial role in the sample, but the antibacterial effect of Example 5 is better than that of a single component, indicating that Scutellaria baicalensis and Forsythia suspensa act in the solid lipid nanoparticles, so that they have a synergistic effect on Propionibacterium acnes and Staphylococcus aureus.

[0067] Cell experiment 1. Materials and methods

[0068] 1. Materials

[0069] B16-F10 cells (Wuhan Pronocell Life Science Co., Ltd.); DMEM medium (Gibco, USA); 0.25% trypsin-EDTA (Gibco, USA); dimethyl sulfoxide (DMSO, Sigma, USA); PBS buffer (Beijing Solebao Technology Co., Ltd.); MTT kit (Mainland China Keygene Co., Ltd.).

[0070] 2. Melanin inhibition experiment

[0071] The NaOH lysis method was used to determine the melanin content in the cells. B16 cells in the logarithmic growth phase were digested, inoculated in a 6-well plate at a concentration of 2×105 cells / well, and cultured overnight in a 37°C, 5% CO2 incubator. After the cells were completely attached, the drug was applied and the culture was continued for 72 hours. The supernatant was discarded, washed twice with PBS, and 200 μL of cell lysis solution was added to each well after collecting the cells with a cell scraper. After fully lysing at 4°C for 40 minutes, centrifuged at 12000rpm for 20 minutes. The supernatant was transferred to an EP tube to determine the protein concentration. 190 μL of 1 mol / L NaOH (containing 10% DMSO) lysis solution was added to the precipitate, and after fully dissolving in an 80°C water bath for 1 hour, the absorbance was measured at 405nm, and the final melanin content was normalized using the relative protein concentration. The calculation formula is:

[0072] Melanin relative content = (OD 405样品 / protein concentration) / (OD 405空白 / protein concentration)×100%

[0073] Each sample was measured three times.

[0074] 3. Cytotoxicity Assay

[0075] Collect cells in logarithmic growth phase, plate the cells to be tested to a density of 5000 cells / well, culture at 37°C and 5% CO2, grow the cells to a cell suspension of 105 cells / ml, replace the medium with DMEM complete medium containing 0.1mg / ml test solution, and add the same volume of DMEM complete medium as the control group. Continue to culture for 24h, add 20μL 0.5% MTT solution (5g / L) to each well, discard the culture medium after 4h of culture, rinse with PBS 2-3 times, add 150μL dimethyl sulfoxide to each well, and shake at low speed on a shaker for 10min to fully dissolve the crystals. Measure the absorbance of each well at 490nm.

[0076] Cell survival rate = (absorbance of the assay well - absorbance of the blank control group) / (absorbance of the control group - absorbance of the blank control group) × 100%.

[0077] 2. Results

[0078] Table 3

[0079] Intracellular melanin content (%) Cell survival rate (%) Comparison 100.00±1.23 100.0±1.6 Example 1 75.75±2.36 120.8±1.7 Example 2 73.26±1.35 119.5±2.6 Example 3 74.23±1.54 125.6±0.9 Example 4 72.15±1.26 123.5±0.8 Example 5 65.09±1.58 127.8±1.7 Comparative Example 1 97.32±1.26 104.6±1.3 Comparative Example 2 90.64±1.87 108.3±1.8 Comparative Example 3 89.35±0.98 106.5±1.2 Comparative Example 4 90.32±1.24 108.3±2.3 Comparative Example 5 98.65±1.89 107.9±1.6 Comparative Example 6 97.89±1.76 106.5±1.4 Comparative Example 7 100.00±0.97 102.1±2.8

[0080] Figure 4 The bar graph of intracellular melanin content obtained by performing cell experiments on blank control, Examples 1-5, and Comparative Examples 1-7 of the present invention. It can be seen from the figure that the intracellular melanin content in Example 5 is the lowest, because the content of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa in Example 5 is more than that in Example 1-4. In addition, by comparing Examples 1-5 and Comparative Examples 1-7, it can be seen that the simultaneous addition of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa is better than the addition of a single content, which also shows from another aspect that Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa have a synergistic effect in inhibiting melanin.

[0081] In addition, it can be seen from Table 3 that the cell survival rates of Examples 1-5 and Comparative Examples 1-7 are higher than those of the blank control, which means that the samples are not toxic to cells. At the same time, the cell survival rates in Examples 1-5 are better than those in Comparative Examples 1-7, which also shows that Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa are helpful for cell growth and reproduction, and also show that Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa also have synergistic effects in cell reproduction and other aspects.

[0082] Animal experiments

[0083] 1. Preparation of gel

[0084] The preparation process of the lipid nanoparticle gel of a specific embodiment 6 is as follows:

[0085] S1. Preparation of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts: weigh 50g of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa respectively, grind them into 150 mesh using a high-speed multifunctional grinder at 34000r / min, soak them in 70% ethanol by mass at a solid-liquid ratio of 1:10 overnight, reflux at above 80°C for 1h, repeat twice, filter with diatomaceous earth until clear, and collect the filtrate; combine the two filtrates, distill under reduced pressure at 60°C in vacuum, concentrate into an extract, and store in a refrigerator.

[0086] S2. Preparation of lipid nanoparticles: Weigh the prescribed amount of distearin and soybean lecithin, add to 5 mL of ethanol, and dissolve by ultrasonication. Then weigh 10 mg of Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa extracts respectively, add to the lipid solution, stir until the extracts are completely dissolved, and heat to 60°C in a water bath to keep warm. Weigh another 50 mL of purified water, heat to 6°C in a water bath to keep warm. Under magnetic stirring at 3000 r / min, quickly add the organic phase containing the extracts into the purified water to disperse to form a white emulsion. Maintain the water bath temperature at 60°C and continue high-speed stirring for 30 min. Volatilize the ethanol, and add purified water at the same temperature to a total volume of 50 mL. Place the solution in a high-speed dispersing homogenizer for homogenization at a speed of 20,000 r / min for 10 min. After homogenization, quickly place the emulsion in an ice water bath to obtain solid lipid nanoparticles.

[0087] S3. Preparation of lipid nanoparticle gel: Weigh 0.5 g of Carbomer 940 and add it to 50 mL of purified water, and continue stirring until it is evenly dispersed; take another 50 mL of solid lipid nanoparticle solution and add it to the Carbomer 940 solution, and stir to mix evenly; during the stirring process, add triethylamine to adjust the pH of the solution to between 6.5 to form a gel solution, thereby obtaining a solid lipid nanoparticle gel.

[0088] A specific preparation process of the common punch and gel of Comparative Example 8 is as follows:

[0089] Weigh 50 mL of distilled water, weigh 6 g of glycerin (moisturizer) and 87.5 mg of sodium bisulfite (stabilizer), add them into the distilled water and stir to dissolve, then weigh 0.75 g of Carbomer 940 and disperse it evenly in the solution, adjust the pH value to 6.5 with triethylamine to form a gel solution, finally add 0.568 g of Angelica dahurica, Scutellaria baicalensis, Forsythia suspensa and 0.170 g of Cnidium monnieri respectively, stir and disperse evenly to obtain a gel, which is stored in the refrigerator for later use.

[0090] 2. In vitro transdermal permeation test

[0091] Male SD rats were fasted for one night before the experiment. The rats’ abdominal hair was cut with electric clippers and then completely removed with depilatory agents. The rats were killed and the abdominal skin was cut. Subcutaneous fat and muscle tissue were removed with tweezers and washed with saline. The rat skin was sandwiched in the middle of the Franz diffusion cell, with the stratum corneum facing the supply cell and the dermis facing the receiving cell. The transdermal area was about 3.14 cm 28.45 mL of 20% PEG 400 in phosphate buffer (pH 7.4) was added to the receiving pool as the receiving solution to meet the sink conditions. The receiving pool was placed at (32 ± 0.5) °C to maintain a constant temperature. A stirring magnet was placed in the receiving pool at a speed of 200 r / min. 1.0 g of Chonghe gel (10 mg / g of Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa) and 1.25 g of solid lipid nanoparticle gel (8 mg / g of Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa) were added to the supply pool for transdermal experiments. 0.5 mL of receiving solution was taken out from the receiving pool at 1, 2, 3, 4, 6, 8, 10, and 12 h (and blank receiving solution of the same temperature and volume was added), centrifuged, and the supernatant was appropriately diluted. The drug content at each time point was determined by HPLC. After the experiment, the mouse skin was removed, washed with saline, cut into pieces, added with 1.0 mL of saline, ground into a slurry, transferred to a 10 mL volumetric flask, added with a small amount of acetonitrile, ultrasonicated for 20 min, fixed to volume, took an appropriate amount of solution and centrifuged, took the supernatant, determined the drug content by HPLC, and calculated the retention of the two gels in the skin.

[0092] 3. Results

[0093] Figure 5 The transdermal Qi-t relationship diagram of Comparative Example 8 (Chonghe gel) and Example 6 (solid lipid nanoparticle gel). Figure 5 The results show that the transdermal absorption of solid lipid nanoparticle gel is significantly improved compared with Chonghe gel. In addition, it can be seen from the figure that the drug retention in the skin of the solid lipid nanoparticle gel group is about 4 times that of the Chonghe gel group, indicating that the solid lipid nanoparticle gel can adhere to the skin surface, play the role of drug storage, increase the drug retention in the skin, and prolong the drug action time.

[0094] Photosensitivity studies

[0095] 1. Materials and Methods

[0096] Drugs: Bergamot lactone, samples prepared in Example 1 and Comparative Examples 1-7

[0097] Animals: Male Kunming rats, weighing 200-230 g, housed under standard conditions for one week.

[0098] Preparation of intragastric administration solution: The samples prepared in Examples 1 and 2-4 were dissolved in water, so that the sample concentration in the prepared intragastric administration sample was 1 g / ml.

[0099] Method: 40 rats were randomly selected and randomly divided into 4 groups, each with 10 rats, namely, Example 1 and Comparative Examples 1-7. After fasting for 12 hours, the rats in each group were gavaged with the gavage samples prepared in each group. At 0, 1, 3, 10, and 24 hours after administration, 0.3 ml of blood was collected from the orbital vein into a centrifuge tube moistened with sodium heparin and centrifuged at 3500 r·min. -1 Centrifuge for 15 minutes, take the upper plasma, accurately draw 100 μL of plasma sample, accurately add 300 μL of acetonitrile to precipitate protein, vortex for 10 minutes, 12000 r·min -1 Centrifuge for 10 min, take the supernatant, and blow dry with a vacuum concentrator; then reconstitute the sample with 100 μL of 75% methanol, vortex for 10 min, and rotate at 12000 r·min. -1 After centrifugation for 10 min, the supernatant was taken for identification by liquid chromatography.

[0100] 2. Results

[0101] Table 4 Results of bergamot lactone content in rats

[0102] Angelica dahurica contains coumarin components, and this type of coumarin component is an important factor causing the photosensitivity of Angelica dahurica. Among them, common coumarin components include imperatorin, isoimperatorin, oxidized imperatorin, white angelica, bergamot lactone, and fennel. The content of bergamot lactone in mice in the Example 1 group was 0.083 mg / ml, while in the Comparative Examples 2-3 groups, after not adding Scutellaria baicalensis and Fructus Forsythiae, the content of bergamot lactone in mice became 0.125 mg / ml and 0.138 mg / ml. The content of bergamot lactone in the Comparative Example 4 group, in which neither Scutellaria baicalensis nor Fructus Forsythiae was added, became 0.159 mg / ml. From the data comparison in Table 4, it can be seen that after adding Scutellaria baicalensis and Fructus Forsythiae, the coumarin components in mice decreased, which may be due to the synergistic effect between the three, which enables mice to absorb coumarin components faster, thereby reducing the photosensitivity of mice.

[0103] In summary, the acne-removing and whitening composition prepared by the present invention uses Angelica dahurica, Scutellaria baicalensis, and Forsythia suspensa as raw materials, and encapsulates them in a solid lipid nanoparticle gel. The obtained gel surface is smooth, non-aggregated, and has good dispersibility. In addition, since carbomer adheres to the skin as a gel matrix, it can reduce the evaporation of water on the skin surface, increase the hydration of the stratum corneum, change the structure of the stratum corneum, and facilitate drug penetration; on the other hand, since the drug is encapsulated in a fat-soluble lipid carrier, the lipophilicity of the drug is increased, and due to the presence of the surfactant soybean lecithin, it can interact with the skin and change the tight binding of the stratum corneum. Based on the combined effect of these two aspects, the permeability and retention of the drug in the skin are increased, and it is expected to improve the therapeutic effect of herbal plant extracts on skin diseases.

[0104] Finally, it should be noted that the above-mentioned embodiments only express the specific implementation of the present invention, and the description is relatively specific and detailed, but it cannot be understood as limiting the scope of the present invention. It should be pointed out that for ordinary technicians in this field, several modifications and improvements can be made without departing from the concept of the present invention, which all belong to the protection scope of the present invention.

Claims

1. A method for preparing a whitening and anti-acne drug, characterized in that The steps are as follows: S1. Weigh 50g of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa respectively, crush them with a high-speed multifunctional grinder, soak them in 70% mass fraction ethanol overnight, reflux them at above 80°C for 1-6h, repeat twice, filter them with diatomaceous earth until the filtrate is clear and collect the filtrate; combine the two filtrates, distill them under reduced pressure at 60°C in vacuum, concentrate them into an extract, and store them in a refrigerator; S2. Weigh the prescribed amount of solid lipids and soybean lecithin, add them to 5mL of ethanol, dissolve them with ultrasound, weigh a certain amount of Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts respectively, add them to the lipid solution, stir until the extracts are completely dissolved, and heat them in a water bath to 60-90°C for insulation; weigh another 50mL of purified water, heat them in a water bath to 6-15°C for insulation; stir them magnetically at 3000r / min, and quickly add the organic phase containing the extracts into the purified water , disperse to form a white emulsion, maintain a water bath temperature of 60-90°C, continue high-speed stirring for 30 minutes, evaporate the ethanol, add purified water at the same temperature to a total volume of 50mL; put the solution into a high-speed dispersing homogenizer for homogenization at a homogenization speed of 20000r / min for 10 minutes, and after the homogenization, quickly put the emulsion into an ice water bath to obtain solid lipid nanoparticles, which are placed in a 4°C refrigerator for storage and standby; S3, weigh 0.5g of Carbomer 940 and add it to 50mL of purified water, continue stirring until uniformly dispersed; take another 50mL of solid lipid nanoparticle solution and add it to the Carbomer 940 solution, stir and mix evenly; adjust the pH value to about 6.5-7.5 to form a gel solution, that is, obtain solid lipid nanoparticle gel; The solid lipid used in step S2: one or more of glyceryl behenate, glyceryl distearate, glyceryl trimyristate, and glyceryl monostearate; In step S2, the amounts of the Angelica dahurica, Scutellaria baicalensis and Forsythia suspensa extracts are 10-30 mg respectively.

2. The method for preparing a whitening and anti-acne drug according to claim 1, characterized in that: In step S1, Angelica dahurica, Scutellaria baicalensis, Forsythia suspensa, etc. are crushed into 70-300 meshes using a high-speed multifunctional crusher at a rotation speed of 34000 r / min.

3. The method for preparing a whitening and anti-acne drug according to claim 2, characterized in that: In step S1, 70% by mass ethanol is used at a material-liquid ratio of 1:10 for soaking overnight.

4. The method for preparing a whitening and anti-acne drug according to claim 1, characterized in that: In step S3, triethylamine is continuously added during the stirring process to adjust the pH of the solution to between 6.5 and 7.

5.

5. The whitening and anti-acne drug prepared by the preparation method according to any one of claims 1 to 4.

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