Composition for treating atopic dermatitis and application thereof

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to a composition for treating atopic dermatitis and application thereof. The composition comprises the following components: pseudo-ginseng extract, artemether, alpine glycoside and oleanolic acid in a mass ratio of (2-4): (2-4): (1-2): (1-2). The composition disclosed by the invention has anti-inflammatory and anti-pruritus effects and can be used for treating atopic dermatitis, eczema and other diseases. The active components of the composition are all from plant sources, the composition has good safety and atopic dermatitis treatment efficacy, the use dosage is low, the components can achieve a synergistic interaction effect, and the composition shows an excellent effect; the composition can be applied to functional skin care products (face cream, emulsion and essence) and external medicines (applying agents and ointments).

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedicine, and particularly relates to a composition for treating atopic dermatitis and application thereof. Background Art

[0002] Atopic dermatitis is an itchy, relapsing, chronic inflammatory skin disease characterized by chronicity, recurrences, and a high incidence in children. Currently, the first-line treatment for atopic dermatitis is glucocorticoid anti-inflammatory drugs. However, topical glucocorticoids increase the risk of skin atrophy, especially in skin wrinkles, and also pose a risk of affecting growth and development, making them particularly detrimental to pediatric patients. In the past two years, inhibitors targeting inflammatory targets such as calcineurin, IL4, IL13, and JAK have been approved for clinical use in atopic dermatitis, improving the current state of atopic dermatitis treatment. However, due to the high cost of these drugs, some patients have drug tolerance issues, making them unsuitable for long-term use in chronic diseases. Therefore, the development of topical anti-inflammatory and anti-pruritic drugs with minimal toxicity and side effects, strong activity, and low cost is of great value in the treatment of atopic dermatitis.

[0003] Ginsenosides are a class of anti-inflammatory compounds found widely in Panax genus plants (such as Panax ginseng, American ginseng, and Panax notoginseng). Rare and diol-type saponins exhibit potent immunomodulatory activity. Numerous studies have demonstrated that ginsenosides promote skin wound healing. Recent research suggests that saponins in the fibrous roots of Panax notoginseng contain more rare saponins, such as ginsenoside Rh1 and notoginsenoside R2, than in the taproot, potentially demonstrating enhanced anti-inflammatory properties. Artemether, an important antimalarial drug and artemisinin ether derivative, is less polar than artemisinin and exhibits improved skin penetration. Studies have shown that artemether is effective in treating atopic dermatitis, but carries significant risks of neurotoxicity and hematotoxicity. Oleanolic acid is a pentacyclic triterpenoid compound widely found in plant extracts. Oleanolic acid is widely recognized for its anti-inflammatory and anti-cancer effects, but due to its hydrophobic nature, it has low bioavailability, which greatly limits its potential for oral administration. This suggests its potential for topical skin administration. Urinoside, an arbutin derivative, is believed to have intestinal barrier repair functions, as well as hypoglycemic, hypolipidemic, and uric acid-lowering, liver-protective effects. However, its efficacy in treating atopic dermatitis has not yet been demonstrated. Therefore, the use of drug combinations to achieve dose reduction and increased efficacy is an important strategy for the application of these drugs. Summary of the Invention

[0004] The present invention proposes a composition consisting of Panax notoginseng extract, artemether, oleanolic acid, and uronoside. These four components synergistically enhance the anti-inflammatory efficacy of each component, demonstrating potential for the treatment of atopic dermatitis. This composition combines good safety with therapeutic efficacy for atopic dermatitis, and can be used at a low dosage. The components achieve synergistic effects, demonstrating excellent results. A search revealed no reports of this combination in the treatment and research of atopic dermatitis.

[0005] In a first aspect, the present invention provides a composition for treating atopic dermatitis, wherein the weight ratio of each component is Panax notoginseng extract: artemether: uronoside: oleanolic acid = (2~4): (2~4): (1~2): (1~2).

[0006] Preferably, the Panax notoginseng extract is a Panax notoginseng root extract. The Panax notoginseng root extract contains 50%-80% total saponins, including 30%-50% ginsenoside Rb1, 5%-20% ginsenoside Rd, 5%-15% ginsenoside Rg1, 2%-10% notoginsenoside R2(s), and 0.5%-2% ginsenoside Rh1.

[0007] Preferably, according to the weight ratio, the optimal ratio of each component is: Panax notoginseng extract: artemether: uronoside: oleanolic acid = 2:2:1:1.

[0008] In a second aspect, the present invention provides a drug for preventing or treating atopic dermatitis, comprising the above-mentioned composition for treating atopic dermatitis and a pharmaceutically acceptable carrier.

[0009] Preferably, the dosage form of the drug is any one of ointment, cream, external solution, tincture, lotion, powder or lyophilized powder.

[0010] In a third aspect, the present invention provides use of the above-mentioned composition for treating atopic dermatitis in cosmetics.

[0011] In a fourth aspect, the present invention provides a cosmetic comprising the above-mentioned composition for treating atopic dermatitis and an excipient acceptable in cosmetics.

[0012] Preferably, the dosage form of the cosmetic is any one of an emulsion, a cream, a gel or an aqueous solution.

[0013] Preferably, the cosmetic comprises the following components in parts by weight: Cetearyl alcohol, 2 parts; Squalane, 2 parts; Polydimethylsiloxane, 1 part; Glycerin, 6 parts; Sodium hyaluronate, 0.1 part; Lecithin, 0.5 parts; soothing anti-inflammatory agent, 1.2 parts; 0.2 parts of p-hydroxyacetophenone water, 87 parts; Wherein, the soothing anti-inflammatory agent is the above-mentioned composition for treating atopic dermatitis.

[0014] Compared with the prior art, the present invention has at least the following beneficial effects: In the epidermal inflammation model of cells, when the individual components were at a final concentration of 10 μg / ml, Panax notoginseng extract, uronoside, and oleanolic acid were unable to significantly inhibit the expression of three markers IL24, IL33, and CCL26. The inhibitory effect of 10 μg / ml artemether was: IL24 was downregulated to 50% of the model group, IL33 was downregulated to 76% of the model group, and CCL26 was downregulated to 89% of the model group. The composition provided by the present invention can significantly inhibit the expression of atopic dermatitis markers, and at the optimal ratio, IL24 was downregulated to 39% of the model group, IL33 was downregulated to 27% of the model group, and CCL26 was downregulated to 64% of the model group. The inhibitory efficacy was significantly stronger than that of the individual components at the same dose. Therefore, the advantage of the composition of the present invention is that the components have a good synergistic effect, which is far superior to the individual components.

[0015] Animal experiments have shown that the composition of the present invention can significantly alleviate pruritus in mice and improve their skin structure and clinical symptoms, with efficacy comparable to that of dexamethasone. Furthermore, at a similar dose, dexamethasone causes weight loss and spleen damage in mice, exhibiting significant immunotoxicity; whereas the composition effectively improves mouse weight without affecting spleen weight, demonstrating a superior safety profile.

[0016] The composition of the present invention has anti-inflammatory and anti-itching effects and can treat diseases such as atopic dermatitis. The active ingredients in this composition are all plant-derived, combining good safety with therapeutic efficacy for atopic dermatitis. Furthermore, the composition can be used at a low dosage, and the ingredients can achieve synergistic effects, demonstrating excellent results. It can be used in functional skin care products (creams, lotions, serums) and topical medications (scalds, ointments). BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 The effect of different ratios of the composition and a single component on the relative expression of the IL24 gene in the IL4 / IL13-induced epidermal inflammation model of the present invention; Figure 2 The effect of different ratios of the composition and a single component on the relative expression of the IL33 gene in the IL4 / IL13-induced epidermal inflammation model of the present invention; Figure 3The effect of different ratios of the composition and a single component on the relative expression of the CCL26 gene in the IL4 / IL13-induced epidermal inflammation model of the present invention; Figure 4 The improvement effect of the composition of the present invention and dexamethasone on the clinical scores of atopic dermatitis mice (n=6); Figure 5 The improvement effect of the composition of the present invention and dexamethasone on the body weight of mice with atopic dermatitis (n=6); Figure 6 Effects of the composition of the present invention and dexamethasone on the spleen coefficient of atopic dermatitis mice (n=6) Figure 7 The improvement effect of the composition of the present invention and dexamethasone on the pruritus symptoms in atopic dermatitis mice (n=6); Figure 8 The improvement effect of the composition of the present invention and dexamethasone on the skin structure of atopic dermatitis mice (n=6). DETAILED DESCRIPTION

[0018] The technical solution of the present invention will be further described below with reference to the accompanying drawings and through specific embodiments. However, the following examples are merely simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0019] Unless otherwise specified, the raw materials and materials used in the examples of the present invention were purchased through general commercial channels.

[0020] The sources of the raw materials, materials, and instruments involved in the following embodiments or application examples are as follows: Experimental reagents: Artemether, B20493, Shanghai Yuanye Biotechnology Co., Ltd.; Urinary rutin, WKQ-0000889, Sichuan Weikeqi Biotechnology Co., Ltd.; Oleanolic acid, A10055, Shanghai Yuanye Biotechnology Co., Ltd.; DMEM high glucose medium, Gibco, Thermo Fisher Scientific Inc. (USA); Fetal bovine serum, Gibco, Thermo Fisher Scientific Inc. (USA); cytokines IL4 (GP20499) and IL13 (GP20560), GlpBio Technology (USA); Dimethyl sulfoxide (DMSO), 14200201, Xilong Science Co., Ltd.; Cetearyl alcohol (CAS: 67762-27-0), S26921, Shanghai Yuanye Biotechnology Co., Ltd.; Squalane (CAS: 111-01-3), S22248, Shanghai Yuanye Biotechnology Co., Ltd.; Polydimethylsiloxane (CAS: 70131-67-8), T22990, Shanghai Yuanye Biotechnology Co., Ltd.; Lecithin (CAS: 8002-43-5), T90784, Shanghai Yuanye Biotechnology Co., Ltd.; p-Hydroxyacetone (CAS: 99-93-4), A10384, Shanghai Yuanye Biotechnology Co., Ltd.

[0021] Experimental instruments: CO2 incubator (Series 2 Water Jacketed, Thermo Fisher, USA); microcentrifuge (Fresco 21, Thermo Fisher, USA); qPCR instrument (Applied Biosystems QuantStudio 3, Thermo Fisher, USA).

[0022] In the present invention, the Panax notoginseng root extract is homemade, and the preparation method is as follows: The fibrous roots of Panax notoginseng were dried and crushed, and then extracted with 85% ethanol under heating and reflux at a solid-liquid ratio of 1:10 (g / ml). The heating temperature was 60-80 degrees Celsius, and the heating and reflux extraction was repeated for 2 hours. The extraction was repeated twice, and the ethanol was removed by vacuum rotary evaporation at 50-60 degrees Celsius and concentrated to obtain a concentrate. The concentrate was adsorbed on AB-8 macroporous resin and eluted in sections with water at 30% and 50% concentrations for 2-7 BV. The 60% ethanol eluate (5-8 BV) was collected and freeze-dried to obtain a Panax notoginseng root extract. The Panax notoginseng root extract contains 50%-80% total saponins by weight, including 30%-50% ginsenoside Rb1, 5%-20% ginsenoside Rd, and 5%-15% ginsenoside Rg1; 2%-10% notoginsenoside R2(s); and 0.5%-2% ginsenoside Rh1.

[0023] The notoginseng root extract used in the embodiments of the present invention has a saponin component content of 60.4% by weight; it mainly contains ginsenoside Rb1 38.2%, ginsenoside Rd 6.2%, ginsenoside Rg1 9.1%, notoginsenoside R2 5.1%; and ginsenoside Rh1 0.72%.

[0024] Statistical analysis All data presented in this paper represent at least three independent replicates, and data are presented as mean values. All data were plotted and statistically analyzed using GraphPad Prism software. Statistical analysis was performed using one-way or multi-way analysis of variance or independent sample t-test. Data were considered significantly different when p ≤ 0.05.

[0025] <Example 1> 2.5 mg of Panax notoginseng root extract, 2.5 mg of artemether, 2.5 mg of uronoside, and 2.5 mg of oleanolic acid were mixed and dissolved in 1 ml of DMSO to prepare the mother solution of Composition 1.

[0026] The mother liquor was diluted 1000 times with culture medium to obtain a working solution of composition 1 for testing, wherein the final concentration of composition 1 contained therein was 10 μg / ml, and the mass ratio of each component was 1:1:1:1; that is, the final concentrations were 2.5 μg / ml of Panax notoginseng root extract, 2.5 μg / ml of artemether, 2.5 μg / ml of uronoside, and 2.5 μg / ml of oleanolic acid.

[0027] In the in vitro epidermal inflammation model, Composition 1 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 61% (see Figure 1 ), IL33 was reduced to 24% (see Figure 2 ), CCL26 was lowered to 72% (see Figure 3 ), the above three genes were significantly downregulated compared with Example 13.

[0028] <Example 2> Prepare a mother solution and working solution of Composition 2. The working solution of Composition 2 was used for testing, containing a final concentration of 10 μg / ml of Composition 2. The steps were the same as in Example 1, except that the mass ratio of the components was 2:1:1:1; that is, the final concentrations of the components were 4 μg / ml of Panax notoginseng root extract, 2 μg / ml of artemether, 2 μg / ml of uronoside, and 2 μg / ml of oleanolic acid.

[0029] In the in vitro epidermal inflammation model, Composition 2 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 63% (see Figure 1 ), IL33 was reduced to 17% (see Figure 2 ), CCL26 was lowered to 73% (see Figure 3 ), the above three genes were significantly downregulated compared with Example 13.

[0030] <Example 3> Prepare a mother solution and working solution of Composition 3. The working solution of Composition 3 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 1:2:1:1; that is, the final concentrations were 2 μg / ml of Panax notoginseng root extract, 4 μg / ml of artemether, 2 μg / ml of uronoside, and 2 μg / ml of oleanolic acid.

[0031] In the in vitro epidermal inflammation model, the working solution of composition 3 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 56% (see Figure 1 ), IL33 was reduced to 45% (see Figure 2 ), CCL26 was lowered to 59% (see Figure 3 ), the above three genes were significantly downregulated compared with Example 13.

[0032] <Example 4> Prepare a mother solution and working solution of Composition 4. The working solution of Composition 4 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 1:1:2:1; that is, the final concentrations were 2 μg / ml of Panax notoginseng root extract, 2 μg / ml of artemether, 4 μg / ml of uronoside, and 2 μg / ml of oleanolic acid.

[0033] In the in vitro epidermal inflammation model, the working solution of composition 4 can significantly inhibit the expression of inflammatory genes IL24 and IL33 but not CCL26 (see Figure 3 ), compared with the model group, IL24 was downregulated to 76% (see Figure 1 ), IL33 down to 75% (see Figure 2 ), the above two genes had no significant difference compared with Example 13.

[0034] <Example 5> Prepare a mother solution and working solution of Composition 5. The working solution of Composition 5 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 1:1:1:2; that is, the final concentrations were 2 μg / ml of Panax notoginseng root extract, 2 μg / ml of artemether, 2 μg / ml of uronoside, and 4 μg / ml of oleanolic acid.

[0035] In the in vitro epidermal inflammation model, the working solution of composition 5 significantly inhibited the expression of the inflammatory gene IL33 but not IL24 (see Figure 1 ) and CCL26 (see Figure 3 ), compared with the model group, IL33 was downregulated to 9%, and IL33 was significantly downregulated compared with Example 13 (see Figure 2 ).

[0036] <Example 6> Prepare a mother solution and working solution of Composition 6. The working solution of Composition 6 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 2:2:1:1; that is, the final concentrations were 3.33 μg / ml of Panax notoginseng root extract, 3.33 μg / ml of artemether, 1.67 μg / ml of uronoside, and 1.67 μg / ml of oleanolic acid.

[0037] In the in vitro epidermal inflammation model, the working solution of composition 6 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 39% (see Figure 1 ), IL33 was reduced to 27% (see Figure 2 ), CCL26 was lowered to 64% (see Figure 3 ), the above three genes were significantly down-regulated compared with Example 13.

[0038] <Example 7> Prepare a mother solution and working solution of Composition 7. The working solution of Composition 7 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 3:1:1:1; that is, the final concentrations were 5 μg / ml of Panax notoginseng root extract, 1.67 μg / ml of artemether, 1.67 μg / ml of uronoside, and 1.67 μg / ml of oleanolic acid.

[0039] In an in vitro epidermal inflammation model, the working solution of composition 7 significantly inhibited the expression of the inflammatory gene IL33 but not IL24 (see Figure 1 ) and CCL26 (see Figure 3 ), compared with the model group, IL33 was downregulated to 21%, and significantly downregulated compared with Example 13 (see Figure 2 ).

[0040] <Example 8> Prepare a mother solution and working solution of Composition 8. The working solution of Composition 8 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 1:3:1:1; that is, the final concentrations were 1.67 μg / ml of Panax notoginseng root extract, 5 μg / ml of artemether, 1.67 μg / ml of uronoside, and 1.67 μg / ml of oleanolic acid.

[0041] In the in vitro epidermal inflammation model, the working solution of composition 8 significantly inhibited the expression of inflammatory genes IL24 and CCL26 but not IL33 (see Figure 2 ), compared with the model group, IL24 was downregulated to 46% (see Figure 1 ), CCL26 was lowered to 58% (see Figure 3 ), the above two genes were significantly downregulated compared with Example 13.

[0042] <Example 9> Prepare a mother solution and working solution of Composition 9. The working solution of Composition 9 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 3:3:1:1; that is, the final concentrations were 3.25 μg / ml of Panax notoginseng root extract, 3.25 μg / ml of artemether, 1.08 μg / ml of uronoside, and 1.08 μg / ml of oleanolic acid.

[0043] In the in vitro epidermal inflammation model, the working solution of Composition 9 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 35% (see Figure 1 ), IL33 was reduced to 47% (see Figure 2 ), CCL26 was lowered to 65% (see Figure 3 ), the above three genes were significantly down-regulated compared with Example 13.

[0044] <Example 10> Prepare a mother solution and working solution of Composition 10. The working solution of Composition 10 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 4:4:1:1; that is, the final concentrations were 4 μg / ml of Panax notoginseng root extract, 4 μg / ml of artemether, 1 μg / ml of uronoside, and 1 μg / ml of oleanolic acid.

[0045] In the in vitro epidermal inflammation model, the working solution of Composition 10 significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26. Compared with the model group, IL24 was downregulated to 60% (see Figure 1 ), IL33 was reduced to 46% (see Figure 2 ), CCL26 was lowered to 72% (see Figure 3 ), the above three genes were significantly down-regulated compared with Example 13.

[0046] <Example 11> Prepare a mother solution and working solution of Composition 11. The working solution of Composition 11 was used for testing, with a final concentration of 30 μg / ml. The steps were the same as in Example 1, except that the mass ratio of the components was 5:5:1:1; that is, the final concentrations were 4.17 μg / ml of Panax notoginseng root extract, 4.17 μg / ml of artemether, 0.83 μg / ml of uronoside, and 0.83 μg / ml of oleanolic acid.

[0047] In an in vitro epidermal inflammation model, the working solution of Composition 11 significantly inhibited the expression of inflammatory genes IL24 and IL33 but not CCL26 (see Figure 3 ), compared with the model group, IL24 was downregulated to 54% (see Figure 1 ), IL33 was reduced to 63% (see Figure 2), among the above two genes, IL24 was significantly downregulated compared with Example 13, and the expression level of IL33 was not significantly different from that of Example 13.

[0048] <Example 12> 100 mg of Panax notoginseng root extract was dissolved in 1 ml of DMSO to prepare the stock solution of Panax notoginseng root extract; the stock solution was diluted 1000, 2000, and 10000 times with culture medium to obtain the working solution of Panax notoginseng root extract for testing, and the final concentrations of Panax notoginseng root extract contained therein were 10 μg / ml, 50 μg / ml, and 100 μg / ml, respectively.

[0049] In the in vitro epidermal inflammation model, 10 μg / ml Panax notoginseng root extract could not significantly inhibit the expression of inflammatory genes, while 50 μg / ml Panax notoginseng root extract significantly inhibited the expression of inflammatory gene IL33, with IL33 downregulated to 37% of the model group. 100 μg / ml Panax notoginseng root extract significantly inhibited the expression of inflammatory genes IL24 and IL33, with IL24 downregulated to 41% and IL33 downregulated to 17% of the model group (see Figure 4 ).

[0050] <Example 13> 10 mg of artemether was dissolved in 1 ml of DMSO to prepare the artemether stock solution; the stock solution was diluted 1000-fold with culture medium to obtain the artemether working solution for testing, which contained an artemether final concentration of 10 μg / ml.

[0051] In the in vitro epidermal inflammation model, 10 μg / ml artemether significantly inhibited the expression of inflammatory genes IL24, IL33, and CCL26, with IL24 downregulated to 50% of the model group (see Figure 1 ), IL33 was downregulated to 76% of the model group (see Figure 2 ), CCL26 was downregulated to 89% of the model group (see Figure 3 ).

[0052] <Example 14> Prepare a stock solution of urogynoside by dissolving 450 mg of urogynoside in 1 ml of DMSO. Dilute the stock solution 1000-fold with culture medium to obtain urogynoside working solution 1 for testing, with a final urogynoside concentration of 450 μg / ml. Dilute the stock solution 45,000-fold with culture medium to obtain urogynoside working solution 2 for testing, with a final urogynoside concentration of 10 μg / ml.

[0053] In the in vitro epidermal inflammation model, 10 μg / ml urocandin could not significantly inhibit the expression of inflammatory genes IL24, IL33, and CCL26, while 450 μg / ml urocandin significantly inhibited the expression of inflammatory genes IL33 and CCL26, but not IL24, with IL33 downregulated to 40% of the model group (see Figure 2 ), CCL26 was downregulated to 79% of the model group (see Figure 3 ).

[0054] <Example 15> 10 mg of oleanolic acid was dissolved in 1 ml of DMSO to prepare the oleanolic acid stock solution. The stock solution was diluted 1000 times with culture medium to obtain the oleanolic acid working solution for testing, which contained a final oleanolic acid concentration of 10 μg / ml.

[0055] In the in vitro epidermal inflammation model, 10 μg / ml oleanolic acid could not significantly inhibit the expression of inflammatory genes IL24, IL33, and CCL26 (see Figure 1-3 ).

[0056] <Example 16> Prepare a mother solution and working solution of Composition 16. The working solution of Composition 16 was used for testing, with a final concentration of 10 μg / ml. The steps were the same as in Example 6, except that the mass ratio of the components was 2:2:1 for Panax notoginseng root extract: artemether: oleanolic acid; the final concentrations were 4 μg / ml for Panax notoginseng root extract, 4 μg / ml for artemether, and 2 μg / ml for oleanolic acid.

[0057] In an in vitro epidermal inflammation model, the working solution of composition 16 significantly inhibited the inflammatory gene IL24 but not IL33 and CCL26 (see Figure 2-3 ), compared with the model group, IL24 was downregulated to 29% (see Figure 1 ), which is significantly lower than that of Example 13.

[0058] <Test Example 1> The process of establishing the in vitro epidermal inflammation model is as follows: Human keratinocyte HaCaT cells were purchased from ATCC. Cells were cultured in DMEM (DMEM-high glucose complete medium) supplemented with 10% FBS and 1% double-antibody at 37°C in a 5% CO2 atmosphere. Experiments were performed using cells in the logarithmic growth phase, and drug stimulation was performed when the cell confluence reached approximately 70%.

[0059] HaCaT cells were co-incubated with 50 ng / ml of IL4 and IL13 for 24 hours to serve as an epidermal inflammation model. For the drug treatment group, a combination of Panax notoginseng root extract, uronoside, artemether, oleanolic acid, and IL4 / IL13 were administered simultaneously. After 24 hours of incubation, qPCR was performed to detect the inflammatory genes IL24, IL33, and CCL26. Results are shown in the table. Figures 1 to 5 And Tables 1~3.

[0060] above Figures 1-3 In the table, # indicates that Example 12, Example 13, or Example 14 showed a significant difference compared with the model group. # indicates p ≤ 0.05; ## indicates p ≤ 0.01; ### indicates p ≤ 0.001. * indicates that Examples 1-11 showed a significant difference compared with Example 13. * indicates p ≤ 0.05; ** indicates p ≤ 0.01; *** indicates p ≤ 0.001.

[0061] Table 1 Regulatory effects of different ratios of combinations and single components on epidermal inflammatory gene expression

[0062] <Test Example 2> SPF-certified male Balb / c mice, 7-8 weeks old, weighing 20-25 g, were used as experimental animals. They were obtained from Beijing Weitonglihua Laboratory Animal Technology Co., Ltd. (animal qualification certificate number: 110011241110592062). Animals were housed in individually ventilated cages, with no more than five animals per cage. The ambient temperature was maintained at 20-26°C, humidity at 35-75%, with a 12-hour light-on / 12-hour dark cycle, and free access to water and food. This experimental protocol and any modifications were reviewed and approved by the Laboratory Animal Welfare and Ethics Committee of Hefei Zhongke Puruisheng Biomedical Technology Co., Ltd. (IACUC-20241101).

[0063] This experiment set up a solvent group, a model group, a positive drug group, a low-dose group of the composition, and a high-dose group of the composition, with 6 mice randomly set up in each group. Acetone and olive oil were mixed in a volume ratio of 4:1 as a blank solvent, and the composition in the above Example 6 was selected as the test drug, that is, the mass ratio of the components of the composition was Panax notoginseng root extract: artemisinin: oleanolic acid: uroguanidine = 2:2:1:1, and two doses of 0.3% and 0.6% (W / V, g / ml) were set as the low-dose group and the high-dose group of the composition respectively. Dexamethasone was selected as the positive drug, and the dose was set to 0.04% (W / V, g / ml) as the positive drug group. The mouse atopic dermatitis model stimulated by MC903 was used, and blank solvent was used for administration as the model group. Non-modeling mice were set up and given blank solvent as the solvent group. The mouse MC903 atopic dermatitis model is specifically as follows: The back of the mouse was depilated, and the sensitizer MC903, or MC903 and the drug (Example 6) were applied to the depilatory area daily. The weight of the mouse was tested daily, the scratching behavior of the mouse was observed, and the skin condition of the test area was observed and the clinical score was recorded. After 14 days, the back of the mouse was photographed, and the mouse was sacrificed and the skin of the test area was collected. The mouse skin was stained with HE to observe the morphology of the epidermis and dermis. The results are shown in Figures 6-8 And Tables 4 to 6. Among them, Figure 7 In the figure, A is the total number of scratching times of the mouse within a fixed time (30 minutes); B is the total duration (seconds) of the mouse scratching behavior within a fixed time (30 minutes). Figure 8 In the figure, A is the HE staining image of mouse skin tissue (20X); B is the statistics of the epidermal thickness (micrometers) of mouse skin.

[0064] Table 2 Effects of the combination and dexamethasone on the clinical scores of atopic dermatitis mice over time

[0065] Table 3 Effects of the combination and dexamethasone on the body weight of atopic dermatitis mice over time

[0066] Table 4 Effects of the combination and dexamethasone on the spleen, epidermis and pruritus symptoms in mice with atopic dermatitis

[0067] The results showed that on the 14th day of treatment, the 0.6% dose of the composition significantly reduced the clinical score of atopic dermatitis in mice, and the results were close to those of the dexamethasone group ( Figure 4 ).

[0068] On the 14th day of treatment, the body weight of mice in the 0.6% dose of the composition group was significantly higher than that of mice in the model group and the dexamethasone group, indicating that this dose of the composition did not cause obvious toxic side effects in mice and was beneficial to the recovery of atopic dermatitis mice ( Figure 5 ).

[0069] On the 14th day of treatment, the spleen index of mice in the 0.6% dose group of the composition was not significantly different from that in the solvent group and the model group, while the spleen index of mice in the dexamethasone group decreased significantly, indicating that the composition at this dose did not produce similar immunotoxicity as dexamethasone ( Figure 6 ).

[0070] On the 14th day of treatment, the number of scratching times and the duration of scratching in the 0.3% and 0.6% dose combination groups and the dexamethasone group were significantly less than those in the model group, indicating that all three drugs can significantly relieve the itching symptoms of mice ( Figure 7 ).

[0071] On the 14th day of treatment, the epidermal thickness of mice in the 0.3% and 0.6% dose combination groups and the dexamethasone group was significantly thinner than that in the model group, and the epidermal structure was tighter, indicating that all three drugs can significantly improve the skin structure of mice and alleviate the epidermal thickening caused by atopic dermatitis ( Figure 8 ).

[0072] <Application Example 1> This application example provides a moisturizing cream with soothing and antipruritic effects. The formula of the moisturizing cream is shown in Table 5 below: Table 5. Formula of moisturizing cream

[0073] Its preparation method is: (1) Add the raw materials of phase A to the oil phase pot, stir and heat to 85℃; (2) Add the raw materials of phase B and phase E to the water phase pot, mix well, and heat to 85°C with stirring; (3) The materials in the water phase pot and the oil phase pot are pumped into the emulsification pot under negative pressure, homogenized for 10 minutes, cooled to 75°C, added with the raw materials of phase D, continued to stir, cooled to 40°C, added with the raw materials of phase C, and mixed evenly to obtain the product.

[0074] The soothing anti-inflammatory agent in Table 5 is prepared by directly weighing Panax notoginseng extract, artemether, uronoside and oleanolic acid according to the proportions of Example 6.

[0075] The applicant declares that the above is only a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention fall within the scope of protection and disclosure of the present invention.

Claims

1. A composition for treating atopic dermatitis, characterized in that: The mass ratio of each component is Panax notoginseng extract:artemisinin:uranin:oleanolic acid = (2~4): (2~4): (1~2): (1~2).

2. The composition for treating atopic dermatitis according to claim 1, wherein The Panax notoginseng extract is derived from Panax notoginseng root fibers.

3. The composition for treating atopic dermatitis according to claim 2, wherein According to weight ratio, the total saponin content of the Panax notoginseng root extract is 50%-80%, including ginsenoside Rb1 30%-50%, ginsenoside Rd 5%-20%, ginsenoside Rg1 5%-15%; notoginseng saponin R2(s) 2%-10%; and ginsenoside Rh1 0.5%-2%.

4. The composition for treating atopic dermatitis according to claim 1, wherein According to the weight ratio, Panax notoginseng extract: artemisinin: urocanthoside: oleanolic acid = 2:2:1:

1.

5. A drug for preventing or treating atopic dermatitis, characterized in that: A composition for treating atopic dermatitis comprising the composition of claims 1 to 4 and a pharmaceutically acceptable carrier.

6. Use of the composition for treating atopic dermatitis according to claims 1 to 4 in cosmetics.

7. A cosmetic, characterized in that: The composition for treating atopic dermatitis comprises the composition according to claims 1 to 4 and an auxiliary material acceptable in cosmetics.

8. A cosmetic according to claim 7, characterized in that: The cosmetic comprises the following components in parts by weight: Cetearyl alcohol, 2 parts; Squalane, 2 parts; Polydimethylsiloxane, 1 part; Glycerin, 6 parts; Sodium hyaluronate, 0.1 part; Lecithin, 0.5 parts; soothing anti-inflammatory agent, 1.2 parts; 0.2 parts of p-hydroxyacetophenone water, 87 parts; Wherein, the soothing anti-inflammatory agent is the composition for treating atopic dermatitis according to claims 1 to 4.

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