A compound cosmetic raw material combination with synergistic anti-inflammatory effect and application thereof

Abstract

The present application provides a composition of panax notoginseng saponins and polygonum multiflorum polysaccharide compounded in a specific ratio, which has synergistic anti-inflammatory effect, better anti-inflammatory activity than the two components used alone, and can significantly inhibit the release of inflammatory factors by macrophages in inflammatory response. The composition involved in the present application can be used as a cosmetic raw material for preparing skin care products with soothing, anti-inflammatory or repairing effects.

Description

Technical Field

[0001] This invention relates to the field of cosmetic raw materials, specifically to the preparation of a composition having a synergistic anti-inflammatory effect by combining total saponins of Panax notoginseng and polysaccharides of Portulaca oleracea, and to the preparation of skin care products with soothing anti-inflammatory and skin barrier repair properties using this composition as a raw material. Background Technology

[0002] Using active ingredients derived from natural plants as cosmetic raw materials is a current hot topic in cosmetic research and development. In my country, approximately 82.64% of cosmetics contain plant extracts, and 80.17% of these cosmetics use a combination of two or more plant extracts. Combining several ingredients with similar effects may produce synergistic effects that enhance each other's activity, or antagonistic effects that weaken each other's activity. However, many formulators, when designing cosmetics, simply use experience to combine each plant extract in a certain proportion, neglecting the potential synergistic or antagonistic interactions between ingredients with similar activities.

[0003] Purslane polysaccharides and Panax notoginseng saponins are two commonly used plant-derived cosmetic ingredients with similar soothing and anti-inflammatory effects. Purslane polysaccharides exert their anti-inflammatory effects by reducing the expression levels of serum pro-inflammatory factors IL-6, IL-17, and TNF-α, and increasing the expression level of the anti-inflammatory factor IL-10. Panax notoginseng saponins can inhibit the phosphorylation of NF-κB p65, prevent p65 translocation into the nucleus, and reduce the expression of the iNOS-NO and NF-κB signaling pathways, thereby exerting their anti-inflammatory effects. Both purslane polysaccharides and Panax notoginseng saponins can reduce the expression of inflammatory factors and exert anti-inflammatory effects; they may have a synergistic effect, but there are currently no research reports on the synergistic effect of their combined use.

[0004] This invention provides a composition of total saponins from Panax notoginseng and polysaccharides from Portulaca oleracea in a specific ratio. This composition exhibits a synergistic anti-inflammatory effect, demonstrating better anti-inflammatory activity than either component used alone, and significantly inhibiting the release of inflammatory factors from macrophages during inflammatory responses. The composition involved in this invention can be used as a cosmetic ingredient to prepare skincare products with soothing, anti-inflammatory, or repairing effects, providing a new approach for designing compound formulations of various cosmetic ingredients with similar activities. Summary of the Invention

[0005] This invention provides a method for preparing a composition by compounding total saponins of Panax notoginseng and purslane polysaccharides in a certain proportion. This compound composition exhibits better anti-inflammatory activity than either component used alone, and can significantly inhibit the release of inflammatory factors by macrophages during inflammatory responses. This compound composition can be used as a cosmetic ingredient in the preparation of cosmetics with soothing, anti-inflammatory, or repairing effects.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] A soothing and anti-inflammatory composition, characterized in that it comprises total saponins of Panax notoginseng and purslane polysaccharides, wherein the weight ratio of total saponins of Panax notoginseng to purslane polysaccharides is 20%-40%: 60%-80%.

[0008] The total saponins of Panax notoginseng contain ginsenoside Rg1, ginsenoside Rb1, and notoginsenoside R1.

[0009] The purslane polysaccharide contains neutral polysaccharides, acidic polysaccharides, and pectin polysaccharides.

[0010] A soothing and anti-inflammatory composition, characterized in that it comprises a pre-mixed solution of total saponins from Panax notoginseng and a pre-mixed solution of purslane polysaccharides, wherein the weight ratio of the pre-mixed solution of total saponins from Panax notoginseng to the pre-mixed solution of purslane polysaccharides is 25%-35%: 65%-75%.

[0011] The pre-prepared solution of total saponins from Panax notoginseng contains 70%-90% propylene glycol, 5%-20% PEG-40 hydrogenated castor oil, and ginsenoside Rg1.

[0012] A mixed solution of 2%-5%, ginsenoside Rb11%-4%, and notoginsenoside R10.1%-1%, wherein the sum of each raw material is 100%.

[0013] The pre-mixed solution of Panax notoginseng saponins is a mixed solution containing 78.5% propylene glycol, 20% PEG-40 hydrogenated castor oil, 13%-4% ginsenoside Rg, 12%-3% ginsenoside Rb, and 10.5% Panax notoginseng saponins, wherein the sum of each raw material is 100%.

[0014] The purslane polysaccharide premix is ​​a mixed solution containing 60%-80% water, 15%-39% 1,3-propanediol, and 0.1%-5% neutral polysaccharides, acidic polysaccharides, and pectin polysaccharides, wherein the sum of all raw materials is 100%.

[0015] The purslane polysaccharide premix is ​​a mixed solution containing 65-70% water, 25-35% 1,3-propanediol, and 0.2%-0.4% neutral polysaccharides, acidic polysaccharides, and pectin polysaccharides, wherein the sum of all raw materials is 100%.

[0016] Furthermore, the following solutions are provided:

[0017] I. Preparation of a soothing and anti-inflammatory compound composition by compounding anti-inflammatory active ingredients

[0018] Table 1. Composition of the compound composition

[0019]

[0020] Furthermore, the pre-prepared solution of Panax notoginseng total saponins is a mixed solution containing propylene glycol (78.5%), PEG-40 hydrogenated castor oil (20%), ginsenoside Rg1 (3%-4%), ginsenoside Rb1 (2%-3%), and Panax notoginseng saponin R1 (0.5%).

[0021] Furthermore, the purslane polysaccharide premix is ​​a mixed solution containing water (65-70%), 1,3-propanediol (25-35%), and neutral polysaccharides, acidic polysaccharides, and pectin polysaccharides (0.2%-0.4%).

[0022] The preparation method of the compound composition includes the following steps:

[0023] (1) Purchase Panax notoginseng total saponins and Portulaca polysaccharide raw materials.

[0024] (2) Weigh out the total saponins of Panax notoginseng and set aside; heat propylene glycol and set aside; heat PEG-40 hydrogenated castor oil and set aside. Add propylene glycol to the hydrogenated castor oil, stir, then add the total saponins of Panax notoginseng and cool to room temperature. The pre-prepared solution of Panax notoginseng saponins is then obtained.

[0025] (3) Weigh out the purslane polysaccharide and set aside. Mix it evenly with 1,3-propanediol, add it to a mixing bowl, add deionized water, and stir thoroughly until completely dissolved. After complete dissolution, it will be a transparent and homogeneous liquid. After complete dissolution, filter out the material to obtain the purslane polysaccharide pre-prepared solution;

[0026] Furthermore, this includes the following steps:

[0027] (1) The total saponins of Panax notoginseng and the polysaccharides of Portulaca oleracea were obtained by purchasing from the market.

[0028] (2) Weigh 5-10g of total Panax notoginseng saponins raw material and set aside; add 65-70mL of propylene glycol to a beaker and heat and stir to 80℃, set aside; add 18-23mL of hydrogenated castor oil to another beaker and heat to dissolve, set aside. Add propylene glycol to the hydrogenated castor oil and stir for 5 minutes, then add the total Panax notoginseng saponins raw material and cool to room temperature. The Panax notoginseng saponin pre-prepared solution is obtained.

[0029] (3) Weigh 0.2-0.4 g of purslane polysaccharide, mix it evenly with 30 ml of 1,3-propanediol, add it to a mixing bowl, add 70 ml of deionized water, and stir thoroughly to dissolve completely (stirring speed 60 r / min, stirring time 30 min). After complete dissolution, it is a transparent and homogeneous liquid. After complete dissolution, filter the material through a 200-mesh filter cloth to obtain the purslane polysaccharide pre-prepared solution.

[0030] (4) The pre-mixed solutions (2) and (3) were subjected to CCK-8 cytotoxicity testing to screen out safe concentration ranges, and the anti-inflammatory efficacy of 16 compound groups was verified. The experiment involved pairwise compounding of pre-mixed solutions (2) at four concentrations (1, 5, 25, 125 μg / mL) and pre-mixed solutions (3) at four concentrations (2, 10, 50, 250 μg / mL). The anti-inflammatory efficacy was assessed by evaluating the inhibition rate of these 16 compound combinations on the release of the inflammatory factor TNF-α from RAW264.7 macrophages induced by lipopolysaccharide. The SynergyScore system was used to quantitatively evaluate the interaction between the two components in the compounding at different concentrations. It was found that when the (2) premixed solution at a concentration of 125 μg / mL was combined with the (3) premixed solution at a concentration of 50 μg / mL or 250 μg / mL, both showed significant inhibitory effects on TNF-α, with inhibition rates of 45.18% and 61.42%, respectively. Their inhibitory effects were not significantly different from those of 100 μg / mL dexamethasone (inhibition rate of 53.17%) (one-way ANOVA, P>0.05); the SynergyScore values ​​were 15.58 and 26.44, respectively. This demonstrates that the combination of the (2) premixed solution at 125 μg / mL with the (3) premixed solution at 50 μg / mL or 250 μg / mL exhibits a synergistic anti-inflammatory effect. The remaining 14 combinations with lower doses only showed additive anti-inflammatory effects and did not exhibit a synergistic effect.

[0031] (5) Based on the experimental conclusion of (4), take 4.5-5.5 kg of (2) premixed solution and 1.5-5.5 kg of (3) premixed solution, mix them evenly, and obtain a compound composition.

[0032] The characteristics of the compound composition involved in this invention are as follows:

[0033] Synergistic anti-inflammatory effects of compound formulations. Natural anti-inflammatory ingredients extracted from plants can exert a good synergistic therapeutic effect on inflammation through combined action on different targets and multiple pathways, achieving the same or even better therapeutic effect with lower drug dosages. The combination of Panax notoginseng and Portulaca oleracea can regulate inflammatory responses through multiple pathways, including leukocyte migration, oxidative stress response, and transcription factor activity. Portulaca oleracea polysaccharides may play a leading role in the release of TNF-α inflammatory factors; while total saponins of Panax notoginseng play a leading role in inhibiting the NF-κBp65 target. When combined, the total saponins of Panax notoginseng and the polysaccharides of Portulaca oleracea do not exert a synergistic effect by jointly intervening in the same target, but rather focus on regulating different targets, affecting different molecular networks, and complementarily regulating the inflammatory response to exert a network synergistic effect. This invention involves pairwise compounding of different concentrations of Panax notoginseng saponin pre-mixed solutions (1, 5, 25, 125 μg / mL) and Portulaca oleracea polysaccharide pre-mixed solutions (2, 10, 50, 250 μg / mL). The anti-inflammatory efficacy was assessed by evaluating the inhibitory rate of these 16 compound combinations on the release of the inflammatory cytokine TNF-α from RAW264.7 macrophages induced by lipopolysaccharide. The Synergy Score system was used to quantitatively evaluate the interaction between the two components in the compounding at different concentrations. It was found that when the pre-prepared solution of Panax notoginseng saponins at a concentration of 125 μg / mL was combined with the pre-prepared solution of Portulaca oleracea polysaccharides at a concentration of 50 μg / mL or 250 μg / mL, both showed significant inhibitory effects on TNF-α, with inhibition rates of 45.18% and 61.42%, respectively. Their inhibitory effects were not significantly different from those of 100 μg / mL dexamethasone (inhibition rate of 53.17%) (P>0.05); the SynergyScore values ​​were 15.58 and 26.44, respectively. This demonstrates that the combined use of the 125 μg / mL pre-prepared solution of total Panax notoginseng saponins and the 50 μg / mL or 250 μg / mL pre-prepared solution of Portulaca oleracea polysaccharides exhibits a synergistic anti-inflammatory effect. The remaining 14 lower-dose combinations only showed additive anti-inflammatory effects and did not exhibit a synergistic effect.

[0034] II. Using compound compositions as active ingredients to prepare moisturizing water with soothing and repairing effects.

[0035] Furthermore, a moisturizing water with soothing and repairing effects is characterized by comprising the aforementioned compound composition.

[0036] Add 1-10% of the compound composition to an aqueous solvent to prepare a moisturizing water.

[0037] The formulation information of other ingredients in the moisturizing water involved in this invention is shown in Table 2.

[0038] Table 2. Information on Moisturizing Water Components

[0039]

[0040] The PHL is a mixed solution of 65% 1,3-propanediol, 30% 1,2-hexanediol, and 5% octanoyl hydroxamic acid.

[0041] The preparation method of the compound composition includes the following steps:

[0042] Furthermore, a moisturizing water is provided, containing 0.5-5 kg ​​of a compound composition, 40-50 kg of water, 0.01-0.025 kg of dipotassium glycyrrhizate, 1-1.5 kg of 4-tert-butylcyclohexanol, 0.75-1 kg of trehalose, 0.015-0.035 kg of tocopherol, 0.01-0.025 kg of hyaluronic acid, 0.04-0.06 kg of acetylglucosamine, 0.045-0.1 kg of ethylhexylglycerin, 0.25-0.4 kg of capryloyl hydroxamic acid, 0.25-0.5 kg of anise extract, 0.015-0.025 kg of glutathione, 0.03-0.05 kg of PE9010, 0.25-0.5 kg of PHL, and 0.005-0.015 kg of fragrance.

[0043] The preparation method of the aforementioned moisturizing water includes the following steps:

[0044] (1) After adding the water according to the formula to the emulsifying pot, add dipotassium glycyrrhizate and 4-tert-butylcyclohexanol to the emulsifying pot and stir until completely dissolved. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0045] (2) Disperse the trehalose, tocopherol, hyaluronic acid, acetylglucosamine and sea fennel extract according to the formula, and then add them to the emulsifying pot described in (1), and stir until they are evenly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0046] (3) Add the compound composition prepared in Example 1, ethylhexylglycerin, capryloyl hydroxamic acid, GE9010, and PHL to the emulsifying pot described in (2) and stir until uniformly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0047] (4) Add the solution obtained from the fragrance to the emulsification pot described in (3), stir and mix for 30 minutes to obtain water. The temperature during this process is room temperature, the stirring speed is 20 rpm, and the vacuum degree is -0.06 MPa.

[0048] (5) The water obtained in (4) is filtered through a 200-mesh filter cloth and discharged to obtain moisturizing water.

[0049] (6) Sensory, physicochemical and microbiological indicators of the moisturizing water prepared in (5) were tested. The product is non-greasy, non-sticky, comfortable to use, and free of foreign matter; it conforms to a specific fragrance; pH value is 5.5-6.5; total bacterial count is ≤500 CFU / g; mold and yeast count is ≤50 CFU / g.

[0050] (7) Use a semi-automatic filling machine to fill the qualified moisturizing water from (6) into the tube, and seal the tube with a sealing machine to obtain soothing and repairing moisturizing water.

[0051] The characteristics of the moisturizing water involved in this invention are as follows:

[0052] 1. Synergistic anti-inflammatory effect of moisturizing water: Panax notoginseng total saponins and purslane polysaccharides affect different molecular networks by intervening in and regulating different target sites, thereby complementarily regulating the inflammatory response and exerting a network synergistic effect.

[0053] 2. Repairing the skin barrier: Tocopherol and acetyl glucosamine repair the skin barrier, promote cell regeneration, have antioxidant effects, protect skin blood vessels, stabilize skin cell membranes, and reduce the expression of inflammatory factors, thereby relieving symptoms such as skin redness and dryness.

[0054] 3. Relieves pain and itching: Sea fennel extract acts on sensory nerve receptors in the skin, relieving pain and itching. It can soothe the "sensory sensation," promote the release of β-endorphin in the skin, and inhibit the release of the neuropeptide CGRP, quickly relieving stinging / itching caused by inflammation.

[0055] III. A cream suitable for people with sensitive skin prepared by using a compound composition as an active ingredient. Further, a cream with soothing and repairing effects is characterized by containing the aforementioned compound composition.

[0056] Furthermore, 1-10% of the compound composition is added to the emulsion solvent to prepare a cream.

[0057] The formulation information of other ingredients in the cream involved in this invention is shown in Table 3.

[0058] Table 3. Cream Ingredient Information

[0059]

[0060] Furthermore, the water is deionized water.

[0061] Furthermore, the butanediol is 1,2-butanediol.

[0062] Furthermore, the sodium hyaluronate is sodium hyaluronate.

[0063] Furthermore, the betaine is betaine.

[0064] Furthermore, the dipotassium glycyrrhizate is dipotassium glycyrrhizate.

[0065] Furthermore, the cetearyl glucoside is cetearyl glucoside.

[0066] Furthermore, the sorbitan oleivate is sorbitan oleivate.

[0067] Furthermore, the caprylic / capric triglyceride is caprylic / capric succinate triglyceride.

[0068] Furthermore, the polydimethylsiloxane is PEG-17 dimethylsiloxane.

[0069] Furthermore, the shea butter is a product of the avocado tree (BUTYROSPERMUM PARKII).

[0070] Furthermore, the jojoba oil is jojoba (Simondia chininsis) seed oil.

[0071] Furthermore, the plant squalane is squalane.

[0072] Furthermore, the prickly pear fruit oil is PRINSEPIAUTILIS oil.

[0073] Furthermore, Besterol-E is a phytosterol oleate.

[0074] Furthermore, the INS is isononyl isononanoate.

[0075] The preparation method of the compound composition includes the following steps:

[0076] Furthermore, a cream for sensitive skin is provided, comprising 0.5-5 kg ​​of a compound composition, 32.5-35 kg of deionized water, 1.5-3 kg of butylene glycol, 0.03-0.04 kg of sodium hyaluronate, 0.5-1.5 kg of betaine, 0.01-0.03 kg of dipotassium glycyrrhizate, 0.25-1 kg of cetearyl glucoside, 0.5-1.5 kg of sorbitan oleate, 1-2.5 kg of caprylic / capric triglycerides, 0.5-1.5 kg of cetearyl alcohol, 0.5-1.5 kg of dimethicone, 1-3 kg of shea butter, 0.4-0.8 kg of jojoba oil, 0.5-1.5 kg of plant squalane, 0.2-0.5 kg of sea buckthorn fruit oil, 0.25-0.75 kg of besterol-E, and 1-2 kg of INS. The preparation method of the cream includes the following steps:

[0077] (1) After adding the water specified in the formula to the emulsifying pot, add butylene glycol, sodium hyaluronate, betaine and dipotassium glycyrrhizate to the emulsifying pot and stir until completely dissolved. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0078] (2) The cetearyl glucoside, sorbitan olive oil ester, caprylic / capric triglyceride, cetearyl alcohol and polydimethylsiloxane are dispersed in the formula and then added to the emulsifying pot described in (1), and stirred until uniformly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0079] (3) Add the formula-prescribed amounts of shea butter, jojoba oil, plant squalane, sea buckthorn oil, besterol-E, and INS to the emulsifying pot described in (2) and stir until evenly mixed. The temperature during this process is room temperature, and the stirring speed is 20 rpm.

[0080] (4) Add the solution obtained in (3) to the emulsifying pot described in (2) and stir for 30 minutes to obtain a cream. The temperature during this process is room temperature, the stirring speed is 20 rpm, and the vacuum degree is -0.06 MPa.

[0081] (5) The cream obtained in (4) is filtered through a 200-mesh filter cloth and discharged to obtain the cream.

[0082] (6) The cream prepared in (5) shall be subjected to sensory, physicochemical and microbiological tests. The appearance shall be white or milky white, glossy, fine in texture and free of foreign matter; it shall conform to the specific fragrance; pH value shall be 5.5 to 6.5; total colony count shall be ≤500 CFU / g; mold and yeast count shall be ≤50 CFU / g.

[0083] (7) Use a semi-automatic filling machine to fill the qualified cream from (7) into the tube, and use a sealing machine to seal the tube to obtain the cream.

[0084] The characteristics of the cream involved in this invention are as follows:

[0085] (1) The synergistic anti-inflammatory effect of the compound combination: The compound combination in Example 1, through intervention and regulation of different target sites, affects different molecular networks, and complementarily regulates the inflammatory response, thereby exerting a network synergistic effect. Dipotassium glycyrrhizate, to assist in anti-inflammation;

[0086] (2) Butylene glycol and sodium hyaluronate protect skin cells by locking in moisture. Betaine enhances skin's moisture activity, improves skin elasticity, and makes skin smoother; it inhibits cell necrosis caused by high osmotic pressure in the external environment and maintains the balance of the stratum corneum. Plant-derived squalane lubricates the skin and reduces dryness and peeling symptoms. It fuses with the skin's lipid film to form a protective layer and promotes the generation of new cells.

[0087] (3) Anti-inflammatory and antibacterial: Shea butter, jojoba oil and sea buckthorn oil moisturize, promote the rehydration of skin stratum corneum cells, and have anti-inflammatory and antibacterial effects. Attached Figure Description

[0088] Figure 1 Effects of Panax notoginseng saponin premix and Portulaca oleracea polysaccharide premix on the viability of RAW264.7 cells

[0089] Figure 2 Contour plots of SynergyScore values ​​after combining different concentrations of Panax notoginseng saponin pre-mixed solution (PNS) with Portulaca oleracea polysaccharide pre-mixed solution (POP).

[0090] Figure 3 Contour plots of SynergyScore values ​​after combining different concentrations of Panax notoginseng saponin pre-mixed solution (PNS) with Portulaca oleracea polysaccharide pre-mixed solution (POP).

[0091] Figure 4 Microscopic morphology of RAW264.7 cells in different groups (20x objective lens)

[0092] A: Blank control group

[0093] B: Negative control group

[0094] C: Positive control group

[0095] D: Test sample group (12.50 μL / mL)

[0096] E: Test sample group (6.25 μL / mL)

[0097] Figure 5 Comparison of the levels of TNF-α secreted by LPS-induced RAW264.7 cells in the test samples

[0098] Figure 6Comparison of LPS-induced IL-1β secretion levels in RAW264.7 cells from test samples

[0099] Figure 7 Comparison of LPS-induced IL-6 secretion levels in RAW264.7 cells by test samples

[0100] Figure 8 Microscopic morphology of RAW264.7 cells in different groups (20x objective lens)

[0101] A: Blank control group

[0102] B: Negative control group

[0103] C: Positive control group

[0104] D: Test sample group (1.00 mg / mL)

[0105] E: Test sample group (0.50 mg / mL)

[0106] Figure 9 Comparison of the levels of TNF-α secreted by LPS-induced RAW264.7 cells in the test samples

[0107] Figure 10 Comparison of LPS-induced IL-1β secretion levels in RAW264.7 cells from test samples

[0108] Figure 11 Comparison of LPS-induced IL-6 secretion levels in RAW264.7 cells by test samples Detailed Implementation

[0109] Example 1: Preparation of the compound composition (10 kg)

[0110] The reagents and instruments used in the following examples are as follows:

[0111] Reagents:

[0112] The raw materials for Panax notoginseng total saponins and Portulaca oleracea polysaccharides were provided by Yunnan Bailan Bio-resources Co., Ltd.

[0113] instrument:

[0114] Mettler Toledo Instruments (Shanghai) Co., Ltd.

[0115] Thermostatic Magnetic Stirrer Hangzhou Instrument & Motor Co., Ltd.

[0116] Low-speed centrifuge Anhui Zhongke Zhongjia Scientific Instruments Co., Ltd.

[0117] This embodiment illustrates the preparation method of the compound composition described in this invention.

[0118] (1) The raw materials of Panax notoginseng total saponins and Portulaca polysaccharide were provided by Yunnan Bailan Bio-resources Co., Ltd.

[0119] (2) Weigh 0.1-0.15 kg of Panax notoginseng total saponins raw material and set aside; add 7.6-7.9 L of propylene glycol to a beaker and heat and stir to 80°C, set aside; add 18-22 L of hydrogenated castor oil to another beaker and heat to dissolve, set aside. Add propylene glycol to the hydrogenated castor oil and stir for 5 min, then add the Panax notoginseng total saponins raw material and cool to room temperature. The Panax notoginseng total saponins pre-prepared solution is obtained.

[0120] (3) Weigh 0.02-0.04 kg of purslane polysaccharide, and then measure 2.5-3.5 L of 1,3-propanediol into a mixing pot and mix well; then add 6.5-7 L of deionized water and stir well (stirring speed 60 r / min, stirring time 30 min). After complete dissolution, it is a transparent and homogeneous liquid, which is the purslane polysaccharide pre-prepared solution.

[0121] (4) Take 4.5-5.5 kg of (2) premixed solution and 1.5-5.5 kg of (3) premixed solution, mix them evenly to obtain a compound composition.

[0122] Example 2: Preparation of a soothing and moisturizing water using a compound combination as the active ingredient (50 kg)

[0123] The reagents and instruments used in the following examples are as follows:

[0124] Reagents:

[0125] Example 1 prepared a compound composition. Deionized water, dipotassium glycyrrhizate, 4-tert-butylcyclohexanol, trehalose, tocopherol, hyaluronic acid, acetylglucosamine, sea fennel extract, ethylhexylglycerin, capryloyl hydroxamic acid, chlorpyrifos, PE9010, PHL, and fragrance were purchased from Yunnan Bailan Biological Resources Development Co., Ltd.

[0126] instrument:

[0127] Laboratory vacuum homogenizer / emulsifier, purchased from Shanghai Huashuo Intelligent Equipment Co., Ltd.; electronic balance, purchased from Mettler Toledo, model PL1002E / 02.

[0128] Sealing machine, Guangzhou Changxin Packaging Materials Co., Ltd. This embodiment illustrates the preparation of moisturizing water using the compound combination described in this invention as the active ingredient.

[0129] Furthermore, a moisturizing water is provided, comprising 3.8 kg of the compound composition of Example 1, 42.5 kg of water, 0.13 kg of dipotassium glycyrrhizate, 1.5 kg of 4-tert-butylcyclohexanol, 0.88 kg of trehalose, 0.03 kg of tocopherol, 0.03 kg of hyaluronic acid, 0.06 kg of acetylglucosamine, 0.5 kg of sea fennel extract, 0.02 kg of glutenin, 0.04 kg of PE9010, 0.38 kg of PHL, and 0.15 kg of fragrance.

[0130] Preparation method:

[0131] (1) After adding the water according to the formula to the emulsifying pot, add dipotassium glycyrrhizate and 4-tert-butylcyclohexanol to the emulsifying pot and stir until completely dissolved. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0132] (2) Disperse the trehalose, tocopherol, hyaluronic acid, acetylglucosamine and sea fennel extract according to the formula, and then add them to the emulsifying pot described in (1), and stir until they are evenly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0133] (3) Add the compound composition prepared in Example 1, ethylhexylglycerin, capryloyl hydroxamic acid, GE9010, and PHL in the amount specified in (2) to the emulsifying pot and stir until uniformly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0134] (4) Add the solution obtained from the fragrance to the emulsification pot described in (3), stir and mix for 30 minutes to obtain water. The temperature during this process is room temperature, the stirring speed is 20 rpm, and the vacuum degree is -0.06 MPa.

[0135] (5) The water obtained in (4) is filtered through a 200-mesh filter cloth and discharged to obtain moisturizing water.

[0136] (6) Sensory, physicochemical and microbiological indicators of the moisturizing water prepared in (5) were tested. The product is non-greasy, non-sticky, comfortable to use, and free of foreign matter; it conforms to a specific fragrance; pH value is 5.5-6.5; total bacterial count is ≤500 CFU / g; mold and yeast count is ≤50 CFU / g.

[0137] (7) Use a semi-automatic filling machine to fill the qualified moisturizing water from (6) into the tube, and seal the tube with a sealing machine to obtain soothing and repairing moisturizing water.

[0138] Example 3: Preparation of a soothing and repairing cream using a compound combination as active ingredients (50 kg)

[0139] Reagents:

[0140] Example 1 prepared a compound composition. Deionized water, butylene glycol, sodium hyaluronate, betaine, dipotassium glycyrrhizate, cetearyl glucoside, sorbitan oleate, caprylic / capric triglyceride, cetearyl alcohol, polydimethylsiloxane, shea butter, jojoba oil, plant squalane, sea buckthorn oil, Besterol-E and INS were purchased from Yunnan Bailan Biological Resources Development Co., Ltd.

[0141] instrument:

[0142] Laboratory vacuum homogenizer / emulsifier, purchased from Shanghai Huashuo Intelligent Equipment Co., Ltd., Model 1

[0143] Electronic balance, purchased from Mettler Toledo, model PL1002E / 02

[0144] Sealing machine, Guangzhou Changxin Packaging Materials Co., Ltd.

[0145] This embodiment illustrates the preparation of a soothing and repairing cream using the compound combination described in this invention as an active ingredient.

[0146] It contains 3.8 kg of compound composition, 33.75 kg of deionized water, 2 kg of butylene glycol, 0.035 kg of sodium hyaluronate, 1 kg of betaine, 0.025 kg of dipotassium glycyrrhizate, 0.5 kg of cetearyl glucoside, 0.75 kg of sorbitan oleate, 1.25 kg of caprylic / capric triglyceride, 0.75 kg of cetearyl alcohol, 0.75 kg of polydimethylsiloxane, 1.75 kg of shea butter, 0.6 kg of jojoba oil, 0.5 kg of plant squalane, 0.4 kg of sea buckthorn oil, 0.5 kg of besterol-E, and 1.5 kg of INS.

[0147] Preparation method:

[0148] (1) After adding the water specified in the formula to the emulsifying pot, add butylene glycol, sodium hyaluronate, betaine and dipotassium glycyrrhizate to the emulsifying pot and stir until completely dissolved. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0149] (2) The cetearyl glucoside, sorbitan olive oil ester, caprylic / capric triglyceride, cetearyl alcohol and polydimethylsiloxane are dispersed in the formula and then added to the emulsifying pot described in (1), and stirred until uniformly mixed. The temperature during this process is room temperature and the stirring speed is 20 rpm.

[0150] (3) Add the formula-prescribed amounts of shea butter, jojoba oil, plant squalane, sea buckthorn oil, besterol-E, and INS to the emulsifying pot described in (2) and stir until evenly mixed. The temperature during this process is room temperature, and the stirring speed is 20 rpm.

[0151] (4) Add the solution obtained in (3) to the emulsification pot described in (2) and stir for 30 minutes to obtain a frosting. The temperature during this process is room temperature, the stirring speed is 20 rpm, and the vacuum degree is -0.06 MPa.

[0152] (5) The cream obtained in (4) is filtered through a 200-mesh filter cloth and discharged to obtain the cream.

[0153] (6) The cream prepared in (5) shall be subjected to sensory, physicochemical and microbiological tests. The appearance shall be white or milky white, glossy, fine in texture and free of foreign matter; it shall conform to the specific fragrance; pH value shall be 5.5 to 6.5; total colony count shall be ≤500 CFU / g; mold and yeast count shall be ≤50 CFU / g.

[0154] (7) Use a semi-automatic filling machine to fill the qualified cream from (7) into the tube, and use a sealing machine to seal the tube to obtain the cream.

[0155] Experimental Example 1: Skin Cytotoxicity Test

[0156] The skin is divided into the epidermis, dermis, and subcutaneous tissue from the outside in. Following the "Implementation Guidelines for Alternative Methods of Cosmetic Evaluation" edited by Cheng Shujun, cytotoxicity tests were performed on human immortalized epidermal cells (HaCaT) in the epidermis and human skin fibroblasts (HSF) in the dermis to evaluate the cytotoxic effects of anti-inflammatory components on human skin cells. When the relative cell viability was ≥80%, the test substance was considered to have no cytotoxicity at that concentration. The two plant extract pre-prepared solutions (2) and (3) prepared in Example 1 were diluted to the specified concentration with culture medium and co-cultured with the two cell types for 24 hours. Then, 100 μL of culture medium and 10 μL of LCK-8 solution were added to each well, and the mixture was incubated for 2 hours. The absorbance (OD) value at 450 nm was measured using a microplate reader. The cell viability of each test group was calculated using the following formula. Each experiment was independently repeated 3 times, and the data were presented as follows: As shown in Tables 4 and 5, when the concentrations of Panax notoginseng extract and Portulaca oleracea extract are below 250 ug / mL, the HSF cell survival rate is >80%; when the concentrations are below 500 ug / mL, the Hacat cell survival rate is >80%, indicating that the test samples have no effect on cell proliferation at these concentrations.

[0157]

[0158] Table 4. Effects of Panax notoginseng saponin pre-prepared solution on the survival rate of HSF and Hacat cells.

[0159]

[0160] Table 5. Effects of Purslane Polysaccharide Premix on the Viability of HSF and Hacat Cells

[0161]

[0162] Example 2: Evaluation of the synergistic anti-inflammatory efficacy of the compound composition

[0163] Test objective

[0164] The anti-inflammatory efficacy of a combination of two soothing ingredients (prepared from Example 1) was evaluated by using a lipopolysaccharide (LPS)-induced model to measure the secretion of the inflammatory factor TNF-α by macrophage cells (RAW264.7).

[0165] principle

[0166] LPS-induced RAW264.7 cells are a classic cell model for studying inflammatory factors. LPS binds to antigen recognition receptors on the surface of macrophages, inducing macrophages to secrete various inflammatory factors, including TNF-α. TNF-α can activate three signaling pathways: caspase, JNK, and the transcription factor NF-κB, thereby achieving its biological functions such as cytotoxicity, antiviral activity, immune regulation, and apoptosis.

[0167] This method evaluates the inhibitory effect of the compound composition on TNF-α secretion by comparing the difference in TNF-α secretion levels in RAW264.7 cells after administration of the negative control and the compound composition. The content was determined using enzyme-linked immunosorbent assay (ELISA). The principle is as follows: the inflammatory factor TNF-α specifically binds to TNF-α antibodies coated on an ELISA plate. Substrate-labeled anti-TNF-α antibodies then bind, and the substrate is catalyzed to generate a colored product. The TNF-α content was calculated by measuring the optical density (OD value) of the TNF-α content and the colored product at a wavelength of 450 nm.

[0168] Experimental Section

[0169] Main materials, reagents and instruments

[0170] The 16-group compound combination of Panax notoginseng total saponins and Portulaca oleracea polysaccharides was prepared by mixing the ingredients prepared in (2) and (3) of Example 1 according to the concentration ratio; TNF-α ELISA kit was purchased from Xinbosheng Biotechnology Co., Ltd.; CCK-8 assay kit and dexamethasone were purchased from Solarbio Science & Technology Co., Ltd.; dimethyl sulfoxide (DMSO) and LPS (from E. coli) were purchased from Merck Life Science, Germany; mouse mononuclear macrophage leukemia cell line RAW264.7 was obtained from Kunming Cell Bank of the Chinese Academy of Sciences, model KCB200603YJ; DMEM medium was purchased from Baisha Biotechnology Co., Ltd.; fetal bovine serum was purchased from Thermo Fisher Scientific Co., Ltd.; phosphate buffered saline (PBS) was purchased from Jiangsu Kaiji Biotechnology Co., Ltd.

[0171] 96-well cell culture plates, 24-well plates, T25 culture flasks, and T75 culture flasks were purchased from Corning Life Sciences Co., Ltd.; an ELISA reader was purchased from Guzi Corporation (USA); a CO2 incubator was purchased from Memert Biotech GmbH (Germany); a 37°C incubator was purchased from Shanghai Senxin Instrument Co., Ltd.; a centrifuge was purchased from Shanghai Scientific Instrument Factory; an automatic autoclave was purchased from China Inspection Southwest Metrology Co., Ltd.; pipettes and pipette tips were purchased from White Shark Biotechnology Co., Ltd.; and a vortex mixer was purchased from Qilinbell Instrument Manufacturing Co., Ltd.

[0172] Experimental methods

[0173] 1. Preparation of test samples

[0174] Stimulant: Accurately weigh 10 mg LPS, dissolve it in 5 mL PBS to prepare a 2 mg / mL LPS stock solution. Aliquot 100 μL into 0.5 mL EP tubes and store at -20°C. Dilute with culture medium to a concentration of 5 μg / mL before use as the stimulant.

[0175] Experimental group: 10 mg of Panax notoginseng saponin pre-prepared solution and purslane polysaccharide pre-prepared solution were accurately weighed and dissolved in 0.5 mL and 1 mL DMSO, respectively, to prepare stock solutions of Panax notoginseng saponin pre-prepared solution and purslane polysaccharide pre-prepared solution with a mass concentration of 20 mg / mL and 10 mg / mL, respectively. 100 μL and 200 μL of each were aliquoted into 0.5 mL EP tubes and stored at -20℃. Before use, the solutions were serially diluted with culture medium to the target concentration for cell viability detection and evaluation of the anti-inflammatory activity of different compound combinations.

[0176] Positive control group: Accurately weigh 5 mg of dexamethasone, dissolve it in 1 mL of DMSO, aliquot 100 μL into 0.5 mL EP tubes, store at -20 °C, and dilute with culture medium to a concentration of 100 μg / mL before use. Add the irritant as a positive control.

[0177] 2. Cell Culture

[0178] RAW264.7 cells were added to DMEM high-glucose culture medium containing 10% fetal bovine serum and cultured in an incubator (37°C, 5% CO2) according to standard procedures. When the cell confluence reached 80-90%, adherent cells were scraped off the bottom of the flask, digested, and passaged. The cells were passaged or the medium was changed 2-3 times per week.

[0179] 3. RAW264.7 Cell Viability Assay

[0180] When the cell confluence reached 80-90%, a cell suspension was obtained using the cell passage method described above, and the cell density was adjusted to 6.5 × 10⁻⁶. 4Cells were added at a concentration of 100 μL / mL to 200 μL of cell suspension in each well of a 96-well plate and incubated for 24 h at 37°C, 5% CO2. The old culture medium was first removed from the 96-well plate, followed by 200 μL of complete culture medium containing different concentrations of the test substance (experimental group) and 200 μL of complete culture medium without the test substance (negative control group) in each well. Each group was divided into three replicates. The plates were incubated at constant temperature for 24 h. The supernatant was discarded, and 100 μL of a mixture of culture medium and 10 μL of CCK-8 solution was added to each well. The plates were incubated for 2 h, and the absorbance (OD) at 450 nm was measured using a microplate reader. Cell viability was calculated for each test group using the following formula. Each experiment was independently repeated three times. Data were presented as follows: This indicates that when cell viability is >80%, the test sample at that concentration is considered to have no effect on the proliferation of RAW264.7 cells.

[0181]

[0182] 4. In vitro cell assays to verify the anti-inflammatory efficacy of the test substance.

[0183] In the above cell culture method, when the cell confluence reaches 80-90%, the cell suspension is taken and the cell density is adjusted to 9 × 10⁶ cells / year. 4 Cells were added at a density of 1000 μL / mL to 24-well plates, and incubated at 37°C (5% CO2) for 24 h. Once the cell deposition rate met the experimental requirements, the model and control groups were replaced with fresh complete culture medium. The positive control group was added with complete culture medium containing 100 μg / mL dexamethasone. The experimental groups were added with culture medium containing pre-prepared Panax notoginseng saponins (1, 5, 25, 125 μg / mL) and pre-prepared Portulaca oleracea polysaccharides (2, 10, 50, 250 μg / mL), and a medium containing 16 different compound combinations. Cells were pretreated for 2 h. Except for the control group, both the model and experimental groups were incubated with LPS (5 μg / mL) in complete culture medium for another 16 h. The supernatant was collected, centrifuged at 3000 rpm for 10 min, and stored. TNF-α levels were detected according to the ELISA kit instructions. Each experiment was independently repeated three times, and data were presented as follows: express.

[0184]

[0185] 5. Synergistic effect analysis

[0186] The synergistic effect of the combined use of Panax notoginseng saponin premix and Portulaca oleracea polysaccharide premix was analyzed using the SynergyFinder analysis tool (https: / / synergyfinder.fimm.fi). This tool is a publicly accessible online platform for drug combination analysis, used for interactive analysis and visualization of multi-drug combination response data. The combined use of drugs was quantified to quantitatively describe the interaction between the two components. A SynergyScore > 10 indicates a synergistic effect; -10 ≤ SynergyScore ≤ 10 indicates an additive effect; and SynergyScore < -10 indicates an antagonistic effect.

[0187] 6. Data Processing

[0188] Experimental data were statistically analyzed using SPSS 17.0 software. Quantitative data were expressed as mean plus or minus standard deviation. This indicates that one-way ANOVA was used to compare data among multiple groups, and P < 0.05 was considered statistically significant.

[0189] Test Results

[0190] 1. Effects of different concentrations of pre-mixed solutions on the viability of RAW264.7 cells

[0191] After 24 hours of intervention with different concentrations of Panax notoginseng saponin pre-prepared solution or Portulaca oleracea polysaccharide pre-prepared solution, the cell viability of RAW264.7 cells was as follows: Figure 1 As shown in the figure, when the concentration of the Panax notoginseng saponin premix was less than 125 μg / mL and the concentration of the Portulaca oleracea polysaccharide premix was less than 250 μg / mL, the cell viability was greater than 80%, indicating that the above concentrations are safe dosage ranges that have no effect on cell survival and proliferation. When the concentration of the Portulaca oleracea polysaccharide premix was between 62.5 and 250 μg / mL, the viability of RAW264.7 cells was >100%, indicating that it promoted cell proliferation. In subsequent tests, this concentration will be selected as the maximum concentration of the Panax notoginseng saponin premix and the Portulaca oleracea polysaccharide premix in the compound combination design. Based on this, after isometric dilution and pairwise compounding, a series of anti-inflammatory activity tests will be carried out.

[0192] 2. Results of anti-inflammatory effects of pre-mixed solutions with different ratios

[0193] (1) Inflammation model

[0194] In this experiment, after 16 hours of LPS stimulation of RAW264.7 cells, the TNF-α content in the model group (LPS stimulation) increased by 133.61 times compared with the blank group (no LPS stimulation), which meets the validity verification requirements of the group standard "Test of Cosmetic Soothing Efficacy - In Vitro TNF-α Inflammatory Factor Content Determination Method for Lipopolysaccharide-Induced Macrophage RAW264.7". This proves that under the experimental conditions, the research model of RAW264.7 cells producing the inflammatory factor TNF-α stimulated by 5 μg / mL LPS is valid.

[0195] (2) Analysis of anti-inflammatory efficacy and synergistic effects

[0196] Different concentrations of Panax notoginseng saponin pre-prepared solution and Portulaca oleracea polysaccharide pre-prepared solution both inhibited LPS-induced TNF-α production from RAW264.7 (see Table 6). According to the T / SHRH034-2021 standard, when the TNF-α inhibition rate is ≥25%, the tested component is judged to have a soothing effect. When Panax notoginseng saponin pre-prepared solution was used alone at concentrations of 25 μg / mL and 125 μg / mL, the TNF-α inhibition rate was greater than 25%; when Portulaca oleracea polysaccharide pre-prepared solution was used alone at a concentration of 250 μg / mL, the TNF-α inhibition rate was also greater than 25%. When the pre-mixed solution of Panax notoginseng saponins (125 μg / mL) and the pre-mixed solution of Portulaca oleracea polysaccharides (250 μg / mL) were combined, the inhibition rate of TNF-α was 61.42%, which was greater than the sum of the inhibition rates of the pre-mixed solutions of Panax notoginseng saponins and Portulaca oleracea polysaccharides used alone at the same dosage. The SynergyScore was 26.44 (see...). Figure 2 When the pre-mixed solution of Panax notoginseng saponins (125 μg / mL) and the pre-mixed solution of Portulaca oleracea polysaccharides (50 μg / mL) were combined, the SynergyScore was 15.58, indicating that the above combination had a synergistic effect. The remaining 14 combinations showed results within the range of -10 ≤ SynergyScore ≤ 10, indicating that these combinations only had an additive effect and no synergistic effect.

[0197] Table 6. Comparison of the inhibition rate (%) of LPS-induced TNF-α release from RAW264.7 cells after pairwise mixing of pre-mixed solutions with different concentrations.

[0198]

[0199] SynergyScore > 10 indicates that the combination has a synergistic effect; -10 ≤ SynergyScore ≤ 10 indicates that the combination has an additive effect; SynergyScore < -10 indicates that the combination has an antagonistic effect.

[0200] Dexamethasone is a commonly used anti-inflammatory drug in clinical practice. In this experiment, dexamethasone (100 μg / mL) was used as the positive control group, and its inhibition rate against TNF-α was 53.17%. Figure 3 It can be seen that both combinations that produced a synergistic effect, 125 μg / mL Panax notoginseng saponin pre-mixed solution + 50 μg / mL Portulaca polysaccharide pre-mixed solution (synergistic group 1) and 125 μg / mL Panax notoginseng saponin pre-mixed solution + 250 μg / mL Portulaca polysaccharide pre-mixed solution (synergistic group 2), significantly reduced (P<0.01) the TNF-α secretion content of LPS-induced RAW264.7 cells, with no statistically significant difference compared with the positive control group (P>0.05).

[0201] Natural anti-inflammatory ingredients extracted from plants can exert a good synergistic therapeutic effect on inflammation through combined action on different targets and multiple pathways, achieving the same or even better therapeutic effect with lower drug dosages. Network pharmacology studies have shown that the combined use of Panax notoginseng and Portulaca oleracea can regulate inflammatory responses through multiple pathways, including leukocyte migration, oxidative stress response, and transcription factor activity. Portulaca oleracea polysaccharides may play a leading role in the release of TNF-α inflammatory factors; while total saponins of Panax notoginseng play a leading role in inhibiting the NF-κBp65 target. This suggests that when the two are combined, total saponins of Panax notoginseng and Portulaca oleracea polysaccharides do not exert a synergistic effect by jointly intervening in the same target, but rather focus on regulating different targets and affecting different molecular networks, complementarily regulating inflammatory responses to exert a network synergistic effect. This may also be the essence of the compound formulation of plant-based ingredients in cosmetics.

[0202] 3. Conclusion

[0203] When a pre-mixed solution of Panax notoginseng saponins at 125 μg / mL is combined with a pre-mixed solution of Portulaca oleracea polysaccharides at 50 μg / mL or 250 μg / mL, it exhibits a synergistic anti-inflammatory effect, effectively inhibiting the release of the inflammatory factor TNF-α from LPS-stimulated macrophages RAW264.7, with an inhibitory effect comparable to that of 100 μg / mL dexamethasone. However, when the pre-mixed solutions of Panax notoginseng saponins and Portulaca oleracea polysaccharides are combined at lower concentrations, they do not exhibit a synergistic anti-inflammatory effect, but only an additive one.

[0204] Experiment 3 evaluates the anti-inflammatory effects of moisturizing water.

[0205] Test objective

[0206] The anti-inflammatory efficacy of the moisturizing water (prepared in Example 2) was evaluated according to the standard method of "T / SHRH034-2021 Test of Soothing Efficacy of Cosmetics - In Vitro TNF-α Inflammatory Factor Content Determination and Lipopolysaccharide-Induced Macrophage RAW264.7 Test Method" (2021 version).

[0207] principle:

[0208] LPS-induced RAW264.7 cells are a classic cell model for studying inflammatory factors. LPS binds to antigen recognition receptors on the surface of macrophages, inducing macrophages to secrete various inflammatory factors such as TNF-α, IL-1β, and IL-6.

[0209] TNF-α can activate three signaling pathways: Caspase protease, JNK, and transcription factor NF-κB, thereby achieving its biological functions such as cytotoxicity, antiviral activity, immune regulation, and apoptosis.

[0210] IL-1β is an important mediator of inflammatory responses. When the body is in an inflammatory state or undergoes an immune response, it can participate in various cellular activities, including cell proliferation, differentiation, and apoptosis. IL-1β can also lead to systemic inflammation by activating endothelial cells, inducing neutrophil mobilization in the bone marrow (leukocytosis), and activating various types of leukocytes, thereby increasing acute-phase reactive proteins such as C-reactive protein.

[0211] IL-6 production follows different patterns in various types of inflammation. In the early stages of infectious inflammation, different pathogens stimulate monocytes and macrophages to produce IL-6 via Toll-like receptors (TLRs) in the PAMP (Physical Pattern of Associative Angiogenesis). In non-infectious inflammation (such as burns or traumatic injuries), damaged cells are stimulated to produce IL-6 via TLRs in the DAMP (Damage-Associated Pattern of Associative Angiogenesis). This acute IL-6 expression plays a central role in host defense by stimulating various cell populations.

[0212] This method evaluates the inhibitory effect of the test sample on the secretion of TNF-α, IL-1β, and IL-6 by comparing the differences in the levels of TNF-α, IL-1β, and IL-6 secreted by RAW264.7 cells after drug administration between the negative control and the test sample. The content was determined using an enzyme-linked immunosorbent assay (ELISA). The specific principle (taking TNF-α as an example) is as follows: the inflammatory factor TNF-α specifically binds to the TNF-α antibody coated on the ELISA plate. Then, the substrate-labeled anti-TNF-α antibody binds, and the substrate is catalyzed to generate a colored product. The TNF-α content is calculated by measuring the optical density (OD value) of the TNF-α content and the colored product at a wavelength of 450 nm.

[0213] Research and testing methods:

[0214] 1. Cells: The mouse mononuclear macrophage leukemia cell line RAW264.7 was obtained from the Kunming Cell Bank of the Chinese Academy of Sciences, with cell type KCB200603YJ selected.

[0215] 2. Culture conditions: DMEM medium, fetal bovine serum, trypsin / EDTA solution (0.25% trypsin solution and 0.02 mol / L EDTA solution mixed 1:1), phosphate buffer (PBS), cell culture flasks.

[0216] 3. Irritant: Lipopolysaccharide (LPS, E. coli source), Sigma-Aldrich, USA, batch number: 0000126449.

[0217] 4. Positive control: Dexamethasone (Soleb D8040).

[0218] 5. Materials and Instruments: 96-well plate; TNF-α ELISA kit, IL-1β ELISA kit, IL-6 ELISA kit ( ELISA kit; CCK-8 cytotoxicity detection kit (Beijing Polymer Biotechnology Co., Ltd. MF128-01); microplate reader; clean bench; dimethyl sulfoxide (DMSO).

[0219] 6. Test Grouping

[0220] Table 7. Experimental group design

[0221]

[0222] 7. Test Procedure

[0223] 7.1 Cell and Test Sample Preparation

[0224] (1) Cell preparation:

[0225] a. Cell resuscitation:

[0226] 1) Preheat the water bath to 37°C, prepare clean disposable PE gloves, and add 9 mL of sterile DMEM culture medium to a sterile centrifuge tube;

[0227] 2) Remove the cells from the liquid nitrogen tank, place them in PE gloves, and quickly immerse them in a water bath. Shake the cryovial to accelerate thawing, ideally until all cells are thawed within 1 minute.

[0228] 3) In a clean bench, add the revived cell solution to a centrifuge tube containing DMEM medium, centrifuge at 1000 rpm / min for 4 minutes, and remove the supernatant after centrifugation.

[0229] 4) Resuspend the cells in 6 mL of DMEM medium containing 10% serum, inoculate them into a T25 culture flask, add medium to 5 mL, and incubate in an incubator.

[0230] b. Cell digestion and passage:

[0231] Cells were digested using trypsin / EDTA (preferably 0.25%). 1 mL of trypsin was used in T25 culture flasks, and 3 mL in T75 culture flasks. Under a microscope, once most cells became rounded and were in suspension, approximately 2-3 times the volume of trypsin was added to DMEM medium containing 10% serum to stop digestion. The cells were then collected in centrifuge tubes and centrifuged at 1000 rpm for 4 minutes. After centrifugation, the supernatant was discarded, and DMEM medium containing 10% serum was added to the centrifuge tubes. The cells were then mixed by pipetting and transferred to two T25 culture flasks.

[0232] (2) Preparation of test sample solution

[0233] Before testing, the test sample is diluted with culture medium to the test dose concentration.

[0234] 7.2 CCK-8 cell viability assay

[0235] The in vitro toxicity of the test samples to RAW264.7 cells was examined using CCK-8 (CellCounting Kit8). The assay was performed at a concentration of 6.5 × 10⁻⁶ cells. 4 Cells were seeded at a density of 200 μL / mL into 96-well plates. After incubation for 24 h, the old culture medium was discarded. Different concentrations of the test substance diluted with culture medium were added to the 96-well plates, and incubated for 24 h. The supernatant was discarded. A mixture of 100 μL culture medium and 10 μL CCK-8 solution was added to each well, and incubated for 2 h. The absorbance at 450 nm was measured using a microplate reader. Cell viability was calculated using a negative control as a baseline. When cell viability was ≥80%, the test sample concentration was considered to have no effect on cell proliferation, i.e., a safe cell concentration.

[0236]

[0237] 7.3 ELISA kit for detecting inflammatory factor levels

[0238] (1) Preparation of irritant solution

[0239] Preparation of LPS stock solution (12.5 μL / mL): Dissolve 10 mg LPS in 5000 μL PBS, vortex for 5 min, let stand for 30 min, then aliquot 100 μL into 0.5 mL centrifuge tubes and freeze at -20 °C.

[0240] Preparation of LPS working solution (5 μg / mL): Take 100 μL of LPS with a concentration of 12.5 μL / mL, add 1900 μL of culture medium to obtain a working solution of 100 μg / mL. After drug administration pretreatment for 2 h, add 106 μL of LPS working solution to a 6-well plate.

[0241] (2) Preparation of positive control solution:

[0242] Preparation of dexamethasone stock solution (100 mg / mL): Dissolve 100 mg of dexamethasone in 1000 μL LDMSO, shake for 2 h to mix thoroughly, dispense into 20 μL / vial, store at -80℃, and filter through a 0.22 μm filter membrane for sterilization before use.

[0243] Preparation of dexamethasone working solution (100 μg / mL): Take 20 μL of dexamethasone stock solution with a concentration of 100 mg / mL, and dilute it to 2 mL with culture medium to obtain dexamethasone working solution 1 with a concentration of 1000 μg / mL. Take 250 μL of dexamethasone working solution 1 with a concentration of 1000 μg / mL, and dilute it to 2.5 mL with culture medium to obtain dexamethasone working solution 2 with a concentration of 100 μg / mL. For administration, transfer 2000 μL of dexamethasone working solution 2 to a 6-well plate.

[0244] (3) Cell inoculation

[0245] Dilute cells to the seeding density using cell culture medium (100,000 cells / mL, achieving 45%–60% confluence after 24 hours). Seed the cells into 6-well plates, 2000 μL per well. After seeding, incubate in a CO2 incubator for 24 hours. Ensure that the cell confluence is within the range of 45%–60% 24 hours after seeding.

[0246] (4) Induction and drug administration

[0247] After the cells reached 45-60% confluence, the old culture medium was discarded and replaced with serum-free medium (high glucose DMEM), and cultured for another 12 hours. The old culture medium in the 6-well plate was then discarded. Culture medium containing a specific concentration of the test substance was added to the test wells, cell culture medium was added to the negative control wells, and culture medium containing dexamethasone was added to the positive control wells. Cell culture medium was added to the blank / solvent control wells, 2000 μL per well. After administration, the plate was incubated at 37°C in a 5% CO2 incubator for 2 hours. After 2 hours, except for the blank / solvent control wells, 106 μL of 100 μg / ml LPS working solution was added to the remaining wells. The plate was gently shaken to mix. The plate was then returned to a 37°C, 5% CO2 incubator and cultured for 24 hours.

[0248] (5) Collection of cell supernatant

[0249] After incubation, collect 2000 μL of cell culture supernatant into a 2.0 mL sterile centrifuge tube, centrifuge at 3000 rpm for 10 min, and then store the supernatant. Store at -20°C.

[0250] (6) ELISA detection

[0251] a. Preparations before testing:

[0252] Taking the TNF-α kit as an example

[0253] 1) Please remove the kit from the refrigerator 20 minutes in advance to allow it to equilibrate to room temperature.

[0254] 2) Dilute the 20× concentrated washing solution with double-distilled water to make 1× working solution. Return any unused portion to 4℃.

[0255] 3) Standards: Centrifuge at 1000 rpm for 1 min before opening. Add 0.5 mL of universal diluent for both standards and samples to the lyophilized standard, let stand for 15 minutes until fully dissolved, then gently mix (concentration 2000 pg / mL). Then dilute as needed (e.g., use the following concentrations for the standard curve: 2000, 1000, 500, 250, 125, 62.5, 31.25, 15.63, 7.81 pg / mL).

[0256] 4) Biotinylated antibody working solution: Dilute 30× concentrated biotinylated antibody to 1× working solution with biotinylated antibody diluent 20 minutes before use, according to the amount required for the current experiment. Use on the same day.

[0257] 5) Enzyme conjugate working solution: Dilute the 30× concentrated enzyme conjugate to 1× working solution with enzyme conjugate diluent 20 minutes before use, according to the amount required for the current experiment. Use on the same day.

[0258] b. Operating steps:

[0259] 1) Remove the required strips from the sealed bag that has been equilibrated to room temperature. Put the unused strips and desiccant back into the aluminum foil bag, press the self-sealing strip firmly, seal the bag, and return it to 4°C.

[0260] 2) Add standard and specimen general diluent to blank wells, and add specimen or standard of different concentrations (100 μL / well) to the other corresponding wells. Seal the reaction wells with sealing tape and incubate at 37°C in the dark for 90 minutes.

[0261] 3) Prepare the biotinylated antibody working solution 20 minutes in advance.

[0262] 4) Wash the plate 5 times.

[0263] 5) Add biotinylated antibody diluent to the blank wells, and add biotinylated antibody working solution (100 μL / well) to the remaining wells. Seal the reaction wells with new sealing tape and incubate at 37°C in the dark for 60 minutes.

[0264] 6) Prepare the enzyme conjugate working solution 20 minutes in advance. Store at room temperature (22-25℃) away from light.

[0265] 7) Wash the plate 5 times.

[0266] 8) Add enzyme conjugate dilution to blank wells and enzyme conjugate working solution (100 μL / well) to the remaining wells. Seal the reaction wells with new sealing tape and incubate at 37°C in the dark for 30 minutes.

[0267] 9) Turn on the power of the ELISA reader, preheat the instrument, and set the detection program.

[0268] 10) Wash the plate 5 times.

[0269] 11) Add 100 μL of chromogenic substrate (TMB) per well and incubate at 37°C in the dark for 15 minutes.

[0270] 12) Add the reaction stop solution (100 μL / well), mix well, and immediately measure the OD. 450nm Value (within 3 minutes).

[0271] c. Result Interpretation:

[0272] The OD value of each standard and specimen should be subtracted from the OD value of the blank well. Plot the standard concentration on the x-axis and the OD value on the y-axis using software to create a curve and select the best fit; a quadratic polynomial equation is recommended for fitting. The concentration of the specimen can be found on the standard curve using its OD value.

[0273] 8. Calculation of TNF-α, IL-1β and IL-6 levels and inhibition rates

[0274] (1) Content

[0275] Standard solutions of TNF-α, IL-1β, and IL-6 were prepared and serially diluted to a series of solutions of known concentrations. The OD values ​​(OD values) at 450 nm were then measured using the ELISA method described above. 450 The regression equation for the standard curve was fitted, and the OD values ​​of each test sample were then used to calculate the standard curve. 450 The values ​​were substituted into the equation to calculate the levels of TNF-α, IL-1β, and IL-6 in the test sample. Three replicates were performed for each group, and the mean and standard deviation (SD) were calculated. SPSS statistical software was used to perform t-tests to compare the TNF-α, IL-1β, and IL-6 levels of each sample group with the negative control group. A p-value < 0.05 was considered statistically significant.

[0276] (2) Inhibition rates of TNF-α, IL-1β and IL-6

[0277] Inhibition rate (%) = (1-T / C)×100%, where T is the average content of TNF-α, IL-1β or IL-6 in each sample group, and C is the average content of TNF-α, IL-1β or IL-6 in the negative control group.

[0278] Research and testing results:

[0279] 1. CCK-8 cell viability assay

[0280] The effects of different doses of the test sample (moisturizing water) on the viability of RAW264.7 cells were tested using the CCK-8 kit, as shown in Table 8.

[0281] Table 8. Comparison of the effects of different concentrations of test samples on the viability of RAW264.7 cells.

[0282]

[0283] The selected test sample concentration should ensure cell viability ≥80%. Therefore, 12.50 μL / mL and 6.25 μL / mL were selected as the concentrations for subsequent inflammatory factor content testing.

[0284] 2. Cell morphology characteristics of RAW264.7

[0285] The morphology of RAW264.7 cells in each group is as follows: Figure 4 As shown, normal RAW264.7 cells are round or oval in shape. Figure 4 A), after 24 hours of LPS stimulation, the cells in the negative group showed morphological changes, increased volume, and stretched into a spindle shape. Figure 4 B). Compared with the negative group, when different concentrations of test samples were co-stimulated with LPS, most cells remained round, and their morphology tended to resemble normal cells. Figure 4 D, 4E).

[0286] 3. Detection of the content and inhibition rate of the inflammatory factor TNF-α

[0287] Table 9. Inhibition of LPS-induced secretion of the inflammatory cytokine TNF-α by the test samples

[0288]

[0289] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0290] T – Average TNF-α content in the sample group

[0291] C – Average TNF-α content in the negative control group

[0292] Figure 5The effects of each test group on the LPS-induced TNF-α secretion level and inhibition rate in RAW264.7 cells were compared with Table 9. Using 12.5 μL / mL and 6.25 μL / mL test samples (moisturizing water), the LPS-induced TNF-α secretion levels in RAW264.7 cells were 1325.06 pg / mL and 1599.81 pg / mL, respectively, significantly lower than the negative control group (2060.50 pg / mL, p<0.05), with TNF-α inhibition rates of 35.69% and 22.36%, respectively. The positive control group using 100 μg / mL dexamethasone showed an LPS-induced TNF-α secretion level of 1278.31 pg / mL in RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with a TNF-α inhibition rate of 37.96%.

[0293] 4. Detection of the content and inhibition rate of the inflammatory factor IL-1β

[0294] Table 10. Inhibition of LPS-induced secretion of the inflammatory cytokine IL-1β by the test samples

[0295]

[0296] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0297] T – Average IL-1β content in the sample group

[0298] C – Average IL-1β level in the negative control group

[0299] Figure 6 The effects of each test group on the IL-1β secretion induced by LPS in RAW264.7 cells and the inhibition rate were compared with Table 10. Using 12.5 μL / mL and 6.25 μL / mL test samples (moisturizing water), the IL-1β secretion levels of LPS-induced RAW264.7 cells were 17.84 pg / mL and 21.41 pg / mL, respectively, significantly lower than the negative control group (95.01 pg / mL, p<0.05), with IL-1β inhibition rates of 81.22% and 77.46%, respectively. The positive control group using 100 μg / mL dexamethasone showed an IL-1β secretion level of 18.36 pg / mL in LPS-induced RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with an IL-1β inhibition rate of 80.68%.

[0300] 5. Detection of the content and inhibition rate of the inflammatory factor IL-6

[0301] Table 11 shows the inhibitory effect of the test samples on the secretion of the inflammatory cytokine IL-6 induced by LPS in RAW264.7 cells.

[0302]

[0303] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0304] T – Average IL-6 content in the sample group

[0305] C – Average IL-6 level in the negative control group

[0306] Figure 7 The effects of each test group on the IL-6 secretion level and inhibition rate induced by LPS in RAW264.7 cells were compared with those in Table 11. Using 12.5 μL / mL and 6.25 μL / mL test samples (moisturizing water), the IL-6 secretion levels induced by LPS in RAW264.7 cells were 147.04 pg / mL and 354.81 pg / mL, respectively, which were lower than the negative control group (544.04 pg / mL, p<0.05), with IL-6 inhibition rates of 70.59% and 38.18%, respectively. The positive control group using 100 μg / mL dexamethasone showed an IL-6 secretion level of 49.78 pg / mL induced by LPS in RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with an IL-1β inhibition rate of 91.12%.

[0307] summary:

[0308] Under the conditions of this experiment, the 12.5 μL / mL test product (moisturizing water) inhibited the secretion of TNF-α, IL-1β, and IL-6 by LPS-induced RAW264.7 cells, with inhibition rates of 35.69%, 81.22%, and 70.59%, respectively; the 6.25 μL / mL test product (moisturizing water) also inhibited the secretion of TNF-α, IL-1β, and IL-6 by LPS-induced RAW264.7 cells, with inhibition rates of 22.36%, 77.46%, and 38.18%, respectively. In conclusion, the test product possesses soothing and anti-inflammatory effects.

[0309] Example 4: Evaluation of the efficacy of creams prepared using compound compositions

[0310] Test objective

[0311] The anti-inflammatory efficacy of the moisturizing water (prepared in Example 3) was evaluated according to the standard method of "T / SHRH034-2021 Cosmetic Soothing Efficacy Test - In Vitro TNF-α Inflammatory Factor Content Determination Lipopolysaccharide Induced Macrophage RAW264.7 Test Method" (2021 version).

[0312] The research and testing methods for the principle are the same as those in Experiment Example 3.

[0313] Research and testing results:

[0314] 1. CCK-8 cell viability assay

[0315] The effects of different doses of the test sample (cream) on the viability of RAW264.7 cells were tested using the CCK-8 kit, as shown in Table 2.

[0316] Table 12. Comparison of the effects of different concentrations of test samples on the viability of RAW264.7 cells.

[0317]

[0318] The selected test sample concentration should ensure cell viability ≥90%. Therefore, 1.00 mg / mL and 0.50 mg / mL were selected as the dose concentrations for subsequent inflammatory factor content testing.

[0319] 2. Cell morphology characteristics of RAW264.7

[0320] The morphology of RAW264.7 cells in each group is as follows: Figure 8 As shown, normal RAW264.7 cells are round or oval in shape. Figure 8 A), after 24 hours of LPS stimulation, the cells in the negative group showed morphological changes, increased volume, and stretched into a spindle shape. Figure 8 B). Compared with the negative group, when different concentrations of test samples were co-stimulated with LPS, most cells remained round, and their morphology tended to resemble normal cells. Figure 8 D, 8E).

[0321] 3. Detection of the content and inhibition rate of the inflammatory factor TNF-α

[0322] Table 13. Inhibition of LPS-induced secretion of the inflammatory cytokine TNF-α by the test samples

[0323]

[0324] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0325] T – Average TNF-α content in the sample group

[0326] C – Average TNF-α content in the negative control group

[0327] Figure 9The effects of each test group on the LPS-induced TNF-α secretion level and inhibition rate in RAW264.7 cells were compared with Table 13. Using 1 mg / mL and 0.5 mg / mL test samples (cream), the LPS-induced TNF-α secretion levels in RAW264.7 cells were 852.67 pg / mL and 1042.33 pg / mL, respectively, significantly lower than the negative control group (1725.77 pg / mL, p<0.05), with TNF-α inhibition rates of 50.39% and 39.60%, respectively. The positive control group using 100 μg / mL dexamethasone showed an LPS-induced TNF-α secretion level of 957.75 pg / mL in RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with a TNF-α inhibition rate of 44.50%.

[0328] 4. Detection of the content and inhibition rate of the inflammatory factor IL-1β

[0329] Table 14 shows the inhibitory effect of the test samples on the secretion of the inflammatory cytokine IL-1β induced by LPS in RAW264.7 cells.

[0330]

[0331] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0332] T – Average IL-1β content in the sample group

[0333] C – Average IL-1β level in the negative control group

[0334] Figure 10 The effects of each test group on the IL-1β secretion induced by LPS in RAW264.7 cells and the inhibition rate were compared with Table 14. Using 1 mg / mL and 0.5 mg / mL test samples (cream), the IL-1β secretion levels in LPS-induced RAW264.7 cells were 9.16 pg / mL and 10.97 pg / mL, respectively, significantly lower than the negative control group (27.99 pg / mL, p<0.05), with IL-1β inhibition rates of 67.26% and 61.88%, respectively. The positive control group using 100 μg / mL dexamethasone showed an IL-1β secretion level of 16.39 pg / mL in LPS-induced RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with an IL-1β inhibition rate of 40.24%.

[0335] 5. Detection of the content and inhibition rate of the inflammatory factor IL-6

[0336] Table 15 shows the inhibitory effect of the test samples on the secretion of the inflammatory cytokine IL-6 induced by LPS in RAW264.7 cells.

[0337]

[0338] Note: Inhibition rate (%) = (1 - T / C) × 100%

[0339] T – Average IL-6 content in the sample group

[0340] C – Average IL-6 level in the negative control group

[0341] Figure 11 The effects of each test group on the LPS-induced IL-6 secretion level and inhibition rate in RAW264.7 cells were compared with Table 15. Using 1 mg / mL and 0.5 mg / mL test samples (cream), the LPS-induced IL-6 secretion levels in RAW264.7 cells were 729.15 pg / mL and 652.22 pg / mL, respectively, lower than the negative control group (777.00 pg / mL, p>0.05), with IL-6 inhibition rates of 16.06% and 6.16%, respectively. The positive control group using 100 μg / mL dexamethasone showed an LPS-induced IL-6 secretion level of 275.81 pg / mL in RAW264.7 cells, also significantly lower than the negative control group (p<0.05), with an IL-1β inhibition rate of 64.50%. Summary:

[0342] Under the conditions of this experiment, the 1 mg / mL test product (cream) inhibited the secretion of TNF-α, IL-1β, and IL-6 by LPS-induced RAW264.7 cells, with inhibition rates of 50.59%, 61.88%, and 16.06%, respectively; the 0.5 mg / mL test product (cream) also inhibited the secretion of TNF-α, IL-1β, and IL-6 by LPS-induced RAW264.7 cells, with inhibition rates of 39.60%, 67.26%, and 6.16%, respectively. In conclusion, the test product has soothing and anti-inflammatory effects. Example 5: Safety Test for Moisturizing Water-Sensitive Skin and Subject Self-Assessment

[0343] Test objective

[0344] Asian adult subjects with sensitive skin used the test sample (moisturizing water) for 28 consecutive days under normal conditions. The efficacy and safety of the test sample in soothing and moisturizing, and whether it met the claims of being suitable for sensitive skin, were evaluated through subject self-assessment and dermatologist assessment.

[0345] Test Results

[0346] 1. Subject self-assessment

[0347] After using the test sample for 28 days, the overall satisfaction rate of the 30 subjects with the product (moisturizing water) was 100.0%.

[0348]

[0349] 14 days after using the sample

[0350]

[0351] 28 days after using the sample

[0352]

[0353]

[0354] 2. Safety assessment by dermatologists

[0355] Summary of adverse reactions:

[0356] project Adverse event ratio Adverse events observed by dermatologists 0％(0 / 30) Adverse events assessed by a dermatologist as related to the trial sample. 0％(0 / 30) Serious adverse events assessed by a dermatologist as related to the trial sample. 0％(0 / 30)

[0357] Subject self-assessment

[0358] Throughout the entire test, none of the 30 participants experienced any discomfort.

[0359] Test Conclusion

[0360] Thirty adult subjects aged 21 to 36 with sensitive skin used the test sample (moisturizing water) for 28 consecutive days under normal conditions. Through self-assessment by the subjects, the results showed that the test sample (moisturizing water) had a soothing and moisturizing effect after 28 days of use.

[0361] Under the supervision of dermatologists, 30 adult subjects aged 21 to 36 with sensitive skin were observed using the test sample continuously for 28 days under normal conditions to evaluate its potential to cause adverse skin reactions. The dermatologists' assessment results showed that none of the 30 subjects experienced adverse reactions after use, and the subjects' self-reported results also showed no discomfort during or after use. These results met the requirements of the test sample (moisturizing lotion)'s claim of being "suitable for sensitive skin."

[0362] Human efficacy evaluation of the cream in Experiment Example 6

[0363] Test objective

[0364] Asian adult female subjects with sensitive skin used the test sample (cream) for 28 consecutive days under normal conditions. The moisture content of the stratum corneum and the transepidermal water loss (TEWL) value of the skin were measured by an instrument probe, and the subjects also conducted self-assessments to evaluate the efficacy of the test sample in moisturizing and repairing, and whether it met the "suitable for sensitive skin" claim.

[0365] Test Results

[0366] 1. Instrument measurement

[0367]

[0368] 2. Subject self-assessment

[0369]

[0370] After using the test sample for 28 days, the overall satisfaction rate of the 31 subjects with the test sample (cream) was 100.0%, and 100.0% of the subjects believed that the product was mild and non-irritating and would not cause allergic reactions.

[0371] Test Conclusion

[0372] Thirty-one Asian adult female subjects aged 41 to 60 with sensitive skin used the test sample for 28 consecutive days under normal conditions. The moisture content of the stratum corneum and the transepidermal water loss (TEWL) value of the skin were measured using an instrument probe. The results showed that the test sample had moisturizing and repairing effects after 28 days of use. According to the subjects' self-assessment, the results showed that the test sample had a soothing effect after 28 days of use and met the claim of being "suitable for sensitive skin".

Claims

1. A soothing anti-inflammatory composition, characterized in that it comprises , including the total saponins of panax notoginseng pre-liquid and the pre-liquid of the polygonum multiflorum polysaccharide, the preparation method is: (1) purchase the total saponins of panax notoginseng and the polygonum multiflorum polysaccharide raw materials; (2) take 0.1-0.15 kg of the total saponins of panax notoginseng raw materials, for standby; in a beaker, add 7.6-7.9 L of propylene glycol and heat stirring to 80 DEG C, for standby; in another beaker, add 18-22 L of hydrogenated castor oil, heat and dissolve, for standby; the propylene glycol is added to the hydrogenated castor oil, stirring 5 min, then add the total saponins of panax notoginseng raw materials, cooling to room temperature; the total saponins of panax notoginseng pre-liquid is obtained; (3) take 0.02-0.04 kg of the polygonum multiflorum polysaccharide raw materials, then take 2.5-3.5 L of 1,3-propanediol and put into a stirring pot, mix evenly; then add 6.5-7 L of deionized water, stir evenly; after complete dissolution, it is a transparent uniform liquid, the polygonum multiflorum polysaccharide pre-liquid is obtained; (4) take 4.5-5.5 kg of (2) the total saponins of panax notoginseng pre-liquid, and take 1.5-5.5 kg of (3) the polygonum multiflorum polysaccharide pre-liquid, mix evenly, a compound composition is obtained; The mass concentration of the total saponins of panax notoginseng pre-liquid and the polygonum multiflorum polysaccharide pre-liquid is 125 μg / mL and 50 μg / mL respectively, or the mass concentration of the total saponins of panax notoginseng pre-liquid and the polygonum multiflorum polysaccharide pre-liquid is 125 μg / mL and 250 μg / mL respectively.

2. The composition of claim 1, wherein It is prepared into cosmetics.

3. The composition of claim 2, wherein The cosmetic is moisturizing water or cream.

4. A moisturizing water, containing 0.5-5 kg of the composition of claim 1, 40-50 kg of water, 0.01-0.025 kg of dipotassium glycyrrhizinate, 1-1.5 kg of 4-tert-butylcyclohexanol, 0.75-1 kg of trehalose, 0.015-0.035 kg of tocopherol, 0.01-0.025 kg of hyaluronic acid, 0.04-0.06 kg of acetylchitosamine, 0.045-0.1 kg of ethylhexylglycerin, 0.25-0.4 kg of capryloylhydroxamic acid sea, 0.25-0.5 kg of anise extract, 0.015-0.025 kg of extremely gain, 0.03-0.05 kg of PE9010, 0.25-0.5 kg of PHL and 0.005-0.015 kg of essence.

5. A cream for sensitive skin, containing 0.5-5 kg of the composition of claim 1, 32.5-35 kg of deionized water, 1.5-3 kg of butanediol, 0.03-0.04 kg of sodium hyaluronate, 0.5-1.5 kg of betaine, 0.01-0.03 kg of dipotassium glycyrrhizinate, 0.25-1 kg of cetyl stearyl glucoside, 0.5-1.5 kg of sorbitan olivate, 1-2.5 kg of caprylic / capric triglyceride, 0.5-1.5 kg of cetyl stearyl alcohol, 0.5-1.5 kg of dimethicone, 1-3 kg of shea butter, 0.4-0.8 kg of jojoba oil, 0.5-1.5 kg of vegetable squalane, 0.2-0.5 kg of pistachio oil, 0.25-0.75 kg of Besterol-E and 1-2 kg of INS.

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